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Sample records for carota ssp carota

  1. Essential-oil composition of Daucus carota ssp. major (Pastinocello Carrot) and nine different commercial varieties of Daucus carota ssp. sativus fruits.

    PubMed

    Flamini, Guido; Cosimi, Elena; Cioni, Pier Luigi; Molfetta, Ilaria; Braca, Alessandra

    2014-07-01

    The chemical composition of the essential oils obtained by hydrodistillation from the pastinocello carrot, Daucus carota ssp. major (Vis.) Arcang. (flowers and achenes), and from nine different commercial varieties of D. carota L. ssp. sativus (achenes) was investigated by GC/MS analyses. Selective breeding over centuries of a naturally occurring subspecies of the wild carrot, D. carota L. ssp. sativus, has produced the common garden vegetable with reduced bitterness, increased sweetness, and minimized woody core. On the other hand, the cultivation of the pastinocello carrot has been abandoned, even if, recently, there has been renewed interest in the development of this species, which risks genetic erosion. The cultivated carrot (D. carota ssp. sativus) and the pastinocello carrot (D. carota ssp. major) were classified as different subspecies of the same species. This close relationship between the two subspecies urged us to compare the chemical composition of their essential oils, to evaluate the differences. The main essential-oil constituents isolated from the pastinocello fruits were geranyl acetate (34.2%), α-pinene (12.9%), geraniol (6.9%), myrcene (4.7%), epi-α-bisabolol (4.5%), sabinene (3.3%), and limonene (3.0%). The fruit essential oils of the nine commercial varieties of D. carota ssp. sativus were very different from that of pastinocello, as also confirmed by multivariate statistical analyses.

  2. Mitochondrial gene diversity associated with the atp9 stop codon in natural populations of wild carrot (Daucus carota ssp. carota).

    PubMed

    Mandel, Jennifer R; McAssey, Edward V; Roland, Katherine M; McCauley, David E

    2012-01-01

    Mitochondrial genomes extracted from the wild populations of Daucus carota have been used as a genetic resource by breeders of cultivated carrot, yet little is known concerning the extent of their diversity in nature. Of special interest is an SNP in the putative stop codon of the mitochondrial gene atp9 that has been associated previously with male-sterile and male-fertile phenotypic variants. In this study, either the sequence or PCR/RFLP genotypes were obtained from the mitochondrial genes atp1, atp9, and cox1 found in D. carota individuals collected from 24 populations in the eastern United States. More than half of the 128 individuals surveyed had a CAA or AAA, rather than TAA, genotype at the position usually thought to function as an atp9 stop codon in this species. We also found no evidence for mitochondrial RNA editing (Cytosine to Uridine) of the CAA stop codon in either floral or leaf tissue. Evidence for intragenic recombination, as opposed to the more common intergenic recombination in plant mitochondrial genomes, in our data set is presented. Indel and SNP variants elsewhere in atp9, and in the other 2 genes surveyed, were nonrandomly associated with the 3 atp9 stop codon variants, though further analysis suggested that multilocus genotypic diversity had been enhanced by recombination. Overall the mitochondrial genetic diversity was only modestly structured among populations with an F(ST) of 0.34.

  3. Carotenoid Biosynthesis in Daucus carota.

    PubMed

    Simpson, Kevin; Cerda, Ariel; Stange, Claudia

    2016-01-01

    Carrot (Daucus carota) is one of the most important vegetable cultivated worldwide and the main source of dietary provitamin A. Contrary to other plants, almost all carrot varieties accumulate massive amounts of carotenoids in the root, resulting in a wide variety of colors, including those with purple, yellow, white, red and orange roots. During the first weeks of development the root, grown in darkness, is thin and pale and devoid of carotenoids. At the second month, the thickening of the root and the accumulation of carotenoids begins, and it reaches its highest level at 3 months of development. This normal root thickening and carotenoid accumulation can be completely altered when roots are grown in light, in which chromoplasts differentiation is redirected to chloroplasts development in accordance with an altered carotenoid profile. Here we discuss the current evidence on the biosynthesis of carotenoid in carrot roots in response to environmental cues that has contributed to our understanding of the mechanism that regulates the accumulation of carotenoids, as well as the carotenogenic gene expression and root development in D. carota. PMID:27485223

  4. Carotenoid Biosynthesis in Daucus carota.

    PubMed

    Simpson, Kevin; Cerda, Ariel; Stange, Claudia

    2016-01-01

    Carrot (Daucus carota) is one of the most important vegetable cultivated worldwide and the main source of dietary provitamin A. Contrary to other plants, almost all carrot varieties accumulate massive amounts of carotenoids in the root, resulting in a wide variety of colors, including those with purple, yellow, white, red and orange roots. During the first weeks of development the root, grown in darkness, is thin and pale and devoid of carotenoids. At the second month, the thickening of the root and the accumulation of carotenoids begins, and it reaches its highest level at 3 months of development. This normal root thickening and carotenoid accumulation can be completely altered when roots are grown in light, in which chromoplasts differentiation is redirected to chloroplasts development in accordance with an altered carotenoid profile. Here we discuss the current evidence on the biosynthesis of carotenoid in carrot roots in response to environmental cues that has contributed to our understanding of the mechanism that regulates the accumulation of carotenoids, as well as the carotenogenic gene expression and root development in D. carota.

  5. New Claims for Wild Carrot (Daucus carota subsp. carota) Essential Oil.

    PubMed

    Alves-Silva, Jorge M; Zuzarte, Mónica; Gonçalves, Maria José; Cavaleiro, Carlos; Cruz, Maria Teresa; Cardoso, Susana M; Salgueiro, Lígia

    2016-01-01

    The essential oil of Daucus carota subsp. carota from Portugal, with high amounts of geranyl acetate (29.0%), α-pinene (27.2%), and 11αH-himachal-4-en-1β-ol (9.2%), was assessed for its biological potential. The antimicrobial activity was evaluated against several Gram-positive and Gram-negative bacteria, yeasts, dermatophytes, and Aspergillus strains. The minimal inhibitory concentration (MIC) and minimal lethal concentration (MLC) were evaluated showing a significant activity towards Gram-positive bacteria (MIC = 0.32-0.64 μL/mL), Cryptococcus neoformans (0.16 μL/mL), and dermatophytes (0.32-0.64 μL/mL). The inhibition of the germ tube formation and the effect of the oil on Candida albicans biofilms were also unveiled. The oil inhibited more than 50% of filamentation at concentrations as low as 0.04 μL/mL (MIC/128) and decreased both biofilm mass and cell viability. The antioxidant capacity of the oil, as assessed by two in chemico methods, was not relevant. Still, it seems to exhibit some anti-inflammatory potential by decreasing nitric oxide production around 20% in LPS-stimulated macrophages, without decreasing macrophages viability. Moreover, the oils safety profile was assessed on keratinocytes, alveolar epithelial cells, macrophages, and hepatocytes. Overall, the oil demonstrated a safety profile at concentrations below 0.64 μL/mL. The present work highlights the bioactive potential of D. carota subsp. carota suggesting its industrial exploitation. PMID:26981143

  6. New Claims for Wild Carrot (Daucus carota subsp. carota) Essential Oil.

    PubMed

    Alves-Silva, Jorge M; Zuzarte, Mónica; Gonçalves, Maria José; Cavaleiro, Carlos; Cruz, Maria Teresa; Cardoso, Susana M; Salgueiro, Lígia

    2016-01-01

    The essential oil of Daucus carota subsp. carota from Portugal, with high amounts of geranyl acetate (29.0%), α-pinene (27.2%), and 11αH-himachal-4-en-1β-ol (9.2%), was assessed for its biological potential. The antimicrobial activity was evaluated against several Gram-positive and Gram-negative bacteria, yeasts, dermatophytes, and Aspergillus strains. The minimal inhibitory concentration (MIC) and minimal lethal concentration (MLC) were evaluated showing a significant activity towards Gram-positive bacteria (MIC = 0.32-0.64 μL/mL), Cryptococcus neoformans (0.16 μL/mL), and dermatophytes (0.32-0.64 μL/mL). The inhibition of the germ tube formation and the effect of the oil on Candida albicans biofilms were also unveiled. The oil inhibited more than 50% of filamentation at concentrations as low as 0.04 μL/mL (MIC/128) and decreased both biofilm mass and cell viability. The antioxidant capacity of the oil, as assessed by two in chemico methods, was not relevant. Still, it seems to exhibit some anti-inflammatory potential by decreasing nitric oxide production around 20% in LPS-stimulated macrophages, without decreasing macrophages viability. Moreover, the oils safety profile was assessed on keratinocytes, alveolar epithelial cells, macrophages, and hepatocytes. Overall, the oil demonstrated a safety profile at concentrations below 0.64 μL/mL. The present work highlights the bioactive potential of D. carota subsp. carota suggesting its industrial exploitation.

  7. New Claims for Wild Carrot (Daucus carota subsp. carota) Essential Oil

    PubMed Central

    Alves-Silva, Jorge M.; Zuzarte, Mónica; Gonçalves, Maria José; Cavaleiro, Carlos; Cruz, Maria Teresa; Cardoso, Susana M.

    2016-01-01

    The essential oil of Daucus carota subsp. carota from Portugal, with high amounts of geranyl acetate (29.0%), α-pinene (27.2%), and 11αH-himachal-4-en-1β-ol (9.2%), was assessed for its biological potential. The antimicrobial activity was evaluated against several Gram-positive and Gram-negative bacteria, yeasts, dermatophytes, and Aspergillus strains. The minimal inhibitory concentration (MIC) and minimal lethal concentration (MLC) were evaluated showing a significant activity towards Gram-positive bacteria (MIC = 0.32–0.64 μL/mL), Cryptococcus neoformans (0.16 μL/mL), and dermatophytes (0.32–0.64 μL/mL). The inhibition of the germ tube formation and the effect of the oil on Candida albicans biofilms were also unveiled. The oil inhibited more than 50% of filamentation at concentrations as low as 0.04 μL/mL (MIC/128) and decreased both biofilm mass and cell viability. The antioxidant capacity of the oil, as assessed by two in chemico methods, was not relevant. Still, it seems to exhibit some anti-inflammatory potential by decreasing nitric oxide production around 20% in LPS-stimulated macrophages, without decreasing macrophages viability. Moreover, the oils safety profile was assessed on keratinocytes, alveolar epithelial cells, macrophages, and hepatocytes. Overall, the oil demonstrated a safety profile at concentrations below 0.64 μL/mL. The present work highlights the bioactive potential of D. carota subsp. carota suggesting its industrial exploitation. PMID:26981143

  8. Genetic and phenological variation of tocochromanol (vitamin E) content in wild (Daucus carota L. var. carota) and domesticated carrot (D. carota L. var. sativa).

    PubMed

    Luby, Claire H; Maeda, Hiroshi A; Goldman, Irwin L

    2014-01-01

    Carrot roots (Daucus carota L. var. sativa) produce tocochromanol compounds, collectively known as vitamin E. However, little is known about their types and amounts. Here we determined the range and variation in types and amounts of tocochromanols in a variety of cultivated carrot accessions throughout carrot postharvest storage and reproductive stages and in wild-type roots (Daucus carota L. var. carota). Of eight possible tocochromanol compounds, we detected and quantified α-, and the combined peak for β- and γ- forms of tocopherols and tocotrienols. Significant variation in amounts of tocochromanol compounds was observed across accessions and over time. Large increases in α-tocopherol were noted during both reproductive growth and the postharvest stages. The variation of tocochromanols in carrot root tissue provides useful information for future research seeking to understand the role of these compounds in carrot root tissue or to breed varieties with increased levels of these compounds.

  9. Genetic and phenological variation of tocochromanol (vitamin E) content in wild (Daucus carota L. var. carota) and domesticated carrot (D. carota L. var. sativa)

    PubMed Central

    Luby, Claire H; Maeda, Hiroshi A; Goldman, Irwin L

    2014-01-01

    Carrot roots (Daucus carota L. var. sativa) produce tocochromanol compounds, collectively known as vitamin E. However, little is known about their types and amounts. Here we determined the range and variation in types and amounts of tocochromanols in a variety of cultivated carrot accessions throughout carrot postharvest storage and reproductive stages and in wild-type roots (Daucus carota L. var. carota). Of eight possible tocochromanol compounds, we detected and quantified α-, and the combined peak for β- and γ- forms of tocopherols and tocotrienols. Significant variation in amounts of tocochromanol compounds was observed across accessions and over time. Large increases in α-tocopherol were noted during both reproductive growth and the postharvest stages. The variation of tocochromanols in carrot root tissue provides useful information for future research seeking to understand the role of these compounds in carrot root tissue or to breed varieties with increased levels of these compounds. PMID:26504534

  10. Essential oils of Daucus carota subsp. carota of Tunisia obtained by supercritical carbon dioxide extraction.

    PubMed

    Marzouki, Hanen; Khaldi, Abdelhamid; Falconieri, Danilo; Piras, Alessandra; Marongiu, Bruno; Molicotti, Paola; Zanetti, Stefania

    2010-12-01

    The essential oils and supercritical CO2 extracts of wild Daucus carota L. subsp. carota from two different sites in Tunisia were investigated. The main components of the essential oil of the flowering and mature umbels with seeds from Sejnane were eudesm-7(11)-en-4-ol (8.2 - 8.5%), carotol (3.5 - 5.2%), sabinene (12.0 -14.5%), a-selinene (7.4 - 8.6) and 11-alpha-(H)-himachal-4-en-1-beta-ol (12.7 - 17.4%), whereas the oils from Tunis were predominantly composed of elemicin (31.5 - 35.3%) and carotol (48.0 - 55.7%). The antimicrobial activity of the essential oils were assayed by using the broth dilution method on Escherichia coli ATCC 35218 and Staphylococcus aureus ATCC 43300, and clinical strains of Candida albicans and C. tropicalis 1011 RM. The MIC values obtained were all > 2.5% (v/v).

  11. Survival and flowering of hybrids between cultivated and wild carrots (Daucus carota) in Danish grasslands.

    PubMed

    Hauser, Thure P; Shim, Sang In

    2007-01-01

    Many crop species are able to hybridize with related weedy or wild relatives, which could lead to transfer of cultivar genes, and among them transgenes, into wild populations. It is not clear, however, whether the hybrids and their descendants are able to survive and reproduce in natural habitats, as inherited cultivar traits may be maladaptive under such conditions. To test this, we produced hybrid (F(1)) seeds by controlled crosses between wild [see text for formula] and cultivated carrots (Daucus carota ssp. carota and ssp. sativa, respectively) and sowed them into three Danish grasslands of different age, in parallel with seeds of wild carrots. Replicate plots were sown in fall and spring. Survival and flowering of the emerging plants were monitored for the following three years. Both hybrid and wild carrots survived and flowered in highest frequency at a recently disturbed site, and much less at two older sites. Hybrids emerged in higher proportions than wild carrots in the first year and survived to similar or slightly lower frequencies at the end of the experiment. Hybrids flowered as frequently or slightly less frequently than wild plants, and developed fewer and smaller umbels. Despite a somewhat lower reproductive potential compared to wild carrots, first generation hybrids between cultivated and wild carrots are likely to survive and produce offspring in natural grasslands in Denmark. This, together with other studies, suggests that cultivar genes may transfer relatively easily into wild carrot populations.

  12. Alternative oxidase involvement in Daucus carota somatic embryogenesis.

    PubMed

    Frederico, António Miguel; Campos, Maria Doroteia; Cardoso, Hélia Guerra; Imani, Jafargholi; Arnholdt-Schmitt, Birgit

    2009-12-01

    Plant alternative oxidase (AOX) is a mitochondrial inner membrane enzyme involved in alternative respiration. The critical importance of the enzyme during acclimation upon stress of plant cells is not fully understood and is still an issue of intensive research and discussion. Recently, a role of AOX was suggested for the ability of plant cells to change easily its fate upon stress. In order to get new insights about AOX involvement in cell reprogramming, quantitative real-time polymerase chain reaction (PCR) and inhibitor studies were performed during cell redifferentiation and developmental stages of Daucus carota L. somatic embryogenesis. Transcript level analysis shows that D. carota AOX genes (DcAOX1a and DcAOX2a) are differentially expressed during somatic embryogenesis. DcAOX1a shows lower expression levels, being mainly down-regulated, whereas DcAOX2a presented a large up-regulation during initiation of the realization phase of somatic embryogenesis. However, when globular embryos start to develop, both genes are down-regulated, being this state transient for DcAOX2a. In addition, parallel studies were performed using salicylhydroxamic acid (SHAM) in order to inhibit AOX activity during the realization phase of somatic embryogenesis. Embryogenic cells growing in the presence of the inhibitor were unable to develop embryogenic structures and its growth rate was diminished. This effect was reversible and concentration dependent. The results obtained contribute to the hypothesis that AOX activity supports metabolic reorganization as an essential part of cell reprogramming and, thus, enables restructuring and de novo cell differentiation.

  13. Genotyping-by-sequencing provides the discriminating power to investigate the subspecies of Daucus carota (Apiaceae)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Premise of study: Premise of study: The taxonomic classification of the subspecies of Daucus carota is unresolved. Intercrosses among traditionally recognized subspecies has been well-documented, as have intercrosses with other Daucus species containing 2n = 18 chromosomes (D. sahariensis and D. syr...

  14. Genotyping-by-sequencing provides insights into the classification of the subspecies of Daucus carota

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Premise of study: Premise of study: The taxonomic classification of the subspecies of Daucus carota is unresolved. Intercrosses among traditionally recognized subspecies has been well-documented, as have intercrosses with other Daucus species containing 2n = 18 chromosomes (D. sahariensis and D. syr...

  15. The next generation of carotenoid studies in carrot (Daucus carota L.)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Orange carrot (Daucus carota L.) is one of the richest sources of naturally occurring ß-carotene while red and yellow carrot varieties contain large quantities of lycopene and lutein. The human body utilizes carotenoids, particularly ß-carotene (provitamin A) as a precursor for the production of ret...

  16. Reassessment of practical subspecies identifications of the USDA Daucus carota L. germplasm collection: Morphological data

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The genus Daucus includes about 20 species. The most widespread and economically important species, Daucus carota L., occurs on almost every continent. Cultivated carrot, subsp. sativus (Hoffm.) Schubl. & G. Martens, has been selected from wild populations that are extremely diverse, especially in t...

  17. Sequential Event Processing: Domain Specificity or Task Specificity? Commentary on Carota and Sirigu

    ERIC Educational Resources Information Center

    Toni, Ivan

    2008-01-01

    The article by Carota and Sirigu addresses a fundamental issue, namely the domain specificity of people's ability to learn and implement sequential structures of events. The authors review theoretical positions and empirical findings related to this issue, providing a useful summary of representative models of sequential event structures, and a…

  18. Daucus carota Pentane-Based Fractions Suppress Proliferation and Induce Apoptosis in Human Colon Adenocarcinoma HT-29 Cells by Inhibiting the MAPK and PI3K Pathways.

    PubMed

    Shebaby, Wassim N; Bodman-Smith, K B; Mansour, Anthony; Mroueh, Mohamad; Taleb, Robin I; El-Sibai, Mirvat; Daher, Costantine F

    2015-07-01

    Daucus carota L. ssp. carota (Apiacea, wild carrot, Queen Anne's lace) has been used in folk medicine throughout the world and recently was shown to possess anticancer and antioxidant activities. This study aims to determine the anticancer activity of the pentane fraction (F1) and the 1:1 pentane:diethyl ether fraction (F2) of the Daucus Carota oil extract (DCOE) against human colon adenocarcinoma cell lines (HT-29 and Caco-2). Treatment of cells with various concentrations of F1 or F2 fractions produced a dose-dependent inhibition of cell proliferation. Flow cytometric analysis indicated that both fractions induced sub-G1 phase accumulation and increased apoptotic cell death. Western blot revealed the activation of caspase-3, PARP cleavage, and a considerable increase in Bax and p53 levels, and a decrease in Bcl-2 level. Treatment of HT-29 cells with either fraction markedly decreased the levels of both phosphorylated Erk and Akt. Furthermore, the combined treatment of F1 or F2 with wortmannin showed no added inhibition of cell survival suggesting an effect of F1 or F2 through the phosphatidyl inositol 3-kinase (PI3K) pathway. This study proposes that DCOE fractions (F1 and F2) inhibit cell proliferation by inducing cell cycle arrest and apoptosis in HT-29 cells through the suppression of mitogen-activated protein kinase (MAPK)/Erk and PI3K/Akt pathways.

  19. [Spontaneous spectinomycin resistance mutations of the chloroplast rrn16 gene in Daucus carota callus lines].

    PubMed

    Filipenko, E A; Sidorchuk, Iu V; Deĭneko, E V

    2011-01-01

    Bioballistic transformation of carrot Daucus carota L. callus cultures with a plasmid containing the aadA (aminoglycoside 3'-adenyltransferase) gene and subsequent selection oftransformants on a selective medium containing spectinomycin (100-500 mg/l) yielded ten callus lines resistant to this antibiotic. PCR analysis did not detect exogenous DNA in the genomes of spectinomycin-resistant calluses. Resistance proved to be due to spontaneous mutations that occurred in two different regions of the chloroplast rrn16 gene, which codes for the 16S rRNA. Six lines displayed the G > T or G > C transverions in position 1012 of the rrn16 gene, and three lines had the A > G transition in position 1138 of the gene. Chloroplast mutations arising during passages of callus cultures in the presence of spectinomycin were described in D. carota for the first time. The cause of spectinomycin resistance was not identified in one line. The mutations observed in the D. carota plastid genome occurred in the region that is involved in the formation of a double-stranded region at the 3' end of the 16S rRNA and coincided in positions with the nucleotide substitutions found in spectinomycin-resistant plants of tobacco Nicotiana tabacum L. and bladderpod Lesquerella fendleri L.

  20. Two new eudesmane-type glucopyranosides from the fruits of Daucus carota L.

    PubMed

    Xu, Jin-Ke; Lu, Xiang-Hong; Zhu, Wei; Yang, Bing-Xian; Zhang, Lin; Tian, Jing-Kui

    2015-01-01

    Two new eudesmane-type glucopyranosides have been isolated from the fruits of Daucus carota L. On the basis of their spectroscopic and chemical evidence, the new compounds were elucidated as daucucarotol-10-O-β-D-glucopyranoside (1) and decahydro-7-[(2-O-β-D-glucopyranosyl)-isopropyl]-1β,4aα-dimethyl-(1α,4α,8aβ)-naphthalenetriol (2). Compounds 1 and 2 showed moderate antitumour activity against human ECA-109 and gave IC50 values of 23.22 and 26.76 μM, respectively.

  1. Identification and characterization of terpene synthases potentially involved in the formation of volatile terpenes in carrot (Daucus carota L.) roots

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Plants produce numerous volatile organic compounds, which are important in determining the quality and nutraceutical properties of fruit and root crops, including the taste and the aroma of carrots (Daucus carota L.). A combined chemical, biochemical and molecular study was conducted to evaluate the...

  2. Development of a high-throughput SNP resource to advance genomic, genetic and breeding research in carrot (Daucus carota L.)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The rapid advancement in high-throughput SNP genotyping technologies along with next generation sequencing (NGS) platforms has decreased the cost, improved the quality of large-scale genome surveys, and allowed specialty crops with limited genomic resources such as carrot (Daucus carota) to access t...

  3. Enantioselective Reduction by Crude Plant Parts: Reduction of Benzofuran-2-yl Methyl Ketone with Carrot ("Daucus carota") Bits

    ERIC Educational Resources Information Center

    Ravia, Silvana; Gamenara, Daniela; Schapiro, Valeria; Bellomo, Ana; Adum, Jorge; Seoane, Gustavo; Gonzalez, David

    2006-01-01

    The use of biocatalysis and biotransformations are important tools in green chemistry. The enantioselective reduction of a ketone by crude plant parts, using carrot ("Daucus carota") as the reducing agent is presented. The experiment introduces an example of a green chemistry procedure that can be tailored to fit in a regular laboratory session.…

  4. Intra-population genetic diversity of cultivated carrot (Daucus carota L.) assessed by analysis of microsatellite markers.

    PubMed

    Maksylewicz, Anna; Baranski, Rafal

    2013-01-01

    Intra-population variation of 18 cultivated carrot (Daucus carota L. ssp. sativus) populations of diverse origins was evaluated using codominant microsatellite (SSR) markers. Using 27 genomic and EST-derived SSR markers, 253 alleles were identified with a mean 9.4 alleles per marker. Most of the alleles (60.5%) were rare i.e., with the frequency ≤ 0.05 while only 3.95% of alleles occurred with frequency > 0.6. EST-derived SSR markers were less polymorphic than genomic SSR markers. Differences in allele occurrence allowed 16 out of 18 populations to be assigned to either the Western or Asian carrot gene pools with high probability. Populations could be also discriminated due to the presence of private alleles (25.3% of all alleles). Most populations had excess of alleles in the homozygous state indicating their inbreeding, although heterozygous loci were common in F1 hybrids. Genetic diversity was due to allelic variation among plants within populations (62% of total variation) and between populations (38%). Accessions originating from continental Asia and Europe had more allelic variants and higher diversity than those from Japan and USA. Also, allelic richness and variability in landraces was higher than in F1 hybrids and open-pollinated cultivars.

  5. Levels of Lycopene β-Cyclase 1 Modulate Carotenoid Gene Expression and Accumulation in Daucus carota

    PubMed Central

    Moreno, Juan Camilo; Pizarro, Lorena; Fuentes, Paulina; Handford, Michael; Cifuentes, Victor; Stange, Claudia

    2013-01-01

    Plant carotenoids are synthesized and accumulated in plastids through a highly regulated pathway. Lycopene β-cyclase (LCYB) is a key enzyme involved directly in the synthesis of α-carotene and β-carotene through the cyclization of lycopene. Carotenoids are produced in both carrot (Daucus carota) leaves and reserve roots, and high amounts of α-carotene and β-carotene accumulate in the latter. In some plant models, the presence of different isoforms of carotenogenic genes is associated with an organ-specific function. D. carota harbors two Lcyb genes, of which DcLcyb1 is expressed in leaves and storage roots during carrot development, correlating with an increase in carotenoid levels. In this work, we show that DcLCYB1 is localized in the plastid and that it is a functional enzyme, as demonstrated by heterologous complementation in Escherichia coli and over expression and post transcriptional gene silencing in carrot. Transgenic plants with higher or reduced levels of DcLcyb1 had incremented or reduced levels of chlorophyll, total carotenoids and β-carotene in leaves and in the storage roots, respectively. In addition, changes in the expression of DcLcyb1 are accompanied by a modulation in the expression of key endogenous carotenogenic genes. Our results indicate that DcLcyb1 does not possess an organ specific function and modulate carotenoid gene expression and accumulation in carrot leaves and storage roots. PMID:23555569

  6. Structure determination of bisacetylenic oxylipins in carrots (Daucus carota L.) and enantioselective synthesis of falcarindiol.

    PubMed

    Schmiech, Ludger; Alayrac, Carole; Witulski, Bernhard; Hofmann, Thomas

    2009-11-25

    Although bisacetylenic oxylipins have been demonstrated to exhibit diverse biological activities, the chemical structures of many representatives of this class of phytochemicals still remain elusive. As carrots play an important role in our daily diet and are known as a source of bisacetylenes, an extract made from Daucus carota L. was screened for bisacetylenic oxylipins, and, after isolation, their structures were determined by means of LC-MS and 1D/2D NMR spectroscopy. Besides the previously reported falcarinol, falcarindiol, and falcarindiol 3-acetate, nine additional bisacetylenes were identified, among which six derivatives are reported for the first time in literature and three compounds were previously not identified in carrots. To determine the absolute stereochemistry of falcarindiol in carrots, the (3R,8R)-, (3R,8S)-, (3S,8R)-, and (3S,8S)-stereoisomers of falcarindiol were synthesized according to a novel 10-step total synthesis involving a Cadiot-Chodkiewicz cross-coupling reaction of (S)- and (R)-trimethylsilanyl-4-dodecen-1-yn-3-ol and (R)- and (S)-5-bromo-1-penten-4-yn-3-ol, respectively. Comparative chiral HPLC analysis of the synthetic stereoisomers with the isolated phytochemical led to the unequivocal assignment of the (Z)-(3R,8S)-configuration for falcarindiol in carrot extracts from Daucus carota L.

  7. Levels of lycopene β-cyclase 1 modulate carotenoid gene expression and accumulation in Daucus carota.

    PubMed

    Moreno, Juan Camilo; Pizarro, Lorena; Fuentes, Paulina; Handford, Michael; Cifuentes, Victor; Stange, Claudia

    2013-01-01

    Plant carotenoids are synthesized and accumulated in plastids through a highly regulated pathway. Lycopene β-cyclase (LCYB) is a key enzyme involved directly in the synthesis of α-carotene and β-carotene through the cyclization of lycopene. Carotenoids are produced in both carrot (Daucus carota) leaves and reserve roots, and high amounts of α-carotene and β-carotene accumulate in the latter. In some plant models, the presence of different isoforms of carotenogenic genes is associated with an organ-specific function. D. carota harbors two Lcyb genes, of which DcLcyb1 is expressed in leaves and storage roots during carrot development, correlating with an increase in carotenoid levels. In this work, we show that DcLCYB1 is localized in the plastid and that it is a functional enzyme, as demonstrated by heterologous complementation in Escherichia coli and over expression and post transcriptional gene silencing in carrot. Transgenic plants with higher or reduced levels of DcLcyb1 had incremented or reduced levels of chlorophyll, total carotenoids and β-carotene in leaves and in the storage roots, respectively. In addition, changes in the expression of DcLcyb1 are accompanied by a modulation in the expression of key endogenous carotenogenic genes. Our results indicate that DcLcyb1 does not possess an organ specific function and modulate carotenoid gene expression and accumulation in carrot leaves and storage roots.

  8. Bioactive C₁₇-Polyacetylenes in Carrots (Daucus carota L.): Current Knowledge and Future Perspectives.

    PubMed

    Dawid, Corinna; Dunemann, Frank; Schwab, Wilfried; Nothnagel, Thomas; Hofmann, Thomas

    2015-10-28

    C17-polyacetylenes (PAs) are a prominent group of oxylipins and are primarily produced by plants of the families Apiaceae, Araliaceae, and Asteraceae, respectively. Recent studies on the biological activity of polyacetylenes have indicated their potential to improve human health due to anticancer, antifungal, antibacterial, anti-inflammatory, and serotogenic effects. These findings suggest targeting vegetables with elevated levels of bisacetylenic oxylipins, such as falcarinol, by breeding studies. Due to the abundant availability, high diversity of cultivars, worldwide experience, and high agricultural yields, in particular, carrot (Daucus carota L.) genotypes are a very promising target vegetable. This paper provides a review on falcarinol-type C17-polyacetylenes in carrots and a perspective on their potential as a future contributor to improving human health and well-being.

  9. Myo-Inositol trisphosphate mobilizes calcium from fusogenic carrot (Daucus carota L. ) protoplasts

    SciTech Connect

    Rincon, M.; Boss, W.F.

    1987-02-01

    To determine whether or not inositol trisphosphate (IP/sub 3/) mobilizes calcium in higher plant cells; they investigated the effect of IP/sub 3/ on Ca/sup 2 +/ fluxes in fusogenic carrot (Daucus carota L.) protoplasts. The protoplasts were incubated in /sup 45/Ca/sup 2 +/-containing medium and the /sup 45/Ca/sup 2 +/ associated with the protoplasts was monitored with time. Addition of IP/sub 3/ (20 micromolar) caused a 17% net loss of the accumulated /sup 45/Ca/sup 2 +/ within 4 minutes. There was a reuptake of /sup 45/Ca/sup 2 +/ and the protoplasts recovered to their initial value by 10 minutes. Phytic acid (IP/sub 6/), also stimulated /sup 45/Ca/sup 2 +/ efflux from the protoplasts. Both the IP/sub 3/- and the IP/sub 6/-induced /sup 45/Ca/sup 2 +/ efflux were inhibited by the calmodulin antagonist, trifluoperazine.

  10. Effect of Daucus carota var. boissieri extracts on immune response of Schistosoma mansoni infected mice.

    PubMed

    Shalaby, N M; Maghraby, A S; el-Hagrassy, A M

    1999-01-01

    Isolation and structure elucidation was carried out of flavonoid constituents found in fractionated extracts of the seeds and green leaves of Daucus carota L. var. boissieri (Apiaceae). The flavonoids are mainly apigenin, luteolin, their glycosidic precursors and 2,4,5-trimethoxybenzaldehyde. Fatty acids, hydrocarbons and sterols were identified by GLC. The effect of various carrot extracts on the immune responses of Schistosoma mansoni infected mice was studied. The rate of reduction in worm infestation in mice injected with some fractions indicated a strong protection. Some extracts induced humoral immune response through raising the IgG level at 2, 4 and 6 weeks post-infection as compared with infected control. The phenotypic analysis of the cellular immune response in spleen and mesenteric lymph nodes was accomplished by direct immunofluorescence. The data showed that some extracts stimulated the blastogenesis of CD4(+)-T splenocytes and mesenteric lymph node cells.

  11. Improvement of texture properties and flavor of frozen dough by carrot (Daucus carota) antifreeze protein supplementation.

    PubMed

    Zhang, Chao; Zhang, Hui; Wang, Li; Gao, Hong; Guo, Xiao Na; Yao, Hui Yuan

    2007-11-14

    The effects of concentrated carrot protein (CCP) containing 15.4% (w/w) carrot (Daucus carota) antifreeze protein on texture properties of frozen dough and volatile compounds of crumb were studied. CCP supplementation lowered the freezable water content of the dough, resulting in some beneficial effects including holding loaf volume steadily and making the dough softer and steadier during frozen storage. Furthermore, SPME-GC-MS analysis showed CCP supplementation did not give any negative influences on volatile compounds of crumb and gave a pleasant aroma felt like Michelia alba DC from trans-caryophyllene simultaneously. Combining our previous results that CCP supplementation improves the fermentation capacity of the frozen dough, CCP could be used as a beneficial additive for frozen dough processing. PMID:17935294

  12. Interruption of Somatic Embryogenesis in Daucus carota L. by 5-Bromodeoxyuridine

    PubMed Central

    Thomas, John C.; Nessler, Craig; Katterman, Frank

    1989-01-01

    Embryogenic Daucus carota L. cells grown in 9 micromolar 2.4-dichlorophenoxyacetic acid are resistant to greater than 5 micromolar 5-bromodeoxyuridine (BrdU). In contrast, 5 micromolar BrdU strongly inhibits somatic embryogenesis within 24 hours after transfer of cells to an auxin-free medium. DNA synthesis rates in control and BrdU-treated cultures are rapid and similar; however, the DNA content does not reach levels as great in the presence of BrdU as in control cultures. BrdU substitutes for thymidine in the DNA in 28% of the available sites 48 hours after auxin removal. Following DNA repair, somatic embryogenesis resumes. BrdU DNA incorporation leads to somatic embryogenesis inhibition and provides an alternative to auxin treatment for the interruption of carrot cell culture differentiation. Images Figure 2 Figure 3 Figure 7 PMID:16666898

  13. Bioactive C₁₇-Polyacetylenes in Carrots (Daucus carota L.): Current Knowledge and Future Perspectives.

    PubMed

    Dawid, Corinna; Dunemann, Frank; Schwab, Wilfried; Nothnagel, Thomas; Hofmann, Thomas

    2015-10-28

    C17-polyacetylenes (PAs) are a prominent group of oxylipins and are primarily produced by plants of the families Apiaceae, Araliaceae, and Asteraceae, respectively. Recent studies on the biological activity of polyacetylenes have indicated their potential to improve human health due to anticancer, antifungal, antibacterial, anti-inflammatory, and serotogenic effects. These findings suggest targeting vegetables with elevated levels of bisacetylenic oxylipins, such as falcarinol, by breeding studies. Due to the abundant availability, high diversity of cultivars, worldwide experience, and high agricultural yields, in particular, carrot (Daucus carota L.) genotypes are a very promising target vegetable. This paper provides a review on falcarinol-type C17-polyacetylenes in carrots and a perspective on their potential as a future contributor to improving human health and well-being. PMID:26451696

  14. Identification and Characterization of Terpene Synthases Potentially Involved in the Formation of Volatile Terpenes in Carrot (Daucus carota L.) Roots.

    PubMed

    Yahyaa, Mosaab; Tholl, Dorothea; Cormier, Guy; Jensen, Roderick; Simon, Philipp W; Ibdah, Mwafaq

    2015-05-20

    Plants produce an excess of volatile organic compounds, which are important in determining the quality and nutraceutical properties of fruit and root crops, including the taste and aroma of carrots (Daucus carota L.). A combined chemical, biochemical, and molecular study was conducted to evaluate the differential accumulation of volatile terpenes in a diverse collection of fresh carrots (D. carota L.). Here, we report on a transcriptome-based identification and functional characterization of two carrot terpene synthases, the sesquiterpene synthase, DcTPS1, and the monoterpene synthase, DcTPS2. Recombinant DcTPS1 protein produces mainly (E)-β-caryophyllene, the predominant sesquiterpene in carrot roots, and α-humulene, while recombinant DcTPS2 functions as a monoterpene synthase with geraniol as the main product. Both genes are differentially transcribed in different cultivars and during carrot root development. Our results suggest a role for DcTPS genes in carrot aroma biosynthesis.

  15. Daucus carota Pentane/Diethyl Ether Fraction Inhibits Motility and Reduces Invasion of Cancer Cells.

    PubMed

    Zgheib, Perla; Daher, Costantine F; Mroueh, Mohamad; Nasrallah, Anita; Taleb, Robin I; El-Sibai, Mirvat

    2014-01-01

    Daucus carota (DC) is a herb used in folklore medicine in Lebanon to treat numerous diseases including cancer. Recent studies in our laboratory on DC oil and its fractions revealed potent anticancer activities in vitro and in vivo. The present study aims to investigate the effect of the most potent DC fraction, pentane/diethyl ether (50:50), on lung, skin, breast and glioblastoma cancer cell motility and invasion. Upon treatment, a pronounced decrease in cancer cell motility was observed in the 4 cell lines. The treatment also led to a decrease in cancer cell invasion and an increased cell adhesion. Additionally, the DC fraction caused a decrease in the activation of the ρ-GTPases Rac and CDC42, a finding that may partially explain the treatment-induced decrease in cell motility. The current study demonstrates a crucial effect of the DC pentane/diethyl ether fraction on cancer cell motility and metastasis, making it a potential candidate for cancer therapy specifically targeting cancer motility and metastasis.

  16. [Influence of arbuscular mycorrhizal inoculation on growth and phoxim residue of carrot (Daucus carota L.)].

    PubMed

    Wang, Fa-Yuan; Chen, Xin; Sun, Xian-Ming; Shi, Zhao-Yong

    2010-12-01

    A pot culture experiment was carried out to study the influence of arbuscular mycorrhizal (AM) fungi on the growth and phoxim residue of carrot (Daucus carota L). Four levels of phoxim (0, 200, 400, 800 mg x L(-1)) and two AM fungal inocula, Glomus intraradices BEG 141(141), Glomus mosseae BEG 167 (167),and one nonmycorrhizal inoculum (CK), were applied to the sterilized soil. The plants were harvested after 5 months of growth and phoxim was irrigated into the root zone 14 d before plant harvest. Although decreasing with the increase of phoxim dosage, root infection rates of all the mycorrhizal plants were higher than 70%. Phoxim showed no significant dose effect on shoot wet weights and root yields, which were all increased by AM inoculation at four phoxim dosages. Phoxim residues in shoots and roots increased with the increase of phoxim dosage, but decreased by AM inoculation. In general, Glomus intraradices BEG 141 showed more pronounced effects on the growth and phoxim residue of carrot than Glomus mosseae BEG 167 did. Our results show a promising potential of AM fungi in carrot production and controlling pesticide residues.

  17. Fluorescent in situ hybridization with arbitrarily amplified DNA fragments differentiates carrot (Daucus carota L.) chromosomes.

    PubMed

    Nowicka, Anna; Grzebelus, Ewa; Grzebelus, Dariusz

    2012-03-01

    Carrot (Daucus carota L.) chromosomes are small and poorly differentiated in size and morphology. Here we demonstrate that fluorescent in situ hybridization (FISH) signals derived from arbitrary PCR probes can be used for chromosome identification in carrot. To prepare probes, we searched for nonpolymorphic products abundantly amplified with arbitrary decamer primers in a group of accessions representing carrot genetic diversity. As a result, 13 fragments ranging in size from 517 to 1758 bp were selected, sequenced, and used as probes for fluorescent in situ hybridization. Four of these probes produced clear and reproducible hybridization signals. The sequences showed similarity to a number of carrot BAC-end sequences, indicating their repetitive character. Three of them were similar to internal portions of gypsy and copia LTR retrotransposons previously identified in plants. Hybridization signals for the four probes were observed as dotted tracks on chromosomes, differing in distribution and intensity. Generally, they were present in pericentromeric and (or) interstitial localizations on chromosome arms. The use of the four probes allowed discrimination of chromosome pairs and construction of more detailed karyotypes and idiograms of carrot.

  18. Calcium transport in vesicles from carrot cells: Stimulation by calmodulin and phosphatidylserine. [Daucus carota cv. Danvers

    SciTech Connect

    Wenling Hsieh; Sze, Heven )

    1991-05-01

    The transport properties of Ca-pumping ATPases from carrot (Daucus carota cv. Danvers) tissue culture cells were studied. ATP dependent Ca transport in vesicles that comigrated with an ER marker, was stimulated 3-4 fold by calmodulin. Cyclopiazonic acid (a specific inhibitor of the sarcoplasmic/endoplasmic reticulum Ca-ATPase) partially inhibited oxalate-stimulated Ca transport activity; however, it had little or not effect on calmodulin-stimulated Ca uptake. The results suggested the presence of two types of Ca ATPases, and ER- and a plasma membrane-type. Incubation of membranes with (gamma{sup 32}P)ATP resulted in the formation of a single acyl ({sup 32}P) phosphoprotein of 120 kDa. Formation of this phosphoprotein was dependent on Ca, and enhanced by La {sup 3+}, characteristic of the plasma membrane CaATPase. Acidic phospholipids, like phosphatidylserine, stimulated Ca transport, similar to their effect on the erythrocyte plasma membrane CaATPase. These results would indicate that the calmodulin-stimulated Ca transport originated in large part from a plasma membrane-type Ca pump of 120 kDa.

  19. Regulation of ascorbic acid biosynthesis and recycling during root development in carrot (Daucus carota L.).

    PubMed

    Wang, Guang-Long; Xu, Zhi-Sheng; Wang, Feng; Li, Meng-Yao; Tan, Guo-Fei; Xiong, Ai-Sheng

    2015-09-01

    Ascorbic acid (AsA), also known as vitamin C, is an essential nutrient in fruits and vegetables. The fleshy root of carrot (Daucus carota L.) is a good source of AsA for humans. However, the metabolic pathways and molecular mechanisms involved in the control of AsA content during root development in carrot have not been elucidated. To gain insights into the regulation of AsA accumulation and to identify the key genes involved in the AsA metabolism, we cloned and analyzed the expression of 21 related genes during carrot root development. The results indicate that AsA accumulation in the carrot root is regulated by intricate pathways, of which the l-galactose pathway may be the major pathway for AsA biosynthesis. Transcript levels of the genes encoding l-galactose-1-phosphate phosphatase and l-galactono-1,4-lactone dehydrogenase were strongly correlated with AsA levels during root development. Data from this research may be used to assist breeding for improved nutrition, quality, and stress tolerance in carrots.

  20. Carrot (Daucus carota L.): Nephroprotective against gentamicin-induced nephrotoxicity in rats

    PubMed Central

    Sodimbaku, Vamsi; Pujari, Latha; Mullangi, Raviteja; Marri, Saisudheer

    2016-01-01

    Objectives: Daucus carota L.(DC) commonly known as carrot, folkorically used as ethnomedicine to treat nephrosis and other urinary disorders. Hence, the present study was aimed to investigate the nephroprotective effects of ethanolic root extract of DC against gentamicin-induced nephrotoxicity in Albino Wistar rats. Methods: Nephrotoxicity in rats was induced by intraperitoneal administration of gentamicin (100 mg/kg/day) for 8 days. Rats of either sex were divided into four groups (n = 6). Group 1 served as control that received normal saline (i.p.) whereas Group 2 (GM) was treated with gentamicin which served as gentamicin-intoxicated group. Group 3–4 (DC200, DC 400) were pretreated with DC at doses of 200 mg/kg and 400 mg/kg (p.o.), respectively, 1 h before the gentamicin intoxication. Following treatment, the nephroprotective effects of DC were evaluated by using serum levels of urea, blood urea nitrogen (BUN), uric acid, and creatinine levels; change in body weight and wet kidney weight along with the histological observations among the experimental groups. Results: Gentamicin intoxication induced elevated serum urea, BUN, uric acid, and creatinine levels which was found to be significantly (P < 0.01) decreased in a dose-dependent manner in groups received DC which was also evidenced by the histological observations. Conclusion: DC showed a significant nephroprotective effect in a dose-dependent manner by ameliorating the gentamicin-induced nephrotoxicity and thus authenticates its ethnomedicinal use. PMID:27127313

  1. Actinorugispora endophytica gen. nov., sp. nov., an actinomycete isolated from Daucus carota.

    PubMed

    Liu, Min-Jiao; Zhu, Wen-Yong; Li, Jie; Zhao, Guo-Zhen; Xiong, Zhi; Park, Dong-Jin; Hozzein, Wael N; Kim, Chang-Jin; Li, Wen-Jun

    2015-08-01

    An actinomycete strain, designated YIM 690008T, was isolated from Daucus carota collected from South Korea and its taxonomic position was investigated by using a polyphasic approach. The strain grew well on most media tested and no diffusible pigment was produced. The aerial mycelium formed wrinkled single spores and short spore chains, some of which were branched. The whole-cell hydrolysates contained meso-diaminopimelic acid, glucose, mannose, ribose, galactose and rhamnose. The predominant menaquinones were MK-10(H4), MK-10(H6), MK-10(H8) and MK-10(H2). The polar lipids were diphosphatidylglycerol, phosphatidylcholine, phosphatidylinositol, phosphatidylinositol mannosides, some unknown phospholipids, glycolipids and polar lipids. The major fatty acids were i-C16 : 0, ai-C17 : 0 and C18 : 1ω9c. The DNA G+C content of the genomic DNA was 63.1 mol%. Phylogenetic analysis indicated that the isolate belongs to the family Nocardiopsaceae. However, based on phenotypic, chemotaxonomic and genotypic data, it was concluded that strain YIM 690008T represents a novel genus and novel species of the family Nocardiopsaceae, for which the name Actinorugispora endophytica gen. nov., sp. nov. (type strain YIM 690008T = DSM 46770T = JCM 30099T = KCTC 29480T) is proposed.

  2. [Influence of arbuscular mycorrhizal inoculation on growth and phoxim residue of carrot (Daucus carota L.)].

    PubMed

    Wang, Fa-Yuan; Chen, Xin; Sun, Xian-Ming; Shi, Zhao-Yong

    2010-12-01

    A pot culture experiment was carried out to study the influence of arbuscular mycorrhizal (AM) fungi on the growth and phoxim residue of carrot (Daucus carota L). Four levels of phoxim (0, 200, 400, 800 mg x L(-1)) and two AM fungal inocula, Glomus intraradices BEG 141(141), Glomus mosseae BEG 167 (167),and one nonmycorrhizal inoculum (CK), were applied to the sterilized soil. The plants were harvested after 5 months of growth and phoxim was irrigated into the root zone 14 d before plant harvest. Although decreasing with the increase of phoxim dosage, root infection rates of all the mycorrhizal plants were higher than 70%. Phoxim showed no significant dose effect on shoot wet weights and root yields, which were all increased by AM inoculation at four phoxim dosages. Phoxim residues in shoots and roots increased with the increase of phoxim dosage, but decreased by AM inoculation. In general, Glomus intraradices BEG 141 showed more pronounced effects on the growth and phoxim residue of carrot than Glomus mosseae BEG 167 did. Our results show a promising potential of AM fungi in carrot production and controlling pesticide residues. PMID:21360902

  3. Abscisic acid regulation of DC8, a carrot embryonic gene. [Daucus carota

    SciTech Connect

    Hatzopoulos, P.; Fong, F.; Sung, Z.R. Texas A M Univ., College Station )

    1990-10-01

    DC8 encodes a hydrophylic 66 kilodalton protein located in the cytoplasm and cell walls of carrot (Daucus carota) embryo and endosperm. During somatic embryogenesis, the levels of DC8 mRNA and protein begin to increase 5 days after removal of auxin. To study the role of abscisic acid (ABA) in the regulation of DC8 gene, fluridone, 1-methyl-3-phenyl,-5(3-trifluoro-methyl-phenyl)-4(1H)-pyridinone, was used to inhibit the endogenous ABA content of the embryos. Fluridone, 50 micrograms per milliliter, effectively inhibits the accumulation of ABA in globular-tage embryos. Western and Northern analysis show that when fluridone is added to the culture medium DC8 protein and mRNA decrease to very low levels. ABA added to fluridone supplemented culture media restores the DC8 protein and mRNA to control levels. Globular-stage embryos contain 0.9 to 1.4 {times} 10{sup {minus}7} molar ABA while 10{sup {minus}6} molar exogenously supplied ABA is the optimal concentration for restoration of DC8 protein accumulation in fluridone-treated embryos. The mRNA level is increased after 15 minutes of ABA addition and reaches maximal levels by 60 minutes. Evidence is presented that, unlike other ABA-regulated genes, DC8 is not induced in nonembryonic tissues via desiccation nor addition of ABA.

  4. Purification and characterization of 3-methylcrotonyl-CoA carboxylase from somatic embryos of Daucus carota.

    PubMed

    Chen, Y; Wurtele, E S; Wang, X; Nikolau, B J

    1993-08-15

    3-Methylcrotonyl-CoA carboxylase, a biotin enzyme, was purified from embryos of Daucus carota. Polyethylene glycol precipitation and monomeric avidin affinity chromatography were used to purify all biotin enzymes from cell-free extracts of embryos. The resulting 3-methylcrotonyl-CoA carboxylase preparation had a specific activity of 745 nmol/min.mg protein, representing a 3725-fold purification of the enzyme and a 135% recovery of activity. Fractionation of the purified biotin-containing proteins by anionic exchange chromatography using Q-Sepharose partially resolved the 3-methylcrotonyl-CoA carboxylase from the other biotin enzymes. 3-Methylcrotonyl-CoA carboxylase has a biotin-containing subunit with a molecular mass of about 78,000 Da and a non-biotin-containing subunit of about 65,000 Da. The native enzyme is 987,000 Da. The optimum pH for activity is between 8.0 and 8.4. The apparent Km values for the substrates 3-methylcrotonyl-CoA, sodium bicarbonate, and ATP are 42 +/- 2 microM, 4.0 +/- 0.9 mM, and 21 +/- 2 microM, respectively. The enzyme is inhibited by acetoacetyl-CoA and palmitoyl-CoA.

  5. Regulation of ascorbic acid biosynthesis and recycling during root development in carrot (Daucus carota L.).

    PubMed

    Wang, Guang-Long; Xu, Zhi-Sheng; Wang, Feng; Li, Meng-Yao; Tan, Guo-Fei; Xiong, Ai-Sheng

    2015-09-01

    Ascorbic acid (AsA), also known as vitamin C, is an essential nutrient in fruits and vegetables. The fleshy root of carrot (Daucus carota L.) is a good source of AsA for humans. However, the metabolic pathways and molecular mechanisms involved in the control of AsA content during root development in carrot have not been elucidated. To gain insights into the regulation of AsA accumulation and to identify the key genes involved in the AsA metabolism, we cloned and analyzed the expression of 21 related genes during carrot root development. The results indicate that AsA accumulation in the carrot root is regulated by intricate pathways, of which the l-galactose pathway may be the major pathway for AsA biosynthesis. Transcript levels of the genes encoding l-galactose-1-phosphate phosphatase and l-galactono-1,4-lactone dehydrogenase were strongly correlated with AsA levels during root development. Data from this research may be used to assist breeding for improved nutrition, quality, and stress tolerance in carrots. PMID:25956452

  6. Daucus carota Pentane/Diethyl Ether Fraction Inhibits Motility and Reduces Invasion of Cancer Cells.

    PubMed

    Zgheib, Perla; Daher, Costantine F; Mroueh, Mohamad; Nasrallah, Anita; Taleb, Robin I; El-Sibai, Mirvat

    2014-01-01

    Daucus carota (DC) is a herb used in folklore medicine in Lebanon to treat numerous diseases including cancer. Recent studies in our laboratory on DC oil and its fractions revealed potent anticancer activities in vitro and in vivo. The present study aims to investigate the effect of the most potent DC fraction, pentane/diethyl ether (50:50), on lung, skin, breast and glioblastoma cancer cell motility and invasion. Upon treatment, a pronounced decrease in cancer cell motility was observed in the 4 cell lines. The treatment also led to a decrease in cancer cell invasion and an increased cell adhesion. Additionally, the DC fraction caused a decrease in the activation of the ρ-GTPases Rac and CDC42, a finding that may partially explain the treatment-induced decrease in cell motility. The current study demonstrates a crucial effect of the DC pentane/diethyl ether fraction on cancer cell motility and metastasis, making it a potential candidate for cancer therapy specifically targeting cancer motility and metastasis. PMID:26088465

  7. Nutritional Content and Antioxidant Properties of Pulp Waste from Daucus carota and Beta vulgaris.

    PubMed

    Shyamala, B N; Jamuna, P

    2010-12-01

    This study reports the chemical composition and antioxidant potential of pulp waste from two vegetables, carrot (Daucus carota) and beetroot (Beta vulgaris). Different in vitro assays used for determining antioxidant potential of extracts of pulp wastes were: 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging capacity, reducing power and total antioxidant activity by phosphomolybdenum method. Total polyphenols, tannins and antioxidative components such as vitamin C, total carotenoids and β-carotene were analysed in the samples. The moisture content of samples ranged from 79 - 84%. The protein content was high in beetroot (13.23 mg/100g) and low in carrot (6.21mg/100g). Total polyphenols were higher in methanol extracts of samples (220-250 mg/100g) compared to ethanol and aqueous extracts. The antioxidant activity determined by the DPPH method exhibited 40% and 78% activity in methanol extracts of carrot and beetroot pulp waste (20 mg) respectively. Overall, the results suggest that carrot and beetroot pulp wastes can be exploited for their nutrients and antioxidant components and used for value addition in food formulations. Hence, these results pave the way for utilisation of bio-wastes from the food industry.

  8. Effects of bioactive compounds from carrots (Daucus carota L.), polyacetylenes, beta-carotene and lutein on human lymphoid leukaemia cells.

    PubMed

    Zaini, Rana G; Brandt, Kirsten; Clench, Malcolm R; Le Maitre, Christine L

    2012-07-01

    New therapies for leukaemia are urgently needed. Carrots have been suggested as a potential treatment for leukaemia in traditional medicine and have previously been studied in other contexts as potential sources of anticancer agents. Indicating that carrots may contain bioactive compounds, which may show potential in leukaemia therapies. This study investigated the effects of five fractions from carrot juice extract (CJE) on human lymphoid leukaemia cell lines, together with five purified bioactive compounds found in Daucus carota L, including: three polyacetylenes (falcarinol, falcarindiol and falcarindiol-3-acetate) and two carotenoids (beta-carotene and lutein). Their effects on induction of apoptosis using Annexin V/PI and Caspase 3 activity assays analysed via flow cytometry and inhibition of cellular proliferation using Cell Titer Glo assay and cell cycle analysis were investigated. Treatment of all three lymphoid leukaemia cell lines with the fraction from carrot extracts which contained polyacetylenes and carotenoids was significantly more cytotoxic than the 4 other fractions. Treatments with purified polyacetylenes also induced apoptosis in a dose and time responsive manner. Moreover, falcarinol and falcarindiol-3-acetate isolated from Daucus carota L were more cytotoxic than falcarindiol. In contrast, the carotenoids showed no significant effect on either apoptosis or cell proliferation in any of the cells investigated. This suggests that polyacetylenes rather than beta-carotene or lutein are the bioactive components found in Daucus carota L and could be useful in the development of new leukemic therapies. Here, for the first time, the cytotoxic effects of polyacetylenes have been shown to be exerted via induction of apoptosis and arrest of cell cycle.

  9. Formation of norisoprenoid flavor compounds in carrot (Daucus carota L.) roots: characterization of a cyclic-specific carotenoid cleavage dioxygenase 1 gene

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Carotenoids are isoprenoid pigments that upon oxidative cleavage lead to the production of norisoprenoids that have profound effect on flavor and aromas of agricultural produce. The biosynthetic pathway to norisoprenoids in carrots (Daucus carota L.) is still widely unknown. We found that geranial i...

  10. Quantification of the relative abundance of plastome to nuclear genome in leaf and root tissues of carrot (Daucus carota L.) using quantitative PCR

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Carrot (Daucus carota L.), is an important horticultural crop with significant health benefits, providing pro-vitamin A carotenoids in the human diet. Carotenoids primarily serve as photoprotectants in leaves during photosynthesis where they accumulate in chloroplasts. Carotenoids can also accumulat...

  11. UDP-Glucose: (1,3)-β-Glucan Synthase from Daucus carota L. 1

    PubMed Central

    Lawson, Stephen G.; Mason, Theresa L.; Sabin, Robert D.; Sloan, Margaret E.; Drake, Richard R.; Haley, Boyd E.; Wasserman, Bruce P.

    1989-01-01

    The membrane-bound UDP-glucose-β-(1,3)-glucan synthase from Daucus carota L. was characterized and a solubilization procedure was developed. The enzyme exhibited maximal activity in the presence of 0.75 millimolar Ca2+, 0.5 millimolar EGTA, and 5 millimolar cellobiose at pH 7.5 and 30°C at 1 millimolar UDPG. Reaction products were confirmed to be (1,3)-linked glucan. Polypeptides of 150, 57, and 43 kilodaltons were labeled with the photoactivatible affinity label 5-azido-uridine 5′-β-[32P] diphosphateglucose. Labeling of the 150 and 57 kilodalton polypeptides was completely protected against by 1 millimolar non-radioactive UDPG suggesting that one or both of these polypeptides may represent the UDPG binding subunit of glucan synthase. Carrot glucan synthase was solubilized with the detergent 3-[(3-cholamidopropyl)dimethylammonio]-1-propane sulfonate (CHAPS) in the absence of divalent cations and chelators; however, the percentage of enzyme which could be solubilized showed variability with membrane source. With microsomal membranes, up to 80% of the enzyme was released with 0.7% CHAPS. Solubilized enzyme was stable for at least 9 hours at 4°C. When more highly purified membrane fractions were isolated from sucrose step gradients a slightly different picture emerged. Activity from the 20/30% interface (Golgi and tonoplast enriched) was readily solubilized and expressed. Activity from the 30/40% interface (plasma membrane enriched) was also solubilized; however, it was necessary to add heat inactivated microsomes to assay mixtures for full activity to be expressed. A requirement for endogenous activators is suggested. Images Figure 2 Figure 5 PMID:16666718

  12. Patterns of Gene Flow between Crop and Wild Carrot, Daucus carota (Apiaceae) in the United States.

    PubMed

    Mandel, Jennifer R; Ramsey, Adam J; Iorizzo, Massimo; Simon, Philipp W

    2016-01-01

    Studies of gene flow between crops and their wild relatives have implications for both management practices for cultivation and understanding the risk of transgene escape. These types of studies may also yield insight into population dynamics and the evolutionary consequences of gene flow for wild relatives of crop species. Moreover, the comparison of genetic markers with different modes of inheritance, or transmission, such as those of the nuclear and chloroplast genomes, can inform the relative risk of transgene escape via pollen versus seed. Here we investigate patterns of gene flow between crop and wild carrot, Daucus carota (Apiaceae) in two regions of the United States. We employed 15 nuclear simple sequence repeat (SSR) markers and one polymorphic chloroplast marker. Further, we utilized both conventional population genetic metrics along with Shannon diversity indices as the latter have been proposed to be more sensitive to allele frequency changes and differentiation. We found that populations in both regions that were proximal to crop fields showed lower levels of differentiation to the crops than populations that were located farther away. We also found that Shannon measures were more sensitive to differences in both genetic diversity and differentiation in our study. Finally, we found indirect evidence of paternal transmission of chloroplast DNA and accompanying lower than expected levels of chloroplast genetic structure amongst populations as might be expected if chloroplast DNA genes flow through both seed and pollen. Our findings of substantial gene flow for both nuclear and chloroplast markers demonstrate the efficiency of both pollen and seed to transfer genetic information amongst populations of carrot. PMID:27603516

  13. Nutritional, physical, and sensory evaluation of hydroponic carrots (Daucus carota L.) from different nutrient delivery systems.

    PubMed

    Gichuhi, P N; Mortley, D; Bromfield, E; Bovell-Benjamin, A C

    2009-01-01

    Carrot (Daucus carota L.) has the highest carotenoid content among foods and is consumed in large quantities worldwide, while at the same time its market demand continues to increase. Carotenoids have also been associated with protective effects against cancer and other chronic diseases. The most predominant carotenoids in carrots are beta- and alpha-carotenes. Moisture, ash, fat, texture, color, carotene content, and consumer acceptance of carrots grown in a hydroponic system with nutrient film technique (NFT) and microporous tube membrane system (MTMS) were evaluated. The moisture contents of the NFT- and MTMS-grown carrots ranged from 86.8 +/- 0.13% to 92.2 +/- 2.25% and 80.9 +/- 0.31% to 91.6 +/- 1.01%, respectively. Fat and ash contents of the carrots were negligible. NFT-grown Oxheart had the most beta-carotene (9900 +/- 20 microg/100 g) while Juwaroot had the least (248 +/- 10 microg/100 g). However, the beta-carotene content of Juwaroot from the NFT batch II carrots was 3842 +/- 6 microg/100 g. MTMS-grown carrots had less variation in the total beta-carotene contents (2434 +/- 89 to 10488 +/- 8 microg/100 g) than those from NFT. Overall, Nantes Touchan (4.8 +/- 2.3) and Nevis-F (7 +/- 1.4) from NFT were the least and most preferred by consumers. Mignon was also acceptable to consumers, and significantly (P < 0.05) more preferred than the other carrots in that NFT batch. MTMS-grown Kinko and Paramex, which were significantly (P < 0.05) more preferred than Nandrin-F and the commercial field-grown carrot, were equally liked by consumers. Nevis-F, Mignon (NFT), Paramex, and Kinko (MTMS) are potentially good cultivars to be included in NASA's food system.

  14. Calcium transport in tonoplast and endoplasmic reticulum vesicles isolated from cultured carrot cells. [Daucus carota Danvers

    SciTech Connect

    Bush, D.R.; Sze, H.

    1986-02-01

    Two active calcium (Ca/sup 2 +/) transport systems have been identified and partially characterized in membrane vesicles isolated from cultured carrot cells (Daucus carota Danvers). Both transport systems required MgATP for activity and were enhanced by 10 millimolar oxalate. Ca/sup 2 +/ transport in membrane vesicles derived from isolated vacuoles equilibrated at 1.10 grams per cubic centimeter and comigrated with Cl/sup -/-stimulated, NO/sub 3//sup -/-inhibited ATPase activity on sucrose density gradients. Ca/sup 2 +/ transport in this system was insensitive to vanadate, but was inhibited by nitrate, carbonyl cyanide-m-chlorophenylhydrazone (CCCP), N,N'-dicyclohexylcarbodiimide (DCCD), and 4,4-diisothiocyano-2,2'-stilbene disulfonic acid (DIDS). The K/sub m/ for MgATP and Ca/sup 2 +/ were 0.1 mM and 21 micromolar, respectively. The predominant Ca/sup 2 +/ transport system detectable in microsomal membrane preparations equilibrated at a density of 1.13 grams per cubic centimeter and comigrated with the endoplasmic reticulum (ER) marker, antimycin A-insensitive NADH-dependent cytochrome c reductase. Ca/sup 2 +/ transport activity and the ER marker also shifted in parallel in ER shifting experiments. This transport system was inhibited by vanadate (I/sub 50/ = 12 micromolar) and was insensitive to nitrate, CCCP, DCCD, and DIDS. Transport exhibited cooperative MgATP dependent kinetics. Ca/sup 2 +/ dependent kinetics were complex with an apparent K/sub m/ ranging from 0.7 to 2 micromolar. We conclude that the vacuolar-derived system is a Ca/sup 2 +//H/sup +/ antiport located on the tonoplast and that the microsomal transport system is a Ca,Mg-ATPase enriched on the ER. These two Ca/sup 2 +/ transport systems are proposed to restore and maintain cytoplasmic Ca/sup 2 +/ homeostasis under changing cellular and environmental conditions.

  15. Combating photooxidative stress in green hairy roots of Daucus carota cultivated under light irradiation.

    PubMed

    Mukherjee, Chiranjit; Sircar, Debabrata; Chatterjee, Moniya; Das, Sampa; Mitra, Adinpunya

    2014-01-15

    The light-dependent generation of active oxygen species, which can disrupt normal metabolic process of plant, is termed as photo-oxidative stress. Plants are equipped with enzymatic and non-enzymatic antioxidative defence system to reduce the effect of such stress. Hairy root culture of Daucus carota when cultivated under continuous illumination (250 μmol m(-2)s(-1)) turned green. To know the reason behind that and photo-oxidative stress response in green hairy roots, activities of several antioxidant enzymes were measured. When compared with normal hairy roots, green hairy roots showed an enhanced superoxide dismutase (SOD) activity. Treatment with a SOD inhibitor diethyldithiocarbamate led to suppression of SOD activity in a concentration-dependent manner in green hairy roots. Interestingly, SOD-suppressed root showed three-fold enhanced caffeic acid glucoside accumulation in the soluble fraction as compared to untreated ones. While ascorbate peroxidase activity showed marginal increase in green hairy roots, a decrease in the activities of guaiacol peroxidase and catalase were observed. SDS-PAGE of crude protein profile from green hairy roots showed a distinct band, which was absent in normal hairy roots. MALDI-TOF-MS/MS analysis of the extracted protein confirmed it as the large subunit of RuBisCO. RT-PCR based expression analysis of betaine aldehyde dehydrogenase showed enhanced transcript levels in green hairy roots as compared to normal hairy roots, whereas reverse trends were observed with the transcripts accumulation for phenylalanine ammonia-lyase and chalcone synthase. These findings corroborate with the in vitro BADH activities in hairy roots, and thus indicate an important role of this stress enzyme in combating photo-oxidative stress in green hairy roots upon continuous light exposure.

  16. Characterisation of polyacetylenes isolated from carrot (Daucus carota) extracts by negative ion tandem mass spectrometry.

    PubMed

    Rai, Dilip K; Brunton, Nigel P; Koidis, Anastasios; Rawson, Ashish; McLoughlin, Padraig; Griffiths, William J

    2011-08-15

    The potential use of negative electrospray ionisation mass spectrometry (ESI-MS) in the characterisation of the three polyacetylenes common in carrots (Daucus carota) has been assessed. The MS scans have demonstrated that the polyacetylenes undergo a modest degree of in-source decomposition in the negative ionisation mode while the positive ionisation mode has shown predominantly sodiated ions and no [M+H](+) ions. Tandem mass spectrometric (MS/MS) studies have shown that the polyacetylenes follow two distinct fragmentation pathways: one that involves cleavage of the C3-C4 bond and the other with cleavage of the C7-C8 bond. The cleavage of the C7-C8 bond generated product ions m/z 105.0 for falcarinol, m/z 105/107.0 for falcarindiol, m/z 147.0/149.1 for falcarindiol-3-acetate. In addition to these product ions, the transitions m/z 243.2 → 187.1 (falcarinol), m/z 259.2 → 203.1 (falcarindiol), m/z 301.2 → 255.2/203.1 (falcarindiol-3-acetate), mostly from the C3-C4 bond cleavage, can form the basis of multiple reaction monitoring (MRM)-quantitative methods which are poorly represented in the literature. The 'MS(3) ' experimental data confirmed a less pronounced homolytic cleavage site between the C11-C12 bond in the falcarinol-type polacetylenes. The optimised liquid chromatography (LC)/MS conditions have achieved a baseline chromatographic separation of the three polyacetylenes investigated within 40 min total run-time.

  17. Patterns of Gene Flow between Crop and Wild Carrot, Daucus carota (Apiaceae) in the United States

    PubMed Central

    Mandel, Jennifer R.; Ramsey, Adam J.; Iorizzo, Massimo; Simon, Philipp W.

    2016-01-01

    Studies of gene flow between crops and their wild relatives have implications for both management practices for cultivation and understanding the risk of transgene escape. These types of studies may also yield insight into population dynamics and the evolutionary consequences of gene flow for wild relatives of crop species. Moreover, the comparison of genetic markers with different modes of inheritance, or transmission, such as those of the nuclear and chloroplast genomes, can inform the relative risk of transgene escape via pollen versus seed. Here we investigate patterns of gene flow between crop and wild carrot, Daucus carota (Apiaceae) in two regions of the United States. We employed 15 nuclear simple sequence repeat (SSR) markers and one polymorphic chloroplast marker. Further, we utilized both conventional population genetic metrics along with Shannon diversity indices as the latter have been proposed to be more sensitive to allele frequency changes and differentiation. We found that populations in both regions that were proximal to crop fields showed lower levels of differentiation to the crops than populations that were located farther away. We also found that Shannon measures were more sensitive to differences in both genetic diversity and differentiation in our study. Finally, we found indirect evidence of paternal transmission of chloroplast DNA and accompanying lower than expected levels of chloroplast genetic structure amongst populations as might be expected if chloroplast DNA genes flow through both seed and pollen. Our findings of substantial gene flow for both nuclear and chloroplast markers demonstrate the efficiency of both pollen and seed to transfer genetic information amongst populations of carrot. PMID:27603516

  18. Patterns of Gene Flow between Crop and Wild Carrot, Daucus carota (Apiaceae) in the United States.

    PubMed

    Mandel, Jennifer R; Ramsey, Adam J; Iorizzo, Massimo; Simon, Philipp W

    2016-01-01

    Studies of gene flow between crops and their wild relatives have implications for both management practices for cultivation and understanding the risk of transgene escape. These types of studies may also yield insight into population dynamics and the evolutionary consequences of gene flow for wild relatives of crop species. Moreover, the comparison of genetic markers with different modes of inheritance, or transmission, such as those of the nuclear and chloroplast genomes, can inform the relative risk of transgene escape via pollen versus seed. Here we investigate patterns of gene flow between crop and wild carrot, Daucus carota (Apiaceae) in two regions of the United States. We employed 15 nuclear simple sequence repeat (SSR) markers and one polymorphic chloroplast marker. Further, we utilized both conventional population genetic metrics along with Shannon diversity indices as the latter have been proposed to be more sensitive to allele frequency changes and differentiation. We found that populations in both regions that were proximal to crop fields showed lower levels of differentiation to the crops than populations that were located farther away. We also found that Shannon measures were more sensitive to differences in both genetic diversity and differentiation in our study. Finally, we found indirect evidence of paternal transmission of chloroplast DNA and accompanying lower than expected levels of chloroplast genetic structure amongst populations as might be expected if chloroplast DNA genes flow through both seed and pollen. Our findings of substantial gene flow for both nuclear and chloroplast markers demonstrate the efficiency of both pollen and seed to transfer genetic information amongst populations of carrot.

  19. Identification and characterization of DcUCGalT1, a galactosyltransferase responsible for anthocyanin galactosylation in purple carrot (Daucus carota L.) taproots.

    PubMed

    Xu, Zhi-Sheng; Ma, Jing; Wang, Feng; Ma, Hong-Yu; Wang, Qiu-Xia; Xiong, Ai-Sheng

    2016-01-01

    Purple carrots (Daucus carota ssp. sativus var. atrorubens Alef.) accumulate large amounts of cyanidin-based anthocyanins in their taproots. Cyanidin can be glycosylated with galactose, xylose, and glucose in sequence by glycosyltransferases resulting in cyanidin 3-xylosyl (glucosyl) galactosides in purple carrots. The first step in the glycosylation of cyanidin is catalysis by UDP-galactose: cyanidin galactosyltransferase (UCGalT) transferring the galactosyl moiety from UDP-galactose to cyanidin. In the present study, a gene from 'Deep purple' carrot, DcUCGalT1, was cloned and heterologously expressed in E. coli BL21 (DE3). The recombinant DcUCGalT1 galactosylated cyanidin to produce cyanidin-3-O-galactoside and showed optimal activity for cyanidin at 30 °C and pH 8.6. It showed lower galactosylation activity for peonidin, pelargonidin, kaempferol and quercetin. It accepted only UDP-galactose as a glycosyl donor when cyanidin was used as an aglycone. The expression level of DcUCGalT1 was positively correlated with anthocyanin biosynthesis in carrots. The enzyme extractions from 'Deep purple' exhibited galactosylation activity for cyanidin, peonidin and pelargonidin, while those from 'Kuroda' (a non-purple cultivar) did not. PMID:27264613

  20. Identification and characterization of DcUCGalT1, a galactosyltransferase responsible for anthocyanin galactosylation in purple carrot (Daucus carota L.) taproots.

    PubMed

    Xu, Zhi-Sheng; Ma, Jing; Wang, Feng; Ma, Hong-Yu; Wang, Qiu-Xia; Xiong, Ai-Sheng

    2016-06-06

    Purple carrots (Daucus carota ssp. sativus var. atrorubens Alef.) accumulate large amounts of cyanidin-based anthocyanins in their taproots. Cyanidin can be glycosylated with galactose, xylose, and glucose in sequence by glycosyltransferases resulting in cyanidin 3-xylosyl (glucosyl) galactosides in purple carrots. The first step in the glycosylation of cyanidin is catalysis by UDP-galactose: cyanidin galactosyltransferase (UCGalT) transferring the galactosyl moiety from UDP-galactose to cyanidin. In the present study, a gene from 'Deep purple' carrot, DcUCGalT1, was cloned and heterologously expressed in E. coli BL21 (DE3). The recombinant DcUCGalT1 galactosylated cyanidin to produce cyanidin-3-O-galactoside and showed optimal activity for cyanidin at 30 °C and pH 8.6. It showed lower galactosylation activity for peonidin, pelargonidin, kaempferol and quercetin. It accepted only UDP-galactose as a glycosyl donor when cyanidin was used as an aglycone. The expression level of DcUCGalT1 was positively correlated with anthocyanin biosynthesis in carrots. The enzyme extractions from 'Deep purple' exhibited galactosylation activity for cyanidin, peonidin and pelargonidin, while those from 'Kuroda' (a non-purple cultivar) did not.

  1. Identification and characterization of DcUCGalT1, a galactosyltransferase responsible for anthocyanin galactosylation in purple carrot (Daucus carota L.) taproots

    PubMed Central

    Xu, Zhi-Sheng; Ma, Jing; Wang, Feng; Ma, Hong-Yu; Wang, Qiu-Xia; Xiong, Ai-Sheng

    2016-01-01

    Purple carrots (Daucus carota ssp. sativus var. atrorubens Alef.) accumulate large amounts of cyanidin-based anthocyanins in their taproots. Cyanidin can be glycosylated with galactose, xylose, and glucose in sequence by glycosyltransferases resulting in cyanidin 3-xylosyl (glucosyl) galactosides in purple carrots. The first step in the glycosylation of cyanidin is catalysis by UDP-galactose: cyanidin galactosyltransferase (UCGalT) transferring the galactosyl moiety from UDP-galactose to cyanidin. In the present study, a gene from ‘Deep purple’ carrot, DcUCGalT1, was cloned and heterologously expressed in E. coli BL21 (DE3). The recombinant DcUCGalT1 galactosylated cyanidin to produce cyanidin-3-O-galactoside and showed optimal activity for cyanidin at 30 °C and pH 8.6. It showed lower galactosylation activity for peonidin, pelargonidin, kaempferol and quercetin. It accepted only UDP-galactose as a glycosyl donor when cyanidin was used as an aglycone. The expression level of DcUCGalT1 was positively correlated with anthocyanin biosynthesis in carrots. The enzyme extractions from ‘Deep purple’ exhibited galactosylation activity for cyanidin, peonidin and pelargonidin, while those from ‘Kuroda’ (a non-purple cultivar) did not. PMID:27264613

  2. Eating Quality of Carrots (Daucus carota L.) Grown in One Conventional and Three Organic Cropping Systems over Three Years.

    PubMed

    Bach, Vibe; Kidmose, Ulla; Kristensen, Hanne L; Edelenbos, Merete

    2015-11-11

    The eating quality of carrots (Daucus carota L.) was investigated to evaluate the impact of cropping systems (one conventional and three organic systems) and growing years (2007, 2008, and 2009) on root size, chemical composition, and sensory quality. The content of dry matter, sugars, polyacetylenes, and terpenes as well as the sensory quality and root size were related to the climate during the three growing years. A higher global radiation and a higher temperature sum in 2009 as compared to 2007 and 2008 resulted in larger roots, higher contents of dry matter, sucrose, total sugars, and total polyacetylenes, and lower contents of terpenes, fructose, and glucose. No differences were found between conventional and organic carrots with regard to the investigated parameters. This result shows that organically grown carrots have the same eating quality as conventionally grown carrots, while being produced in a more sustainable way.

  3. Modeling the transfer of arsenic from soil to carrot (Daucus carota L.)--a greenhouse and field-based study.

    PubMed

    Ding, Changfeng; Zhou, Fen; Li, Xiaogang; Zhang, Taolin; Wang, Xingxiang

    2015-07-01

    Reliable empirical models describing arsenic (As) transfer in soil-plant systems are needed to estimate the human As burden from dietary intake. A greenhouse experiment was conducted in parallel with a field trial located at three sites through China to develop and validate soil-plant transfer models to predict As concentrations in carrot (Daucus carota L.). Stepwise multiple linear regression relationships were based on soil properties and the pseudo total (aqua regia) or available (0.5 M NaHCO3) soil As fractions. Carrot As contents were best predicted by the pseudo total soil As concentrations in combination with soil pH and Fe oxide, with the percentage of variation explained being up to 70 %. The constructed prediction model was further validated and improved to avoid overprotection using data from the field trial. The final obtained model is of great practical relevance to the prediction of As uptake under field conditions. PMID:25750050

  4. Studies on the introduction and mobility of the maize Activator element in Arabidopsis thaliana and Daucus carota.

    PubMed Central

    Van Sluys, M A; Tempé, J; Fedoroff, N

    1987-01-01

    We have co-transformed carrot (Daucus carota) and Arabidopsis thaliana with an Agrobacterium tumefaciens non-tumorigenic T-DNA carrying the maize transposable element Activator (Ac) and an Agrobacterium rhizogenes Ri T-DNA. We present evidence that the Ac element transposes in transformed root or root-derived callus cultures of both species. We show that fertile plants can be regenerated from transformed, root-derived callus cultures of Arabidopsis, demonstrating the utility of the Ri plasmid for introducing the maize Ac element into plants. We also present evidence that Ac elements that excise from the transforming T-DNA early after transformation continue to be mobile in carrot root cultures. Images Fig. 2. Fig. 3. Fig. 4. Fig. 5. Fig. 6. Fig. 7. PMID:2832144

  5. Modeling the transfer of arsenic from soil to carrot (Daucus carota L.)--a greenhouse and field-based study.

    PubMed

    Ding, Changfeng; Zhou, Fen; Li, Xiaogang; Zhang, Taolin; Wang, Xingxiang

    2015-07-01

    Reliable empirical models describing arsenic (As) transfer in soil-plant systems are needed to estimate the human As burden from dietary intake. A greenhouse experiment was conducted in parallel with a field trial located at three sites through China to develop and validate soil-plant transfer models to predict As concentrations in carrot (Daucus carota L.). Stepwise multiple linear regression relationships were based on soil properties and the pseudo total (aqua regia) or available (0.5 M NaHCO3) soil As fractions. Carrot As contents were best predicted by the pseudo total soil As concentrations in combination with soil pH and Fe oxide, with the percentage of variation explained being up to 70 %. The constructed prediction model was further validated and improved to avoid overprotection using data from the field trial. The final obtained model is of great practical relevance to the prediction of As uptake under field conditions.

  6. [Effect of blanching and hygroscopic coating on quality of fresh cut carrots (Daucus carota var. chantenay) during storage].

    PubMed

    Uquiche Carrasco, Edgar; Cisneros-Zevallos, Luis

    2002-06-01

    The effect of blanching at low temperatures (solution 1% acid citric, 50 degrees C for 30 seconds) and the application of glycerol as humectant (3% p/p, 20 seconds) to preserve the quality of fresh-cut carrots (Daucus carota) were studied as individual or combined treatments. Four treatments were evaluated: a control by dipping samples for 30 seconds in distilled water (T1); blanching (T2); glycerol application (T3); and blanching plus glycerol application (T4). Total carotenoids content, color, soluble solids and weight loss were monitored during storage. Results showed no differences between treatments in carotenoids content (p > 0.10) and soluble solids (p > 0.05). However, differences were observed between treatments in weight loss (p < 0.05) and color change (p < 0.05). Blanched samples (T2 and T4) showed small changes in orange color intensity compared to treatments T1 and T3 (p < 0.05).

  7. Eating Quality of Carrots (Daucus carota L.) Grown in One Conventional and Three Organic Cropping Systems over Three Years.

    PubMed

    Bach, Vibe; Kidmose, Ulla; Kristensen, Hanne L; Edelenbos, Merete

    2015-11-11

    The eating quality of carrots (Daucus carota L.) was investigated to evaluate the impact of cropping systems (one conventional and three organic systems) and growing years (2007, 2008, and 2009) on root size, chemical composition, and sensory quality. The content of dry matter, sugars, polyacetylenes, and terpenes as well as the sensory quality and root size were related to the climate during the three growing years. A higher global radiation and a higher temperature sum in 2009 as compared to 2007 and 2008 resulted in larger roots, higher contents of dry matter, sucrose, total sugars, and total polyacetylenes, and lower contents of terpenes, fructose, and glucose. No differences were found between conventional and organic carrots with regard to the investigated parameters. This result shows that organically grown carrots have the same eating quality as conventionally grown carrots, while being produced in a more sustainable way. PMID:26513153

  8. Enhanced accumulation of phytosterols and phenolic compounds in cyclodextrin-elicited cell suspension culture of Daucus carota.

    PubMed

    Miras-Moreno, Begoña; Almagro, Lorena; Pedreño, M A; Sabater-Jara, Ana Belén

    2016-09-01

    In this work, suspension-cultured cells of Daucus carota were used to evaluate the effect of β-cyclodextrins on the production of isoprenoid and phenolic compounds. The results showed that the phytosterols and phenolic compounds were accumulated in the extracellular medium (15100μgL(-1) and 477.46μgL(-1), respectively) in the presence of cyclodextrins. Unlike the phytosterol and phenolic compound content, β-carotene (1138.03μgL(-1)), lutein (25949.54μgL(-1)) and α-tocopherol (8063.82μgL(-1)) chlorophyll a (1625.13μgL(-1)) and b (9.958 (9958.33μgL(-1)) were mainly accumulated inside the cells. Therefore, cyclodextrins were able to induce the cytosolic mevalonate pathway, increasing the biosynthesis of phytosterols and phenolic compounds, and accumulate them outside the cells. However, in the absence of these cyclic oligosaccharidic elicitors, carrot cells mainly accumulated carotenoids through the methylerythritol 4-phosphate pathway. Therefore, the use of cyclodextrins would allow the extracellular accumulation of both phytosterols and phenolic compounds by diverting the carbon flux towards the cytosolic mevalonate/phenylpropanoid pathway. PMID:27457992

  9. Quantifying biochemical quality parameters in carrots (Daucus carota L.) - FT-Raman spectroscopy as efficient tool for rapid metabolite profiling.

    PubMed

    Krähmer, Andrea; Böttcher, Christoph; Rode, Andrea; Nothnagel, Thomas; Schulz, Hartwig

    2016-12-01

    Application of FT-Raman spectroscopy for simultaneous quantification of carotenoids, carbohydrates, polyacetylenes and phenylpropanoids with high bioactive potential was investigated in storage roots of Daucus carota. Within single FT-Raman experiment carbohydrates, carotenoids, and polyacetylenes could be reliably quantified with high coefficients of determination of R(2)>0.91. The most abundant individual representatives of each compound class could be quantified with comparably high quality resulting in R(2)=0.97 and 0.96 for α-carotene and β-carotene, in R(2)=0.90 for falcarindiol (FaDOH), R(2)=0.99, 0.98 and 0.96 for fructose, glucose and sucrose. In contrast, application of FT-Raman spectroscopy for quantification of two laserine-type phenylpropanoids was investigated but failed due to low concentration and Raman response. Furthermore, evaluation of metabolic profiles by principle component analysis (PCA) revealed metabolic variety of carrot root composition depending on root color and botanical relationship. PMID:27374560

  10. Transfer model of lead in soil-carrot (Daucus carota L.) system and food safety thresholds in soil.

    PubMed

    Ding, Changfeng; Li, Xiaogang; Zhang, Taolin; Wang, Xingxiang

    2015-09-01

    Reliable empirical models describing lead (Pb) transfer in soil-plant systems are needed to improve soil environmental quality standards. A greenhouse experiment was conducted to develop soil-plant transfer models to predict Pb concentrations in carrot (Daucus carota L.). Soil thresholds for food safety were then derived inversely using the prediction model in view of the maximum allowable limit for Pb in food. The 2 most important soil properties that influenced carrot Pb uptake factor (ratio of Pb concentration in carrot to that in soil) were soil pH and cation exchange capacity (CEC), as revealed by path analysis. Stepwise multiple linear regression models were based on soil properties and the pseudo total (aqua regia) or extractable (0.01 M CaCl2 and 0.005 M diethylenetriamine pentaacetic acid) soil Pb concentrations. Carrot Pb contents were best explained by the pseudo total soil Pb concentrations in combination with soil pH and CEC, with the percentage of variation explained being up to 93%. The derived soil thresholds based on added Pb (total soil Pb with the geogenic background part subtracted) have the advantage of better applicability to soils with high natural background Pb levels. Validation of the thresholds against data from field trials and literature studies indicated that the proposed thresholds are reasonable and reliable.

  11. Reinvestigation of the bitter compounds in carrots (Daucus carota L.) by using a molecular sensory science approach.

    PubMed

    Schmiech, Ludger; Uemura, Daisuke; Hofmann, Thomas

    2008-11-12

    In order to reinvestigate the key molecules inducing bitter off-taste of carrots ( Daucus carota L.), a sensory-guided fractionation approach was applied to bitter carrot extracts. Besides the previously reported bitter compounds, 6-methoxymellein (1), falcarindiol (2), falcarinol (3), and falcarindiol-3-acetate (4), the following compounds were identified for the first time as bitter compounds in carrots with low bitter recognition thresholds between 8 and 47 micromol/L: vaginatin (5), isovaginatin (6), 2-epilaserine oxide (7), laserine oxide (8), laserine (14), 2-epilaserine (15), 6,8-O-ditigloyl- (9), 6-O-angeloyl-, 8-O-tigloyl- (10), 6-O-tigloyl-, 8-O-angeloyl- (11), and 6-, 8-O-diangeloyl-6 ss,8alpha,11-trihydroxygermacra-1(10) E,4 E-diene (12), as well as 8-O-angeloyl-tovarol (13) and alpha-angeloyloxy-latifolone (16). Among these bitter molecules, compounds 9, 10, 13, and 16 were not previously identified in carrots and compounds 6, 11, and 12 were yet not reported in the literature.

  12. Quantifying biochemical quality parameters in carrots (Daucus carota L.) - FT-Raman spectroscopy as efficient tool for rapid metabolite profiling.

    PubMed

    Krähmer, Andrea; Böttcher, Christoph; Rode, Andrea; Nothnagel, Thomas; Schulz, Hartwig

    2016-12-01

    Application of FT-Raman spectroscopy for simultaneous quantification of carotenoids, carbohydrates, polyacetylenes and phenylpropanoids with high bioactive potential was investigated in storage roots of Daucus carota. Within single FT-Raman experiment carbohydrates, carotenoids, and polyacetylenes could be reliably quantified with high coefficients of determination of R(2)>0.91. The most abundant individual representatives of each compound class could be quantified with comparably high quality resulting in R(2)=0.97 and 0.96 for α-carotene and β-carotene, in R(2)=0.90 for falcarindiol (FaDOH), R(2)=0.99, 0.98 and 0.96 for fructose, glucose and sucrose. In contrast, application of FT-Raman spectroscopy for quantification of two laserine-type phenylpropanoids was investigated but failed due to low concentration and Raman response. Furthermore, evaluation of metabolic profiles by principle component analysis (PCA) revealed metabolic variety of carrot root composition depending on root color and botanical relationship.

  13. Morphological markers for the detection of introgression from cultivated into wild carrot (Daucus carota L.) reveal dominant domestication traits.

    PubMed

    Grebenstein, C; Kos, S P; de Jong, T J; Tamis, W L M; de Snoo, G R

    2013-05-01

    Hybridisation and subsequent introgression have recently received much attention in the context of genetically modified crops. But crop-wild hybrid detection in the field can be difficult, as most domestication traits seem to be recessive, and the hybrid phenotype may also depend on the direction of the cross or environmental factors. Our aim was to develop a reliable set of morphological markers that differ between two wild and 13 cultivated carrots (Daucus carota L.) and to evaluate their inheritance in hybrid lines. We then examined these morphological markers in four F1 hybrids obtained by fertilising plants from the two wild accessions with pollen from two common carrot cultivars. Of the 16 traits that differed between the two carrot subspecies, three took intermediate values in the hybrids, eight resembled the cultivar parent (dominant domestication traits), two resembled the wild parent (domestication traits recessive), and three were not significant or growth condition-dependent. Root:shoot ratio was seven times higher for cultivars than for wild plants, while still attaining equivalent total dry weight, which shows that dry matter production by the shoot is much higher in cultivars than in wild plants. High root:shoot ratios were also present in the hybrids. While we found no maternal effects, the type of cultivar used for pollination had an impact on hybrid characteristics. The morphological markers developed here provide insights into the mode of inheritance of ecologically relevant traits and can be useful for pre-screening wild populations for hybrid detection prior to genetic analysis.

  14. Alternative oxidase (AOX) and phenolic metabolism in methyl jasmonate-treated hairy root cultures of Daucus carota L.

    PubMed

    Sircar, Debabrata; Cardoso, Hélia G; Mukherjee, Chiranjit; Mitra, Adinpunya; Arnholdt-Schmitt, Birgit

    2012-05-01

    Methyl-jasmonate (MJ)-treated hairy roots of Daucus carota L. were used to study the influence of alternative oxidase (AOX) in phenylpropanoid metabolism. Phenolic acid accumulation, as well as total flavonoids and lignin content of the MJ-treated hairy roots were decreased by treatment with salicylhydroxamic acid (SHAM), a known inhibitor of AOX. The inhibitory effect of SHAM was concentration dependent. Treatment with propyl gallate (PG), another inhibitor of AOX, also had a similar inhibitory effect on accumulation of phenolic acid, total flavonoids and lignin. The transcript levels of two DcAOX genes (DcAOX2a and DcAOX1a) were monitored at selected post-elicitation time points. A notable rise in the transcript levels of both DcAOX genes was observed preceding the MJ-induced enhanced accumulation of phenolics, flavonoids and lignin. An appreciable increase in phenylalanine ammonia-lyase (PAL) transcript level was also observed prior to enhanced phenolics accumulation. Both DcAOX genes showed differential transcript accumulation patterns after the onset of elicitation. The transcript levels of DcAOX1a and DcAOX2a attained peak at 6hours post elicitation (hpe) and 12hpe, respectively. An increase in the transcript levels of both DcAOX genes preceding the accumulation of phenylpropanoid-derivatives and lignin showed a positive correlation between AOX activity and phenylpropanoid biosynthesis. The results provide important new insight about the influence of AOX in phenylpropanoid biosynthesis.

  15. Purple carrot (Daucus carota L.) polyacetylenes decrease lipopolysaccharide-induced expression of inflammatory proteins in macrophage and endothelial cells.

    PubMed

    Metzger, Brandon T; Barnes, David M; Reed, Jess D

    2008-05-28

    Carrots ( Daucus carota L.) contain phytochemicals including carotenoids, phenolics, polyacetylenes, isocoumarins, and sesquiterpenes. Purple carrots also contain anthocyanins. The anti-inflammatory activity of extracts and phytochemicals from purple carrots was investigated by determining attenuation of the response to lipopolysaccharide (LPS). A bioactive chromatographic fraction (Sephadex LH-20) reduced LPS inflammatory response. There was a dose-dependent reduction in nitric oxide production and mRNA of pro-inflammatory cytokines (IL-6, IL-1beta, TNF-alpha) and iNOS in macrophage cells. Protein secretions of IL-6 and TNF-alpha were reduced 77 and 66% in porcine aortic endothelial cells treated with 6.6 and 13.3 microg/mL of the LH-20 fraction, respectively. Preparative liquid chromatography resulted in a bioactive subfraction enriched in the polyacetylene compounds falcarindiol, falcarindiol 3-acetate, and falcarinol. The polyacetylenes were isolated and reduced nitric oxide production in macrophage cells by as much as 65% without cytotoxicity. These results suggest that polyacetylenes, not anthocyanins, in purple carrots are responsible for anti-inflammatory bioactivity.

  16. Enhanced accumulation of phytosterols and phenolic compounds in cyclodextrin-elicited cell suspension culture of Daucus carota.

    PubMed

    Miras-Moreno, Begoña; Almagro, Lorena; Pedreño, M A; Sabater-Jara, Ana Belén

    2016-09-01

    In this work, suspension-cultured cells of Daucus carota were used to evaluate the effect of β-cyclodextrins on the production of isoprenoid and phenolic compounds. The results showed that the phytosterols and phenolic compounds were accumulated in the extracellular medium (15100μgL(-1) and 477.46μgL(-1), respectively) in the presence of cyclodextrins. Unlike the phytosterol and phenolic compound content, β-carotene (1138.03μgL(-1)), lutein (25949.54μgL(-1)) and α-tocopherol (8063.82μgL(-1)) chlorophyll a (1625.13μgL(-1)) and b (9.958 (9958.33μgL(-1)) were mainly accumulated inside the cells. Therefore, cyclodextrins were able to induce the cytosolic mevalonate pathway, increasing the biosynthesis of phytosterols and phenolic compounds, and accumulate them outside the cells. However, in the absence of these cyclic oligosaccharidic elicitors, carrot cells mainly accumulated carotenoids through the methylerythritol 4-phosphate pathway. Therefore, the use of cyclodextrins would allow the extracellular accumulation of both phytosterols and phenolic compounds by diverting the carbon flux towards the cytosolic mevalonate/phenylpropanoid pathway.

  17. Cadmium accumulation in the edible parts of different cultivars of radish, Raphanus sativus L., and carrot, Daucus carota var. sativa, grown in a Cd-contaminated soil.

    PubMed

    Zheng, Rui-Lun; Li, Hua-Fen; Jiang, Rong-Feng; Zhang, Fu-Suo

    2008-07-01

    Cadmium accumulation among 12 cultivars of radish (Raphanus sativus) and 10 cultivars of carrot (Daucus carota var. sativa) was studied in a Cd-contaminated soil. The Cd concentration in the edible parts of radish and carrot ranged from 0.04 to 0.14 and 0.14 to 0.19 mg kg(-1) fresh weight, respectively. All the tested carrot cultivars and 33% of the tested radish cultivars exceeded the Chinese allowable limit for Cd. The study showed a greater scope for selecting radish cultivars than for carrot to avoid the excess of the Cd limit when grown on lightly contaminated soils.

  18. Variation of the chemical composition and antimicrobial activity of the essential oils of natural populations of Tunisian Daucus carota L. (Apiaceae).

    PubMed

    Rokbeni, Nesrine; M'rabet, Yassine; Dziri, Salma; Chaabane, Hedia; Jemli, Marwa; Fernandez, Xavier; Boulila, Abdennacer

    2013-12-01

    The essential oils of Daucus carota L. (Apiaceae) seeds sampled from ten wild populations spread over northern Tunisia were characterized by GC-FID and GC/MS analyses. In total, 36 compounds were identified in the D. carota seed essential oils, with a predominance of sesquiterpene hydrocarbons in most samples (22.63-89.93% of the total oil composition). The main volatile compounds identified were β-bisabolene (mean content of 39.33%), sabinene (8.53%), geranyl acetate (7.12%), and elemicin (6.26%). The volatile composition varied significantly across the populations, even for oils of populations harvested in similar areas. The chemometric principal component analysis and the hierarchical clustering identified four groups, each corresponding to a composition-specific chemotype. The in vitro antimicrobial activity of the isolated essential oils was preliminarily evaluated, using the disk-diffusion method, against one Gram-positive (Staphylococcus aureus) and two Gram-negative bacteria (Escherichia coli and Salmonella typhimurium), as well as against a pathogenic yeast (Candida albicans). All tested essential oils exhibited interesting antibacterial and antifungal activities against the assayed microorganisms.

  19. Sensory and health properties of steamed and boiled carrots (Daucus carota ssp. sativus).

    PubMed

    Bongoni, Radhika; Stieger, Markus; Dekker, Matthijs; Steenbekkers, Bea; Verkerk, Ruud

    2014-11-01

    This study examined the influences of domestic processing conditions applied by consumers on firmness, colour and amount of phytochemicals and liking and sensory attributes intensity rating of carrots. The aim was to identify a cooking method and time that yields carrots with higher amount of β-carotene while maintaining consumer liking. Instrumentally measured firmness and colour showed comparable degradation trends between cooking methods. While boiling showed a significant decrease in the amount β-carotene after 20 min (-19%), steaming maintained the amount (+40%). Cooking method did not show a significant effect on liking and intensity ratings for the majority of the sensory attributes. Medium firm carrots were liked the most and low firm carrots the least. This study demonstrates that for optimum liking, carrots should be in the range of medium firmness. This can be obtained through either cooking methods but steamed carrots possess a higher amount of β-carotene and maintains liking.

  20. Prediction model for cadmium transfer from soil to carrot (Daucus carota L.) and its application to derive soil thresholds for food safety.

    PubMed

    Ding, Changfeng; Zhang, Taolin; Wang, Xingxiang; Zhou, Fen; Yang, Yiru; Huang, Guifeng

    2013-10-30

    At present, soil quality standards used for agriculture do not fully consider the influence of soil properties on cadmium (Cd) uptake by crops. This study aimed to develop prediction models for Cd transfer from a wide range of Chinese soils to carrot (Daucus carota L.) using soil properties and the total or available soil Cd content. Path analysis showed soil pH and organic carbon (OC) content were the two most significant properties exhibiting direct effects on Cd uptake factor (ratio of Cd concentration in carrot to that in soil). Stepwise multiple linear regression analysis also showed that total soil Cd, pH, and OC were significant variables contributing to carrot Cd concentration, explaining 90% of the variance across the 21 soils. Soil thresholds for carrot (cultivar New Kuroda) cropping based on added or total Cd were then derived from the food safety standard and were presented as continuous or scenario criteria.

  1. Accumulation of p-hydroxybenzoic acid in hairy roots of Daucus carota 2: confirming biosynthetic steps through feeding of inhibitors and precursors.

    PubMed

    Sircar, Debabrata; Mitra, Adinpunya

    2009-09-01

    Biosynthesis of hydroxybenzoates even at enzymatic level is poorly understood. In this report, effect of feeding of putative biosynthetic precursors and pathway-specific enzyme inhibitors of early phenylpropanoid pathway on p-hydroxybenzoic acid accumulation in chitosan-elicited hairy roots of Daucus carota was studied. Three selective metabolic inhibitors of plant phenylpropanoid pathway, namely, aminooxyacetic acid (AOAA), piperonylic acid (PIP) and 3,4-methylenedioxycinnamic acid (MDCA), which are known to inhibit phenylalanine ammonia-lyase (PAL), cinnamate-4-hydroxylase (C4H) and 4-coumarate-CoA ligase (4CL) respectively, the three early enzymes of phenylpropanoid metabolism, were chosen with the anticipation that selective inhibition of these enzymes in vivo may provide information on the metabolic route to p-hydroxybenzoic acid formation. Supplementation of AOAA (0.2-1.0 mM) and PIP (0.2-1.0 mM) resulted in the reduced accumulation of p-hydroxybenzoic acid in the wall-bound fraction. However, addition of MDCA (0.2-1.25 mM), did not suppress p-hydroxybenzoic acid accumulation but suppressed lignin and total flavonoid accumulation, suggesting that 4CL enzyme activity is not required for p-hydroxybenzoic acid formation. Feeding of elicited hairy roots with phenylalanine, coumaric acid and p-hydroxybenzaldehyde had a stimulatory effect on p-hydroxybenzoic acid accumulation; however, maximum stimulatory effect was shown by p-hydroxybenzaldehyde. This suggests that p-hydroxybenzaldehyde might be the immediate precursor in p-hydroxybenzoic acid biosynthesis. Finally, in vitro conversion of p-coumaric acid to p-hydroxybenzoic acid with p-hydroxybenzaldehyde as intermediate using cell-free extract provided an unequivocal support for CoA-independent and non-beta-oxidative route of p-hydroxybenzoic acid biosynthesis in Daucus carota.

  2. Induction of extracellular defense-related proteins in suspension cultured-cells of Daucus carota elicited with cyclodextrins and methyl jasmonate.

    PubMed

    Sabater-Jara, Ana B; Almagro, Lorena; Pedreño, María A

    2014-04-01

    Suspension cultured-cells (SCC) of Daucus carota were used to evaluate the effect of methyl jasmonate and cyclodextrins, separately or in combination, on the induction of defense responses, particularly the accumulation of pathogenesis-related proteins. A comparative study of the extracellular proteome (secretome) between control and elicited carrot SCC pointed to the presence of amino acid sequences homologous to glycoproteins which have inhibitory activity against the cell-wall-degrading enzymes secreted by pathogens and/or are induced when carrot cells are exposed to a pathogen elicitor. Other amino acid sequences were homologous to Leucine-Rich Repeat domain-containing proteins, which play an essential role in defense against pathogens, as well as in the recognition of microorganisms, making them important players in the innate immunity of this plant. Also, some tryptic peptides were shown to be homologous to a thaumatin-like protein, showing high specificity to abiotic stress and to different reticuline oxidase-like proteins that displayed high levels of antifungal activity, suggesting that methyl jasmonate and cyclodextrins could play a role in mediating defense-related gene product expression in SCC of D. carota. Apart from these elicitor-inducible proteins, we observed the presence of PR-proteins in both control and elicited carrot SCC, suggesting that their expression is mainly constitutive. These PR-proteins are putative class IV chitinases, which also have inhibitory activity against pathogen growth and the class III peroxidases that participate in response to environmental stress (e.g. pathogen attack and oxidative), meaning that they are involved in defense responses triggered by both biotic and abiotic factors.

  3. Influence of Saline Irrigation on Growth, Ion Accumulation and Partitioning, and Leaf Gas Exchange of Carrot (Daucus carota L.)

    PubMed Central

    GIBBERD, MARK R.; TURNER, NEIL C.; STOREY, RICHARD

    2002-01-01

    Like those of many horticultural crop species, the growth and leaf gas exchange responses of carrot (Daucus carota L.) to salinity are poorly understood. In this study ion accumulation in root tissues (periderm, xylem and phloem tissues) and in leaves of different ages was assessed for carrot plants grown in the field with a low level of salinity (5·8 mm Na+ and 7·5 mm Cl–) and in a glasshouse with salinity ranging from 1–80 mm. At low levels of salinity (1–7·5 mm), in both the field and glasshouse, carrot leaves accumulated high concentrations of Cl– (140–200 mm); these appear to be the result of a high affinity for Cl– uptake and a low retention of Cl– in the root system. However, Cl– uptake is under tight control, with an 80‐fold increase in external salinity resulting in only a 1·5‐fold change in the Cl– concentration of the shoot and no increase in the Cl– concentration of the root xylem tissue. In contrast to Cl–, shoot Na+ concentrations were comparatively low (30–40 mm) but increased by seven‐fold when salinity was increased by 80‐fold. Growth over the 56‐d treatment period in the glasshouse was insensitive to salinity less than 20 mm, but at higher concentrations the yield of carrot tap roots declined by 7 % for each 10 mm increase in salinity. At low levels of salinity the accumulation of high concentrations of Cl– (150 mm) in carrot laminae did not appear to limit leaf gas exchange. However, photosynthesis and stomatal conductance were reduced by 38 and 53 %, respectively, for plants grown at a salinity of 80 mm compared with those grown at 1 mm. Salinity‐induced reductions in both pi and carbon isotope discrimination (Δ) were small (2·5 Pa and 1·4 ‰, respectively, at 80 mm) indicating that the reduction in photosynthesis was only marginally influenced by CO2 supply. At a salinity of 80 mm the photosynthetic capacity was reduced, with a 30 % reduction in the CO2‐saturated rate of

  4. Carrot (Daucus carota L.).

    PubMed

    Wally, Owen S D; Punja, Zamir K

    2015-01-01

    Plants are susceptible to infection by a broad range of fungal pathogens. A range of proteins have been evaluated that can enhance tolerance to these pathogens by heterologous expression in transgenic carrot tissues. The protocols for carrot transformation with Arabidopsis NPR1 (Non-Expressor of Pathogenesis-Related Proteins 1) are described in this chapter, using the herbicide resistance gene bar, which encodes phosphinothricin acetyltransferase, as a selectable marker. In this protocol, petiole segments (0.5-1.0 cm long) from aseptically grown carrot seedlings are exposed to Agrobacterium tumefaciens strain LBA4404 for 10-30 min and cocultivated for 2-3 days. Herbicide selection is then imposed for 8-12 weeks on a series of different tissue culture media until embryogenic calli are produced. The transfer of the embryogenic calli to hormone-free medium results in embryo development which eventually gives rise to transgenic plantlets. Embryogenic calli can also be propagated in suspension cultures. This protocol has yielded transgenic carrot plants with defined T-DNA inserts at the rate of between 1 and 3 Southern-positive independent events out of 100.

  5. Differential Contribution of the First Two Enzymes of the MEP Pathway to the Supply of Metabolic Precursors for Carotenoid and Chlorophyll Biosynthesis in Carrot (Daucus carota).

    PubMed

    Simpson, Kevin; Quiroz, Luis F; Rodriguez-Concepción, Manuel; Stange, Claudia R

    2016-01-01

    Carotenoids and chlorophylls are photosynthetic pigments synthesized in plastids from metabolic precursors provided by the methylerythritol 4-phosphate (MEP) pathway. The first two steps in the MEP pathway are catalyzed by the deoxyxylulose 5-phosphate synthase (DXS) and reductoisomerase (DXR) enzymes. While DXS has been recently shown to be the main flux-controlling step of the MEP pathway, both DXS and DXR enzymes have been proven to be able to promote an increase in MEP-derived products when overproduced in diverse plant systems. Carrot (Daucus carota) produces photosynthetic pigments (carotenoids and chlorophylls) in leaves and in light-exposed roots, whereas only carotenoids (mainly α- and β-carotene) accumulate in the storage root in darkness. To evaluate whether DXS and DXR activities influence the production of carotenoids and chlorophylls in carrot leaves and roots, the corresponding Arabidopsis thaliana genes were constitutively expressed in transgenic carrot plants. Our results suggest that DXS is limiting for the production of both carotenoids and chlorophylls in roots and leaves, whereas the regulatory role of DXR appeared to be minor. Interestingly, increased levels of DXS (but not of DXR) resulted in higher transcript abundance of endogenous carrot genes encoding phytoene synthase, the main rate-determining enzyme of the carotenoid pathway. These results support a central role for DXS on modulating the production of MEP-derived precursors to synthesize carotenoids and chlorophylls in carrot, confirming the pivotal relevance of this enzyme to engineer healthier, carotenoid-enriched products. PMID:27630663

  6. Apparent inhibition of. beta. -fructosidase secretion by tunicamycin may be explained by breakdown of the unglycosylated protein during secretion. [Daucus carota

    SciTech Connect

    Faye, L. ); Chrispeels, M.J. )

    1989-03-01

    Suspension-cultured carrot (Daucus carota) cells synthesize and secrete {beta}-fructosidase, a glycoprotein with asparagine-linked glycans. Treatment of the cells with tunicamycin completely inhibits the apparent secretion of {beta}-fructosidase as measured by the accumulation of the {sup 35}S-labelled protein in the cell wall or the culture medium. In the past, such a result has been interpreted as an inhibition of secretion by tunicamycin, but we suggest another explanation based on the following results. In the presence of tunicamycin, unglycosylated {beta}-fructosidase is synthesized and is associated with an endoplasmic-reticulum-rich microsomal fraction. Pulse-chase experiments show that the unglycosylated {beta}-fructosidase does not remain in the cells and appears to be secreted in the same way as glycosylated {beta}-fructosidase; however, no radioactive, unglycosylated {beta}-fructosidase accumulates extracellularly (cell wall or medium). Protoplasts obtained from carrot cells secrete {beta}-fructosidase protein and activity, and treatment of the protoplasts with tunicamycin results in the synthesis of unglycosylated {beta}-fructosidase. In the presence of tunicamycin, there is no accumulation of {beta}-fructosidase activity or unglycosylated {beta}-fructosidase polypeptide in the protoplast incubation medium. These results are consistent with the interpretation that the glycans of {beta}-fructosidase are necessary for its stability, and that in these suspension-cultured cells, the unglycosylated enzyme is degraded during the last stage(s) of secretion, or immediately after its arrival in the wall.

  7. A novel chimeric MOMP antigen expressed in Escherichia coli, Arabidopsis thaliana, and Daucus carota as a potential Chlamydia trachomatis vaccine candidate.

    PubMed

    Kalbina, Irina; Wallin, Anita; Lindh, Ingrid; Engström, Peter; Andersson, Sören; Strid, Ke

    2011-12-01

    The major outer membrane protein (MOMP) of Chlamydia trachomatis is a highly antigenic and hydrophobic transmembrane protein. Our attempts to express the full-length protein in a soluble form in Escherichia coli and in transgenic plants failed. A chimeric gene construct of C. trachomatis serovar E MOMP was designed in order to increase solubility of the MOMP protein but with retained antigenicity. The designed construct was successfully expressed in E. coli, in Arabidopsis thaliana, and in Daucus carota. The chimeric MOMP expressed in and purified from E. coli was used as antigen for production of antibodies in rabbits. The anti-chimeric MOMP antibodies recognized the corresponding protein in both E. coli and in transgenic plants, as well as in inactivated C. trachomatis elementary bodies. Transgenic Arabidopsis and carrots were characterized for the number of MOMP chimeric genetic inserts and for protein expression. Stable integration of the transgene and the corresponding protein expression were demonstrated in Arabidopsis plants over at least six generations. Transgenic carrots showed a high level of expression of the chimeric MOMP - up to 3% of TSP.

  8. Chlorogenic acid biosynthesis: characterization of a light-induced microsomal 5-O-(4-coumaroyl)-D-quinate/shikimate 3'-hydroxylase from carrot (Daucus carota L. ) cell suspension cultures

    SciTech Connect

    Kuehnl, T.K.; Koch, U.; Heller, W.; Wellmann, E.

    1987-10-01

    Microsomal preparations from carrot (Daucus carota L.) cell suspension cultures catalyze the formation of trans-5-O-caffeoyl-D-quinate (chlorogenate) from trans-5-O-(4-coumaroyl)-D-quinate. trans-5-O-(4-Coumaroyl)shikimate is converted to about the same extent to trans-5-O-caffeoylshikimate. trans-4-O-(4-Coumaroyl)-D-quinate, trans-3-O-(4-coumaroyl)-D-quinate, trans-4-coumarate, and cis-5-O-(4-coumaroyl)-D-quinate do not act as substrates. The reaction is strictly dependent on molecular oxygen and on NADPH as reducing cofactor. NADH and ascorbic acid cannot substitute for NADPH. Cytochrome c, Tetcyclacis, and carbon monoxide inhibit the reaction suggesting a cytochrome P-450-dependent mixed-function monooxygenase. Competition experiments as well as induction and inhibition phenomena indicate that there is only one enzyme species which is responsible for the hydroxylation of the 5-O-(4-coumaric) esters of both D-quinate and shikimate. The activity of this enzyme is greatly increased by in vivo irradiation of the cells with blue/uv light. We conclude that the biosynthesis of the predominant caffeic acid conjugates in carrot cells occurs via the corresponding 4-coumaric acid esters. Thus, in this system, 5-O-(4-coumaroyl)-D-quinate can be seen as the final intermediate in the chlorogenic acid pathway.

  9. Differences in protodermal cell wall structure in zygotic and somatic embryos of Daucus carota (L.) cultured on solid and in liquid media.

    PubMed

    Dobrowolska, Izabela; Majchrzak, Oliwia; Baldwin, Timothy C; Kurczynska, Ewa U

    2012-01-01

    The ultrastructure, cuticle, and distribution of pectic epitopes in outer periclinal walls of protodermal cells of Daucus carota zygotic and somatic embryos from solid and suspension culture were investigated. Lipid substances were present as a continuous layer in zygotic and somatic embryos cultured on solid medium. Somatic embryos from suspension cultures were devoid of cuticle. The ultrastructure of the outer walls of protodermis of embryos was similar in zygotic and somatic embryos from solid culture. Fibrillar material was observed on the surface of somatic embryos. In zygotic embryos, in cotyledons and root pectic epitopes recognised by the antibody JIM5 were observed in all cell walls. In hypocotyls of these embryos, these pectic epitopes were not present in the outer periclinal and anticlinal walls of the protodermis. In somatic embryos from solid media, distribution of pectic epitopes recognised by JIM5 was similar to that described for their zygotic counterparts. In somatic embryos from suspension culture, pectic epitopes recognised by JIM5 were detected in all cell walls. In the cotyledons and hypocotyls, a punctate signal was observed on the outside of the protodermis. Pectic epitopes recognised by JIM7 were present in all cell walls independent of embryo organs. In zygotic embryos, this signal was punctate; in somatic embryos from both cultures, this signal was uniformly distributed. In embryos from suspension cultures, a punctate signal was detected outside the surface of cotyledon and hypocotyl. These data are discussed in light of current models for embryogenesis and the influence of culture conditions on cell wall structure.

  10. Influence of variation in soil copper on the yield and nutrition of carrots grown in microplots on two organic soils. [Daucus carota

    SciTech Connect

    Mathur, S.P.; Belanger, A.

    1987-01-01

    Carrots (Daucus carota L. cv. Gold Pak 128) were grown in microplots of two organic soils at site A (a peat), and site B (a muck) in the summer of 1984. The soil surface (0 to 20 cm) varied in total Cu from 13 to 1659, and 81 to 1745 ..mu..g/g at sites A and B, respectively, mainly due to three levels of applications of CuSO/sub 4/ x 5H/sub 2/O in 1978. Neither the yield nor the nutrition (P, K, Ca, Mg, Fe, Mn, Zn, Cu, Mo and B) of the carrot crop was significantly adversely affected by the copper applications or the resultant total soil copper levels at both sites A and B. There were decreases in some foliar nutrient levels due to dilution effects attributable to the significant positive correlations between both root and leaf yields and total soil copper at site B. At both sites A and B, the copper additions appeared to have increased the availability of soil Mn, in accord with earlier evidence.

  11. Formation of norisoprenoid flavor compounds in carrot (Daucus carota L.) roots: characterization of a cyclic-specific carotenoid cleavage dioxygenase 1 gene.

    PubMed

    Yahyaa, Mosaab; Bar, Einat; Dubey, Neeraj Kumar; Meir, Ayala; Davidovich-Rikanati, Rachel; Hirschberg, Joseph; Aly, Radi; Tholl, Dorothea; Simon, Philipp W; Tadmor, Yaakov; Lewinsohn, Efraim; Ibdah, Mwafaq

    2013-12-18

    Carotenoids are isoprenoid pigments that upon oxidative cleavage lead to the production of norisoprenoids that have profound effect on flavor and aromas of agricultural products. The biosynthetic pathway to norisoprenoids in carrots (Daucus carota L.) is still largely unknown. We found the volatile norisoprenoids farnesylacetone, α-ionone, and β-ionone accumulated in Nairobi, Rothild, and Purple Haze cultivars but not in Yellowstone and Creme de Lite in a pattern reflecting their carotenoid content. A cDNA encoding a protein with carotenoid cleavage dioxygenase activity, DcCCD1, was identified in carrot and was overexpressed in Escherichia coli strains previously engineered to produce different carotenoids. The recombinant DcCCD1 enzyme cleaves cyclic carotenes to generate α- and β-ionone. No cleavage products were found when DcCCD1 was co-expressed in E. coli strains accumulating non-cyclic carotenoids, such as phytoene or lycopene. Our results suggest a role for DcCCD1 in carrot flavor biosynthesis.

  12. Differential Contribution of the First Two Enzymes of the MEP Pathway to the Supply of Metabolic Precursors for Carotenoid and Chlorophyll Biosynthesis in Carrot (Daucus carota)

    PubMed Central

    Simpson, Kevin; Quiroz, Luis F.; Rodriguez-Concepción, Manuel; Stange, Claudia R.

    2016-01-01

    Carotenoids and chlorophylls are photosynthetic pigments synthesized in plastids from metabolic precursors provided by the methylerythritol 4-phosphate (MEP) pathway. The first two steps in the MEP pathway are catalyzed by the deoxyxylulose 5-phosphate synthase (DXS) and reductoisomerase (DXR) enzymes. While DXS has been recently shown to be the main flux-controlling step of the MEP pathway, both DXS and DXR enzymes have been proven to be able to promote an increase in MEP-derived products when overproduced in diverse plant systems. Carrot (Daucus carota) produces photosynthetic pigments (carotenoids and chlorophylls) in leaves and in light-exposed roots, whereas only carotenoids (mainly α- and β-carotene) accumulate in the storage root in darkness. To evaluate whether DXS and DXR activities influence the production of carotenoids and chlorophylls in carrot leaves and roots, the corresponding Arabidopsis thaliana genes were constitutively expressed in transgenic carrot plants. Our results suggest that DXS is limiting for the production of both carotenoids and chlorophylls in roots and leaves, whereas the regulatory role of DXR appeared to be minor. Interestingly, increased levels of DXS (but not of DXR) resulted in higher transcript abundance of endogenous carrot genes encoding phytoene synthase, the main rate-determining enzyme of the carotenoid pathway. These results support a central role for DXS on modulating the production of MEP-derived precursors to synthesize carotenoids and chlorophylls in carrot, confirming the pivotal relevance of this enzyme to engineer healthier, carotenoid-enriched products. PMID:27630663

  13. Differential Contribution of the First Two Enzymes of the MEP Pathway to the Supply of Metabolic Precursors for Carotenoid and Chlorophyll Biosynthesis in Carrot (Daucus carota)

    PubMed Central

    Simpson, Kevin; Quiroz, Luis F.; Rodriguez-Concepción, Manuel; Stange, Claudia R.

    2016-01-01

    Carotenoids and chlorophylls are photosynthetic pigments synthesized in plastids from metabolic precursors provided by the methylerythritol 4-phosphate (MEP) pathway. The first two steps in the MEP pathway are catalyzed by the deoxyxylulose 5-phosphate synthase (DXS) and reductoisomerase (DXR) enzymes. While DXS has been recently shown to be the main flux-controlling step of the MEP pathway, both DXS and DXR enzymes have been proven to be able to promote an increase in MEP-derived products when overproduced in diverse plant systems. Carrot (Daucus carota) produces photosynthetic pigments (carotenoids and chlorophylls) in leaves and in light-exposed roots, whereas only carotenoids (mainly α- and β-carotene) accumulate in the storage root in darkness. To evaluate whether DXS and DXR activities influence the production of carotenoids and chlorophylls in carrot leaves and roots, the corresponding Arabidopsis thaliana genes were constitutively expressed in transgenic carrot plants. Our results suggest that DXS is limiting for the production of both carotenoids and chlorophylls in roots and leaves, whereas the regulatory role of DXR appeared to be minor. Interestingly, increased levels of DXS (but not of DXR) resulted in higher transcript abundance of endogenous carrot genes encoding phytoene synthase, the main rate-determining enzyme of the carotenoid pathway. These results support a central role for DXS on modulating the production of MEP-derived precursors to synthesize carotenoids and chlorophylls in carrot, confirming the pivotal relevance of this enzyme to engineer healthier, carotenoid-enriched products.

  14. Differential Contribution of the First Two Enzymes of the MEP Pathway to the Supply of Metabolic Precursors for Carotenoid and Chlorophyll Biosynthesis in Carrot (Daucus carota).

    PubMed

    Simpson, Kevin; Quiroz, Luis F; Rodriguez-Concepción, Manuel; Stange, Claudia R

    2016-01-01

    Carotenoids and chlorophylls are photosynthetic pigments synthesized in plastids from metabolic precursors provided by the methylerythritol 4-phosphate (MEP) pathway. The first two steps in the MEP pathway are catalyzed by the deoxyxylulose 5-phosphate synthase (DXS) and reductoisomerase (DXR) enzymes. While DXS has been recently shown to be the main flux-controlling step of the MEP pathway, both DXS and DXR enzymes have been proven to be able to promote an increase in MEP-derived products when overproduced in diverse plant systems. Carrot (Daucus carota) produces photosynthetic pigments (carotenoids and chlorophylls) in leaves and in light-exposed roots, whereas only carotenoids (mainly α- and β-carotene) accumulate in the storage root in darkness. To evaluate whether DXS and DXR activities influence the production of carotenoids and chlorophylls in carrot leaves and roots, the corresponding Arabidopsis thaliana genes were constitutively expressed in transgenic carrot plants. Our results suggest that DXS is limiting for the production of both carotenoids and chlorophylls in roots and leaves, whereas the regulatory role of DXR appeared to be minor. Interestingly, increased levels of DXS (but not of DXR) resulted in higher transcript abundance of endogenous carrot genes encoding phytoene synthase, the main rate-determining enzyme of the carotenoid pathway. These results support a central role for DXS on modulating the production of MEP-derived precursors to synthesize carotenoids and chlorophylls in carrot, confirming the pivotal relevance of this enzyme to engineer healthier, carotenoid-enriched products.

  15. Effect of endogenous ascorbic acid oxidase activity and stability on vitamin C in carrots (Daucus carota subsp. sativus) during thermal treatment.

    PubMed

    Leong, Sze Ying; Oey, Indrawati

    2012-10-15

    The purpose of this research was to study the effect of endogenous ascorbic acid oxidase (AAO) on vitamin C in carrots (Daucus carota subsp. sativus), namely Nantes, Egmont Gold and baby carrots during thermal treatment. Enzyme-substrate reaction kinetics of AAO were described using Michaelis-Menten equation. The estimated K(m) and V(max) values of AAO ranged from 50.34 to 63.54 μM and 23.70 to 26.82 μmol/min, respectively. Nantes carrots had the lowest AAO activity. On the other hand, Egmont Gold had the highest V(max). AAO activity in all carrot cultivars was stable up to 50 °C and inactivated above 50 °C. Irreversible thermal inactivation of AAO followed first order kinetics (55-70 °C) and the estimated activation energy of the three carrot cultivars situated between 114.33 and 191.45 kJ/mol. Regarding vitamin C stability, thermal treatment at 60-70 °C has resulted in total conversion of l-AA to DHAA due to residual AAO activity; a complete AAO inactivation was found in 80 °C-treated carrots with high vitamin C retention predominantly in l-AA form, up to 90%. On average, the carrots had a total vitamin C content amounting from 368.24 to 379.87 μg/g dry matter and the Nantes carrots had the highest vitamin C content. The effectiveness of rapid inactivation of endogenous AAO via heating (>80 °C, 10 min) prior to matrix disruption gave protection to l-AA towards enzymatic oxidation, thus resulted in a higher vitamin C content and stability in carrots.

  16. Genome-wide analysis of AP2/ERF transcription factors in carrot (Daucus carota L.) reveals evolution and expression profiles under abiotic stress.

    PubMed

    Li, Meng-Yao; Xu, Zhi-Sheng; Huang, Ying; Tian, Chang; Wang, Feng; Xiong, Ai-Sheng

    2015-12-01

    AP2/ERF is a large transcription factor family that regulates plant physiological processes, such as plant development and stress response. Carrot (Daucus carota L.) is an important economical crop with a genome size of 480 Mb; the draft genome sequencing of this crop has been completed by our group. However, little is known about the AP2/ERF factors in carrot. In this study, a total of 267 putative AP2/ERF factors were identified from the whole-genome sequence of carrot. These AP2/ERF proteins were phylogenetically clustered into five subfamilies based on their similarity to the amino acid sequences from Arabidopsis. The distribution and comparative genome analysis of the AP2/ERF factors among plants showed the AP2/ERF factors had expansion during the evolutionary process, and the AP2 domain was highly conserved during evolution. The number of AP2/ERF factors in land plants expanded during their evolution. A total of 60 orthologous and 145 coorthologous AP2/ERF gene pairs between carrot and Arabidopsis were identified, and the interaction network of orthologous genes was constructed. The expression patterns of eight AP2/ERF family genes from each subfamily (DREB, ERF, AP2, and RAV) were related to abiotic stresses. Yeast one-hybrid and β-galactosidase activity assays confirmed the DRE and GCC box-binding activities of DREB subfamily genes. This study is the first to identify and characterize the AP2/ERF transcription factors in carrot using whole-genome analysis, and the findings may serve as references for future functional research on the transcription factors in carrot.

  17. Effect of pulsed electric field treatment on enzyme kinetics and thermostability of endogenous ascorbic acid oxidase in carrots (Daucus carota cv. Nantes).

    PubMed

    Leong, Sze Ying; Oey, Indrawati

    2014-03-01

    The objective of this research was to study the enzyme kinetics and thermostability of endogenous ascorbic acid oxidase (AAO) in carrot purée (Daucus carota cv. Nantes) after being treated with pulsed electric field (PEF) processing. Various PEF treatments using electric field strength between 0.2 and 1.2kV/cm and pulsed electrical energy between 1 and 520kJ/kg were conducted. The enzyme kinetics and the kinetics of AAO thermal inactivation (55-70°C) were described using Michaelis-Menten model and first order reaction model, respectively. Overall, the estimated Vmax and KM values were situated in the same order of magnitude as the untreated carrot purée after being exposed to pulsed electrical energy between 1 and 400kJ/kg, but slightly changed at pulsed electrical energy above 500kJ/kg. However, AAO presented different thermostability depending on the electric field strength applied. After PEF treatment at the electric field strength between 0.2 and 0.5kV/cm, AAO became thermolabile (i.e. increase in inactivation rate (k value) at reference temperature) but the temperature dependence of k value (Ea value) for AAO inactivation in carrot purée decreased, indicating that the changes in k values were less temperature dependent. It is obvious that PEF treatment affects the temperature stability of endogenous AAO. The changes in enzyme kinetics and thermostability of AAO in carrot purée could be related to the resulting carrot purée composition, alteration in intracellular environment and the effective concentration of AAO released after being subjected to PEF treatment.

  18. Genome-wide analysis of AP2/ERF transcription factors in carrot (Daucus carota L.) reveals evolution and expression profiles under abiotic stress.

    PubMed

    Li, Meng-Yao; Xu, Zhi-Sheng; Huang, Ying; Tian, Chang; Wang, Feng; Xiong, Ai-Sheng

    2015-12-01

    AP2/ERF is a large transcription factor family that regulates plant physiological processes, such as plant development and stress response. Carrot (Daucus carota L.) is an important economical crop with a genome size of 480 Mb; the draft genome sequencing of this crop has been completed by our group. However, little is known about the AP2/ERF factors in carrot. In this study, a total of 267 putative AP2/ERF factors were identified from the whole-genome sequence of carrot. These AP2/ERF proteins were phylogenetically clustered into five subfamilies based on their similarity to the amino acid sequences from Arabidopsis. The distribution and comparative genome analysis of the AP2/ERF factors among plants showed the AP2/ERF factors had expansion during the evolutionary process, and the AP2 domain was highly conserved during evolution. The number of AP2/ERF factors in land plants expanded during their evolution. A total of 60 orthologous and 145 coorthologous AP2/ERF gene pairs between carrot and Arabidopsis were identified, and the interaction network of orthologous genes was constructed. The expression patterns of eight AP2/ERF family genes from each subfamily (DREB, ERF, AP2, and RAV) were related to abiotic stresses. Yeast one-hybrid and β-galactosidase activity assays confirmed the DRE and GCC box-binding activities of DREB subfamily genes. This study is the first to identify and characterize the AP2/ERF transcription factors in carrot using whole-genome analysis, and the findings may serve as references for future functional research on the transcription factors in carrot. PMID:25971861

  19. Projected Dietary Intake of Zinc, Copper, and Cerium from Consumption of Carrot (Daucus carota) Exposed to Metal Oxide Nanoparticles or Metal Ions.

    PubMed

    Ebbs, Stephen D; Bradfield, Scott J; Kumar, Pawan; White, Jason C; Ma, Xingmao

    2016-01-01

    The expanding production and use of engineered nanomaterials (ENMs) have raised concerns about the potential risk of those materials to food safety and human health. In a prior study, the accumulation of Zn, Cu, and Ce from ZnO, CuO, or CeO2, respectively, was examined in carrot (Daucus carota L.) grown in sand culture in comparison to accumulation from exposure to equivalent concentrations of ionic Zn(2+), Cu(2+), or Ce(4+). The fresh weight concentration data for peeled and unpeeled carrots were used to project dietary intake of each metal by seven age-mass classes from child to adult based on consumption of a single serving of carrot. Dietary intake was compared to the oral reference dose (oral RfD) for chronic toxicity for Zn or Cu and estimated mean and median oral RfD values for Ce based on nine other rare earth elements. Reverse dietary intake calculations were also conducted to estimate the number of servings of carrot, the mass of carrot consumed, or the tissue concentration of Zn, Cu, or Ce that would cause the oral RfD to be exceeded upon consumption. The projections indicated for Zn and Cu, the oral RfD would be exceeded in only a few highly unrealistic scenarios of exceedingly high Zn or Cu concentrations in the substrate from ZnO or CuO or consumption of excessive amounts of unpeeled carrot. The implications associated with the presence of Ce in the carrot tissues depended upon whether the mean or median oral RfD value from the rare earth elements was used as a basis for comparison. The calculations further indicated that peeling carrots reduced the projected dietary intake by one to two orders of magnitude for both ENM- and ionic-treated carrots. Overall in terms of total metal concentration, the results suggested no specific impact of the ENM form on dietary intake. The effort here provided a conservative view of the potential dietary intake of these three metals that might result from consumption of carrots exposed to nanomaterials (NMs) and how

  20. A plasma membrane-type Ca[sup 2+]-ATPase of 120 kilodaltons on the endoplasmic reticulum from carrot (Daucus carota) cells

    SciTech Connect

    Chen, F.H.; Ratterman, D.M.; Sze, H. )

    1993-06-01

    Cytosolic Ca[sup 2+] levels are regulated in part by Ca[sup 2+]-pumping ATPases that export Ca[sup 2+] from the cytoplasm; The types and properties of Ca[sup 2+] pumps in plants are not well understood. The kinetic properties of a 120-kD phosphoenzyme (PE) intermediate formed during the reaction cycle of a Ca[sup 2+]-ATPase from suspension-cultured carrot (Daucus carota) cells are characterized. Only one Ca[sup 2+]-dependent phosphoprotein was formed when carrot membrane vesicles were incubated with [[gamma]-[sup 32]P]ATP. Formation of this 120-kD phosphoprotein was inhibited by vanadate, enhanced by La[sup 3+], and decreased by hydroxylamine, confirming its identification as an intermediate of a phosphorylated-type Ca[sup 2+]-translocating ATPase. The 120-kD Ca[sup 2+]-ATPase was most abundant in endoplasmic reticulum-enriched fractions, in which the Ca[sup 2+]-ATPase was estimated to be 0.1% of membrane protein. Direct quantitation of Ca[sup 2+]-dependent phosphoprotein was used to examine the kinetics of PE formation. PE formation exhibited a K[sub m] for Ca[sup 2+] of 1 to 2 [mu]m and a K[sub m] for ATP of 67 nm. Relative affinities of substrates, determined by competition experiments, were 0.075 [mu]m for ATP, 1 [mu]m for ADP, 100 [mu]m for ITP, and 250 [mu]m for GTP. Thapsigargin and cyclopiazonic acid, specific inhibitors of animal sarcoplasmic/endoplasmic reticulum Ca[sup 2+]-ATPase, had no effect on PE formation; erythrosin B inhibited with 50% inhibition at <0.1 [mu]m. Calmodulin (1 [mu]m) stimulated PE formation by 25%. The results indicate that the carrot 120-kD Ca[sup 2+]-ATPase is similar but not identical to animal plasma membrane-type Ca[sup 2+]-ATPase and yet is located on endomembranes, such as the endoplasmic reticulum. This type of Ca[sup 2+] pump may reside on the cortical endoplasmic reticulum, thought to play a major role in anchoring the cytoskeleton and in facilitating secretion. 34 refs., 9 figs., 3 tabs.

  1. Projected Dietary Intake of Zinc, Copper, and Cerium from Consumption of Carrot (Daucus carota) Exposed to Metal Oxide Nanoparticles or Metal Ions.

    PubMed

    Ebbs, Stephen D; Bradfield, Scott J; Kumar, Pawan; White, Jason C; Ma, Xingmao

    2016-01-01

    The expanding production and use of engineered nanomaterials (ENMs) have raised concerns about the potential risk of those materials to food safety and human health. In a prior study, the accumulation of Zn, Cu, and Ce from ZnO, CuO, or CeO2, respectively, was examined in carrot (Daucus carota L.) grown in sand culture in comparison to accumulation from exposure to equivalent concentrations of ionic Zn(2+), Cu(2+), or Ce(4+). The fresh weight concentration data for peeled and unpeeled carrots were used to project dietary intake of each metal by seven age-mass classes from child to adult based on consumption of a single serving of carrot. Dietary intake was compared to the oral reference dose (oral RfD) for chronic toxicity for Zn or Cu and estimated mean and median oral RfD values for Ce based on nine other rare earth elements. Reverse dietary intake calculations were also conducted to estimate the number of servings of carrot, the mass of carrot consumed, or the tissue concentration of Zn, Cu, or Ce that would cause the oral RfD to be exceeded upon consumption. The projections indicated for Zn and Cu, the oral RfD would be exceeded in only a few highly unrealistic scenarios of exceedingly high Zn or Cu concentrations in the substrate from ZnO or CuO or consumption of excessive amounts of unpeeled carrot. The implications associated with the presence of Ce in the carrot tissues depended upon whether the mean or median oral RfD value from the rare earth elements was used as a basis for comparison. The calculations further indicated that peeling carrots reduced the projected dietary intake by one to two orders of magnitude for both ENM- and ionic-treated carrots. Overall in terms of total metal concentration, the results suggested no specific impact of the ENM form on dietary intake. The effort here provided a conservative view of the potential dietary intake of these three metals that might result from consumption of carrots exposed to nanomaterials (NMs) and how

  2. Projected Dietary Intake of Zinc, Copper, and Cerium from Consumption of Carrot (Daucus carota) Exposed to Metal Oxide Nanoparticles or Metal Ions

    PubMed Central

    Ebbs, Stephen D.; Bradfield, Scott J.; Kumar, Pawan; White, Jason C.; Ma, Xingmao

    2016-01-01

    The expanding production and use of engineered nanomaterials (ENMs) have raised concerns about the potential risk of those materials to food safety and human health. In a prior study, the accumulation of Zn, Cu, and Ce from ZnO, CuO, or CeO2, respectively, was examined in carrot (Daucus carota L.) grown in sand culture in comparison to accumulation from exposure to equivalent concentrations of ionic Zn2+, Cu2+, or Ce4+. The fresh weight concentration data for peeled and unpeeled carrots were used to project dietary intake of each metal by seven age-mass classes from child to adult based on consumption of a single serving of carrot. Dietary intake was compared to the oral reference dose (oral RfD) for chronic toxicity for Zn or Cu and estimated mean and median oral RfD values for Ce based on nine other rare earth elements. Reverse dietary intake calculations were also conducted to estimate the number of servings of carrot, the mass of carrot consumed, or the tissue concentration of Zn, Cu, or Ce that would cause the oral RfD to be exceeded upon consumption. The projections indicated for Zn and Cu, the oral RfD would be exceeded in only a few highly unrealistic scenarios of exceedingly high Zn or Cu concentrations in the substrate from ZnO or CuO or consumption of excessive amounts of unpeeled carrot. The implications associated with the presence of Ce in the carrot tissues depended upon whether the mean or median oral RfD value from the rare earth elements was used as a basis for comparison. The calculations further indicated that peeling carrots reduced the projected dietary intake by one to two orders of magnitude for both ENM- and ionic-treated carrots. Overall in terms of total metal concentration, the results suggested no specific impact of the ENM form on dietary intake. The effort here provided a conservative view of the potential dietary intake of these three metals that might result from consumption of carrots exposed to nanomaterials (NMs) and how peeling

  3. Competitive inhibition and selectivity enhancement by Ca in the uptake of inorganic elements (Be, Na, Mg, K, Ca, Sc, Mn, Co, Zn, Se, Rb, Sr, Y, Zr, Ce, Pm, Gd, Hf) by carrot (Daucus carota cv. U.S. harumakigosun).

    PubMed

    Ozaki, Takuo; Ambe, Shizuko; Abe, Tomoko; Francis, Arokiasamy J

    2005-01-01

    We investigated the uptake of inorganic elements (Be, Na, Mg, K, Ca, Sc, Mn, Co, Zn, Se, Rb, Sr, Y, Zr, Ce, Pm, Gd, and Hf) and the effect of Ca on their uptake in carrots (Daucus carota cv. U.S. harumakigosun) by the radioactive multitracer technique. The experimental results suggested that Na, Mg, K, and Rb competed for the functional groups outside the cells in roots with Ca but not for the transporter-binding sites on the plasma membrane of the root cortex cells. In contrast, Y, Ce, Pm, and Gd competed with Ca for the transporters on the plasma membrane. The selectivity, which was defined as the value obtained by dividing the concentration ratio of an elemental pair, K/Na, Rb/Na, Be/Sr, and Mg/Sr, in the presence of 0.2 and 2 ppm Ca by that of the corresponding elemental pair in the absence of Ca in the solution was estimated. The selectivity of K and Rb in roots was increased in the presence of Ca. The selectivity of Be in roots was not affected, whereas the selectivity of Mg was increased by Ca. These observations suggest that the presence of Ca in the uptake solution enhances the selectivity in the uptake of metabolically important elements against unwanted elements. PMID:15695849

  4. Production of the p24 capsid protein from HIV-1 subtype C in Arabidopsis thaliana and Daucus carota using an endoplasmic reticulum-directing SEKDEL sequence in protein expression constructs.

    PubMed

    Lindh, Ingrid; Wallin, Anita; Kalbina, Irina; Sävenstrand, Helena; Engström, Peter; Andersson, Sören; Strid, Ake

    2009-07-01

    An optimized gene expression construct was designed in order to increase the accumulation of the HIV-1 subtype C p24 protein in Arabidopsis thaliana and carrot (Daucus carota) plants. An ER retention signal was introduced into the genetic construct generating a p24 protein containing a SEKDEL amino acid sequence at its C-terminus. Mature A. thaliana plants and carrot cells were transformed using Agrobacterium tumefaciens carrying the improved pGreen0229/p24_SEKDEL vector. Several transgenic plant lines were obtained from both plant species by growth on selective medium and confirmed by PCR. Transformed lines were analyzed for p24 protein content by western blotting using anti-p24-specific antibodies and by Southern blotting to establish the number of copies of the insert in the plant nuclear genome. To estimate the accumulation levels of p24 protein in the plants, ELISA was run using soluble plant extracts. By comparing these results with our previous findings, the ER retention signal increased the level of p24 protein fivefold in the A. thaliana plants. In carrot taproot, the content of p24_SEKDEL protein was approximately half of that in Arabidopsis on a fresh weight basis and was stable in planta for several months. However, on a total soluble protein basis, carrots produced considerable higher levels of the p24_SEKDEL protein than Arabidopsis.

  5. Investigating the performance of in situ quantitative nuclear magnetic resonance analysis and applying the method to determine the distribution of saccharides in various parts of carrot roots (Daucus carota L.).

    PubMed

    Bauchard, Elsa; This, Hervé

    2015-01-01

    In order to explore the performance of the analytical method called in situ quantitative nuclear magnetic resonance spectroscopy - is q NMR - the distribution of glucose, fructose and sucrose in various parts of a carrot root (Daucus carota L.) - primary xylem, secondary xylem, phloem, cortex; top part and lower part - was determined. The influence on the quality of spectra of drying samples before analysis was studied, as well as the influence of the length of strips of tissue used in analysis. Finally samples as small as 240 mm(3) could be studied directly, with minimum prior treatment (only drying), along with deuterated water for locking and a sealed capillary tube containing a solution of 0.5% of the sodium salt of (trimethylsilyl)propionic-2,2,3,3-d4 acid, used both as an internal reference and for quantification. With optimized parameters, the coefficients of variation for measurements were observed to have an average value of 0.038, with a standard deviation of 0.047.

  6. Dissimilarity of contemporary and historical gene flow in a wild carrot (Daucus carota) metapopulation under contrasting levels of human disturbance: implications for risk assessment and management of transgene introgression

    PubMed Central

    Rong, Jun; Xu, Shuhua; Meirmans, Patrick G.; Vrieling, Klaas

    2013-01-01

    Background and Aims Transgene introgression from crops into wild relatives may increase the resistance of wild plants to herbicides, insects, etc. The chance of transgene introgression depends not only on the rate of hybridization and the establishment of hybrids in local wild populations, but also on the metapopulation dynamics of the wild relative. The aim of the study was to estimate gene flow in a metapopulation for assessing and managing the risks of transgene introgression. Methods Wild carrots (Daucus carota) were sampled from 12 patches in a metapopulation. Eleven microsatellites were used to genotype wild carrots. Genetic structure was estimated based on the FST statistic. Contemporary (over the last several generations) and historical (over many generations) gene flow was estimated with assignment and coalescent methods, respectively. Key Results The genetic structure in the wild carrot metapopulation was moderate (FST = 0·082) and most of the genetic variation resided within patches. A pattern of isolation by distance was detected, suggesting that most of the gene flow occurred between neighbouring patches (≤1 km). The mean contemporary gene flow was 5 times higher than the historical estimate, and the correlation between them was very low. Moreover, the contemporary gene flow in roadsides was twice that in a nature reserve, and the correlation between contemporary and historical estimates was much higher in the nature reserve. Mowing of roadsides may contribute to the increase in contemporary gene flow. Simulations demonstrated that the higher contemporary gene flow could accelerate the process of transgene introgression in the metapopulation. Conclusions Human disturbance such as mowing may alter gene flow patterns in wild populations, affecting the metapopulation dynamics of wild plants and the processes of transgene introgression in the metapopulation. The risk assessment and management of transgene introgression and the control of weeds need to

  7. Localization of Daucus carota NMCP1 to the nuclear periphery: the role of the N-terminal region and an NLS-linked sequence motif, RYNLRR, in the tail domain.

    PubMed

    Kimura, Yuta; Fujino, Kaien; Ogawa, Kana; Masuda, Kiyoshi

    2014-01-01

    Recent ultrastructural studies revealed that a structure similar to the vertebrate nuclear lamina exists in the nuclei of higher plants. However, plant genomes lack genes for lamins and intermediate-type filament proteins, and this suggests that plant-specific nuclear coiled-coil proteins make up the lamina-like structure in plants. NMCP1 is a protein, first identified in Daucus carota cells, that localizes exclusively to the nuclear periphery in interphase cells. It has a tripartite structure comprised of head, rod, and tail domains, and includes putative nuclear localization signal (NLS) motifs. We identified the functional NLS of DcNMCP1 (carrot NMCP1) and determined the protein regions required for localizing to the nuclear periphery using EGFP-fused constructs transiently expressed in Apium graveolens epidermal cells. Transcription was driven under a CaMV35S promoter, and the genes were introduced into the epidermal cells by a DNA-coated microprojectile delivery system. Of the NLS motifs, KRRRK and RRHK in the tail domain were highly functional for nuclear localization. Addition of the N-terminal 141 amino acids from DcNMCP1 shifted the localization of a region including these NLSs from the entire nucleus to the nuclear periphery. Using this same construct, the replacement of amino acids in RRHK or its preceding sequence, YNL, with alanine residues abolished localization to the nuclear periphery, while replacement of KRRRK did not affect localization. The sequence R/Q/HYNLRR/H, including YNL and the first part of the sequence of RRHK, is evolutionarily conserved in a subclass of NMCP1 sequences from many plant species. These results show that NMCP1 localizes to the nuclear periphery by a combined action of a sequence composed of R/Q/HYNLRR/H, NLS, and the N-terminal region including the head and a portion of the rod domain, suggesting that more than one binding site is implicated in localization of NMCP1.

  8. [Granulated carrots (Daucus carota) in dog nutrition].

    PubMed

    Zentek, J; Meyer, H

    1993-01-01

    In feeding and balance trials with dogs (4) dried, ground carrots were tested in addition (30%) to a basic dry food (rice, corn gluten, fish meal and supplements). The palatability of both mixtures was good: fecal water losses were increased after feeding the carrots. Apparent digestibilities of organic and inorganic matter, especially sodium and potassium (not phosphorus), decreased. Digestibility of carrots was calculated to reach 70% with 1.2 MJ of digestible energy per 100 g dry matter. Reduced fecal pH (6.9-->6.1), higher lactate concentrations and increasing excretion of volatile fatty acids were seen as results of the stimulated bacterial metabolism in the colon. Diurnal pattern of breath hydrogen exhalation was similar in both periods: renal excretions of phenol, indican, and nitrogen were significantly reduced after supplementation with carrots as compared to control period.

  9. Molecular phylogeny of Daucus (Apiaceae)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    We studied the phylogeny of 22 accessions of Daucus: D. broteri, D. capillifolius, D. carota, D. carota subsp. carota, D. carota subsp. commutatus, D. carota subsp. commutatus, D. carota subsp. drepanensis, D. carota subsp. gadecaei, D. carota subsp. gummifer, D. carota subsp. halophilius, D. carota...

  10. Pervasive Mitochondrial Sequence Heteroplasmy in Natural Populations of Wild Carrot, Daucus carota spp. carota L.

    PubMed

    Mandel, Jennifer R; McCauley, David E

    2015-01-01

    Exceptions to the generally accepted rules that plant mitochondrial genomes are strictly maternally inherited and that within-individual sequence diversity in those genomes, i.e., heteroplasmy, should be minimal are becoming increasingly apparent especially with regard to sequence-level heteroplasmy. These findings raise questions about the potential significance of such heteroplasmy for plant mitochondrial genome evolution. Still studies quantifying the amount and consequences of sequence heteroplasmy in natural populations are rare. In this study, we report pervasive sequence heteroplasmy in natural populations of wild carrot, a close relative of the cultivated crop. In order to assay directly for this heteroplasmy, we implemented a quantitative PCR assay that can detect and quantify intra-individual SNP variation in two mitochondrial genes (Cox1 and Atp9). We found heteroplasmy in > 60% of all wild carrot populations surveyed and in > 30% of the 140 component individuals that were genotyped. Heteroplasmy ranged from a very small proportion of the total genotype (e.g., 0.995:0.005) to near even mixtures (e.g., 0.590:0.410) in some individuals. These results have important implications for the role of intra-genomic recombination in the generation of plant mitochondrial genome genotypic novelty. The consequences of such recombination are evident in the results of this study through analysis of the degree of linkage disequilibrium (LD) between the SNP sites at the two genes studied.

  11. Morphogenetic responses of cultured totipotent cells of carrot /Daucus carota var. carota/ at zero gravity

    NASA Technical Reports Server (NTRS)

    Krikorian, A. D.; Steward, F. C.

    1978-01-01

    An experiment designed to test whether embryos capable of developing from isolated somatic carrot cells could do so under conditions of weightlessness in space was performed aboard the unmanned Soviet biosatellite Kosmos 782 under the auspices of the joint United States-Soviet Biological Satellite Mission. Space flight and weightlessness seem to have had no adverse effects on the induction of embryoids or on the development of their organs. A portion of the crop of carrot plantlets originated in space and grown to maturity were not morphologically different from controls.

  12. Pervasive Mitochondrial Sequence Heteroplasmy in Natural Populations of Wild Carrot, Daucus carota spp. carota L.

    PubMed

    Mandel, Jennifer R; McCauley, David E

    2015-01-01

    Exceptions to the generally accepted rules that plant mitochondrial genomes are strictly maternally inherited and that within-individual sequence diversity in those genomes, i.e., heteroplasmy, should be minimal are becoming increasingly apparent especially with regard to sequence-level heteroplasmy. These findings raise questions about the potential significance of such heteroplasmy for plant mitochondrial genome evolution. Still studies quantifying the amount and consequences of sequence heteroplasmy in natural populations are rare. In this study, we report pervasive sequence heteroplasmy in natural populations of wild carrot, a close relative of the cultivated crop. In order to assay directly for this heteroplasmy, we implemented a quantitative PCR assay that can detect and quantify intra-individual SNP variation in two mitochondrial genes (Cox1 and Atp9). We found heteroplasmy in > 60% of all wild carrot populations surveyed and in > 30% of the 140 component individuals that were genotyped. Heteroplasmy ranged from a very small proportion of the total genotype (e.g., 0.995:0.005) to near even mixtures (e.g., 0.590:0.410) in some individuals. These results have important implications for the role of intra-genomic recombination in the generation of plant mitochondrial genome genotypic novelty. The consequences of such recombination are evident in the results of this study through analysis of the degree of linkage disequilibrium (LD) between the SNP sites at the two genes studied. PMID:26295342

  13. Carryover of perfluorooctanoic acid (PFOA) and perfluorooctane sulfonate (PFOS) from soil to plant and distribution to the different plant compartments studied in cultures of carrots (Daucus carota ssp. Sativus), potatoes (Solanum tuberosum), and cucumbers (Cucumis Sativus).

    PubMed

    Lechner, Mareike; Knapp, Holger

    2011-10-26

    A vegetation study was carried out to investigate the carryover of Perfluorooctanoic Acid (PFOA) and Perfluorooctane Sulfonate (PFOS) from soil mixed with contaminated sewage sludge to potato, carrot, and cucumber plants. Analysis was done by liquid-extraction using acetonitrile with dispersive SPE cleanup and subsequent HPLC-MS/MS. In order to assess the transfer potential from soil, transfer factors (TF) were calculated for the different plant compartments: TF = [PFC](plant (wet substance))/[PFC](soil (dry weight)). The highest TF were found for the vegetative plant compartments with average values for PFOS below those for PFOA: cucumber, 0.17 (PFOS), 0.88 (PFOA); potato, 0.36 (PFOS), 0.40 (PFOA); carrot, 0.38 (PFOS), 0.53 (PFOA). Transfer of PFOA and PFOS into potato peelings (average values of TF: PFOA 0.03, PFOS 0.04) exceeded the carryover to the peeled tubers (PFOA 0.01, PFOS < 0.01). In carrots, this difference did not occur (average values of TF: PFOA 0.04, PFOS 0.04). Transfer of PFOS into the unpeeled cucumbers was low and comparable to that of peeled potatoes (TF < 0.01). For PFOA, it was higher (TF: 0.03).

  14. Carryover of perfluorooctanoic acid (PFOA) and perfluorooctane sulfonate (PFOS) from soil to plant and distribution to the different plant compartments studied in cultures of carrots (Daucus carota ssp. Sativus), potatoes (Solanum tuberosum), and cucumbers (Cucumis Sativus).

    PubMed

    Lechner, Mareike; Knapp, Holger

    2011-10-26

    A vegetation study was carried out to investigate the carryover of Perfluorooctanoic Acid (PFOA) and Perfluorooctane Sulfonate (PFOS) from soil mixed with contaminated sewage sludge to potato, carrot, and cucumber plants. Analysis was done by liquid-extraction using acetonitrile with dispersive SPE cleanup and subsequent HPLC-MS/MS. In order to assess the transfer potential from soil, transfer factors (TF) were calculated for the different plant compartments: TF = [PFC](plant (wet substance))/[PFC](soil (dry weight)). The highest TF were found for the vegetative plant compartments with average values for PFOS below those for PFOA: cucumber, 0.17 (PFOS), 0.88 (PFOA); potato, 0.36 (PFOS), 0.40 (PFOA); carrot, 0.38 (PFOS), 0.53 (PFOA). Transfer of PFOA and PFOS into potato peelings (average values of TF: PFOA 0.03, PFOS 0.04) exceeded the carryover to the peeled tubers (PFOA 0.01, PFOS < 0.01). In carrots, this difference did not occur (average values of TF: PFOA 0.04, PFOS 0.04). Transfer of PFOS into the unpeeled cucumbers was low and comparable to that of peeled potatoes (TF < 0.01). For PFOA, it was higher (TF: 0.03). PMID:21905714

  15. Effect of acidification on carrot (Daucus carota) juice cloud stability.

    PubMed

    Schultz, Alison K; Barrett, Diane M; Dungan, Stephanie R

    2014-11-26

    Effects of acidity on cloud stability in pasteurized carrot juice were examined over the pH range of 3.5-6.2. Cloud sedimentation, particle diameter, and ζ potential were measured at each pH condition to quantify juice cloud stability and clarification during 3 days of storage. Acidification below pH 4.9 resulted in a less negative ζ potential, an increased particle size, and an unstable cloud, leading to juice clarification. As the acidity increased, clarification occurred more rapidly and to a greater extent. Only a weak effect of ionic strength was observed when sodium salts were added to the juice, but the addition of calcium salts significantly reduced the cloud stability.

  16. Inositol trisphosphate metabolism in carrot (Daucus carota L. ) cells

    SciTech Connect

    Memon, A.R.; Rincon, M.; Boss, W.F. )

    1989-10-01

    The metabolism of exogenously added D-myo-(1-{sup 3}H)inositol 1,4,5-trisphosphate (IP{sub 3}) has been examined in microsomal membrane and soluble fractions of carrot cells grown in suspension culture. When ({sup 3}H)IP{sub 3} was added to a microsomal membrane fraction, ({sup 3}H)IP{sub 2} was the primary metabolite consisting of approximately 83% of the total recovered ({sup 3}H) by electrophoresis. ({sup 3}H)IP was only 6% of the ({sup 3}H) recovered, and 10% of the ({sup 3}H)IP{sub 3} was not further metabolized. In contrast, when ({sup 3}H)IP{sub 3} was added to the soluble fraction, approximately equal amounts of ({sup 3}H)IP{sub 2} and ({sup 3}H)IP were recovered. Ca{sup 2+} (100 micromolar) tended to enhance IP{sub 3} dephosphorylation but inhibited the IP{sub 2} dephosphorylation in the soluble fraction by about 20%. MoO{sub 4}{sup 2{minus}} (1 millimolar) inhibited the dephosphorylation of IP{sub 3} by the microsomal fraction and the dephosphorylation of IP{sub 2} by the soluble fraction. MoO{sub 4}{sup 2{minus}}, however, did not inhibit the dephosphorylation of IP{sub 3} by the soluble fraction. Li{sup +} (10 and 50 millimolar) had no effect on IP{sub 3} metabolism in either the soluble or membrane fraction; however, Li{sup +} (50 millimolar) inhibited IP{sub 2} dephosphorylation in the soluble fraction about 25%.

  17. Characterization of inositol phosphates in carrot (Daucus carota L. ) cells

    SciTech Connect

    Rincon, M.; Chen, Q.; Boss, W.F. )

    1989-01-01

    We have shown previously that inositol-1,4,5-trisphosphate (IP{sub 3}) stimulates an efflux of {sup 45}Ca{sup 2+} from fusogenic carrot protoplasts. In light of these results, we suggested that IP{sub 3} might serve as a second messenger for the mobilization of intracellular Ca{sup 2+} in higher plant cells. To determine whether or not IP{sub 3} and other inositol phosphates were present in the carrot cells, the cells were labeled with myo-(2-{sup 3}H)inositol for 18 hours and extracted with ice-cold 10% trichloroacetic acid. The inositol metabolites were separated by anion exchange chromatography and by paper electrophoresis. We found that ({sup 3}H)inositol metabolites coeluted with inositol bisphosphate (IP{sub 2}) and IP{sub 3} when separated by anion exchange chromatography. However, we could not detect IP{sub 2} or IP{sub 3} when the inositol metabolites were analyzed by paper electrophoresis even though the polyphosphoinositides, which are the source of IP{sub 2} and IP{sub 3}, were present in these cells. Thus, ({sup 3}H)inositol metabolites other than IP{sub 2} and IP{sub 3} had coeluted on the anion exchange columns. The data indicate that either IP{sub 3} is rapidly metabolized or that it is not present at a detectable level in the carrot cells.

  18. Expression of rabies virus G protein in carrots (Daucus carota).

    PubMed

    Rojas-Anaya, Edith; Loza-Rubio, Elizabeth; Olivera-Flores, Maria Teresa; Gomez-Lim, Miguel

    2009-12-01

    Antigens derived from various pathogens can readily be synthesized at high levels in plants in their authentic forms. Such antigens administered orally can induce an immune response and, in some cases, result in protection against a subsequent challenge. We here report the expression of rabies virus G protein into carrots. The G gene was subcloned into the pUCpSSrabG vector and then used to transform carrot embryogenic cells by particle bombardment. The carrot cells were selected in liquid medium, a method previously unreported. The presence of the transgene was verified by PCR, and by RT-PCR. By western blot, G protein transgene was identified in 93.3% of adult carrot roots. The G protein was quantified by densitometric analysis (range 0.4-1.2%). The expressed protein was antigenic in mice. This confirms that the carrot is an adequate system for antigen expression.

  19. Biofortification of Carrot (Daucus carota L.) with Iodine and Selenium in a Field Experiment.

    PubMed

    Smoleń, Sylwester; Skoczylas, Łukasz; Ledwożyw-Smoleń, Iwona; Rakoczy, Roksana; Kopeć, Aneta; Piątkowska, Ewa; Bieżanowska-Kopeć, Renata; Koronowicz, Aneta; Kapusta-Duch, Joanna

    2016-01-01

    The low content of iodine (I) and selenium (Se) forms available to plants in soil is one of the main causes of their insufficient transfer in the soil-plant-consumer system. Their deficiency occurs in food in the majority of human and farm animal populations around the world. Both elements are classified as beneficial elements. However, plant response to simultaneous fertilization with I and Se has not been investigated in depth. The study (conducted in 2012-2014) included soil fertilization of carrot cv. "Kazan F1" in the following combinations: (1) Control; (2) KI; (3) KIO3; (4) Na2SeO4; (5) Na2SeO3; (6) KI+Na2SeO4; (7) KIO3+Na2SeO4; (8) KI+Na2SeO3; (9) KIO3+Na2SeO3. I and Se were applied twice: before sowing and as top-dressing in a total dose of 5 kg I⋅ha(-1) and 1 kg Se⋅ha(-1). No negative effects of I and Se fertilization were noted with respect to carrot yield. Higher accumulation and the uptake by leaves and storage roots of I and Se were obtained after the application of KI than KIO3, as well as of Na2SeO4 than Na2SeO3, respectively. Transfer factor values for leaves and roots were about a dozen times higher for Se than for I. Selenomethionine content in carrot was higher after fertilization with Na2SeO4 than with Na2SeO3. However, it was the application of Na2SeO3, KI+Na2SeO3 and KIO3+Na2SeO3 that resulted in greater evenness within the years and a higher share of Se from selenomethionine in total Se in carrot plants. Consumption of 100 g f.w. of carrots fertilized with KI+Na2SeO3 and KIO3+Na2SeO3 can supply approximately or slightly exceed 100% of the Recommended Daily Allowance for I and Se. Moreover, the molar ratio of I and Se content in carrot fertilized with KI+Na2SeO3 and KIO3+Na2SeO3 was the best among the research plots. PMID:27303423

  20. Antioxidant phytochemicals and antioxidant capacity of biofortified carrots (Daucus carota L.) of various colors.

    PubMed

    Sun, Ting; Simon, Philipp W; Tanumihardjo, Sherry A

    2009-05-27

    Antioxidants and antioxidant capacity of seven colored carrots were determined. Five anthocyanins, chlorogenic acid, caffeic acid, and four carotenoids were quantified by HPLC. Total phenolic content was determined according to the Folin-Ciocalteu method. Antioxidant capacities of the hydrophilic and hydrophobic fractions were determined by using the 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and 2,2'-diphenyl-1-picrylhydrazyl (DPPH) methods. The relative antioxidant capacity index was determined. Anthocyanins were the major antioxidants in purple-yellow and purple-orange carrots, and chlorogenic acid was a major antioxidant in all carrots. Carotenoids did not contribute to total antioxidant capacity, but correlated with antioxidant capacity of hydrophobic extracts. Both the DPPH and ABTS assays showed that the hydrophilic extract had higher antioxidant capacity than the hydrophobic extract. Purple-yellow carrots had the highest antioxidant capacity, followed by purple-orange carrots, and the other carrots did not significantly differ. This information is useful for consumers and may help horticulturists develop carrots with higher antioxidant capacity.

  1. Content of carotenoids in roots of seventeen cultivars of Daucus carota L.

    PubMed

    Mech-Nowak, Aleksandra; Swiderski, Adam; Kruczek, Michał; Luczak, Irena; Kostecka-Gugała, Anna

    2012-01-01

    The aim of this study was to compare the content of carotenoids in seventeen cultivars of carrots grown in Poland. Conventional orange cultivars with rarely grown were compared: white, yellow and purple with yellow core cultivars. To determine the content of carotenoids, extracts from lyophilized carrot roots were prepared and analyzed by spectrophotometric as well as HPLC methods with DAD detector. The highest content of carotenoids was found in cultivars: 'Korund F(1)' (48 mg/100g of fresh weight) and 'Salsa F(1)' (36 mg/100g of fresh weight). The antioxidant properties of selected cultivars were compared using the DPPH method.

  2. Polyacetylenes from carrots (Daucus carota) improve glucose uptake in vitro in adipocytes and myotubes.

    PubMed

    El-Houri, Rime B; Kotowska, Dorota; Christensen, Kathrine B; Bhattacharya, Sumangala; Oksbjerg, Niels; Wolber, Gerhard; Kristiansen, Karsten; Christensen, Lars P

    2015-07-01

    A dichloromethane (DCM) extract of carrot roots was found to stimulate insulin-dependent glucose uptake (GU) in adipocytes in a dose dependent manner. Bioassay-guided fractionation of the DCM extract resulted in the isolation of the polyacetylenes falcarinol and falcarindiol. Both polyacetylenes were able to significantly stimulate basal and/or insulin-dependent GU in 3T3-L1 adipocytes and porcine myotube cell cultures in a dose-dependent manner. Falcarindiol increased peroxisome proliferator-activated receptor (PPAR)γ-mediated transactivation significantly at concentrations of 3, 10 and 30 μM, while PPARγ-mediated transactivation by falcarinol was only observed at 10 μM. Docking studies accordingly indicated that falcarindiol binds to the ligand binding domain of PPARγ with higher affinity than falcarinol and that both polyacetylenes exhibit characteristics of PPARγ partial agonists. Falcarinol was shown to inhibit adipocyte differentiation as evident by gene expression studies and Oil Red O staining, whereas falcarindiol did not inhibit adipocyte differentiation, which indicates that these polyacetylenes have distinct modes of action. The results of the present study suggest that falcarinol and falcarindiol may represent scaffolds for novel partial PPARγ agonists with possible antidiabetic properties.

  3. Polyacetylene diversity and bioactivity in orange market and locally grown colored carrots (Daucus carota L.).

    PubMed

    Metzger, Brandon T; Barnes, David M

    2009-12-01

    Carrots contain a wide array of phytochemicals such as carotenoids, phenolics, alpha-tocopherol, and polyacetylenes. Carrots are most known for their pro-vitamin A carotenoids but also contain other phytochemicals with documented health benefits. The phytochemicals in colored carrots present a challenge and opportunity due to the wide diversity of potent bioactive compounds. Two commercial carrots, 1 wild carrot, and 13 colored carrot varieties were characterized phytochemically. The carrots were screened in an anti-inflammatory model of lipopolysaccharide-induced nitric oxide production. Deep Purple carrot had the highest concentration of total polyacetylenes, alpha-tocopherol, and total phenolics. Commercial fresh market and baby orange carrots both had high concentrations of pro-vitamin A carotenoids. Purple carrots had higher antioxidant capacity values due to their anthocyanin content. Only seven carrot varieties had inhibitory activity (IC(25) = 257-1321 microg/mL) in macrophage cells. Among the varieties tested during the selected growing season, Deep Purple had the highest polyacetylene content and other important antioxidant phytochemicals. Further work is needed to identify other potential anti-inflammatory phytochemicals in colored carrots on the basis of this research.

  4. Effects of harvesting date and storage on the amounts of polyacetylenes in carrots, Daucus carota.

    PubMed

    Kjellenberg, Lars; Johansson, Eva; Gustavsson, Karl-Erik; Olsson, Marie E

    2010-11-24

    The amounts of three main polyacetylenes in carrots; falcarinol, falcarindiol, and falcarindiol-3-acetate, were determined by HPLC, during three seasons, in carrots harvested several times per season and at different locations in Sweden. The amounts of falcarindiol first decreased from a relatively high level and then increased later in the harvest season. The amounts of falcarindiol-3-acetate showed similar variations, whereas the amounts of falcarinol did not exhibit any significant variation during the harvest season. During storage the amount of polyacetylenes leveled off, increasing in samples initially low and decreasing in samples initially high in polyacetylenes. The amounts of all polyacetylenes varied significantly due to external factors and between stored and fresh samples. This variation opens up possibilities to achieve a chemical composition of polyacetylenes at harvest that minimizes the risk of bitter off-taste and maximizes the positive health effects reported in connection with polyacetylenes in carrots.

  5. Bioactive chemicals from carrot (Daucus carota) juice extracts for the treatment of leukemia.

    PubMed

    Zaini, Rana; Clench, Malcolm R; Le Maitre, Christine L

    2011-11-01

    Overwhelming evidence indicates that consumption of fruits and vegetables with antioxidant properties correlates with reduced risk for cancers, including leukemia. Carrots contain beneficial agents, such as β-carotene and polyacetylenes, which could be effective in the treatment of leukemia. This study investigated the effect of carrot juice extracts on myeloid and lymphoid leukemia cell lines together with normal hematopoietic stem cells. Leukemia cell lines and nontumor control cells were treated with carrot juice extracts for up to 72 hours in vitro. Induction of apoptosis was investigated by using annexin V/propidium iodide staining followed by flow cytometric analysis, and results were confirmed by using 4'-6-diamidino-2-phenylindole morphology. Effects on cellular proliferation were investigated via cell cycle analysis and cell counts. Treatment of leukemia cell lines with carrot juice extract induced apoptosis and inhibited progression through the cell cycle. Lymphoid cell lines were affected to a greater extent than were myeloid cell lines, and normal hematopoietic stem cells were less sensitive than most cell lines. This study has shown that extracts from carrots can induce apoptosis and cause cell cycle arrest in leukemia cell lines. The findings suggest that carrots may be an excellent source of bioactive chemicals for the treatment of leukemia.

  6. Effect of pectin methylesterase on carrot (Daucus carota) juice cloud stability.

    PubMed

    Schultz, Alison K; Anthon, Gordon E; Dungan, Stephanie R; Barrett, Diane M

    2014-02-01

    To determine the effect of residual enzyme activity on carrot juice cloud, 0 to 1 U/g pectin methylesterase (PME) was added to pasteurized carrot juice. Cloud stability and particle diameters were measured to quantify juice cloud stability and clarification for 56 days of storage. All levels of PME addition resulted in clarification; higher amounts had a modest effect in causing more rapid clarification, due to a faster increase in particle size. The cloud initially exhibited a trimodal distribution of particle sizes. For enzyme-containing samples, particles in the smallest-sized mode initially aggregated to merge with the second peak over 5-10 days. This larger population then continued to aggregate more slowly over longer times. This observation of a more rapid destabilization process initially, followed by slower subsequent changes in the cloud, was also manifested in measurements of sedimentation extent and in turbidity tests. Optical microscopy showed that aggregation created elongated, fractal particle structures over time.

  7. Suboptimal temperature favors reserve formation in biennial carrot (Daucus carota) plants.

    PubMed

    González, María V; Sadras, Victor O; Equiza, María A; Tognetti, Jorge A

    2009-09-01

    In response to suboptimal temperatures, temperate annual plants often increase root:shoot ratios, build-up carbohydrates and display typical morphological and anatomical changes. We know less about the responses of biennials such as carrot. As a model plant, carrot has the additional feature of two functionally and morphologically distinct root parts: the taproot, which stores carbohydrate and other compounds, and the fibrous root system involved in acquisition of water and nutrients. Here, we analyze the effects of temperature (12 vs 25°C) on growth, carbohydrate accumulation and whole-plant morphology in two carrot cultivars. Our working hypothesis is that suboptimal temperature favors active formation of reserve structures, rather than passive accumulation of storage carbohydrates. In comparison with plants grown at 25°C, plants grown at 12°C had: (1) higher fibrous root:shoot ratio (13%) , (2) thicker (10-15%) and smaller (up to two- to three-fold) leaves, (3) lower leaf cuticular permeance (two- to four-fold), (4) higher taproot:shoot ratio (two-fold), (5) higher phloem:xylem ratios in taproot (two- to six-fold), (6) unchanged percentage dry matter content (%DMC) in leaves, petioles or fibrous roots and (7) higher %DMC in taproot (20%). However, %DMC of individual taproot tissues (phloem and xylem) was unaffected by temperatures and was consistently higher in the phloem (up to 30%). Therefore, the higher %DMC of whole taproots at 12°C was attributed solely to the increased development of phloem tissue. Carrot, therefore, shares many of the most conspicuous elements of temperate plant responses to low temperatures. Consistently with our hypothesis, however, carrots grown at suboptimal temperature promoted reserve structures, rather than the increase in carbohydrate concentration typical of most temperate annual species and woody perennials.

  8. Carotenoid profiles and consumer sensory evaluation of specialty carrots (Daucus carota, L.) of various colors.

    PubMed

    Surles, Rebecca L; Weng, Ning; Simon, Philipp W; Tanumihardjo, Sherry A

    2004-06-01

    Five different colored carrots were analyzed for their carotenoid profile and underwent sensory evaluation to determine consumer acceptance (n = 96). Four major carotenoids were identified and quantified by use of HPLC methods. High beta-carotene orange carrots were found to contain the greatest concentration of total carotenoids. Except for the white, all the carrots are a significant source of bioavailable carotenoids. Sensory evaluation showed the high beta-carotene orange and white carrots to be favored over the yellow, red, and purple carrots in both blind and nonblind treatments (P < 0.01). However, all the carrots were well accepted by the consumer panel. With this information, carrot growers should be encouraged to cultivate specialty carrots to provide sources of both vitamin A precursors and phytochemicals.

  9. REMOVED: Protective effect of Daucus carota against UV-DMBA induced skin cancer in mice.

    PubMed

    Natarajamurthy, Sindhuja H; Dharmesh, Shylaja M

    2014-08-01

    This article has been removed: please see Elsevier Policy on Article Withdrawal (http://www.elsevier.com/locate/withdrawalpolicy). This article has been removed at the request of "Due to a processing error during the review process, this paper was erroneously published online prematurely. The Publisher apologizes to readers and the authors for any inconvenience caused."

  10. Bioaccessibility of Polyphenols from Plant-Processing Byproducts of Black Carrot (Daucus carota L.).

    PubMed

    Kamiloglu, Senem; Capanoglu, Esra; Bilen, Fatma Damla; Gonzales, Gerard Bryan; Grootaert, Charlotte; Van de Wiele, Tom; Van Camp, John

    2016-03-30

    Plant-processing byproducts of black carrot represent an important disposal problem for the industry; however, they are also promising sources of polyphenols, especially anthocyanins. The present study focused on the changes in polyphenols from black carrot, peel, and pomace during in vitro gastrointestinal digestion. Total phenolic content (TPC), total monomeric anthocyanin content (TMAC), and total antioxidant capacity (TAC) were determined using spectrophotometric methods, whereas identification and quantification of polyphenols were carried out using UPLC-ESI-MS(E) and HPLC-DAD, respectively. TPC, TMAC, and TAC significantly decreased (23-82%) as a result of in vitro gastrointestinal digestion. Nevertheless, the amount of pomace anthocyanins released at all stages of in vitro gastrointestinal digestion was higher than black carrot anthocyanins, suggesting that pomace may be a better source of bioaccessible anthocyanins. Overall, the current study highlighted black carrot byproducts as substantial sources of polyphenols, which may be used to enrich food products.

  11. Molecular characterization and pathogenicity of a carrot (Daucus carota) infecting begomovirus and associated betasatellite from India.

    PubMed

    Kumar, Jitendra; Gunapati, Samatha; Singh, Sudhir P; Gadre, Rekha; Sharma, Naresh C; Tuli, Rakesh

    2013-10-24

    The yellow mosaic pattern and shortening of leaf petiole are common disease symptoms associated with begomovirus infection in carrot. DNA from field infected carrot leaves was analyzed by rolling circle amplification and sequencing. The results established the presence of ageratum enation virus (AEV), which is referred to here as ageratum enation virus-carrot (AEV-Car). Symptomatic ageratum (Ageratum conyzoides) plants, growing adjacent to the carrot fields, also showed the presence of AEV (AEV-Age). Ageratum yellow leaf curl betasatellite (AYLCB) was also detected in the AEV infected carrot and ageratum samples. AEV-Car and AEV-Age are 95-97% identical in their DNA sequences, represents groups of isolates from the respective plant hosts (carrot and ageratum). Agroinoculation using infectious clones of AEV-Car plus AYLCB or AEV-Age plus AYLCB in carrot, ageratum, tobacco (Nicotiana tabacum) and tomato (Solanum lycopersicum) produced yellow mosaic and curling symptoms in leaves of inoculated plants. Agroinoculation of the two isolates together, along with the betasatellite (AEV-Car plus AEV-Age plus AYLCB) resulted in the enhancement of symptoms in comparison to the plants inoculated with single isolate. Plants with more severe symptoms showed a higher level of viral DNA accumulation, suggesting synergistic interactions between the two isolates of AEV.

  12. Biofortification of Carrot (Daucus carota L.) with Iodine and Selenium in a Field Experiment.

    PubMed

    Smoleń, Sylwester; Skoczylas, Łukasz; Ledwożyw-Smoleń, Iwona; Rakoczy, Roksana; Kopeć, Aneta; Piątkowska, Ewa; Bieżanowska-Kopeć, Renata; Koronowicz, Aneta; Kapusta-Duch, Joanna

    2016-01-01

    The low content of iodine (I) and selenium (Se) forms available to plants in soil is one of the main causes of their insufficient transfer in the soil-plant-consumer system. Their deficiency occurs in food in the majority of human and farm animal populations around the world. Both elements are classified as beneficial elements. However, plant response to simultaneous fertilization with I and Se has not been investigated in depth. The study (conducted in 2012-2014) included soil fertilization of carrot cv. "Kazan F1" in the following combinations: (1) Control; (2) KI; (3) KIO3; (4) Na2SeO4; (5) Na2SeO3; (6) KI+Na2SeO4; (7) KIO3+Na2SeO4; (8) KI+Na2SeO3; (9) KIO3+Na2SeO3. I and Se were applied twice: before sowing and as top-dressing in a total dose of 5 kg I⋅ha(-1) and 1 kg Se⋅ha(-1). No negative effects of I and Se fertilization were noted with respect to carrot yield. Higher accumulation and the uptake by leaves and storage roots of I and Se were obtained after the application of KI than KIO3, as well as of Na2SeO4 than Na2SeO3, respectively. Transfer factor values for leaves and roots were about a dozen times higher for Se than for I. Selenomethionine content in carrot was higher after fertilization with Na2SeO4 than with Na2SeO3. However, it was the application of Na2SeO3, KI+Na2SeO3 and KIO3+Na2SeO3 that resulted in greater evenness within the years and a higher share of Se from selenomethionine in total Se in carrot plants. Consumption of 100 g f.w. of carrots fertilized with KI+Na2SeO3 and KIO3+Na2SeO3 can supply approximately or slightly exceed 100% of the Recommended Daily Allowance for I and Se. Moreover, the molar ratio of I and Se content in carrot fertilized with KI+Na2SeO3 and KIO3+Na2SeO3 was the best among the research plots.

  13. Biofortification of Carrot (Daucus carota L.) with Iodine and Selenium in a Field Experiment

    PubMed Central

    Smoleń, Sylwester; Skoczylas, Łukasz; Ledwożyw-Smoleń, Iwona; Rakoczy, Roksana; Kopeć, Aneta; Piątkowska, Ewa; Bieżanowska-Kopeć, Renata; Koronowicz, Aneta; Kapusta-Duch, Joanna

    2016-01-01

    The low content of iodine (I) and selenium (Se) forms available to plants in soil is one of the main causes of their insufficient transfer in the soil-plant-consumer system. Their deficiency occurs in food in the majority of human and farm animal populations around the world. Both elements are classified as beneficial elements. However, plant response to simultaneous fertilization with I and Se has not been investigated in depth. The study (conducted in 2012–2014) included soil fertilization of carrot cv. “Kazan F1” in the following combinations: (1) Control; (2) KI; (3) KIO3; (4) Na2SeO4; (5) Na2SeO3; (6) KI+Na2SeO4; (7) KIO3+Na2SeO4; (8) KI+Na2SeO3; (9) KIO3+Na2SeO3. I and Se were applied twice: before sowing and as top-dressing in a total dose of 5 kg I⋅ha-1 and 1 kg Se⋅ha-1. No negative effects of I and Se fertilization were noted with respect to carrot yield. Higher accumulation and the uptake by leaves and storage roots of I and Se were obtained after the application of KI than KIO3, as well as of Na2SeO4 than Na2SeO3, respectively. Transfer factor values for leaves and roots were about a dozen times higher for Se than for I. Selenomethionine content in carrot was higher after fertilization with Na2SeO4 than with Na2SeO3. However, it was the application of Na2SeO3, KI+Na2SeO3 and KIO3+Na2SeO3 that resulted in greater evenness within the years and a higher share of Se from selenomethionine in total Se in carrot plants. Consumption of 100 g f.w. of carrots fertilized with KI+Na2SeO3 and KIO3+Na2SeO3 can supply approximately or slightly exceed 100% of the Recommended Daily Allowance for I and Se. Moreover, the molar ratio of I and Se content in carrot fertilized with KI+Na2SeO3 and KIO3+Na2SeO3 was the best among the research plots. PMID:27303423

  14. Major Cytogenetic Landmarks and Karyotype Analysis in Carrot (Daucus carota L.) and Other Apiaceace

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Karyotyping can be helpful for understanding species evolution and relationships. Cytological studies in Apiaceae have provided information on the chromosome number and morphology of several crops. However, karyological data of their wild relatives are scarce. In addition, the number of chromosomes ...

  15. Patterns of gene flow between crop and wild carrot, Daucus carota (Apiaceae) in the United States

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Studies of gene flow between crops and their wild relatives have implications for both management practices for farming and breeding as well as understanding the risk of transgene escape. These types of studies may also yield insight into population dynamics and the evolutionary consequences of gene...

  16. Polyacetylenes from carrots (Daucus carota) improve glucose uptake in vitro in adipocytes and myotubes.

    PubMed

    El-Houri, Rime B; Kotowska, Dorota; Christensen, Kathrine B; Bhattacharya, Sumangala; Oksbjerg, Niels; Wolber, Gerhard; Kristiansen, Karsten; Christensen, Lars P

    2015-07-01

    A dichloromethane (DCM) extract of carrot roots was found to stimulate insulin-dependent glucose uptake (GU) in adipocytes in a dose dependent manner. Bioassay-guided fractionation of the DCM extract resulted in the isolation of the polyacetylenes falcarinol and falcarindiol. Both polyacetylenes were able to significantly stimulate basal and/or insulin-dependent GU in 3T3-L1 adipocytes and porcine myotube cell cultures in a dose-dependent manner. Falcarindiol increased peroxisome proliferator-activated receptor (PPAR)γ-mediated transactivation significantly at concentrations of 3, 10 and 30 μM, while PPARγ-mediated transactivation by falcarinol was only observed at 10 μM. Docking studies accordingly indicated that falcarindiol binds to the ligand binding domain of PPARγ with higher affinity than falcarinol and that both polyacetylenes exhibit characteristics of PPARγ partial agonists. Falcarinol was shown to inhibit adipocyte differentiation as evident by gene expression studies and Oil Red O staining, whereas falcarindiol did not inhibit adipocyte differentiation, which indicates that these polyacetylenes have distinct modes of action. The results of the present study suggest that falcarinol and falcarindiol may represent scaffolds for novel partial PPARγ agonists with possible antidiabetic properties. PMID:25970571

  17. Bioaccessibility of Polyphenols from Plant-Processing Byproducts of Black Carrot (Daucus carota L.).

    PubMed

    Kamiloglu, Senem; Capanoglu, Esra; Bilen, Fatma Damla; Gonzales, Gerard Bryan; Grootaert, Charlotte; Van de Wiele, Tom; Van Camp, John

    2016-03-30

    Plant-processing byproducts of black carrot represent an important disposal problem for the industry; however, they are also promising sources of polyphenols, especially anthocyanins. The present study focused on the changes in polyphenols from black carrot, peel, and pomace during in vitro gastrointestinal digestion. Total phenolic content (TPC), total monomeric anthocyanin content (TMAC), and total antioxidant capacity (TAC) were determined using spectrophotometric methods, whereas identification and quantification of polyphenols were carried out using UPLC-ESI-MS(E) and HPLC-DAD, respectively. TPC, TMAC, and TAC significantly decreased (23-82%) as a result of in vitro gastrointestinal digestion. Nevertheless, the amount of pomace anthocyanins released at all stages of in vitro gastrointestinal digestion was higher than black carrot anthocyanins, suggesting that pomace may be a better source of bioaccessible anthocyanins. Overall, the current study highlighted black carrot byproducts as substantial sources of polyphenols, which may be used to enrich food products. PMID:26262673

  18. The SSP Syndrome

    ERIC Educational Resources Information Center

    Radakovitz, Richard

    1975-01-01

    Describes a cure for the high school chemistry student with Second Semester Poisoning (SSP), the student disease of apathy which occurs in the spring. Remedy: an outdoor (preferably) demonstration of a series of controlled combustion experiments, with accompanying explanations of the chemistry involved. (MLH)

  19. Genomic identification of WRKY transcription factors in carrot (Daucus carota) and analysis of evolution and homologous groups for plants.

    PubMed

    Li, Meng-Yao; Xu, Zhi-Sheng; Tian, Chang; Huang, Ying; Wang, Feng; Xiong, Ai-Sheng

    2016-01-01

    WRKY transcription factors belong to one of the largest transcription factor families. These factors possess functions in plant growth and development, signal transduction, and stress response. Here, we identified 95 DcWRKY genes in carrot based on the carrot genomic and transcriptomic data, and divided them into three groups. Phylogenetic analysis of WRKY proteins from carrot and Arabidopsis divided these proteins into seven subgroups. To elucidate the evolution and distribution of WRKY transcription factors in different species, we constructed a schematic of the phylogenetic tree and compared the WRKY family factors among 22 species, which including plants, slime mold and protozoan. An in-depth study was performed to clarify the homologous factor groups of nine divergent taxa in lower and higher plants. Based on the orthologous factors between carrot and Arabidopsis, 38 DcWRKY proteins were calculated to interact with other proteins in the carrot genome. Yeast two-hybrid assay showed that DcWRKY20 can interact with DcMAPK1 and DcMAPK4. The expression patterns of the selected DcWRKY genes based on transcriptome data and qRT-PCR suggested that those selected DcWRKY genes are involved in root development, biotic and abiotic stress response. This comprehensive analysis provides a basis for investigating the evolution and function of WRKY genes. PMID:26975939

  20. Ultraviolet-C light effect on physicochemical, bioactive, microbiological, and sensorial characteristics of carrot (Daucus carota) beverages.

    PubMed

    Hernández-Carranza, Paola; Ruiz-López, Irving Israel; Pacheco-Aguirre, Francisco Manuel; Guerrero-Beltrán, José Ángel; Ávila-Sosa, Raúl; Ochoa-Velasco, Carlos Enrique

    2016-09-01

    The aim of this research was to evaluate the effect of ultraviolet-C light on physicochemical, bioactive, microbial, and sensory characteristics of carrot beverages. Beverages were formulated with different concentrations of carrot juice (60, 80, and 100% [v/v]) and treated with ultraviolet-C light at different flow rates (0, 0.5, 3.9, and 7.9 mL s(-1)) and times (5, 10, 15, 20, and 30 min), equivalent to ultraviolet-C dosages of 13.2, 26.4, 39.6, 52.8, and 79.2 J cm(-2) Total soluble solids, pH, and titratable acidity were not affected by the ultraviolet-C light treatment. Ultraviolet-C light significantly affected (p < 0.05) color parameters of pure juice; however, at low concentration of juice, total color change was slightly affected (ΔE = 2.0 ± 0.7). Phenolic compounds (4.1 ± 0.1, 5.2 ± 0.2, and 8.6 ± 0.3 mg of GAE 100 mL(-1) of beverage with 60, 80, and 100% of juice, respectively) and antioxidant capacity (6.1 ± 0.4, 8.5 ± 0.4, and 9.4 ± 0.3 mg of Trolox 100 mL(-1) of beverage with 60, 80, and 100% of juice, respectively) of carrot beverages were not affected by ultraviolet-C light treatment. Microbial kinetics showed that mesophiles were mostly reduced at high flow rates in carrot beverages with 60% of juice. Maximum logarithmic reductions for mesophiles and total coliforms were 3.2 ± 0.1 and 2.6 ± 0.1, respectively, after 30 min of ultraviolet-C light processing. Beverages were well accepted (6-7) by judges who did not perceive the difference between untreated and Ultraviolet-C light treated beverages. PMID:26893153

  1. Enhanced disease resistance in transgenic carrot (Daucus carota L.) plants over-expressing a rice cationic peroxidase.

    PubMed

    Wally, O; Punja, Z K

    2010-10-01

    Plant class III peroxidases are involved in numerous responses related to pathogen resistance including controlling hydrogen peroxide (H(2)O(2)) levels and lignin formation. Peroxidases catalyze the oxidation of organic compounds using H(2)O(2) as an oxidant. We examined the mechanisms of disease resistance in a transgenic carrot line (P23) which constitutively over-expresses the rice cationic peroxidase OsPrx114 (previously known as PO-C1) and which exhibits enhanced resistance to necrotrophic foliar pathogens. OsPrx114 over-expression led to a slight enhancement of constitutive transcript levels of pathogenesis-related (PR) genes. These transcript levels were dramatically increased in line P23 compared to controls [GUS construct under the control of 35S promoter (35S::GUS)] when tissues were treated with cell wall fragments of the fungal pathogen Sclerotinia sclerotiorum (SS-walls), and to a lesser extent with 2,6-dichloroisonicotinic acid. There was no basal increase in basal H(2)O(2) levels in tissues of the line P23. However, during an oxidative burst response elicited by SS-walls, H(2)O(2) accumulation was reduced in line P23 despite, typical media alkalinization associated with oxidative burst responses was observed, suggesting that OsPrx114 was involved in rapid H(2)O(2) consumption during the oxidative burst response. Tap roots of line P23 had increased lignin formation in the outer periderm tissues, which was further increased during challenge inoculation with Alternaria radicina. Plant susceptibility to a biotrophic pathogen, Erysiphe heraclei, was not affected. Disease resistance to necrotrophic pathogens in carrot as a result of OsPrx114 over-expression is manifested through increased PR transcript accumulation, rapid removal of H(2)O(2) during oxidative burst response and enhanced lignin formation.

  2. In vitro bioaccessibility of β-carotene, Ca, Mg and Zn in landrace carrots (Daucus carota, L.).

    PubMed

    Zaccari, Fernanda; Cabrera, María Cristina; Ramos, Ana; Saadoun, Ali

    2015-01-01

    Four landrace carrots ("Becaria", "CRS", "González" and "Rodríguez") and two marketable cultivars (Kuroda and Brasilia), raw and steamed, were characterised by the total content of β-carotene Ca, Mg and Zn, in vitro bioaccessibility and by colour and were evaluated to determine the effect of particle size in nutrient bioaccessibility. Steaming increased the content of β-carotene extracted from "CRS" and Brasilia (29% and 75%) and decreased the content of β-carotene extracted from "CRS" by 23% in "Rodríguez." In addition, steaming caused a loss of Ca (21%) but did not change the amount of Mg and Zn. The bioaccessibility of β-carotene in raw and pulped carrots was very low (<0.5%). Furthermore, steaming and a smaller particle size increased the bioaccessibility of β-carotene by 3-16 times. Additionally, cooking increased the in vitro bioaccessibility of Ca and Zn but had no effect on Mg. Moreover, homogenisation increased the bioaccessibility by 20% in Ca, 17% in Mg, and 10% in Zn compared to pulping.

  3. Clarification and pasteurisation effects on monomeric anthocyanins and percent polymeric colour of black carrot (Daucus carota L.) juice.

    PubMed

    Türkyılmaz, Meltem; Yemiş, Oktay; Ozkan, Mehmet

    2012-09-15

    Black carrots (BCs) are a rich source of stable anthocyanins (ACNs). The purpose of this study was to evaluate the effects of clarification and pasteurisation on ACNs of black carrot juice (BCJ). Monomeric ACNs, ACN profile and percent polymeric colour were determined during processing of BCJ. While depectinisation and bentonite treatments resulted in 7% and 20% increases in monomeric ACN content of BCJ, respectively, gelatine-kieselsol treatment and pasteurisation resulted in 10% and 3-16% reduction. Percent polymeric colour decreased after clarification, but substantially increased in samples subjected to heat. ACNs of BCJ samples were identified by HPLC-MS. Unclarified BCJ contained cyanidin-3-galactoside-xyloside-glucoside-ferulic acid as the major ACN, followed by cyanidin-3-galactoside-xyloside-glucoside-coumaric acid, and cyanidin-3-galactoside-xyloside-glucoside. After depectinisation, two more ACNs (cyanidin-3-galactoside-xyloside and cyanidin-3-galactoside-xyloside-glucoside-sinapic acid) were also identified. These results indicated that depectinisation and bentonite treatment had positive effect on the colour of BCJ, while gelatin-kieselsol treatment and pasteurisation had negative effect.

  4. Effect of decontamination agents on the microbial population, sensorial quality, and nutrient content of grated carrots (Daucus carota L.).

    PubMed

    Vandekinderen, Isabelle; Van Camp, John; Devlieghere, Frank; Veramme, Kim; Denon, Quenten; Ragaert, Peter; De Meulenaer, Bruno

    2008-07-23

    Several decontamination agents including water, sodium hypochlorite, peroxyacetic acid, neutral electrolyzed oxidizing water, and chlorine dioxide gas were tested for their effectiveness to reduce the natural microflora on grated carrots. Microbial reductions of the total aerobic count obtained after the different treatments varied between 0.11 and 3.29 log colony-forming units (cfu)/g. Whether or not a decontamination step induced significant changes in the sensory attributes of grated carrots is highly dependent on the type and concentration of disinfectant. To maintain the nutritional value, the influence of the decontamination agents on carotenoid content, alpha-tocopherol content, total phenols, and antioxidant capacity was studied. Besides the part of the nutrients that was leached away from the cutting areas by water, the nutrient losses caused by adding sanitizers were rather limited. Compared with the untreated carrots alpha-tocopherol content was, however, significantly reduced when 250 ppm of peroxyacetic acid (-80%) or 200 ppm of sodium hypochlorite (-59%) was used. Additional losses in carotenoid content were caused by contact with chlorine dioxide gas (-9%). On the condition of an optimized decontamination process toward time and concentration, the microbial quality of fresh-cut carrots could be improved without negatively influencing their sensory quality and nutrient content.

  5. Effect of storage and cooking on beta-carotene isomers in carrots ( Daucus carota L. cv. 'Stefano').

    PubMed

    Imsic, Michael; Winkler, Sonja; Tomkins, Bruce; Jones, Rod

    2010-04-28

    Carrots are one of the highest dietary sources of beta-carotene and are naturally high in the (all-E)-beta-carotene isomer, which has higher bioavailability, provitamin A activity, and antioxidant capacity compared to Z (cis) isomers. The objectives of the present study were to investigate the effects of storage temperature, time, and cooking (boiling for 15 min) on the levels of carotene isomers in 'Stefano' carrots. Storing carrots at either 4 degrees C to simulate long-term storage or 20 degrees C to simulate marketing practices resulted in increases in (all-E)-beta-carotene of 20.3% after 3 days at 4 degrees C and 34.4% after 14 days at 20 degrees C, respectively. The levels of Z isomers in raw carrots were low with (13Z)-beta-carotene and (9Z)-beta-carotene accounting for less that 1.8% of the total beta-carotene present. Levels of (9Z)-beta-carotene decreased during storage at either temperature, whereas storage at 4 degrees C resulted in a 109% increase in (13Z)-beta-carotene after 56 days. Cooking significantly increased the levels of (13Z)-beta-carotene and (9Z)-beta-carotene and resulted in the production of (15Z)-beta-carotene, which was absent in raw carrots. Storage at 4 degrees C for 15 days or more prior to cooking reduced the susceptibility of (all-E)-beta-carotene to thermal isomerization during cooking, resulting in lower levels of all three Z-beta-carotene isomers being generated, while storage at 20 degrees C for up to 21 days resulted in significantly higher levels of (all-E)-beta-carotene before and after cooking but had no effect on Z-isomer production during cooking. Consequently, we conclude that, for the greatest health benefit, fresh carrots can be stored for up to 21 days at 20 degrees C or at 4 degrees C for up to 56 days without significant reduction in (all-E)-beta-carotene and should be consumed raw or boiled for less than 15 min to limit Z-beta-carotene isomer formation.

  6. Ultraviolet-C light effect on physicochemical, bioactive, microbiological, and sensorial characteristics of carrot (Daucus carota) beverages.

    PubMed

    Hernández-Carranza, Paola; Ruiz-López, Irving Israel; Pacheco-Aguirre, Francisco Manuel; Guerrero-Beltrán, José Ángel; Ávila-Sosa, Raúl; Ochoa-Velasco, Carlos Enrique

    2016-09-01

    The aim of this research was to evaluate the effect of ultraviolet-C light on physicochemical, bioactive, microbial, and sensory characteristics of carrot beverages. Beverages were formulated with different concentrations of carrot juice (60, 80, and 100% [v/v]) and treated with ultraviolet-C light at different flow rates (0, 0.5, 3.9, and 7.9 mL s(-1)) and times (5, 10, 15, 20, and 30 min), equivalent to ultraviolet-C dosages of 13.2, 26.4, 39.6, 52.8, and 79.2 J cm(-2) Total soluble solids, pH, and titratable acidity were not affected by the ultraviolet-C light treatment. Ultraviolet-C light significantly affected (p < 0.05) color parameters of pure juice; however, at low concentration of juice, total color change was slightly affected (ΔE = 2.0 ± 0.7). Phenolic compounds (4.1 ± 0.1, 5.2 ± 0.2, and 8.6 ± 0.3 mg of GAE 100 mL(-1) of beverage with 60, 80, and 100% of juice, respectively) and antioxidant capacity (6.1 ± 0.4, 8.5 ± 0.4, and 9.4 ± 0.3 mg of Trolox 100 mL(-1) of beverage with 60, 80, and 100% of juice, respectively) of carrot beverages were not affected by ultraviolet-C light treatment. Microbial kinetics showed that mesophiles were mostly reduced at high flow rates in carrot beverages with 60% of juice. Maximum logarithmic reductions for mesophiles and total coliforms were 3.2 ± 0.1 and 2.6 ± 0.1, respectively, after 30 min of ultraviolet-C light processing. Beverages were well accepted (6-7) by judges who did not perceive the difference between untreated and Ultraviolet-C light treated beverages.

  7. Combined effects of gamma irradiation and modified atmosphere packaging on bacterial resistance in grated carrots ( Daucus carota)

    NASA Astrophysics Data System (ADS)

    Lacroix, M.; Lafortune, R.

    2004-09-01

    The present study was conducted to evaluate the efficiency of gamma irradiation combined with modified atmosphere packaging as an alternative treatment to ensure the innocuity and the shelf life extension of pre-cured vegetables. Grated carrots were inoculated with Escherichia coli (10 6 CFU/g) and packed under air or under MAP condition (60% O 2, 30% CO 2 and 10% N 2). The packages were then, gamma irradiated at doses from 0.15 to 0.9 kGy and stored at 4±1°C. E. coli counts were periodically evaluated during 50 days of storage. Results showed that at day 1, an irradiation treatment at a dose of 0.15 kGy reduced by 3 and 4 log the microbial level representing a level of 3 and 2 log CFU/g when samples were irradiated under air and under MAP respectively. However, a level of 3 log CFU/g was detected in both treated samples after 7 days of storage. When samples were irradiated at doses ⩾0.3 kGy no E.coli were detected during the whole storage in samples treated under MAP. However, when samples were treated under air, a level of 1-2 log CFU/g of E.coli was detected after 5 days of storage.

  8. Genomic identification of WRKY transcription factors in carrot (Daucus carota) and analysis of evolution and homologous groups for plants.

    PubMed

    Li, Meng-Yao; Xu, Zhi-Sheng; Tian, Chang; Huang, Ying; Wang, Feng; Xiong, Ai-Sheng

    2016-03-15

    WRKY transcription factors belong to one of the largest transcription factor families. These factors possess functions in plant growth and development, signal transduction, and stress response. Here, we identified 95 DcWRKY genes in carrot based on the carrot genomic and transcriptomic data, and divided them into three groups. Phylogenetic analysis of WRKY proteins from carrot and Arabidopsis divided these proteins into seven subgroups. To elucidate the evolution and distribution of WRKY transcription factors in different species, we constructed a schematic of the phylogenetic tree and compared the WRKY family factors among 22 species, which including plants, slime mold and protozoan. An in-depth study was performed to clarify the homologous factor groups of nine divergent taxa in lower and higher plants. Based on the orthologous factors between carrot and Arabidopsis, 38 DcWRKY proteins were calculated to interact with other proteins in the carrot genome. Yeast two-hybrid assay showed that DcWRKY20 can interact with DcMAPK1 and DcMAPK4. The expression patterns of the selected DcWRKY genes based on transcriptome data and qRT-PCR suggested that those selected DcWRKY genes are involved in root development, biotic and abiotic stress response. This comprehensive analysis provides a basis for investigating the evolution and function of WRKY genes.

  9. Genomic identification of WRKY transcription factors in carrot (Daucus carota) and analysis of evolution and homologous groups for plants

    PubMed Central

    Li, Meng-Yao; Xu, Zhi-Sheng; Tian, Chang; Huang, Ying; Wang, Feng; Xiong, Ai-Sheng

    2016-01-01

    WRKY transcription factors belong to one of the largest transcription factor families. These factors possess functions in plant growth and development, signal transduction, and stress response. Here, we identified 95 DcWRKY genes in carrot based on the carrot genomic and transcriptomic data, and divided them into three groups. Phylogenetic analysis of WRKY proteins from carrot and Arabidopsis divided these proteins into seven subgroups. To elucidate the evolution and distribution of WRKY transcription factors in different species, we constructed a schematic of the phylogenetic tree and compared the WRKY family factors among 22 species, which including plants, slime mold and protozoan. An in-depth study was performed to clarify the homologous factor groups of nine divergent taxa in lower and higher plants. Based on the orthologous factors between carrot and Arabidopsis, 38 DcWRKY proteins were calculated to interact with other proteins in the carrot genome. Yeast two-hybrid assay showed that DcWRKY20 can interact with DcMAPK1 and DcMAPK4. The expression patterns of the selected DcWRKY genes based on transcriptome data and qRT-PCR suggested that those selected DcWRKY genes are involved in root development, biotic and abiotic stress response. This comprehensive analysis provides a basis for investigating the evolution and function of WRKY genes. PMID:26975939

  10. A study on the effect of few eco-friendly manures on the growth attributes of carrot (Daucus carota L.).

    PubMed

    Vijayakumari, B; Yadav, R Hiranmai; Sowmya, M

    2009-01-01

    A pot culture study was carried out at Avinashilingam Deemed University, Coimbatore to assess the effect of few organic manures (herbal microfertilizer, humic acid, farm yard manure (FYM), biofertilizer) and NPK on the growth attributes of carrot. Among the parameters observed, the germination percentage was highest in FYM treated pots on 7 DAS (Days After Sowing) and in NPK treated pots on 21, 28 and 30 DAS. The parameters, such as root length, crown length and vigour index, were studied on 30, 60, 90 and 120 DAS. The root length on 60 and 120 DAS was increased by FYM, whereas on 30 and 90 DAS, increase was observed in NPK treated plants. The crown length was increased by NPK on 30 and 120 DAS, micro herbal fertilizer showed an increase of crown length on 60 DAS and by FYM on 90 DAS. Highest vigour was in FYM treated plants on 30 DAS, in herbal micro fertilizer on 60 DAS in NPK treatment on 90 and 120 DAS. PMID:21114148

  11. In vitro bioaccessibility of β-carotene, Ca, Mg and Zn in landrace carrots (Daucus carota, L.).

    PubMed

    Zaccari, Fernanda; Cabrera, María Cristina; Ramos, Ana; Saadoun, Ali

    2015-01-01

    Four landrace carrots ("Becaria", "CRS", "González" and "Rodríguez") and two marketable cultivars (Kuroda and Brasilia), raw and steamed, were characterised by the total content of β-carotene Ca, Mg and Zn, in vitro bioaccessibility and by colour and were evaluated to determine the effect of particle size in nutrient bioaccessibility. Steaming increased the content of β-carotene extracted from "CRS" and Brasilia (29% and 75%) and decreased the content of β-carotene extracted from "CRS" by 23% in "Rodríguez." In addition, steaming caused a loss of Ca (21%) but did not change the amount of Mg and Zn. The bioaccessibility of β-carotene in raw and pulped carrots was very low (<0.5%). Furthermore, steaming and a smaller particle size increased the bioaccessibility of β-carotene by 3-16 times. Additionally, cooking increased the in vitro bioaccessibility of Ca and Zn but had no effect on Mg. Moreover, homogenisation increased the bioaccessibility by 20% in Ca, 17% in Mg, and 10% in Zn compared to pulping. PMID:25053069

  12. Increased Nicotiana tabacum fitness through positive regulation of carotenoid, gibberellin and chlorophyll pathways promoted by Daucus carota lycopene β-cyclase (Dclcyb1) expression.

    PubMed

    Moreno, J C; Cerda, A; Simpson, K; Lopez-Diaz, I; Carrera, E; Handford, M; Stange, C

    2016-04-01

    Carotenoids, chlorophylls and gibberellins are derived from the common precursor geranylgeranyl diphosphate (GGPP). One of the enzymes in carotenoid biosynthesis is lycopene β-cyclase (LCYB) that catalyzes the conversion of lycopene into β-carotene. In carrot, Dclcyb1 is essential for carotenoid synthesis in the whole plant. Here we show that when expressed in tobacco, increments in total carotenoids, β-carotene and chlorophyll levels occur. Furthermore, photosynthetic efficiency is enhanced in transgenic lines. Interestingly, and contrary to previous observations where overexpression of a carotenogenic gene resulted in the inhibition of the synthesis of gibberellins, we found raised levels of active GA4 and the concommitant increases in plant height, leaf size and whole plant biomass, as well as an early flowering phenotype. Moreover, a significant increase in the expression of the key carotenogenic genes, Ntpsy1, Ntpsy2 and Ntlcyb, as well as those involved in the synthesis of chlorophyll (Ntchl), gibberellin (Ntga20ox, Ntcps and Ntks) and isoprenoid precursors (Ntdxs2 and Ntggpps) was observed. These results indicate that the expression of Dclcyb1 induces a positive feedback affecting the expression of isoprenoid gene precursors and genes involved in carotenoid, gibberellin and chlorophyll pathways leading to an enhancement in fitness measured as biomass, photosynthetic efficiency and carotenoid/chlorophyll composition. PMID:26893492

  13. Analysis of the Thermal Degradation of the Individual Anthocyanin Compounds of Black Carrot (Daucus carota L.): A New Approach Using High-Resolution Proton Nuclear Magnetic Resonance Spectroscopy.

    PubMed

    Iliopoulou, Ioanna; Thaeron, Delphine; Baker, Ashley; Jones, Anita; Robertson, Neil

    2015-08-12

    The black carrot dye is a mixture of cyanidin molecules, the nuclear magnetic resonance (NMR) spectrum of which shows a highly overlapped aromatic region. In this study, the (1)H NMR (800 MHz) aromatic chemical shifts of the mixture were fully assigned by overlaying them with the characterized (1)H NMR chemical shifts of the separated compounds. The latter were isolated using reverse-phase high-performance liquid chromatography (RP-HPLC), and their chemical shifts were identified using (1)H and two-dimensional (2D) correlation spectroscopy (COSY) NMR spectroscopy. The stability of the black carrot mixture to heat exposure was investigated at pH 3.6, 6.8, and 8.0 by heat-treating aqueous solutions at 100 °C and the powdered material at 180 °C. From integration of high-resolution (1)H NMR spectra, it was possible to follow the relative degradation of each compound, offering advantages over the commonly used ultraviolet/visible (UV/vis) and HPLC approaches. UV/vis spectroscopy and CIE color measurements were used to determine thermally induced color changes, under normal cooking conditions.

  14. Anthocyanins in purple-orange carrots (Daucus carota L.) do not influence the bioavailability of beta-carotene in young women.

    PubMed

    Arscott, Sara A; Simon, Philipp W; Tanumihardjo, Sherry A

    2010-03-10

    Purple carrots contain anthocyanins in addition to the provitamin A carotenoids in typical orange carrots. Simultaneous consumption of these phytochemicals in carrots may affect the bioavailability of carotenoids. The bioavailability of beta-carotene in humans was assessed from an acute feeding of orange (OC) and purple (PC) carrots with white (WC) as a control. Carrot smoothies were served to female subjects (n = 5, aged 21-26 years) for breakfast after 1 week on a low carotenoid diet and overnight fast. OC and PC smoothies were equalized to 10.3 mg of all-trans beta-carotene. Plasma beta-carotene was measured for 144 h following treatments. Peak plasma concentrations of OC and PC treatments did not differ. The PC treatment 0-144 h area-under-the-curve for beta-carotene was 76% of the OC treatment (P < 0.05). However, when the first 24 h were compared, OC and PC treatments did not differ, suggesting that anthocyanins in purple carrots do not affect the absorption of beta-carotene postprandially. PMID:20131807

  15. Metabolic fingerprinting reveals differences between shoots of wild and cultivated carrot (Daucus carota L.) and suggests maternal inheritance or wild trait dominance in hybrids.

    PubMed

    Grebenstein, C; Choi, Y H; Rong, J; de Jong, T J; Tamis, W L M

    2011-08-01

    Differences between the metabolic content of cultivars and their related wild species not only have implications for breeding and food quality, but also for the increasingly studied area of crop to wild introgression. Wild and cultivated western carrots belong to the same outcrossing species and hybridize under natural conditions. The metabolic fingerprinting of Dutch wild carrot and of western orange carrot cultivar shoots using (1)H NMR showed only quantitative differences in chemical content, indicating relatively low divergence after domestication. Main differences reside in the primary metabolite content and in the concentrations of chlorogenic acid and feruloyl quinic acid in the shoots of the different carrot types. Wild×cultivar hybrids cannot be distinguished from wild plants based on the metabolome, suggesting maternal, maternal environment, or dominance effects, and indicating high hybrid fitness in wild conditions. Considering these similarities, introgression is a real possibility in carrots, but understanding its consequences would require further studies using backcrosses in a multiple environments.

  16. Combined effects of coating, modified atmosphere packaging, and gamma irradiation on quality maintenance of ready-to-use carrots (Daucus carota).

    PubMed

    Lafortune, R; Caillet, S; Lacroix, M

    2005-02-01

    The objective of this study was to evaluate the effect of an edible coating combined with modified atmosphere (MA; 60% O2, 30% CO2, and 10% N2) packaging and gamma irradiation on the microbiological stability and physicochemical quality of minimally processed carrots. A coating based on calcium caseinate and whey protein isolates was used. Coated and uncoated peeled minicarrots were packed under the MA or air (78.1% N2, 20.9% O2, and 0.036% CO2), irradiated at 0.5 or 1 kGy, and stored at 4 +/- 1 degrees C for 21 days. Samples were evaluated periodically for aerobic plates counts (APCs) and physicochemical properties (firmness, white discoloration, and whiteness index). Gamma irradiation did not significantly affect the physicochemical properties of the carrots (P > 0.05). Microbiological analysis revealed that for uncoated carrots irradiation at 0.5 and 1 kGy under air and MA reduced the APCs by 3.5 and 4 log CFU/g and by 4 and 4.5 log CFU/g, respectively. For coated carrots, irradiation at 0.5 and 1 kGy under air and MA reduced the APCs by 4 and 4.5 log CFU/g and by 3 and 4.25 log CFU/g, respectively. The coating was able to protect carrots against dehydration during storage under air. Coating and irradiation at 1 kGy were also able to protect carrot firmness during storage under air. MA packaging retarded whitening of uncoated carrots but had a detrimental effect on firmness. The edible coating used in this study did not significantly inhibit (P > 0.05) microbial growth on carrots.

  17. [Obtaining of hairy-root, callus and suspension carrot culture (Daucus carota L.) able to accumulate human interferon alpha-2b].

    PubMed

    Luchakivskaia, Iu S; Olevinskaia, Z M; Kishchenko, E M; Spivak, N Ia; Kuchuk, N V

    2012-01-01

    Here we report the obtaining of suspension, callus and hairy root culture initiated from carrot plants of Nantskaya and Perfektzya variety with the highest level of recombinant human interferon alpha-2b accumulation exhibited the highest level of plant protein extract antiviral activity (up to 12.8 x 10(3) IU/mg TSP). The antiviral activity of callus extracts was significantly lower comparing to the activity of plant extracts from parent organisms. However, the antiviral activity level of suspension culture extracts (up to 4.42 x 10(3) IU/mg TSP) and Ri-root ones (up to 4.42 x 10(3) IU/mg TSP) appeared to be comparable to analogical data of antiviral activity of transgenic carrot leaf extracts, this way the described cultures could be possibly used for comparatively speedy obtaining of recombinant therapeutic protein for curing and preventing of virus diseases.

  18. Combined effects of antimicrobial coating, modified atmosphere packaging, and gamma irradiation on Listeria innocua present in ready-to-use carrots (Daucus carota).

    PubMed

    Caillet, S; Millette, M; Salmiéri, S; Lacroix, M

    2006-01-01

    The objective of this study was to evaluate the effect of an edible antimicrobial coating combined with modified atmosphere (MA) packaging (60% O2, 30% CO2, and 10% N2) and gamma irradiation on peeled minicarrots inoculated with Listeria innocua. Carrots were inoculated with L. innocua (10(3) CFU/g) and then coated with an antimicrobial coating based on calcium caseinate containing trans-cinnamaldehyde. The same formulation without trans-cinnamaldehyde was used as an inactive coating. Coated and uncoated carrots were packed under the MA or under air, irradiated at 0.25 or 0.5 kGy, and stored at 4 +/- 1 degrees C for 21 days. Samples were evaluated periodically for enumeration of L. innocua. Unirradiated carrots stored under air had the highest concentrations of L. innocua after 21 days of storage: 2.23 CFU/g in the uncoated samples and 2.26 CFU/ g in samples coated with the inactive coating. These results suggest that the inactive coating did not have any antimicrobial effect against L. innocua. However, the addition of the antimicrobial coating resulted in a 1.29-log reduction in the concentration of L. innocua in carrots packed under air after 21 days of storage and a 1.08-log reduction in carrots packed under MA after 7 days of storage. After 7 days of storage, no L. innocua was detected in samples treated at 0.5 kGy under air or in samples treated at 0.25 kGy under MA. A complete inhibition of L. innocua was also observed during all storage periods in uncoated and coated samples treated at 0.5 kGy under MA. These results indicate that the combination of irradiation and MA conditions play an important role in the radiosensitization of L. innocua.

  19. Isolation, Purification and Characterization of Two Laccases from Carrot (Daucus carota L.) and Their Response to Abiotic and Metal Ions Stresses.

    PubMed

    Ma, Jing; Xu, Zhi-Sheng; Wang, Feng; Xiong, Ai-Sheng

    2015-12-01

    Laccases, which belong to the blue copper oxidase enzyme family, oxidize many organic and inorganic compounds. The laccase-encoding genes DcLac1 and DcLac2 were isolated from the economically important tuberous root carrot, and their proteins were successfully expressed and purified using the Escherichia coli expression system BL21(DE3). DcLac1 and DcLac2 had molecular masses of approximately 64 and 61.9 kDa, respectively. With 2,2'-azinobis-(3-ethylbenzthiazoline-6-sulfonate acid) as the substrate, DcLac1 and DcLac2 had K m values of 3.9043 and 1.255 mM, respectively, and V max values of 54.0832 and 81.7996 μM mg(-1) min(-1), respectively. Moreover, DcLac1 and DcLac2 had optimal pH values of 2.8 and 2.6, respectively, and optimal temperatures of 45 and 40 °C, respectively. The activities of the two enzymes were promoted by Ca(2+), Mg(2+), Cu(2+), and Na(+) but inhibited by Fe(2+), Zn(2+), Mn(2+), K(+), SDS, and EDTA. Expression profiles showed that the two DcLac genes had almost identical responses to high and low temperature stresses but different responses to salt, drought, and metal stresses. This study provided insights into the characteristics and tolerance response mechanisms of laccase in carrot.

  20. The influence of the fungal pathogen Mycocentrospora acerina on the proteome and polyacetylenes and 6-methoxymellein in organic and conventionally cultivated carrots (Daucus carota) during post harvest storage.

    PubMed

    Louarn, Sébastien; Nawrocki, Arkadiusz; Edelenbos, Merete; Jensen, Dan F; Jensen, Ole N; Collinge, David B; Jensen, Birgit

    2012-01-01

    Many carrots are discarded during post harvest cold storage due to development of fungal infections, caused by, e.g., Mycocentrospora acerina (liquorice rot). We compared the susceptibility of carrots grown under conventional and organic agricultural practices. In one year, organically cultivated carrots showed 3× to 7× more symptoms than conventionally cultivated, when studying naturally occurring disease at 4 and 6 months, respectively. On the other hand, we have developed a bioassay for infection studies of M. acerina on carrots and observed that organic roots were more susceptible after one month of storage than conventional ones, but no differences were apparent after four or six months storage. Levels of polyacetylenes (falcarinol, falcarindiol and falcarindiol-3-acetate) did not change, whereas the isocoumarin phytoalexin (6-methoxymellein) accumulated in infected tissue as well as in healthy tissue opposite the infection. The proteomes of carrot and M. acerina were characterized, the intensity of 33 plant protein spots was significantly changed in infected roots including up regulation of defence and stress response proteins but also a decrease of proteins involved in energy metabolism. This combined metabolic and proteomic study indicates that roots respond to fungal infection through altered metabolism: simultaneous induction of 6-methoxymellein and synthesis of defence related proteins.

  1. Anthocyanins in purple-orange carrots (Daucus carota L.) do not influence the bioavailability of beta-carotene in young women.

    PubMed

    Arscott, Sara A; Simon, Philipp W; Tanumihardjo, Sherry A

    2010-03-10

    Purple carrots contain anthocyanins in addition to the provitamin A carotenoids in typical orange carrots. Simultaneous consumption of these phytochemicals in carrots may affect the bioavailability of carotenoids. The bioavailability of beta-carotene in humans was assessed from an acute feeding of orange (OC) and purple (PC) carrots with white (WC) as a control. Carrot smoothies were served to female subjects (n = 5, aged 21-26 years) for breakfast after 1 week on a low carotenoid diet and overnight fast. OC and PC smoothies were equalized to 10.3 mg of all-trans beta-carotene. Plasma beta-carotene was measured for 144 h following treatments. Peak plasma concentrations of OC and PC treatments did not differ. The PC treatment 0-144 h area-under-the-curve for beta-carotene was 76% of the OC treatment (P < 0.05). However, when the first 24 h were compared, OC and PC treatments did not differ, suggesting that anthocyanins in purple carrots do not affect the absorption of beta-carotene postprandially.

  2. Fractionation of Plant Bioactives from Black Carrots (Daucus carota subspecies sativus varietas atrorubens Alef.) by Adsorptive Membrane Chromatography and Analysis of Their Potential Anti-Diabetic Activity.

    PubMed

    Esatbeyoglu, Tuba; Rodríguez-Werner, Miriam; Schlösser, Anke; Liehr, Martin; Ipharraguerre, Ignacio; Winterhalter, Peter; Rimbach, Gerald

    2016-07-27

    Black and purple carrots have attracted interest as colored extracts for coloring food due to their high content of anthocyanins. This study aimed to investigate the polyphenol composition of black carrots. Particularly, the identification and quantification of phenolic compounds of the variety Deep Purple carrot (DPC), which presents a very dark color, was performed by HPLC-PDA and HPLC-ESI-MS(n) analyses. The separation of polyphenols from a DPC XAD-7 extract into an anthocyanin fraction (AF) and co-pigment fraction (CF; primarily phenolic acids) was carried out by membrane chromatography. Furthermore, possible anti-diabetic effects of the DPC XAD-7 extract and its AF and CF were determined. DPC samples (XAD-7, CF, and AF) inhibited α-amylase and α-glucosidase in a dose-dependent manner. Moreover, DPC XAD-7 and chlorogenic acid, but not DPC CF and DPC AF, caused a moderate inhibition of intestinal glucose uptake in Caco-2 cells. However, DPC samples did not affect glucagon-like peptide-1 (GLP-1) secretion and dipeptidyl peptidase IV (DPP-4) activity. Overall, DPC exhibits an inhibitory effect on α-amylase and α-glucosidase activity and on cellular glucose uptake indicating potential anti-diabetic properties.

  3. The impact of fermentation with exopolysaccharide producing lactic acid bacteria on rheological, chemical and sensory properties of pureed carrots (Daucus carota L.).

    PubMed

    Juvonen, Riikka; Honkapää, Kaisu; Maina, Ndegwa H; Shi, Qiao; Viljanen, Kaarina; Maaheimo, Hannu; Virkki, Liisa; Tenkanen, Maija; Lantto, Raija

    2015-08-17

    Fermentation with lactic acid bacteria (LAB) offers a natural means to modify technological and nutritional properties of foods and food ingredients. This study explored the impact of fermentation with different exopolysaccharide (EPS) producing LAB on rheological, chemical and sensory properties of puréed carrots in water, as a vegetable model, with the focus on texture formation. The screening of 37 LAB strains for starter selection revealed 16 Lactobacillus, Leuconostoc and Weissella strains capable of EPS (dextran, levan, and/or β-glucan) production in the carrot raw material. Fermentations with five out of six selected EPS producers modified perceived texture of the liquid carrot model (p<0.05). The formation of low-branched dextran correlated with perceived thickness, whereas the production of β-glucan correlated with perceived elasticity. Low-branched dextran producing Weissella confusa and Leuconostoc lactis strains produced thick texture accompanied by pleasant odour and flavour. The fermentation with the selected EPS-producing LAB strains is a promising clean label approach to replace hydrocolloid additives as texturizers in vegetable containing products, not only carrot.

  4. Broad-spectrum disease resistance to necrotrophic and biotrophic pathogens in transgenic carrots (Daucus carota L.) expressing an Arabidopsis NPR1 gene.

    PubMed

    Wally, Owen; Jayaraj, Jayaraman; Punja, Zamir K

    2009-12-01

    The development of transgenic plants highly resistant to a range of pathogens using traditional signal gene expression strategies has been largely ineffective. Modification of systemic acquired resistance (SAR) through the overexpression of a controlling gene such as NPR1 (non-expressor of PR genes) offers an attractive alternative for augmenting the plants innate defense system. The Arabidopsis (At) NPR1 gene was successfully introduced into 'Nantes Coreless' carrot under control of a CaMV 35S promoter and two independent transgenic lines (NPR1-I and NPR1-XI) were identified by Southern and Northern blot hybridization. Both lines were phenotypically normal compared with non-transformed carrots. Northern analysis did not indicate constitutive or spontaneous induction in carrot cultures of SAR-related genes (DcPR-1, 2, 4, 5 or DcPAL). The duration and intensity of expression of DcPR-1, 2 and 5 genes were greatly increased compared with controls when the lines were treated with purified cell wall fragments of Sclerotinia sclerotiorum as well as with 2,6-dichloroisonicotinic acid. The two lines were challenged with the necrotrophic pathogens Botrytis cinerea, Alternaria radicina and S. sclerotiorum on the foliage and A. radicina on the taproots. Both lines exhibited 35-50% reduction in disease symptoms on the foliage and roots when compared with non-transgenic controls. Leaves challenged with the biotrophic pathogen Erysiphe heraclei or the bacterial pathogen Xanthomonas hortorum exhibited 90 and 80% reduction in disease development on the transgenic lines, respectively. The overexpression of the SAR controlling master switch in carrot tissues offers the ability to control a wide range of different pathogens, for which there is currently little genetic resistance available.

  5. Influence of cultivar and harvest year on the volatile profiles of leaves and roots of carrots (Daucus carota spp. sativus Hoffm.).

    PubMed

    Ulrich, Detlef; Nothnagel, Thomas; Schulz, Hartwig

    2015-04-01

    The focus of the present work centers on the diversity of volatile patterns of carrots. In total 15 main volatiles were semiquantified in leaves and roots using isolation by headspace solid phase microextraction followed by gas chromatography with FID and MS detection. Significant differences in the main number of compounds were detected between the cultivars as well as the years. Genotype-environment interactions (G × E) are discussed. The most abundant metabolites, β-myrcene (leaves) and terpinolene (roots), differ in the sum of all interactions (cultivar × harvest year) by a factor of 22 and 62, respectively. A statistical test indicates significant metabolic differences between cultivars for nine volatiles in leaves and 10 in roots. In contrast to others the volatiles α-pinene, γ-terpinene, limonene, and myristicine in leaves as well as β-pinene, humulene, and bornyl acetate in roots are relatively stable over years. A correlation analysis shows no strict clustering regarding root color. While the biosynthesis in leaves and roots is independent between these two organs for nine of the 15 volatiles, a significant correlation of the myristicine content between leaves and roots was determined, which suggests the use of this compound as a bitter marker in carrot breeding.

  6. Increased Nicotiana tabacum fitness through positive regulation of carotenoid, gibberellin and chlorophyll pathways promoted by Daucus carota lycopene β-cyclase (Dclcyb1) expression.

    PubMed

    Moreno, J C; Cerda, A; Simpson, K; Lopez-Diaz, I; Carrera, E; Handford, M; Stange, C

    2016-04-01

    Carotenoids, chlorophylls and gibberellins are derived from the common precursor geranylgeranyl diphosphate (GGPP). One of the enzymes in carotenoid biosynthesis is lycopene β-cyclase (LCYB) that catalyzes the conversion of lycopene into β-carotene. In carrot, Dclcyb1 is essential for carotenoid synthesis in the whole plant. Here we show that when expressed in tobacco, increments in total carotenoids, β-carotene and chlorophyll levels occur. Furthermore, photosynthetic efficiency is enhanced in transgenic lines. Interestingly, and contrary to previous observations where overexpression of a carotenogenic gene resulted in the inhibition of the synthesis of gibberellins, we found raised levels of active GA4 and the concommitant increases in plant height, leaf size and whole plant biomass, as well as an early flowering phenotype. Moreover, a significant increase in the expression of the key carotenogenic genes, Ntpsy1, Ntpsy2 and Ntlcyb, as well as those involved in the synthesis of chlorophyll (Ntchl), gibberellin (Ntga20ox, Ntcps and Ntks) and isoprenoid precursors (Ntdxs2 and Ntggpps) was observed. These results indicate that the expression of Dclcyb1 induces a positive feedback affecting the expression of isoprenoid gene precursors and genes involved in carotenoid, gibberellin and chlorophyll pathways leading to an enhancement in fitness measured as biomass, photosynthetic efficiency and carotenoid/chlorophyll composition.

  7. Fractionation of Plant Bioactives from Black Carrots (Daucus carota subspecies sativus varietas atrorubens Alef.) by Adsorptive Membrane Chromatography and Analysis of Their Potential Anti-Diabetic Activity.

    PubMed

    Esatbeyoglu, Tuba; Rodríguez-Werner, Miriam; Schlösser, Anke; Liehr, Martin; Ipharraguerre, Ignacio; Winterhalter, Peter; Rimbach, Gerald

    2016-07-27

    Black and purple carrots have attracted interest as colored extracts for coloring food due to their high content of anthocyanins. This study aimed to investigate the polyphenol composition of black carrots. Particularly, the identification and quantification of phenolic compounds of the variety Deep Purple carrot (DPC), which presents a very dark color, was performed by HPLC-PDA and HPLC-ESI-MS(n) analyses. The separation of polyphenols from a DPC XAD-7 extract into an anthocyanin fraction (AF) and co-pigment fraction (CF; primarily phenolic acids) was carried out by membrane chromatography. Furthermore, possible anti-diabetic effects of the DPC XAD-7 extract and its AF and CF were determined. DPC samples (XAD-7, CF, and AF) inhibited α-amylase and α-glucosidase in a dose-dependent manner. Moreover, DPC XAD-7 and chlorogenic acid, but not DPC CF and DPC AF, caused a moderate inhibition of intestinal glucose uptake in Caco-2 cells. However, DPC samples did not affect glucagon-like peptide-1 (GLP-1) secretion and dipeptidyl peptidase IV (DPP-4) activity. Overall, DPC exhibits an inhibitory effect on α-amylase and α-glucosidase activity and on cellular glucose uptake indicating potential anti-diabetic properties. PMID:27362825

  8. Influence of thermal processing on hydrolysis and stability of folate poly-gamma-glutamates in broccoli (Brassica oleracea var. italica), carrot (Daucus carota) and tomato (Lycopersicon esculentum).

    PubMed

    Munyaka, Ann Wambui; Verlinde, Philippe; Mukisa, Ivan Muzira; Oey, Indrawati; Van Loey, Ann; Hendrickx, Marc

    2010-04-14

    The folate poly-gamma-glutamate profile, their concentrations, and hydrolysis by endogenous gamma-glutamyl hydrolase (GGH) were evaluated in broccoli, carrot and tomato. Further studies on the effect of time and temperature on folate poly-gamma-glutamate hydrolysis and stability were carried out in broccoli since this vegetable showed the highest long-chain and total folate poly-gamma-glutamate concentration. The evolution of l-ascorbic acid, total phenols and Trolox equivalent antioxidant capacity (TEAC) values was evaluated in parallel. Upon thermal inactivation of GGH prior to crushing, it was observed that broccoli, carrot and tomato contained poly-gamma-glutamates with one to seven glutamate residues but differed in the predominant poly-gamma-glutamates. Crushing of raw broccoli, carrot and tomato resulted in significant poly-gamma-glutamate profile changes in broccoli and carrot (indicating GGH-catalyzed hydrolysis) but not in tomato. In this study, the actual crushing of raw broccoli matrix had a greater effect on folate poly-gamma-glutamate hydrolysis than incubation conditions (0-30 min at 25-55 degrees C). During treatments at 25-140 degrees C, folate retention was higher at 80 and 100 degrees C than at the other temperatures. A similar trend in thermal stability was observed for folates, vitamin C, total phenols and TEAC value, an indication that conditions that result in endogenous antioxidants degradation might also result in folate degradation.

  9. Influence of antimicrobial compounds and modified atmosphere packaging on radiation sensitivity of Listeria monocytogenes present in ready-to-use carrots (Daucus carota).

    PubMed

    Caillet, S; Millette, M; Turgis, M; Salmieri, S; Lacroix, M

    2006-01-01

    Radiosensitization of Listeria monocytogenes was determined in the presence of trans-cinnamaldehyde, Spanish oregano, winter savory, and Chinese cinnamon on peeled minicarrots packed under air or under a modified atmosphere (60% O2, 30% CO2, and 10% N2). Samples were inoculated with L. monocytogenes HPB 2812 serovar 1/2a (106 CFU/g) and were coated separately with each active compound (0.5%, wt/wt) before being packaged under air or the modified atmosphere and irradiated at doses from 0.07 to 2.4 kGy. Results indicated that the bacterium was more resistant to irradiation under air in the absence of active compound. The dose required to reduce L. monocytogenes population by 1 log CFU (D10) was 0.36 kGy for samples packed under air and 0.17 kGy for those packed under the modified atmosphere. The active compounds evaluated in this study had an effect on the radiation sensitivity of L. monocytogenes on carrots. The most efficient compound was trans-cinnamaldehyde, where a mean 3.8-fold increase in relative radiation sensitivity was observed for both atmospheres compared with the control. The addition of winter savory and Chinese cinnamon produced a similar increase in relative radiation sensitivity but only when samples where packed under modified atmosphere conditions.

  10. Increased Nicotiana tabacum fitness through positive regulation of carotenoid, gibberellin and chlorophyll pathways promoted by Daucus carota lycopene β-cyclase (Dclcyb1) expression

    PubMed Central

    Moreno, J.C.; Cerda, A.; Simpson, K.; Lopez-Diaz, I.; Carrera, E; Handford, M.; Stange, C.

    2016-01-01

    Carotenoids, chlorophylls and gibberellins are derived from the common precursor geranylgeranyl diphosphate (GGPP). One of the enzymes in carotenoid biosynthesis is lycopene β-cyclase (LCYB) that catalyzes the conversion of lycopene into β-carotene. In carrot, Dclcyb1 is essential for carotenoid synthesis in the whole plant. Here we show that when expressed in tobacco, increments in total carotenoids, β-carotene and chlorophyll levels occur. Furthermore, photosynthetic efficiency is enhanced in transgenic lines. Interestingly, and contrary to previous observations where overexpression of a carotenogenic gene resulted in the inhibition of the synthesis of gibberellins, we found raised levels of active GA4 and the concommitant increases in plant height, leaf size and whole plant biomass, as well as an early flowering phenotype. Moreover, a significant increase in the expression of the key carotenogenic genes, Ntpsy1, Ntpsy2 and Ntlcyb, as well as those involved in the synthesis of chlorophyll (Ntchl), gibberellin (Ntga20ox, Ntcps and Ntks) and isoprenoid precursors (Ntdxs2 and Ntggpps) was observed. These results indicate that the expression of Dclcyb1 induces a positive feedback affecting the expression of isoprenoid gene precursors and genes involved in carotenoid, gibberellin and chlorophyll pathways leading to an enhancement in fitness measured as biomass, photosynthetic efficiency and carotenoid/chlorophyll composition. PMID:26893492

  11. Isolation, Purification and Characterization of Two Laccases from Carrot (Daucus carota L.) and Their Response to Abiotic and Metal Ions Stresses.

    PubMed

    Ma, Jing; Xu, Zhi-Sheng; Wang, Feng; Xiong, Ai-Sheng

    2015-12-01

    Laccases, which belong to the blue copper oxidase enzyme family, oxidize many organic and inorganic compounds. The laccase-encoding genes DcLac1 and DcLac2 were isolated from the economically important tuberous root carrot, and their proteins were successfully expressed and purified using the Escherichia coli expression system BL21(DE3). DcLac1 and DcLac2 had molecular masses of approximately 64 and 61.9 kDa, respectively. With 2,2'-azinobis-(3-ethylbenzthiazoline-6-sulfonate acid) as the substrate, DcLac1 and DcLac2 had K m values of 3.9043 and 1.255 mM, respectively, and V max values of 54.0832 and 81.7996 μM mg(-1) min(-1), respectively. Moreover, DcLac1 and DcLac2 had optimal pH values of 2.8 and 2.6, respectively, and optimal temperatures of 45 and 40 °C, respectively. The activities of the two enzymes were promoted by Ca(2+), Mg(2+), Cu(2+), and Na(+) but inhibited by Fe(2+), Zn(2+), Mn(2+), K(+), SDS, and EDTA. Expression profiles showed that the two DcLac genes had almost identical responses to high and low temperature stresses but different responses to salt, drought, and metal stresses. This study provided insights into the characteristics and tolerance response mechanisms of laccase in carrot. PMID:26626349

  12. Analysis of the Thermal Degradation of the Individual Anthocyanin Compounds of Black Carrot (Daucus carota L.): A New Approach Using High-Resolution Proton Nuclear Magnetic Resonance Spectroscopy.

    PubMed

    Iliopoulou, Ioanna; Thaeron, Delphine; Baker, Ashley; Jones, Anita; Robertson, Neil

    2015-08-12

    The black carrot dye is a mixture of cyanidin molecules, the nuclear magnetic resonance (NMR) spectrum of which shows a highly overlapped aromatic region. In this study, the (1)H NMR (800 MHz) aromatic chemical shifts of the mixture were fully assigned by overlaying them with the characterized (1)H NMR chemical shifts of the separated compounds. The latter were isolated using reverse-phase high-performance liquid chromatography (RP-HPLC), and their chemical shifts were identified using (1)H and two-dimensional (2D) correlation spectroscopy (COSY) NMR spectroscopy. The stability of the black carrot mixture to heat exposure was investigated at pH 3.6, 6.8, and 8.0 by heat-treating aqueous solutions at 100 °C and the powdered material at 180 °C. From integration of high-resolution (1)H NMR spectra, it was possible to follow the relative degradation of each compound, offering advantages over the commonly used ultraviolet/visible (UV/vis) and HPLC approaches. UV/vis spectroscopy and CIE color measurements were used to determine thermally induced color changes, under normal cooking conditions. PMID:26160425

  13. The impact of fermentation with exopolysaccharide producing lactic acid bacteria on rheological, chemical and sensory properties of pureed carrots (Daucus carota L.).

    PubMed

    Juvonen, Riikka; Honkapää, Kaisu; Maina, Ndegwa H; Shi, Qiao; Viljanen, Kaarina; Maaheimo, Hannu; Virkki, Liisa; Tenkanen, Maija; Lantto, Raija

    2015-08-17

    Fermentation with lactic acid bacteria (LAB) offers a natural means to modify technological and nutritional properties of foods and food ingredients. This study explored the impact of fermentation with different exopolysaccharide (EPS) producing LAB on rheological, chemical and sensory properties of puréed carrots in water, as a vegetable model, with the focus on texture formation. The screening of 37 LAB strains for starter selection revealed 16 Lactobacillus, Leuconostoc and Weissella strains capable of EPS (dextran, levan, and/or β-glucan) production in the carrot raw material. Fermentations with five out of six selected EPS producers modified perceived texture of the liquid carrot model (p<0.05). The formation of low-branched dextran correlated with perceived thickness, whereas the production of β-glucan correlated with perceived elasticity. Low-branched dextran producing Weissella confusa and Leuconostoc lactis strains produced thick texture accompanied by pleasant odour and flavour. The fermentation with the selected EPS-producing LAB strains is a promising clean label approach to replace hydrocolloid additives as texturizers in vegetable containing products, not only carrot. PMID:26001525

  14. Influence of Boron Nutrition on Net Uptake and Efflux of (32)P and (14)C-Glucose in Helianthus annuus Roots and Cell Cultures of Daucus carota.

    PubMed

    Goldbach, H

    1985-04-01

    (32)P and (14)C-glucose uptake were reduced under B deficiency in both Daucus cell suspensions and Helianthus roots. Similarly, efflux rates were found to be smaller in B deficient material. Efflux rates tended to be more affected than net uptake of both (32)P and glucose. This may explain why sunflower roots showed a higher glucose net uptake immediately after transferring from a B sufficient to a B deficient nutrient solution. The data confirm earlier findings of B as an essential element for membrane function and integrity. B-deficiency effects could be reversed by the addition of B within less than one hour. Cell suspensions reacted similarly to roots with respect to B deficiency and may thus be suitable for further research on B deficiency effects.

  15. The development-specific ssp1 and ssp2 genes of Sclerotinia sclerotiorum encode lectins with distinct yet compensatory regulation.

    PubMed

    Li, Moyi; Rollins, Jeffrey A

    2010-06-01

    The Ssp1 development-specific protein is the most abundant soluble protein in sclerotia and apothecia of Sclerotinia sclerotiorum. Although closely associated with these developmental stages, the functions of the Ssp1 protein and its paralog, Ssp2, are not known. In this study, protein structure prediction analysis revealed that Ssp1 and Ssp2 are structurally similar to fucose-specific lectins. In an effort to understand the function of these abundant, development-specific proteins, a homokaryotic ssp1 deletion mutant was generated. The resulting mutant (Deltassp1) displays a wild-type growth and development phenotype in culture but produces approximately 50% fewer sclerotia in cultures supplemented with hygromycin. Genetic complementation with a wild-type copy of ssp1 restores normal sclerotium formation in the presence of hygromycin. This suggests that Ssp1 might play a role in resistance to glycoside-containing antibiotics encountered in the environment. Although a slight delay in carpogenic germination was observed, no additional effects of ssp1 loss-of-function were found in regards to apothecial morphology or fecundity. When the expression of ssp2 was examined in the Deltassp1 mutant, it was found to be expressed earlier in sclerotial development and its encoded protein accumulated to higher levels in both sclerotia and apothecia. These findings suggest regulatory compensation for loss of Ssp1 coupled with potential functional redundancy among lectins accumulating in sclerotia and apothecia. PMID:20350614

  16. Composition and chemical variability of Mentha suaveolens ssp. suaveolens and M. suaveolens ssp. insularis from Corsica.

    PubMed

    Sutour, Sylvain; Bradesi, Pascale; Casanova, Joseph; Tomi, Félix

    2010-04-01

    A detailed analysis of two essential oils from individual plants of Mentha suaveolens ssp. suaveolens growing wild in Corsica was carried out by combination of GC(RI), GC/MS, and (13)C-NMR analyses. One oil sample is characterized by the pre-eminence of piperitenone oxide, and the second is dominated by piperitenone. In contrast, it was reported that the essential oil of M. suaveolens ssp. insularis, an endemic species to Corsica and Sardinia, contained pulegone and cis-cis-p-menthenolide as main components. A principal-component analysis (PCA) carried out on the composition of the essential oil of 59 individual plants of M. suaveolens sp. allowed the classification into three well-defined groups. All the oil samples from Mentha suaveolens ssp. insularis belonged to the same group, while the oils from M. suaveolens ssp. suaveolens were distributed in the two other groups. The composition of the essential oil isolated from aerial parts of M. suaveolens ssp. has been shown to be an additional tool to differentiate the botanically close subspecies suaveolens and insularis.

  17. The Huygens Surface Science Package (SSP)

    NASA Astrophysics Data System (ADS)

    Zarnecki, John C.; Leese, Mark; Hathi, Brijen; SSP Consortium, The

    2015-04-01

    The Huygens Surface Science Package (SSP), one of the six instruments selected by ESA, was the only one whose prime purpose was to study the surface of Titan in-situ. Designed at a time when even the basic nature of the surface (i.e. solid ice or liquid hydrocarbons) was unknown, it consisted of 9 separate compact and relatively simple sensors designed to span all likely surface scenarios. The Huygens landing on a "dry lake bed" allowed some of the sensors to be fully exercised. The results and their subsequent interpretations will be presented as well as perspectives from nearly 25 years since the original instrument selection.

  18. Starvation-induced expression of SspA and SspB: the effects of a null mutation in sspA on Escherichia coli protein synthesis and survival during growth and prolonged starvation.

    PubMed

    Williams, M D; Ouyang, T X; Flickinger, M C

    1994-03-01

    Maxicell labelling and two-dimensional gel electrophoresis (2-D PAGE) have identified the proteins encoded by sspA and sspB (SspA, SspB) as proteins D27.1 and A25.8, respectively, in the Escherichia coli gene-protein database. SspA expression increases with decreasing growth rate and is induced by glucose, nitrogen, phosphate or amino acid starvation. The promoter, Pssp, is similar to gearbox promoters. Inactivation of SspA (sspA::neo) blocks sspB expression. [35S]-methionine-labelled proteins synthesized during growth and during stationary phase are different in delta sspA strains compared to sspA+ strains. This difference is enhanced during extended stationary phase (24-72 h). Long-term (10 d) viability of arginine-starved isogenic strains shows that sspA+ cultures remain viable significantly longer than delta sspA mutants. 2-D PAGE of proteins expressed during exponential growth shows that expression of at least 11 proteins is altered in delta sspA strains. A functional relA gene is required for sspA to affect protein synthesis.

  19. β-2-himachalen-6-ol: A novel anticancer sesquiterpene unique to the Lebanese wild carrot.

    PubMed

    Taleb, Robin I; Najm, Paul; Shebaby, Wassim; Boulos, Joelle C; Demirdjian, Sally; Hariri, Essa; El-Sibai, Mirvat; Daher, Costantine; Mroueh, Mohamad

    2016-08-22

    Daucus carota ssp. carota, also known as wild carrot, is a commonly used herb in Lebanese folk medicine to treat several ailments including cancer. Previous studies in our laboratories showed that the Daucus carota oil extract (DCOE) and subsequent fractions exhibit antioxidant, anti-inflammatory and anti-cancer activities. In this study, we report the isolation and identification of the major compound responsible for the anti-cancer activity of DCOE along with the mechanism of action involved. GC-MS and NMR spectroscopy revealed the identity of the major compound as β-2-himachalen-6-ol, a novel sesquiterpene unique to the Lebanese wild carrot. β-2-Himachalen-6-ol demonstrated potent anti-cancer activity against B16F-10, Caco-2, MB-MDA-231, A549 and SF-268 cancer cells (IC50 13-4µg/ml; 58-18µM), with SF-268 cells being the most sensitive. The sesquiterpene was shown to induce cell death through apoptosis (flow cytometry), decrease 2D cell motility (wound healing assay) and 3D invasion, as well as increase cell adhesion in SF-268 cells. Additionally, β-2-himachalen-6-ol showed very low toxicity in mice with an LD50>6000mg/kg body weight. In conclusion, the present data demonstrate that β-2-himachalen-6-ol is a potential multi-mechanistic chemotherapeutic drug with high potency and safety. PMID:27240746

  20. School Teams up for SSP Functional Models

    NASA Astrophysics Data System (ADS)

    Pignolet, G.; Lallemand, R.; Celeste, A.; von Muldau, H.

    2002-01-01

    Space Solar Power systems appear increasingly as one of the major solutions to the upcoming global energy crisis, by collecting solar energy in space where this is most easy, and sending it by microwave beam to the surface of the planet, where the need for controlled energy is located. While fully operational systems are still decades away, the need for major development efforts is with us now. Yet, for many decision-makers and for most of the public, SSP often still sounds like science fiction. Six functional demonstration systems, based on the Japanese SPS-2000 concept, have been built as a result of a cooperation between France and Japan, and they are currently used extensively, in Japan, in Europe and in North America, for executive presentations as well as for public exhibitions. There is demand for more models, both for science museums and for use by energy dedicated groups, and a senior high school in La Reunion, France, has picked up the challenge to make the production of such models an integrated practical school project for pre-college students. In December 2001, the administration and the teachers of the school have evaluated the feasibility of the project and eventually taken the go decision for the school year 2002- 2003, when for education purposes a temporary "school business company" will be incorporated with the goal to study and manufacture a limited series of professional quality SSP demonstration models, and to sell them world- wide to institutions and advocacy groups concerned with energy problems and with the environment. The different sections of the school will act as the different services of an integrated business : based on the current existing models, the electronic section will redesign the energy management system and the microwave projector module, while the mechanical section of the school will adapt and re-conceive the whole packaging of the demonstrator. The French and foreign language sections will write up a technical manual for

  1. [Flavonoids of Artemisia campestris, ssp. glutinosa].

    PubMed

    Hurabielle, M; Eberle, J; Paris, M

    1982-10-01

    Four flavanones (pinostrobin, pinocembrin, sakuranetin and naringenin), one dihydroflavonol (7-methyl aromadendrin) and one flavone (hispidulin) have been isolated from Artemisia campestris L. ssp. glutinosa Gay and identified by spectroscopic methods. Artemisia campestris L. sous-espèce glutinosa Gay est une Composée Anthémidée largement répandue sur les sables du littoral méditerranéean et abondante dans certaines régions d'Espagne et d'Italie. Dans le cadre d'une étude chimiotaxonomique du genre Artemisia Tourn., nous nous sommes intéressés à l'analyse des flavonoïdes, composés jamais décrits, à notre connaissance, dans cette espèce d' Artemisia. Les sommités fleuries d' Artemisia campestris sous-espèce glutinosa, séchées et pulvérisées, sont dégraissées à l'ether de pétrole et épuisées par le chloroforme. Le fractionnement de l'extrait chloroformique, par chromatographie sur colonne de silice, et la purification de certaines fractions conduisent à l'isolement de six génines flavoniques, à l'etat pur. L' étude des spectres UV, des spectres de masse et des spectres de RMN [1,2] et la comparaison avec des échantillons authentiques permettent de proposer, pour ces flavonoïdes, les structures de la pinostrobine [3], de la pinocembrine [4], de la sakuranétine, de la naringénine [5] (flavanones), de la méthyl-7-aromadendrine, [6, 7] (dihydroflavonol) et de l'hispiduline [8, 9] (flavone); quatre de ces génines sont méthylées. Parmi ces flavonoïdes, la pinostrobine n'a jamais été décrite, à notre connaissance, dans la famille des Composées; la pinocembrine, la sakuranétine et la naringénine ont déjà été signalées chez quelques Astéracées et Eupatoriées [10], et l'hispiduline dans la tribu des Anthémidées ( Santolina chamaecyparissus L.) [8]. Seule, la méthyl-7-aromadendrine semble décrite, à ce jour, dans le genre Artemisia Tourn. [7].

  2. Galanthindole: a new indole alkaloid from Galanthus plicatus ssp. byzantinus.

    PubMed

    Unver, Nehir; Kaya, G Irem; Werner, Christa; Verpoorte, Robert; Gözler, Belkis

    2003-09-01

    A new indole alkaloid, galanthindole, was isolated from Galanthus plicatus ssp. byzantinus (Amaryllidaceae), a plant native to northwestern Turkey. Incorporating a non-fused indole ring, galanthindole may represent the prototype of a new subgroup of the Amaryllidaceae alkaloids. Two other bases, (+)-11-hydroxyvittatine and hordenine, are also reported from the same plant.

  3. Stress-Induced Accumulation of DcAOX1 and DcAOX2a Transcripts Coincides with Critical Time Point for Structural Biomass Prediction in Carrot Primary Cultures (Daucus carota L.).

    PubMed

    Campos, M Doroteia; Nogales, Amaia; Cardoso, Hélia G; Kumar, Sarma R; Nobre, Tânia; Sathishkumar, Ramalingam; Arnholdt-Schmitt, Birgit

    2016-01-01

    Stress-adaptive cell plasticity in target tissues and cells for plant biomass growth is important for yield stability. In vitro systems with reproducible cell plasticity can help to identify relevant metabolic and molecular events during early cell reprogramming. In carrot, regulation of the central root meristem is a critical target for yield-determining secondary growth. Calorespirometry, a tool previously identified as promising for predictive growth phenotyping has been applied to measure the respiration rate in carrot meristem. In a carrot primary culture system (PCS), this tool allowed identifying an early peak related with structural biomass formation during lag phase of growth, around the 4th day of culture. In the present study, we report a dynamic and correlated expression of carrot AOX genes (DcAOX1 and DcAOX2a) during PCS lag phase and during exponential growth. Both genes showed an increase in transcript levels until 36 h after explant inoculation, and a subsequent down-regulation, before the initiation of exponential growth. In PCS growing at two different temperatures (21°C and 28°C), DcAOX1 was also found to be more expressed in the highest temperature. DcAOX genes' were further explored in a plant pot experiment in response to chilling, which confirmed the early AOX transcript increase prior to the induction of a specific anti-freezing gene. Our findings point to DcAOX1 and DcAOX2a as being reasonable candidates for functional marker development related to early cell reprogramming. While the genomic sequence of DcAOX2a was previously described, we characterize here the complete genomic sequence of DcAOX1. PMID:26858746

  4. Development and in-house validation of allergen-specific ELISA tests for the quantification of Dau c 1.01, Dau c 1.02 and Dau c 4 in carrot extracts (Daucus carota).

    PubMed

    Foetisch, Kay; Dahl, Lotte; Jansen, Baerbel; Becker, Wolf-Meinhard; Lidholm, Jonas; van Ree, Ronald; Broll, Hermann; Kaul, Susanne; Vieths, Stefan; Holzhauser, Thomas

    2011-01-01

    Even though carrot allergy is common in Europe, the amount of different allergens in carrots is still unknown due to a lack of methods for quantitative allergen measurements. The current study aimed at the development of quantitative ELISA tests for the known carrot allergens, namely Dau c 1.01, Dau c 1.02, and Dau c 4 in pure carrot extracts. Monoclonal antibodies targeting the major carrot allergen isoforms Dau c 1.01 and Dau c 1.02 were generated and combined in sandwich ELISA with rabbit antisera against Api g 1, the celery homologue of Dau c 1. A competitive ELISA for the carrot profilin Dau c 4 was based on a polyclonal rabbit antiserum. The three ELISA tests were allergen-specific and displayed detection limits between 0.4 and 6 ng allergen/ml of carrot extract. The mean coefficient of variation (CV) as a means of intraassay variability of the Dau c 1.01, Dau c 1.02 and Dau c 4 ELISA tests was 8.1%, 6.9%, and 11.9%, and the mean interassay CV 13.3%, 37.1% and 15.6%, respectively. Target recovery ranged between 93 and 113%. In conclusion, the specific, accurate and reproducible quantification of three important carrot allergens may help to identify less allergenic carrot varieties, as well as to standardize the amount of allergens in extracts used for carrot allergy diagnosis.

  5. Quantitative studies and sensory analyses on the influence of cultivar, spatial tissue distribution, and industrial processing on the bitter off-taste of carrots (Daucus carota l.) and carrot products.

    PubMed

    Czepa, Andreas; Hofmann, Thomas

    2004-07-14

    Although various reports pointed to 6-methoxymellein (1) as a key player imparting the bitter taste in carrots, activity-guided fractionation experiments recently gave evidence that not this isocoumarin but bisacetylenic oxylipins contribute mainly to the off-taste. Among these, (Z)-heptadeca-1,9-dien-4,6-diyn-3-ol (2), (Z)-3-acetoxy-heptadeca-1,9-dien-4,6-diyn-8-ol (3), and (Z)-heptadeca-1,9-dien-4,6-diyn-3,8-diol (falcarindiol, 4) have been successfully identified. In the present study, an analytical procedure was developed enabling an accurate quantitation of 1-4 in carrots and carrot products. To achieve this, (E)-heptadeca-1,9-dien-4,6-diyn-3,8-diol was synthesized as a suitable internal standard for the quantitative analysis of the bisacetylenes. On the basis of taste activity values, calculated as the ratio of the concentration and the human sensory threshold of a compound, a close relationship between the concentration of 4 and the intensity of the bitter off-taste in carrots, carrot puree, and carrot juice was demonstrated, thus showing that compound 4 might offer a new analytical measure for an objective evaluation of the quality of carrot products. Quantitative analysis on the intermediate products in industrial carrot processing revealed that removing the peel as well as green parts successfully decreased the concentrations in the final carrot puree by more than 50%.

  6. Influence of field attack by carrot psyllid (Trioza apicalis Förster) on sensory quality, antioxidant capacity and content of terpenes, falcarindiol and 6-methoxymellein of carrots (Daucus carota L.).

    PubMed

    Seljåsen, Randi; Vogt, Gjermund; Olsen, Elisabeth; Lea, Per; Høgetveit, Lars Arne; Tajet, Torgeir; Meadow, Richard; Bengtsson, Gunnar B

    2013-03-20

    The effect of different degrees of attack by carrot psyllid (Trioza apicalis) on quality parameters of carrots was studied in field experiments for two years. Treatments were different degrees of physical insect protection by floating row cover. An increasing attack level of psyllids showed an enhancement effect on the antioxidant capacity (ORAC), content of falcarindiol, 6-methoxymellein, and terpenes, and scores for bitter taste, chemical flavor, terpene flavor, and toughness. Carrot psyllid attack decreased the yield, total sugar, fructose, glucose, and sensory attributes sweet taste, color hue, color strength, crispiness, and juiciness. Carrot plants at 8-10 weeks of age tolerated attack by psyllids at low levels (2% leaves with curling or discoloration).

  7. Stress-Induced Accumulation of DcAOX1 and DcAOX2a Transcripts Coincides with Critical Time Point for Structural Biomass Prediction in Carrot Primary Cultures (Daucus carota L.).

    PubMed

    Campos, M Doroteia; Nogales, Amaia; Cardoso, Hélia G; Kumar, Sarma R; Nobre, Tânia; Sathishkumar, Ramalingam; Arnholdt-Schmitt, Birgit

    2016-01-01

    Stress-adaptive cell plasticity in target tissues and cells for plant biomass growth is important for yield stability. In vitro systems with reproducible cell plasticity can help to identify relevant metabolic and molecular events during early cell reprogramming. In carrot, regulation of the central root meristem is a critical target for yield-determining secondary growth. Calorespirometry, a tool previously identified as promising for predictive growth phenotyping has been applied to measure the respiration rate in carrot meristem. In a carrot primary culture system (PCS), this tool allowed identifying an early peak related with structural biomass formation during lag phase of growth, around the 4th day of culture. In the present study, we report a dynamic and correlated expression of carrot AOX genes (DcAOX1 and DcAOX2a) during PCS lag phase and during exponential growth. Both genes showed an increase in transcript levels until 36 h after explant inoculation, and a subsequent down-regulation, before the initiation of exponential growth. In PCS growing at two different temperatures (21°C and 28°C), DcAOX1 was also found to be more expressed in the highest temperature. DcAOX genes' were further explored in a plant pot experiment in response to chilling, which confirmed the early AOX transcript increase prior to the induction of a specific anti-freezing gene. Our findings point to DcAOX1 and DcAOX2a as being reasonable candidates for functional marker development related to early cell reprogramming. While the genomic sequence of DcAOX2a was previously described, we characterize here the complete genomic sequence of DcAOX1.

  8. Stress-Induced Accumulation of DcAOX1 and DcAOX2a Transcripts Coincides with Critical Time Point for Structural Biomass Prediction in Carrot Primary Cultures (Daucus carota L.)

    PubMed Central

    Campos, M. Doroteia; Nogales, Amaia; Cardoso, Hélia G.; Kumar, Sarma R.; Nobre, Tânia; Sathishkumar, Ramalingam; Arnholdt-Schmitt, Birgit

    2016-01-01

    Stress-adaptive cell plasticity in target tissues and cells for plant biomass growth is important for yield stability. In vitro systems with reproducible cell plasticity can help to identify relevant metabolic and molecular events during early cell reprogramming. In carrot, regulation of the central root meristem is a critical target for yield-determining secondary growth. Calorespirometry, a tool previously identified as promising for predictive growth phenotyping has been applied to measure the respiration rate in carrot meristem. In a carrot primary culture system (PCS), this tool allowed identifying an early peak related with structural biomass formation during lag phase of growth, around the 4th day of culture. In the present study, we report a dynamic and correlated expression of carrot AOX genes (DcAOX1 and DcAOX2a) during PCS lag phase and during exponential growth. Both genes showed an increase in transcript levels until 36 h after explant inoculation, and a subsequent down-regulation, before the initiation of exponential growth. In PCS growing at two different temperatures (21°C and 28°C), DcAOX1 was also found to be more expressed in the highest temperature. DcAOX genes’ were further explored in a plant pot experiment in response to chilling, which confirmed the early AOX transcript increase prior to the induction of a specific anti-freezing gene. Our findings point to DcAOX1 and DcAOX2a as being reasonable candidates for functional marker development related to early cell reprogramming. While the genomic sequence of DcAOX2a was previously described, we characterize here the complete genomic sequence of DcAOX1. PMID:26858746

  9. Choleretic activity of Gentiana lutea ssp. symphyandra in rats.

    PubMed

    Oztürk, N; Herekman-Demir, T; Oztürk, Y; Bozan, B; Başer, K H

    1998-08-01

    Effects of an ethanolic extract prepared from G. lutea ssp. symphyandra roots on the bile production and liver in rats were investigated. Bile flows of rats which were treated by a single i.p. dose of CCl(4) 24 h prior to experiments were measured after the cannulation of bile duct under urethane anaesthesia. After an equilibration period of 1 h, the lyophilized extract were administered intraduodenally (500 mg/kg i.p.), while control animals received physiological saline only. To monitor the effect of multiple dose therapy, rats received the same dose of G. lutea ssp. symphyandra extract for 3 days (2 days prior to CCl(4) administration) and their bile flows were measured after the cannulation. In all groups, bile samples were collected for 3 h with 15 min intervals. After the completion of bile flow experiment, rat livers were removed and put in neutral formaldehyde solution (10%) for the histological examination. According to results obtained, multiple dose treatment of rats with the plant extract normalized the decreased bile flow due CCl(4), whereas single dose therapy was ineffective on the impaired bile flow. These data indicate that the extract prepared from Gentiana lutea ssp. symphyandra roots has a hepatoprotective activity.

  10. Space Solar Power Technical Interchange Meeting 2: SSP TIM 2

    NASA Technical Reports Server (NTRS)

    Sanders, Jim; Hawk, Clark W.

    1998-01-01

    The 2nd Space Solar Power Technical Interchange Meeting (SSP TIM 2) was conducted September 21st through 24th with the first part consisting of a Plenary session. The summary results of this Plenary session are contained in part one of this report. The attendees were then organized into Working Breakout Sessions and Integrated Product Team (IPT) Sessions for the purpose of conducting in-depth discussions in specific topic areas and developing a consensus as to appropriate study plans and actions to be taken. The Second part covers the Plenary Summary Session, which contains the summary results of the Working Breakout Sessions and IPT Sessions. The appendix contains the list of attendees. The ob'jective was to provide an update for the study teams and develop plans for subsequent study activities. This SSP TIM 2 was initiated and the results reported electronically over the Internet. The International Space Station (ISS) could provide the following opportunities for conducting research and technology (R&T) which are applicable to SSP: (1) Automation and Robotics, (2) Advanced Power Generation, (3) Advanced Power Management & Distribution (PMAD), (4) Communications Systems and Networks, (5) Energy Storage, (6) In Space Propulsion (ISP), (7) Structural Dynamics and Control, and Assembly and (8) Wireless Power Transmission.

  11. Multiple origins of cultivated grapevine (Vitis vinifera L. ssp. sativa) based on chloroplast DNA polymorphisms.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The domestication of the Eurasian grape (Vitis vinifera ssp. sativa) from its wild ancestor (Vitis vinifera ssp. sylvestris) has long been claimed to have occurred in Transcaucasia where its greatest genetic diversity is found and where very early archaeological evidence, including grape pips and ar...

  12. ssp genes and spore osmotolerance in Bacillus thuringiensis israelensis and Bacillus sphaericus.

    PubMed

    Cucchi, A; Sanchez de Rivas, C

    1995-10-01

    It was shown previously that spores and vegetative cells of Bacillus sphaericus (Bf) and Bacillus thuringiensis israelensis (Bti) are very sensitive to osmotic variations. Since spore osmotolerance has been associated with their SASP (small acid soluble spore proteins) content coded by ssp genes, hybridization assays were performed with sspE and sspA genes from B. subtilis as probes and showed that Bti and Bf strains could lack an sspE-like gene. The B. subtilis sspE gene was then introduced into Bti 4Q2 strain; spores were obtained and showed a 65 to 650 times higher level of osmotolerance to NaCl, without affecting other important properties: hypoosmotic resistance in vegetative cells, spore UV resistance, and larvicidal activity against diptera larvae. PMID:7549769

  13. Nonindependent domestication of the two rice subspecies, Oryza sativa ssp. indica and ssp. japonica, demonstrated by multilocus microsatellites.

    PubMed

    Gao, Li-Zhi; Innan, Hideki

    2008-06-01

    The origins of the Asian cultivated rice Oryza sativa from its wild ancestor O. rufipogon have been debated for decades. The question mainly concerns whether it originated monophyletically or polyphyletically. To shed light on the origins and demographic history of rice domestication, we genotyped a total of 92 individual plants from the two O. sativa subspecies and O. rufipogon for 60 microsatellites. An approximate Bayesian method was applied to estimate demographic parameters for O. rufipogon vs. O. sativa ssp. indica and O. rufipogon vs. O. sativa ssp. japonica. We showed that the japonica subspecies suffered a more severe bottleneck than the indica subspecies and thus a greater loss of genetic variation during its domestication. Across microsatellite loci there is a significant positive correlation in the reduction of genetic diversity between the two subspecies. The results suggest that completely independent domestication of indica and japonica subspecies may not explain our data and that there is at least partial sharing of their ancestral populations and/or recent gene flow between them. PMID:18505887

  14. Potential SSP Perfluorooctanoic Acid Related Fluoropolymer Materials Obsolescence

    NASA Technical Reports Server (NTRS)

    Segars, Matt G.

    2006-01-01

    The Shuttle Environmental Assurance Initiative (SEA) has identified a potential for the Space Shuttle Program (SSP) to incur materials obsolescence issues due to agreements between the fluoro-chemical industry and the United States Environmental Protection Agency (USEPA) to participate in a Global Stewardship Program for perfluorooctanoic acid (PFOA). This presentation will include discussions of the chemistry, regulatory drivers, affected types of fluoropolymer and fluoroelastomer products, timeline for reformulations, and methodology for addressing the issue. It will cover the coordination of assessment efforts with the International Space Station and Head Quarters Air Force Space Command, along with some examples of impacted materials. The presentation is directed at all members of the international aerospace community concerned with identifying potential environmentally driven materials obsolescence issues.

  15. Evaluation of RO modules for the SSP ETC/LSS.

    NASA Technical Reports Server (NTRS)

    Jasionowski, W. J.; Bambenek, R. A.

    1973-01-01

    During the past eight years the NASA Manned Spacecraft Center has supported the development of an Integrated Water and Waste Management System for use in the Space Station Prototype (SSP) Environmental Thermal Control/Life-Support System (ETC/LSS). This system includes the reverse osmosis (RO) process for recycling wash water and the compression distillation process for recovering useable water from urine, urinal flush water, humidity condensate, commode flush water and the wash water concentrated by RO. This paper summarizes the experimental work performed during the past four years to select the best commercially available RO module for this system and to also define which surfactants and germicides are most compatible with the selected module.

  16. Seed germination characteristics of Chrysothamnus nauseosus ssp. viridulus (Astereae, Asteraceae)

    SciTech Connect

    Khan, M.A.; Sankhla, N.; Weber, D.J.; McArthur, E.D.

    1987-04-30

    Rubber rabbitbrush (Chrysothamnus nauseosus (Pallas) Britt. ssp. viridulus) may prove to be a source of high-quality cis-isoprene rubber, but its establishment is limited by a lack of information on seed germination. Consequently, seeds were germinated at alternating temperatures (5-15, 5-25, 15-25, and 20-30 C) in light and dark as well as constant temperatures (15-40 C with 5-C increments) to determine temperature response. Seeds were also germinated in solutions of polyethylene glycol 6000, salinity regimes at all the above-mentioned temperatures to determine salinity and temperature interaction. The hormones GA/sub 3/ and kinetin were used to study their effect on overcoming salt- and temperature-induced germination inhibition. Seeds of C. nauseosus ssp. viridulus were very sensitive to low temperature. Best germination was achieved at 25 and 30 C, but these seeds also germinated at a higher temperature (35 C). The seeds of rabbit brush germinated at both constant and alternating temperatures. Light appears to play little or no role in controlling germination of the seeds of rubber rabbitbrush. However, seeds of rabbitbrush were sensitive to salinity, and seed germination was progressively inhibited by increase in salt concentration, although a few seeds still germinated at the highest saline level. Progressively higher concentrations of polyethylene glycol also progressively inhibited germination. Suppression of seed germination induced by high salt concentrations and high temperatures can be partially alleviated by the application of either GA/sub 3/ or kinetin. 34 references, 5 figures, 3 tables.

  17. Bosom Buddies: The Symbiotic Relationship Between Infants and Bifidobacterium longum ssp. longum and ssp. infantis. Genetic and Probiotic Features.

    PubMed

    Arboleya, Silvia; Stanton, Catherine; Ryan, C Anthony; Dempsey, Eugene; Ross, Paul R

    2016-01-01

    The intestinal microbiota is a complex community that plays an important role in human health from the initial steps of its establishment. Its microbial composition has been suggested to result from selective pressures imposed by the host and is modulated by competition among its members. Bifidobacterium longum is one of the most abundant species of the Bifidobacterium genus in the gut microbiota of healthy breast-fed infants and adults. The recent advancements of 'omics techniques have facilitated the genetic and functional studies of different gut microbiota members. They have revealed the complex genetic pathways used to metabolize different compounds that likely contribute to the competitiveness and persistence of B. longum in the colon. The discovery of a genomic island in B. longum ssp. infantis that encodes specific enzymes for the metabolism of human milk oligosaccharides suggests a specific ecological adaptation. Moreover, B. longum is widely used as probiotic, and beneficial effects in infant health have been reported in several studies. PMID:26934170

  18. An abstract class loader for the SSP and its implementation in TL.

    SciTech Connect

    Wickstrom, Gregory Lloyd; Winter, Victor Lono; Fraij, Fares; Roach, Steve; Beranek, Jason

    2004-08-01

    The SSP is a hardware implementation of a subset of the JVM for use in high consequence embedded applications. In this context, a majority of the activities belonging to class loading, as it is defined in the specification of the JVM, can be performed statically. Static class loading has the net result of dramatically simplifying the design of the SSP as well as increasing its performance. Due to the high consequence nature of its applications, strong evidence must be provided that all aspects of the SSP have been implemented correctly. This includes the class loader. This article explores the possibility of formally verifying a class loader for the SSP implemented in the strategic programming language TL. Specifically, an implementation of the core activities of an abstract class loader is presented and its verification in ACL2 is considered.

  19. Staphylococcus saprophyticus surface-associated protein (Ssp) is associated with lifespan reduction in Caenorhabditis elegans.

    PubMed

    Szabados, Florian; Mohner, Amelie; Kleine, Britta; Gatermann, Sören G

    2013-10-01

    Staphylococcal lipases have been proposed as pathogenicity factors. In Staphylococcus saprophyticus the surface-associated protein (Ssp) has been previously characterized as a cell wall-associated true lipase. A S. saprophyticus Δssp::ermB mutant has been described as less virulent in an in vivo model of urinary tract infection compared with its wild-type. This is the first report showing that S. saprophyticus induced a lifespan reduction in Caenorhabditis elegans similar to that of S. aureus RN4220. In two S. saprophyticus Δssp::ermB mutants lifespan reduction in C. elegans was partly abolished. In order to attribute virulence to the lipase activity itself and distinguish this phenomenon from the presence of the Ssp-protein, the conserved active site of the lipase was modified by site-directed ligase-independent mutagenesis and lipase activity-deficient mutants were constructed. These results indicate that the Ssp is associated with pathogenicity in C. elegans and one could speculate that the lipase activity itself is responsible for this virulence.

  20. Cytogenetic Diversity of Simple Sequences Repeats in Morphotypes of Brassica rapa ssp. chinensis

    PubMed Central

    Zheng, Jin-shuang; Sun, Cheng-zhen; Zhang, Shu-ning; Hou, Xi-lin; Bonnema, Guusje

    2016-01-01

    A significant fraction of the nuclear DNA of all eukaryotes is comprised of simple sequence repeats (SSRs). Although these sequences are widely used for studying genetic variation, linkage mapping and evolution, little attention had been paid to the chromosomal distribution and cytogenetic diversity of these sequences. In this paper, we report the distribution characterization of mono-, di-, and tri-nucleotide SSRs in Brassica rapa ssp. chinensis. Fluorescence in situ hybridization was used to characterize the cytogenetic diversity of SSRs among morphotypes of B. rapa ssp. chinensis. The proportion of different SSR motifs varied among morphotypes of B. rapa ssp. chinensis, with tri-nucleotide SSRs being more prevalent in the genome of B. rapa ssp. chinensis. We determined the chromosomal locations of mono-, di-, and tri-nucleotide repeat loci. The results showed that the chromosomal distribution of SSRs in the different morphotypes is non-random and motif-dependent, and allowed us to characterize the relative variability in terms of SSR numbers and similar chromosomal distributions in centromeric/peri-centromeric heterochromatin. The differences between SSR repeats with respect to abundance and distribution indicate that SSRs are a driving force in the genomic evolution of B. rapa species. Our results provide a comprehensive view of the SSR sequence distribution and evolution for comparison among morphotypes B. rapa ssp. chinensis. PMID:27507974

  1. Cytogenetic Diversity of Simple Sequences Repeats in Morphotypes of Brassica rapa ssp. chinensis.

    PubMed

    Zheng, Jin-Shuang; Sun, Cheng-Zhen; Zhang, Shu-Ning; Hou, Xi-Lin; Bonnema, Guusje

    2016-01-01

    A significant fraction of the nuclear DNA of all eukaryotes is comprised of simple sequence repeats (SSRs). Although these sequences are widely used for studying genetic variation, linkage mapping and evolution, little attention had been paid to the chromosomal distribution and cytogenetic diversity of these sequences. In this paper, we report the distribution characterization of mono-, di-, and tri-nucleotide SSRs in Brassica rapa ssp. chinensis. Fluorescence in situ hybridization was used to characterize the cytogenetic diversity of SSRs among morphotypes of B. rapa ssp. chinensis. The proportion of different SSR motifs varied among morphotypes of B. rapa ssp. chinensis, with tri-nucleotide SSRs being more prevalent in the genome of B. rapa ssp. chinensis. We determined the chromosomal locations of mono-, di-, and tri-nucleotide repeat loci. The results showed that the chromosomal distribution of SSRs in the different morphotypes is non-random and motif-dependent, and allowed us to characterize the relative variability in terms of SSR numbers and similar chromosomal distributions in centromeric/peri-centromeric heterochromatin. The differences between SSR repeats with respect to abundance and distribution indicate that SSRs are a driving force in the genomic evolution of B. rapa species. Our results provide a comprehensive view of the SSR sequence distribution and evolution for comparison among morphotypes B. rapa ssp. chinensis.

  2. A modified PCR-SSP method for the identification of ABO blood group antigens.

    PubMed

    Downing, J; Darke, C

    2003-08-01

    The ABO blood group antigens are carbohydrate molecules synthesized by the glycosyltransferases encoded by the ABO gene on chromosome 9. Kidney transplantation across the ABO barrier generally leads to rapid humoral graft rejection due to the presence of naturally occurring antibodies to the A and B antigens. We have developed a method for ABO typing our cadaveric organ donors by the polymerase chain reaction using sequence-specific primers (PCR-SSP). The method uses 12 primers in eight PCR mixtures and is performed under the same conditions as our routine HLA-A, B, C PCR-SSP typing. The PCR-SSP-based types of 166 regular blood donors and 148 cadaveric organ donors all showed total concordance with their serologically assigned ABO groups. Six individuals possessing the ABO A subgroups (A3, Ax and Aend) all typed as A1 by PCR-SSP, as expected. PCR-SSP is an appropriate method for ABO typing of cadaveric organ donors and, importantly, enables both ABO and HLA typing to be performed on the same DNA material.

  3. Cytogenetic Diversity of Simple Sequences Repeats in Morphotypes of Brassica rapa ssp. chinensis.

    PubMed

    Zheng, Jin-Shuang; Sun, Cheng-Zhen; Zhang, Shu-Ning; Hou, Xi-Lin; Bonnema, Guusje

    2016-01-01

    A significant fraction of the nuclear DNA of all eukaryotes is comprised of simple sequence repeats (SSRs). Although these sequences are widely used for studying genetic variation, linkage mapping and evolution, little attention had been paid to the chromosomal distribution and cytogenetic diversity of these sequences. In this paper, we report the distribution characterization of mono-, di-, and tri-nucleotide SSRs in Brassica rapa ssp. chinensis. Fluorescence in situ hybridization was used to characterize the cytogenetic diversity of SSRs among morphotypes of B. rapa ssp. chinensis. The proportion of different SSR motifs varied among morphotypes of B. rapa ssp. chinensis, with tri-nucleotide SSRs being more prevalent in the genome of B. rapa ssp. chinensis. We determined the chromosomal locations of mono-, di-, and tri-nucleotide repeat loci. The results showed that the chromosomal distribution of SSRs in the different morphotypes is non-random and motif-dependent, and allowed us to characterize the relative variability in terms of SSR numbers and similar chromosomal distributions in centromeric/peri-centromeric heterochromatin. The differences between SSR repeats with respect to abundance and distribution indicate that SSRs are a driving force in the genomic evolution of B. rapa species. Our results provide a comprehensive view of the SSR sequence distribution and evolution for comparison among morphotypes B. rapa ssp. chinensis. PMID:27507974

  4. Ssp1 CaMKK: A Sensor of Actin Polarization That Controls Mitotic Commitment through Srk1 in Schizosaccharomyces pombe

    PubMed Central

    Giménez-Zaragoza, David; López-Avilés, Sandra; Yance-Chávez, Tula; Montserrat, Marta; Pujol, M. Jesús; Bachs, Oriol; Aligue, Rosa

    2015-01-01

    Background Calcium/calmodulin-dependent protein kinase kinase (CaMKK) is required for diverse cellular functions. Mammalian CaMKK activates CaMKs and also the evolutionarily-conserved AMP-activated protein kinase (AMPK). The fission yeast Schizosaccharomyces pombe CaMKK, Ssp1, is required for tolerance to limited glucose through the AMPK, Ssp2, and for the integration of cell growth and division through the SAD kinase Cdr2. Results Here we report that Ssp1 controls the G2/M transition by regulating the activity of the CaMK Srk1. We show that inhibition of Cdc25 by Srk1 is regulated by Ssp1; and also that restoring growth polarity and actin localization of ssp1-deleted cells by removing the actin-monomer-binding protein, twinfilin, is sufficient to suppress the ssp1 phenotype. Conclusions These findings demonstrate that entry into mitosis is mediated by a network of proteins, including the Ssp1 and Srk1 kinases. Ssp1 connects the network of components that ensures proper polarity and cell size with the network of proteins that regulates Cdk1-cyclin B activity, in which Srk1 plays an inhibitory role. PMID:26575035

  5. A cellular study of teosinte Zea mays ssp. parviglumis (Poaceae) caryopsis development showing several processes conserved in maize

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Although recent molecular studies elucidate the genetic background leading to changed morphology of maize female inflorescence and the structure of the caryopsis during the domestication of maize (Zea mays ssp. mays) from its wild progenitor teosinte (Zea mays ssp. parviglumis), the mechanisms under...

  6. Genetic Interactions among AMPK Catalytic Subunit Ssp2 and Glycogen Synthase Kinases Gsk3 and Gsk31 in Schizosaccharomyces Pombe.

    PubMed

    Qingyun; Ma, Yan; Kato, Toshiaki; Furuyashiki, Tomoyuki

    2016-01-01

    In Schizosaccharomyces pombe, Ssp2, an ortholog of AMP-activated protein kinase (AMPK), is critical for cell growth at restrictive temperatures and under glucose depletion as well as sexual differentiation under nitrogen depletion. To identify genes genetically related to Ssp2, we performed a genetic screening to search for the genes whose overexpression rescued the growth defects in Δssp2 cells at restrictive temperatures, and identified 35 cosmids as multicopy suppressor genes. In Southern blot analyses, 22 out of these cosmids were hybridized to an ssp2+ probe. Using nucleotide sequencing, we identified the gsk3+ gene in one of the cosmids, and the remaining 12 cosmids were hybridized to a gsk3+ probe. Overexpression of the gsk3+ gene or the gsk31+ gene, another GSK3 member, rescues defective growth of Δssp2 cells at restrictive temperatures and under glucose depletion as well as sexual differentiation under nitrogen depletion. Δgsk3Δgsk31 double knockout cells, but neither Δgsk3 nor Δgsk31 single knockout cells, phenocopy Δssp2 cells. The deletion of the gsk3+ or gsk31+ gene augments the phenotypes of Δssp2 cells. These findings suggest that Gsk3 and Gsk31 are critical and interact with Ssp2 in multiple cellular functions. PMID:27604537

  7. Analysis of COPII Vesicles Indicates a Role for the Emp47-Ssp120 Complex in Transport of Cell Surface Glycoproteins.

    PubMed

    Margulis, Neil G; Wilson, Joshua D; Bentivoglio, Christine M; Dhungel, Nripesh; Gitler, Aaron D; Barlowe, Charles

    2016-03-01

    Coat protein complex II (COPII) vesicle formation at the endoplasmic reticulum (ER) transports nascent secretory proteins forward to the Golgi complex. To further define the machinery that packages secretory cargo and targets vesicles to Golgi membranes, we performed a comprehensive proteomic analysis of purified COPII vesicles. In addition to previously known proteins, we identified new vesicle proteins including Coy1, Sly41 and Ssp120, which were efficiently packaged into COPII vesicles for trafficking between the ER and Golgi compartments. Further characterization of the putative calcium-binding Ssp120 protein revealed a tight association with Emp47 and in emp47Δ cells Ssp120 was mislocalized and secreted. Genetic analyses demonstrated that EMP47 and SSP120 display identical synthetic positive interactions with IRE1 and synthetic negative interactions with genes involved in cell wall assembly. Our findings support a model in which the Emp47-Ssp120 complex functions in transport of plasma membrane glycoproteins through the early secretory pathway.

  8. Proteomic analysis of bronchoalveolar lavage fluid proteins from mice infected with Francisella tularensis ssp novicida

    PubMed Central

    Varnum, Susan M.; Webb-Robertson, Bobbie-Jo M.; Pounds, Joel G.; Moore, Ronald J.; Smith, Richard D.; Frevert, Charles W.; Skerrett, Shawn J.; Wunschel, David

    2012-01-01

    Francisella tularensis causes the zoonosis tularemia in humans and is one of the most virulent bacterial pathogens. We utilized a global proteomic approach to characterize protein changes in bronchoalveolar lavage fluid from mice exposed to one of three organisms, F. tularensis ssp. novicida, an avirulent mutant of F. tularensis ssp. novicida (F.t. novicida-ΔmglA); and Pseudomonas aeruginosa. The composition of BALF proteins was altered following infection, including proteins involved in neutrophil activation, oxidative stress and inflammatory responses. Components of the innate immune response were induced including the acute phase response and the complement system, however the timing of their induction varied. Francisella tularensis ssp. novicida infected mice do not appear to have an effective innate immune response in the first hours of infection, however within 24 hours they show an upregulation of innate immune response proteins. This delayed response is in contrast to P. aeruginosa infected animals which show an early innate immune response. Likewise, F.t. novicida-ΔmglA infection initiates an early innate immune response, however this response is dimished by 24 hours. Finally, this study identifies several candidate biomarkers, including Chitinase 3-like-1 (CHI3L1 or YKL-40) and peroxiredoxin 1, that are associated with F. tularensis ssp. novicida but not P. aeruginosa infection. PMID:22663564

  9. Occurrence of aphidborne viruses in southernmost South American populations of Fragaria chiloensis ssp. chiloensis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Wild and cultivated Fragaria chiloensis ssp. chiloensis (Fcc) plants were collected at different locations in southern Chile in order to determine the current viral status of this native strawberry. The following aphidborne viruses (ABVs): Strawberry mild yellow edge virus (SMYEV), Strawberry mottle...

  10. The discovery of resistant sources of spring barley, Hordeum vulgare ssp. spontaneum, and unique greenbug biotypes

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The genetic sources for host-plant resistance to the greenbug (Schiazphis graminum Ronani) in barley (Hordeum vulgare ssp. spontaneum) are limited in that only two single dominant genes Rsg1 and Rsg2 are available for resistance to greenbug biotypes. We evaluated four new barley lines from the Wild...

  11. Identification of Functional Candidates amongst Hypothetical Proteins of Treponema pallidum ssp. pallidum

    PubMed Central

    Naqvi, Ahmad Abu Turab; Shahbaaz, Mohd; Ahmad, Faizan; Hassan, Md. Imtaiyaz

    2015-01-01

    Syphilis is a globally occurring venereal disease, and its infection is propagated through sexual contact. The causative agent of syphilis, Treponema pallidum ssp. pallidum, a Gram-negative sphirochaete, is an obligate human parasite. Genome of T. pallidum ssp. pallidum SS14 strain (RefSeq NC_010741.1) encodes 1,027 proteins, of which 444 proteins are known as hypothetical proteins (HPs), i.e., proteins of unknown functions. Here, we performed functional annotation of HPs of T. pallidum ssp. pallidum using various database, domain architecture predictors, protein function annotators and clustering tools. We have analyzed the sequences of 444 HPs of T. pallidum ssp. pallidum and subsequently predicted the function of 207 HPs with a high level of confidence. However, functions of 237 HPs are predicted with less accuracy. We found various enzymes, transporters, binding proteins in the annotated group of HPs that may be possible molecular targets, facilitating for the survival of pathogen. Our comprehensive analysis helps to understand the mechanism of pathogenesis to provide many novel potential therapeutic interventions. PMID:25894582

  12. INFLUENCE OF MOWING ARTEMISIA TRIDENTATA SSP. WYOMINGENSIS ON WINTER HABITAT FOR WILDLIFE

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Mowing is commonly implemented to Artemisia tridentata ssp. wyomingensis (Beetle & A. Young) S.L. Welsh (Wyoming big sagebrush) plant communities to improve wildlife habitat, increase forage production for livestock, and create fuel breaks for fire suppression. However, information detailing the in...

  13. Transfer of soft kernel texture from Triticum aestivum to durum wheat, Triticum turgidum ssp. durum

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Durum wheat (Triticum turgidum ssp. durum) is a leading cereal grain whose primary use is the production of semolina and then pasta. Its rich culinary relationship to humans is related, in part, to its very hard kernel texture. This very hard texture is due to the loss of the Puroindoline genes whi...

  14. Proteomic analysis of bronchoalveolar lavage fluid proteins from mice infected with Francisella tularensis ssp novicida

    SciTech Connect

    Varnum, Susan M.; Webb-Robertson, Bobbie-Jo M.; Pounds, Joel G.; Moore, Ronald J.; Smith, Richard D.; Frevert, Charles; Skerret, Shawn J.; Wunschel, David S.

    2012-07-06

    Francisella tularensis causes the zoonosis tularemia in humans and is one of the most virulent bacterial pathogens. We utilized a global proteomic approach to characterize protein changes in bronchoalveolar lavage fluid from mice exposed to one of three organisms, F. tularensis ssp. novicida, an avirulent mutant of F. tularensis ssp. novicida (F.t. novicida-ΔmglA); and Pseudomonas aeruginosa. The composition of BALF proteins was altered following infection, including proteins involved in neutrophil activation, oxidative stress and inflammatory responses. Components of the innate immune response were induced including the acute phase response and the complement system, however the timing of their induction varied. Francisella tularensis ssp. novicida infected mice do not appear to have an effective innate immune response in the first hours of infection, however within 24 hours they show an upregulation of innate immune response proteins. This delayed response is in contrast to P. aeruginosa infected animals which show an early innate immune response. Likewise, F.t. novicida-ΔmglA infection initiates an early innate immune response, however this response is dimished by 24 hours. Finally, this study identifies several candidate biomarkers, including Chitinase 3-like-1 (CHI3L1 or YKL-40) and peroxiredoxin 1, that are associated with F. tularensis ssp. novicida but not P. aeruginosa infection.

  15. Summary of Recent Results from NASA's Space Solar Power (SSP) Programs and the Current Capabilities of Microwave WPT Technology

    NASA Technical Reports Server (NTRS)

    McSpadden, James; Mankins, John C.; Howell, Joe T. (Technical Monitor)

    2002-01-01

    The concept of placing enormous solar power satellite (SPS) systems in space represents one of a handful of new technological options that might provide large-scale, environmentally clean base load power into terrestrial markets. In the US, the SPS concept was examined extensively during the late 1970s by the U.S. Department of Energy (DOE) and the National Aeronautics and Space Administration (NASA). More recently, the subject of space solar power (SSP) was reexamined by NASA from 1995-1997 in the "fresh look" study, and during 1998 in an SSP "concept definition study". As a result of these efforts, in 1999-2000, NASA undertook the SSP Exploratory Research and Technology (SERT) program which pursued preliminary strategic technology research and development to enable large, multi-megawatt SSP systems and wireless power transmission (WPT) for government missions and commercial markets (in-space and terrestrial). During 2001-2002, NASA has been pursuing an SSP Concept and Technology Maturation (SCTM) program follow-on to the SERT, with special emphasis on identifying new, high-leverage technologies that might advanced the feasibility of future SSP systems. In addition, in 2001, the U.S. National Research Council (NRC) released a major report providing the results of a peer review of NASA's SSP strategic research and technology (R&T) road maps. One of the key technologies needed to enable the future feasibility of SSP/SPS is that of wireless power transmission. Advances in phased array antennas and rectennas have provided the building blocks for a realizable WPT system. These key components include the dc-RF converters in the transmitter, the retrodirective beam control system, and the receiving rectenna. Each subject is briefly covered, and results from the SERT program that studied a 5.8 GHz SPS system are presented. This paper presents a summary results from NASA's SSP efforts, along with a summary of the status of microwave WPT technology development.

  16. BcMF21 is important for pollen development and germination in Brassica campestris ssp. chinensis.

    PubMed

    Jiang, Jingjing; Yu, Youjian; Dong, Heng; Yao, Lina; Zhang, Zhixian; Cao, Jiashu

    2014-01-01

    Brassica campestris Male Fertility 21 (BcMF21) was previously isolated from the flower buds of Chinese cabbage (Brassica campestris L. ssp. chinensis Makino, syn. B. rapa ssp. chinensis) and expressed specifically in tapetum and microspores during the meiosis stage and the uninucleate stage of microspore development. Here, we used antisense RNA technology to knock down the expression level of BcMF21 in B. campestris and analyzed the phenotype of the transgenic plants. Alexander staining and scanning electron microscope revealed sterility and exine deformities in the mature pollen grains of BcMF21 antisense RNA transgenic plants. The germination furrow of the BcMF21 antisense RNA transgenic pollen was covered by lipid like materials. The pollen tubes burst and could not grow normally in vitro. Therefore, we presented here BcMF21 might be an important gene for pollen development and germination.

  17. Effects of Gentiana lutea ssp. symphyandra on the central nervous system in mice.

    PubMed

    Oztürk, Nilgün; Başer, K Hüsnü Can; Aydin, Süleyman; Oztürk, Yusuf; Caliş, Ihsan

    2002-11-01

    A methanolic extact of Gentiana lutea ssp. symphyandra roots has been investigated for its possible effects on the central nervous system of mice. At doses of 250 and 500 mg/kg (i.p.), the methanol extract of Gentiana roots caused a significant increase in the swimming endurance test and exhibited slight analgesic activity, but no lethality in mice suggesting some activity on the central nervous system. However, there was no indication of sedation or muscular fatigue at the doses employed. HPLC analysis showed that three secoiridoid compounds, gentiopicroside, swertiamarine and sweroside were present and may have been responsible for the CNS effects of the methanol extract of Gentiana lutea ssp. symphyandra roots.

  18. SSP Technology Investigation of a High-Voltage DC-DC Converter

    NASA Technical Reports Server (NTRS)

    Pappas, J. A.; Grady, W. M.; George, Patrick J. (Technical Monitor)

    2002-01-01

    The goal of this project was to establish the feasibility of a high-voltage DC-DC converter based on a rod-array triggered vacuum switch (RATVS) for the Space Solar Power system. The RATVS has many advantages over silicon and silicon-carbide devices. The RATVS is attractive for this application because it is a high-voltage device that has already been demonstrated at currents in excess of the requirement for an SSP device and at much higher per-device voltages than existing or near-term solid state switching devices. The RATVS packs a much higher specific power rating than any solid-state device and it is likely to be more tolerant of its surroundings in space. In addition, pursuit of an RATVS-based system would provide NASA with a nearer-term and less expensive power converter option for the SSP.

  19. Lactic acid production by mixed cultures of Kluyveromyces marxianus, Lactobacillus delbrueckii ssp. bulgaricus and Lactobacillus helveticus.

    PubMed

    Plessas, S; Bosnea, L; Psarianos, C; Koutinas, A A; Marchant, R; Banat, I M

    2008-09-01

    Lactic acid production using Kluyveromyces marxianus (IFO 288), Lactobacillus delbrueckii ssp. bulgaricus (ATCC 11842) and Lactobacillus helveticus (ATCC 15009) individually or as mixed culture on cheese whey in stirred or static fermentation conditions was evaluated. Lactic acid production, residual sugar and cell biomass were the main features examined. Increased lactic acid production was observed, when mixed cultures were used in comparison to individual ones. The highest lactic acid concentrations were achieved when K. marxianus yeast was combined with L. delbrueckii ssp. bulgaricus, and when all the strains were used revealing possible synergistic effects between the yeast and the two lactic acid bacteria. The same synergistic effects were further observed and verified when the mixed cultures were applied in sourdough fermentations, proving that the above microbiological system could be applied in the food fermentations where high lactic acid production is sought.

  20. Acute poisoning of friesian heifers by Solanum macrocarpon L. ssp dasyphyllum.

    PubMed

    Bizimenyera, E S

    2003-08-01

    Solanum macrocarpon (African eggplant) is a tropical plant widely cultivated as a delicious vegetable; the non-edible wild variety called Solanum macrocarpon L. ssp dasyphyllum (the wild African eggplant) bears thorns or spikes on the stem and leaves. Thirteen yearling heifers on a dairy farm in Uganda suffered acute poisoning after eating berries of S. macrocarpon L. ssp dasyphyllum. There was sudden onset of anorexia, copious salivation, severe dysentery and passage of red urine. The animals also had central nervous derangement (incordination, walking blindly) and exudative dermatitis. Four heifers died. Necropsy lesions were icterus, hemorrhages, gastroenteritis, lympadenomegally, and friable and bronze colored livers and kidneys. The rumen and reticulum contained masses of the plant seeds. This is the first report of cattle poisoning by this plant.

  1. Applicability of Phytoextraction with Arabidopsis halleri ssp. gemmifera to Remediate Cd-contaminated Andisols

    NASA Astrophysics Data System (ADS)

    Kameyama, Koji; Tani, Shigeru; Sugawara, Reiko; Ishikawa, Yuichi

    The objective of this study was to investigate the applicability of phytoextraction with a Cd-hyperaccumulator plant (Arabidopsis halleri ssp. gemmifera) to remediate Cd-contaminated Andisols. Cd absorption potentials of this plant for Andisols were examined in pot experiments. Sequentially, phytoextraction durations for remediation of Cd-contaminated Andisols were calculated from the experimental data. The results were as follows: (1) Cd concentrations in the plant shoots ranged from 170-750 mgṡkg-1. (2) Cd absorption of the plant for Andisols with ALC (Autoclaved Lightweight aerated Concrete) was less than for Andisols without ALC. However, the plants absorbed the same amount of soil Cd extracted by 0.01 M HCl with or without ALC. (3) Calculations suggest that the applicability of phytoextraction with this plant is high for slightly contaminated Andisols. Therefore, phytoextraction with Arabidopsis halleri ssp. gemmifera may be a viable option for the remediation of Cd-contaminated Andisols.

  2. Chemical, technological, and nutritional characteristics of two lines of "farro" (Triticum turgidum ssp. dicoccum).

    PubMed

    Acquistucci, Rita; Aureli, Gabriella; Codianni, Pasquale; Colonna, Michaela; Galterio, Giovanni

    2004-06-01

    In recent years, the renewed interest for foods with a natural image has increased the demand for dry pasta produced from "hulled" wheat such as the Triticum turgidum ssp. dicoccum, also known as "farro". In order to contribute to the general knowledge, two lines of farro were considered in this study. To have a comparison, an old cultivar of Triticum turgidum ssp. durum (Senatore Cappelli) in addition to a commercial semolina were also examined. All semolina samples were used to produce pasta samples. Results showed some differences among pasta samples that seem to be due not to the presence of specific protein subunits but especially to the quantitative ratio between the different subunits. Results also reconfirmed the role played by the drying technology that is able to affect the sensory characteristics of pasta products. PMID:15285114

  3. Chemical Composition of Different Botanical Origin Honeys Produced by Sicilian Black Honeybees (Apis mellifera ssp. sicula).

    PubMed

    Mannina, Luisa; Sobolev, Anatoly P; Di Lorenzo, Arianna; Vista, Silvia; Tenore, Gian Carlo; Daglia, Maria

    2015-07-01

    In 2008 a Slow Food Presidium was launched in Sicily (Italy) for an early warning of the risk of extinction of the Sicilian native breed of black honeybee (Apis mellifera L. ssp sicula). Today, the honey produced by these honeybees is the only Sicilian honey produced entirely by the black honeybees. In view of few available data regarding the chemical composition of A. mellifera ssp. sicula honeys, in the present investigation the chemical compositions of sulla honey (Hedysarum coronarium L.) and dill honey (Anethum graveolens L.) were studied with a multimethodological approach, which consists of HPLC-PDA-ESI-MSn and NMR spectroscopy. Moreover, three unifloral honeys (lemon honey (obtained from Citrus limon (L.) Osbeck), orange honey (Citrus arantium L.), and medlar honey (Eriobotrya japonica (Thunb.) Lindl)), with known phenol and polyphenol compositions, were studied with NMR spectroscopy to deepen the knowledge about sugar and amino acid compositions.

  4. Cytokinesis in yeast meiosis depends on the regulated removal of Ssp1p from the prospore membrane.

    PubMed

    Maier, Peter; Rathfelder, Nicole; Finkbeiner, Martin G; Taxis, Christof; Mazza, Massimiliano; Le Panse, Sophie; Haguenauer-Tsapis, Rosine; Knop, Michael

    2007-04-01

    Intracellular budding is a developmentally regulated type of cell division common to many fungi and protists. In Saccaromyces cerevisiae, intracellular budding requires the de novo assembly of membranes, the prospore membranes (PSMs) and occurs during spore formation in meiosis. Ssp1p is a sporulation-specific protein that has previously been shown to localize to secretory vesicles and to recruit the leading edge protein coat (LEP coat) proteins to the opening of the PSM. Here, we show that Ssp1p is a multidomain protein with distinct domains important for PI(4,5)P(2) binding, binding to secretory vesicles and inhibition of vesicle fusion, interaction with LEP coat components and that it is subject to sumoylation and degradation. We found non-essential roles for Ssp1p on the level of vesicle transport and an essential function of Ssp1p to regulate the opening of the PSM. Together, our results indicate that Ssp1p has a domain architecture that resembles to some extent the septin class of proteins, and that the regulated removal of Ssp1p from the PSM is the major step underlying cytokinesis in yeast sporulation.

  5. Evaluating the Strange Situation Procedure (SSP) to Assess the Bond between Dogs and Humans

    PubMed Central

    Rehn, Therese; McGowan, Ragen T. S.; Keeling, Linda J.

    2013-01-01

    The Strange Situation Procedure (SSP) is increasingly being used to study attachment between dogs and humans. It has been developed from the Ainsworth Strange Situation Procedure, which is used extensively to investigate attachment between children and their parents. In this experiment, 12 female beagle dogs were tested in two treatments to identify possible order effects in the test, a potential weakness in the SSP. In one treatment (FS), dogs participated together with a ‘familiar person’ and a ‘stranger’. In a control treatment (SS), the same dogs participated together with two unfamiliar people, ‘stranger A’ and ‘stranger B’. Comparisons were made between episodes within as well as between treatments. As predicted in FS, dogs explored more in the presence of the familiar person than the stranger. Importantly, they also explored more in the presence of stranger A (who appeared in the same order as the familiar person and followed the same procedure) than stranger B in SS. Furthermore, comparisons between treatments, where a familiar person was present in FS and stranger A was present in SS, showed no differences in exploration. In combination, these results indicate that the effect of a familiar person on dogs' exploratory behaviour, a key feature when assessing secure attachment styles, could not be tested reliably due to the order in which the familiar person and the stranger appear. It is proposed that in the future only counterbalanced versions of the SSP are used. Alternatively, since dogs reliably initiated more contact with the familiar person compared to the strangers, it is suggested that future studies on attachment in dogs towards humans should focus either on the behaviour of the dog in those episodes of the SSP when the person returns, or on reunion behaviour in other studies, specially designed to address dog-human interactions at this time. PMID:23437277

  6. Extinction risk assessment for the species survival plan (SSP) population of the Bali mynah (Leucopsar rothschildi).

    PubMed

    Earnhardt, Joanne M; Thompson, Steven D; Faust, Lisa J

    2009-05-01

    The Bali mynah Species Survival Plan (SSP), an Association of Zoos and Aquariums program, strives to maintain the genetic and demographic health of its population, avoid unplanned changes in size, and minimize the risk of population extinction. The SSP population meets current demographic and genetic objectives with a population size of 209 birds at 61 institutions and 96% genetic diversity (GD) retained from the source population. However, participating institutions have expressed concerns regarding space allocation, target population size (TPS), breeding restrictions, inbreeding depression, and harvest in relation to future population availability and viability. Based on these factors, we assess five questions with a quantitative risk assessment, specifically a population viability analysis (PVA) using ZooRisk software. Using an individual-based stochastic model, we project potential population changes under different conditions (e.g. changes in TPS and genetic management) to identify the most effective management actions. Our projections indicate that under current management conditions, population decline and extinction are unlikely and that although GD will decline over 100 years the projected loss does not exceed levels acceptable to population managers (less than 90% GD retained). Model simulations indicate that the combination of two genetic management strategies (i.e. priority breeding based on mean kinship and inbreeding avoidance) benefits the retention of GD and reduces the accumulation of inbreeding. The current TPS (250) is greater than necessary to minimize the risk of extinction for the SSP population but any reduction in TPS must be accompanied by continued application of genetic management. If carefully planned, birds can be harvested for transfer to Bali for a reintroduction program without jeopardizing the SSP population.

  7. Antinociceptive effect of some extracts from Ajuga chamaecistus Ging. ssp. tomentella (Boiss.) Rech. f. aerial parts

    PubMed Central

    2014-01-01

    Background The genus Ajuga is used for the treatment of joint pain, gout, and jaundice in traditional Iranian medicine (TIM). Ajuga chamaecistus ssp. tomentella is an exclusive subspecies of Ajuga chamaecistus in the flora of Iran. The aim of this study was to evaluate antinociceptive properties of some extracts from aerial parts of A. chamaecistus ssp. tomentella. Methods Antinociceptive activities of total water and 80% methanol extracts, hexane, diethyl ether and n-butanolic partition fractions of the methanolic extract were analyzed using the formalin test in mice. Indomethacin (10 mg/kg) and normal saline were employed as positive and negative controls, respectively. Results Oral administration of all extracts (200, 400 and 600 mg/kg) 30 min before formalin injection had no effect against the acute phase (0–5 min after formalin injection) of the formalin-induced licking time, but hexane fraction (200 mg/kg) caused a significant effect (p < 0.001) on the chronic phase (15–60 min after formalin injection). Total water and diethyl ether extracts at a dose of 400 mg/kg showed a very significant analgesic activity on the chronic phase (p < 0.001 and p < 0.01, respectively). Conclusions The results of this study suggest that the extracts of A. chamaecistus ssp. tomentella have an analgesic property that supports traditional use of Ajuga genus for joint pain and other inflammatory diseases. PMID:25022284

  8. Simultaneous genotyping of HPA-17w to -21w by PCR-SSP in Chinese Cantonese.

    PubMed

    Zhou, Haojie; Ding, Haoqiang; Chen, Yangkai; Li, Xiaofan; Ye, Xin; Nie, Yongmei

    2015-01-01

    Studies have reported the polymorphism of human platelet antigen (HPA)-17w, -18w, -19w, -20w, and -21w. However, the distribution of these five antigens in Chinese Cantonese is still unknown. In this study, we designed new sequence-specific primers for HPA-19w to -21w and used published primers for HPA-17w and -18w to develop a polymerase chain reaction with the sequence-specific primers (PCR-SSP) method for simultaneously genotyping HPA-17w to -21w. A total of 820 unrelated Cantonese apheresis platelet donors in Guangzhou were involved in this study. Among the five HPAs, complete a/a homozygosity was observed for HPA-17w to -20w with an allele frequency of 1.0000. For HPA-21w, nine individuals (9/820, 1.10%) were found to be HPA-21a/bw heterozygous and the allele frequencies of HPA-21a and HPA-21bw were 0.9945 (1631/1640) and 0.0055 (9/1640), respectively. The reliability of the PCR-SSP method was determined by comparing with the genotyping results by DNA sequencing, and no inconsistencies were observed between the two methods. This study provides a reliable PCR-SSP method for simultaneously genotyping HPA-17w to -21w and could improve HPA-matched platelet transfusion in Chinese Cantonese.

  9. SLA typing using the PCR-SSP method and establishment of the SLA homozygote line in pedigreed SNU miniature pigs.

    PubMed

    Yeom, Su-Cheong; Park, Chung-Gyu; Lee, Byeong-Chun; Lee, Wang-Jae

    2010-04-01

    Seoul National University (SNU) miniature pigs represent a closed colony with 24 founder pigs and a well preserved pedigree. Characterization using mRNA sequence analysis was conducted for 6 swine leukocyte antigen (SLA) loci in parental or founder pigs, and 17 defined alleles were detected. Based on these complete coding sequences, 17 sequence specific primers (SSPs) were designed for polymorphic sites. To validate the specificity of each allele SSP, the PCR-SSP was conducted with defined allele clones as templates. PCR-SSP was conducted with the hot start polymerase and touch-down PCR. The parental or found SNU miniature pigs showed overall SLA class I and II heterozygotes. Using the established PCR-SSP method, we conducted SLA typing for breeding stock including 2 pedigreed pigs and identified the novel SLA class II homozygote haplotye (DRA*0201, DRB1*0403, DQA*0102 and DQB1*0701) and 2 SLA homozygote pig lines: SLA class I Hp-3.0 and class II Hp-0.3, and SLA class I Hp-2.0 and class II Hp-0.2. We thought that our PCR-SSP SLA typing method could be applicable for new SLA homozygote line establishment by assignment and scheduled breeding.

  10. Investigation of biological activity of polar extracts isolated from Phlomis crinita Cav ssp. mauritanica Munby.

    PubMed

    Limem-Ben Amor, Ilef; Skandrani, Ines; Boubaker, Jihed; Ben Sghaïer, Mohamed; Neffati, Aicha; Bhouri, Wissem; Bouhlel, Ines; Chouchane, Nabil; Kilani, Soumaya; Guedon, Emmanuel; Ghoul, Mohamed; Ghedira, Kamel; Chekir-Ghedira, Leila

    2009-01-01

    The lyophilized infusion, the methanol, the ethyl acetate, and the total oligomer flavonoid (TOF)-enriched extracts prepared from the dried leaves of Phlomis crinita Cav. ssp. mauritanica Munby were investigated for the contents of flavonoids, tannins, coumarines and steroids. Antibacterial activity was investigated toward five bacterial strains. An inhibitory effect was observed against Staphyllococcus aureus and Enterococcus feacalis, and the minimal inhibitory concentrations ranged from 2.5 to 5 mg/mL of extract. The tested extracts exhibit an important free radical scavenging activity toward the 1,1-diphenyl 2-picrylhydrazyl (DPPH) free radical; with IC(50) values of 30.5, 6, 32, and 31.5 microg/mL, respectively, in the presence of lyophilized infusion, the TOF, the methanol, and the ethyl acetate extracts. Genotoxic and antigenotoxic properties of the different extracts were studied by using the SOS chromotest with Escherichia coli PQ37. The lyophilized infusion and TOF extracts obtained from P. crinita ssp. mauritanica showed no genotoxicity, whereas methanol and ethyl acetate extracts are considered as marginally genotoxic. On the other hand, we showed that each extract inhibited the mutagenicity induced by aflatoxin B1 (AFB1) (10 microg/assay) and nifuroxazide (NF) (10 microg/assay). The ethyl acetate extract showed the strongest level of protection toward the genotoxicity induced by both directly and indirectly genotoxic NF and AFB1. These tests proved that the lyophilized infusion possesses an antiradical activity likewise, it showed no genotoxic effect; that is why we choose this extract to assess its antiulcerogenic activity by using an ethanol-induced ulcerogenesis model in the rat. This test demonstrates that 300 mg/kg of a P. crinita ssp. mauritanica lyophilized infusion was more effective than the reference compound, cimetidine. PMID:19514937

  11. Establishing Artemisia tridentata ssp wyomingensis on mined lands: Science and economics

    SciTech Connect

    Schuman, G.E.; Vicklund, L.E.; Belden, S.E.

    2005-12-01

    In 1996, the Wyoming Department of Environmental Quality enacted regulations governing the reestablishment of woody shrubs on mined lands. The regulation required that an average density of one shrub m{sup -2} be reestablished on at least 20% of the disturbed land area and that the shrub composition must include dominant premine species. In Wyoming, and much of the Northern Great Plains, that meant that Artemisia tridentata Nutt. ssp. wyomingensis (Beetle and Young) (Wyoming big sagebrush) had to be reestablished on mined lands. Artemisia tridentata Nutt. ssp. wyomingensis had proven difficult to reestablish on mined lands because of poor quality seed, seed dormancy and a poor understanding of the seedbed ecology of this species. Research in the last two decades has produced significant knowledge in the area of direct-seed establishment of Artemisia tridentata Nutt. ssp. wyomingensis on mined lands. Our research has shown that reducing grass seeding rates will reduce competition and result in larger sagebrush plants that are more likely to survive and provide greater structural diversity to the plant community. Economic analyses demonstrated that big sagebrush can be established at a cost of $0.01-0.05 per seedling using direct seeding methods compared to transplanting nursery grown seedlings, estimated to cost $0.72-$1.65 per seedling (depending on size) to grow and from $1.30-$2.40 to plant (flat land to 2:1 slopes). An adequate level of precipitation will be necessary to ensure successful establishment of this species no matter what method of propagation is selected and direct seeding gives greater opportunity for success because of the demonstrated longevity of the seed to germinate 3-5 years after the initial seeding.

  12. Antifungal activity of Lactobacillus paracasei ssp. tolerans isolated from a sourdough bread culture.

    PubMed

    Hassan, Yousef I; Bullerman, Lloyd B

    2008-01-15

    Lactic acid bacteria were isolated from four different sourdough bread cultures previously investigated for antifungal activity. A total of 116 isolates were obtained and screened for antifungal activity against a battery of molds. The most inhibitory isolate obtained was identified by API 50 CHL and 16s ribosomal RNA genotyping and found to be Lactobacillus paracasei ssp. tolerans. This isolate completely inhibited the growth of Fusarium proliferatum M 5689, M 5991 and Fusarium graminearum R 4053 compared to controls in a dual agar plate assay.

  13. Chemical composition and possible in vitro phytotoxic activity of Helichrsyum italicum (Roth) Don ssp. italicum.

    PubMed

    Mancini, Emilia; De Martino, Laura; Marandino, Aurelio; Scognamiglio, Maria Rosa; De Feo, Vincenzo

    2011-09-08

    The chemical composition of the essential oil of Helichrysum italicum (Roth) Don ssp. italicum, collected in the National Park of Cilento and Diano Valley, Southern Italy, was studied by means of GC and GC/MS. Forty four compounds of 45 constituents were identified in the oil, mainly oxygenated sesquiterpenes. The essential oil was evaluated for its potential in vitro phytotoxic activity against germination and early radicle elongation of radish and garden cress. The radicle elongation of radish was significantly inhibited at the highest doses tested, while germination of both seeds was not affected.

  14. A Global Water Resources Assessment under RCP, SSP, and CMIP5 Scenarios

    NASA Astrophysics Data System (ADS)

    Hanasaki, N.; Fujimori, S.

    2012-12-01

    A number of reports have been published on climate change impact assessment on global water resources, but earlier studies need to be updated and refined due to three reasons. First, most of earlier studies were based on an old set of IPCC scenarios consists of SRES (green house gas (GHG) emission and socio-economic scenarios) and CMIP3 (climate scenarios consistent with SRES). A new set of IPCC scenarios is being released (Moss et al., 2008) that consists of RCP (GHG emission scenario), SSP (socio-economic scenario), and CMIP5 (climate scenarios consistent with RCP). In order to take the latest achievements in climate modeling, impact assessments should be based on the new scenario. Second, most of earlier studies focused more on the change in water availability (e.g. runoff and discharge), less for change in water use (agricultural, industrial, domestic water use). Because SSP consists of five scenarios delineating substantially different world, water use scenarios should be developed with care reflecting the difference among them. Third, most of earlier studies assessed water availability and use at annual time resolution. This may overlook seasonal and inter-annual shortage of water due to variability in water availability and use. Here we present a novel assessment on global water resources using a global water resources model called H08 (Hanasaki et al., 2008a,b; 2010). H08 simulates natural water cycle and major human activities, such as water withdrawals and reservoir operation. It estimates water availability and use at daily time interval, which enables to take sub-annual water shortage into account. We first developed water use scenarios for agricultural (irrigation), industrial, and domestic water withdrawal that are consistent with five SSP scenarios. Next, we set up a matrix of scenario combination of RCP, SSP, and CMIP5 for insightful global water resources assessment. Finally we conducted H08 simulation using these scenarios and assessed water

  15. Accumulation of biologically active furanocoumarins in Ruta graveolens ssp. divaricata (Tenore) Gams in vitro culture.

    PubMed

    Ekiert, H; Abou-Mandour, A A; Czygan, F Ch

    2005-01-01

    The study was designed to investigate dynamics of accumulation of five linear furanocoumarins and umbelliferone in stationary liquid cultures of Ruta graveolens ssp. divaricata (Tenore) Gams during 6-week growth cycles. The contents of individual metabolites in biomass increased 1.8-3.5 times while their total content rose 2.3 times. Maximum contents of xanthotoxin, bergapten and isopimpinellin (112.3, 76.2 and 84.0mg/100g d.w., respectively) and maximum total content of all metabolites (283.4 mg/100 g d.w.), obtained on 35th culture day, are interesting from practical point of view.

  16. Lineages of Streptococcus equi ssp. equi in the Irish equine industry

    PubMed Central

    2013-01-01

    Background Streptococcus equi ssp. equi is the causative agent of ‘Strangles’ in horses. This is a debilitating condition leading to economic loss, yard closures and cancellation of equestrian events. There are multiple genotypes of S. equi ssp. equi which can cause disease, but to date there has been no systematic study of strains which are prevalent in Ireland. This study identified and classified Streptococcus equi ssp. equi strains isolated from within the Irish equine industry. Results Two hundred veterinary isolates were subjected to SLST (single locus sequence typing) based on an internal sequence from the seM gene of Streptococcus equi ssp equi. Of the 171 samples which successfully gave an amplicon, 162 samples (137 Irish and 24 UK strains) gave robust DNA sequence information. Analysis of the sequences allowed division of the isolates into 19 groups, 13 of which contain at least 2 isolates and 6 groups containing single isolates. There were 19 positions where a DNA SNP (single nucleotide polymorphism) occurs, and one 3 bp insertion. All groups had multiple (2–8) SNPs. Of the SNPs 17 would result in an amino acid change in the encoded protein. Interestingly, the single isolate EI8, which has 6 SNPs, has the three base pair insertion which is not seen in any other isolate, this would result in the insertion of an Ile residue at position 62 in that protein sequence. Comparison of the relevant region in the determined sequences with the UK Streptococcus equi seM MLST database showed that Group B (15 isolates) and Group I (2 isolates), as well as the individual isolates EI3 and EI8, are unique to Ireland, and some groups are most likely of UK origin (Groups F and M), but many more probably passed back and forth between the two countries. Conclusions The strains occurring in Ireland are not clonal and there is a considerable degree of sequence variation seen in the seM gene. There are two major clades causing infection in Ireland and these strains are

  17. Isolation and characterization of a novel BcMF14 gene from Brassica campestris ssp. chinensis.

    PubMed

    Li, Yanyan; Nie, Chuanpeng; Cao, Jiashu

    2011-03-01

    A putative RALF (rapid alkalinization factor)-like gene (GenBank accession number EF523517), named BcMF14, was isolated from Chinese cabbage (Brassica campestris L. ssp. chinensis Makino, syn. B. rapa ssp. chinensis) by rapid amplification of cDNA ends (RACE) based on a cDNA-AFLP differential fragment exclusively expressed in fertile line. Semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) discovered that BcMF14 was prominently expressed in stage four and five flower buds of fertile line, no expression in vegetative structures or in sterility line. Detailed RT-PCR illuminated its strong expression in stamens. Successful suppression of BcMF14 gene expression greatly reduced the normal pollen grains. The frequency of abnormal pollen grains was 48.95% in the mutant with many shriveled pollen grains with irregular shape and some larger ones with deep hollows along the germination ditch. Pollen germination was stopped because of the severely twisted pollen tubes. These results demonstrate a potential role of the BcMF14 gene in the development of male gametogenesis in Chinese cabbage.

  18. Congenital vestibular disease in captive Sumatran tigers (Panthera tigris ssp. sumatrae) in Australasia.

    PubMed

    Wheelhouse, Jaimee L; Hulst, Frances; Beatty, Julia A; Hogg, Carolyn J; Child, Georgina; Wade, Claire M; Barrs, Vanessa R

    2015-11-01

    The Sumatran tiger (Panthera tigris ssp. sumatrae) is a critically endangered species in the wild. To ensure that demographic and genetic integrity are maintained in the longer term, those Sumatran tigers held in captivity are managed as a global population under a World Association of Zoos and Aquariums Global Species Management Plan (GSMP). A retrospective study, including segregation and pedigree analysis, was conducted to investigate potential cases of congenital vestibular disease (CVD) in captive Sumatran tigers in Australasian zoos using medical and husbandry records, as well as video footage obtained from 50 tigers between 1975 and 2013. Data from the GSMP Sumatran tiger studbook were made available for pedigree and segregation analysis. Fourteen cases of CVD in 13 Sumatran tiger cubs and one hybrid cub (Panthera tigris ssp. sumatrae × Panthera tigris) were identified. Vestibular signs including head tilt, circling, ataxia, strabismus and nystagmus were observed between birth and 2 months of age. These clinical signs persisted for a median of 237 days and had resolved by 2 years of age in all cases. Pedigree analysis revealed that all affected tigers were closely related and shared a single common ancestor in the last four generations. A genetic cause for the disease is suspected and, based on pedigree and segregation analysis, an autosomal dominant mode of inheritance is likely. Further investigations to determine the world-wide prevalence and underlying pathology of this disorder are warranted. PMID:26403953

  19. Congenital vestibular disease in captive Sumatran tigers (Panthera tigris ssp. sumatrae) in Australasia.

    PubMed

    Wheelhouse, Jaimee L; Hulst, Frances; Beatty, Julia A; Hogg, Carolyn J; Child, Georgina; Wade, Claire M; Barrs, Vanessa R

    2015-11-01

    The Sumatran tiger (Panthera tigris ssp. sumatrae) is a critically endangered species in the wild. To ensure that demographic and genetic integrity are maintained in the longer term, those Sumatran tigers held in captivity are managed as a global population under a World Association of Zoos and Aquariums Global Species Management Plan (GSMP). A retrospective study, including segregation and pedigree analysis, was conducted to investigate potential cases of congenital vestibular disease (CVD) in captive Sumatran tigers in Australasian zoos using medical and husbandry records, as well as video footage obtained from 50 tigers between 1975 and 2013. Data from the GSMP Sumatran tiger studbook were made available for pedigree and segregation analysis. Fourteen cases of CVD in 13 Sumatran tiger cubs and one hybrid cub (Panthera tigris ssp. sumatrae × Panthera tigris) were identified. Vestibular signs including head tilt, circling, ataxia, strabismus and nystagmus were observed between birth and 2 months of age. These clinical signs persisted for a median of 237 days and had resolved by 2 years of age in all cases. Pedigree analysis revealed that all affected tigers were closely related and shared a single common ancestor in the last four generations. A genetic cause for the disease is suspected and, based on pedigree and segregation analysis, an autosomal dominant mode of inheritance is likely. Further investigations to determine the world-wide prevalence and underlying pathology of this disorder are warranted.

  20. Effect of plant age on PCB accumulation by Cucurbita pepo ssp. pepo.

    PubMed

    Low, Jennifer E; Whitfield Aslund, Melissa L; Rutter, Allison; Zeeb, Barbara A

    2010-01-01

    A greenhouse experiment was conducted to investigate polychlorinated biphenyl (PCB) uptake and translocation from soil over time in pumpkin plants (Cucurbita pepo ssp. pepo cv. Howden). Plants were grown in weathered soil collected from a former industrial site contaminated with Aroclor 1248 (mean [PCB](soil) = 6.5 mg kg(-1)). Plants were harvested five times over 42 d and analyzed for total PCB concentration in the root and shoot tissues. The concentration of PCBs in the root was not significantly different between harvests (mean [PCB](root) = 21.5 mg kg(-1)). The concentration of PCBs in the shoots was also relatively stable over time (mean [PCB](shoot) = 3.5 mg kg(-1)) despite increases in shoot biomass (fresh weight of 4.3 g at Day 12 to 59 g at Day 42). This suggests that PCBs were continuously accumulated throughout the growth period. The trends found in this study indicate the optimal time to harvest C. pepo ssp pepo plants to maximize PCB phytoextraction is when the plant shoot has reached its maximum biomass.

  1. Evaluation of in vitro antioxidant activity of Sida rhombifolia (L.) ssp. retusa (L.).

    PubMed

    Dhalwal, Kamlesh; Deshpande, Yogesh S; Purohit, Ajit P

    2007-12-01

    Sida rhombifolia (L.) ssp. retusa (L.) is widely used in Ayurvedic medicine for the treatment of fever as well as a diuretic. The comparative antioxidant potentials of ethanol extract of roots, stems, leaves, and whole plant were studied. Estimation of total polyphenolic content and high-performance thin-layer chromatography profile were determined. Further inhibition of oxygen-derived free radicals, viz., assays for free radical scavenging, reducing power, superoxide anion scavenging, nitric oxide scavenging, and anti-lipid peroxidation, were performed. All the antioxidant activities were compared with standard antioxidants such as butylated hydroxyanisole and alpha-tocopherol acetate. Extracts were found to be good scavengers of the 1,1-diphenyl-2-picrylhydrazyl radical in the order root > leaves > whole plant > stem with 50% inhibitory concentrations of 546.1, 852.8, 983.8, and 1,222.5 microg/mL, respectively. All extracts of this plant showed effective free radical scavenging activity, reducing power, and superoxide scavenging activity. Only root extract inhibited lipid peroxidation in rat liver and brain homogenate. All these antioxidant properties were concentration dependent. In addition, total polyphenolic contents of all the extracts were determined as gallic acid equivalents. The highest antioxidant activity was observed in root extract. The results obtained from the current study indicate that S. rhombifolia ssp. retusa is a potential source of natural antioxidants.

  2. Sida rhombifolia ssp. retusa seed extract inhibits DEN induced murine hepatic preneoplasia and carbon tetrachloride hepatotoxicity.

    PubMed

    Poojari, Radhika; Gupta, Sanjay; Maru, Girish; Khade, Bharat; Bhagwat, Sanjay

    2009-01-01

    Sida rhombifolia ssp. retusa is a well established drug in the Ayurvedic system of medicine used for antirheumatism and antiasthmatism. Inhibitory effects of S. rhombifolia ssp. retusa seed extract on DEN induced hepatocellular preneoplastic foci and carbon tetrachloride (CCl4) induced hepatotoxicity was investigated in rats. Rats received DEN, 1ppm/g b.w. in drinking water for 6 weeks or CCl(4), 0.7 ml/kg i.p. once a week for 4 weeks and seed extract 50 mg, 100 mg/kg b.w. orally prior, during and after exposure to DEN/CCl4 for 20 or 5 weeks, respectively. Treatment with seed extract significantly inhibited the increase in DEN/CCl(4) induced activities of pre-cancerous marker enzymes; gamma-glutamyl transpeptidase, glutathione-S-transferase, hepatotoxicity marker enzymes; glutamate pyruvate transaminase, glutamate oxaloacetate transaminase and alkaline phosphatase as well as lipid peroxidase. Depleted glutathione, protein and albumin levels were restored. Also, histopathological and transmission electron microscopic studies showed prevention of cellular degenerative changes. The chemopreventive and hepatoprotective potentials of seed extract are due to free radical scavenging activity and restoration of cellular structural integrity.

  3. Systems Analysis of Early Host Gene Expression Provides Clues for Transient Mycobacterium avium ssp avium vs. Persistent Mycobacterium avium ssp paratuberculosis Intestinal Infections

    PubMed Central

    Khare, Sangeeta; Drake, Kenneth L.; Lawhon, Sara D.; Nunes, Jairo E. S.; Figueiredo, Josely F.; Rossetti, Carlos A.; Gull, Tamara; Everts, Robin E.; Lewin, Harris. A.; Adams, Leslie Garry

    2016-01-01

    It has long been a quest in ruminants to understand how two very similar mycobacterial species, Mycobacterium avium ssp. paratuberculosis (MAP) and Mycobacterium avium ssp. avium (MAA) lead to either a chronic persistent infection or a rapid-transient infection, respectively. Here, we hypothesized that when the host immune response is activated by MAP or MAA, the outcome of the infection depends on the early activation of signaling molecules and host temporal gene expression. To test our hypothesis, ligated jejuno-ileal loops including Peyer’s patches in neonatal calves were inoculated with PBS, MAP, or MAA. A temporal analysis of the host transcriptome profile was conducted at several times post-infection (0.5, 1, 2, 4, 8 and 12 hours). When comparing the transcriptional responses of calves infected with the MAA versus MAP, discordant patterns of mucosal expression were clearly evident, and the numbers of unique transcripts altered were moderately less for MAA-infected tissue than were mucosal tissues infected with the MAP. To interpret these complex data, changes in the gene expression were further analyzed by dynamic Bayesian analysis. Bayesian network modeling identified mechanistic genes, gene-to-gene relationships, pathways and Gene Ontologies (GO) biological processes that are involved in specific cell activation during infection. MAP and MAA had significant different pathway perturbation at 0.5 and 12 hours post inoculation. Inverse processes were observed between MAP and MAA response for epithelial cell proliferation, negative regulation of chemotaxis, cell-cell adhesion mediated by integrin and regulation of cytokine-mediated signaling. MAP inoculated tissue had significantly lower expression of phagocytosis receptors such as mannose receptor and complement receptors. This study reveals that perturbation of genes and cellular pathways during MAP infection resulted in host evasion by mucosal membrane barrier weakening to access entry in the ileum

  4. Mycoplasma hominis ssp. associated endocarditis with myocardial necrosis in an alpaca (Vicugna pacos) in Manitoba in 2011

    PubMed Central

    Tomczyk, Krzysztof M.; Copeland, Shelagh; Postey, Rosemary; Ngeleka, Musangu

    2015-01-01

    Severe endocarditis with myonecrosis, moderate to severe pleural and pericardial effusions, and mild ascites were found on necropsy in 3 alpacas. Mycoplasma hominis ssp. was detected on polymerase chain reaction (PCR) of fresh affected endocardial tissue in 1 alpaca. PMID:25694661

  5. Molecular genetic tagging of Beta vulgaris ssp. maritima-derived resistance to the sugar beet cyst nematode, Heterodera schachtii

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Resistance in commercial sugar beet hybrids to the sugar beet cyst nematode (SBCN) principally has been based on the Hs1 gene from the wild beet Beta procumbens, yet incorporation of this resistance has been detrimental to crop yield in nematode-free fields. Accessions of B. vulgaris ssp maritima w...

  6. Poplar root exudates contain compounds that induce the expression of MiSSP7 in Laccaria bicolor.

    PubMed

    Plett, Jonathan M; Martin, Francis

    2012-01-01

    Communication between organisms is crucial for their survival, especially for sessile organisms such as plants that depend upon interactions with mutualistic organisms to maximize their nutrient acquisition. This communication can take the form of the exchange of volatile compounds, metabolites or effectors - small protein signals secreted from the colonizing cell that change the biology of the host cell.  We recently characterized the first mutualistic effector protein from an ectomycorrhizal fungus, a small secreted protein named MiSSP7 encoded by Laccaria bicolor.  Ectomycorrhizal fungi are soil-borne mutualistic organisms whose hyphae wrap around host roots and grow into the root apoplastic space where the fungus exchanges nutrients such as nitrogen and phosphorus in return for plant derived sugars.  The MiSSP7 protein is induced by root exudates and is highly expressed throughout the root colonization process.  Its presence was responsible for alterations to the plant transcriptomic profile, a mechanism by which MiSSP7 may aid in the formation of the symbiotic interface. Here we further discuss the implications of these findings and, further, we demonstrate that the production of MiSSP7 is induced by two flavonoids, rutin and quercitrin, a class of compounds normally found within the exudates of plant roots.  We also consider the interesting similarities between the mechanisms of effector induction and action between pathogenic and mutualistic fungi.  

  7. Growth Responses of Plantago major L. ssp. pleiosperma (Pilger) to Changes in Mineral Supply : Evidence for Regulation by Cytokinins.

    PubMed

    Kuiper, D

    1988-07-01

    Plants of an inbred line of Plantago major ssp. pleiosperma were subjected to an alteration in mineral supply. Observed responses of growth rate and shoot to root ratio are thought to be induced by changes in endogenous cytokinin concentration and not by mineral concentration in plant tissue.

  8. Identification of a Small Molecule That Modifies MglA/SspA Interaction and Impairs Intramacrophage Survival of Francisella tularensis

    PubMed Central

    Wrench, Algevis P.; Gardner, Christopher L.; Gonzalez, Claudio F.; Lorca, Graciela L.

    2013-01-01

    The transcription factors MglA and SspA of Francisella tularensis form a heterodimer complex and interact with the RNA polymerase to regulate the expression of the Francisella pathogenicity island (FPI) genes. These genes are essential for this pathogen’s virulence and survival within host cells. In this study, we used a small molecule screening to identify quinacrine as a thermal stabilizing compound for F. tularensis SCHU S4 MglA and SspA. A bacterial two-hybrid system was used to analyze the in vivo effect of quinacrine on the heterodimer complex. The results show that quinacrine affects the interaction between MglA and SspA, indicated by decreased β-galactosidase activity. Further in vitro analyses, using size exclusion chromatography, indicated that quinacrine does not disrupt the heterodimer formation, however, changes in the alpha helix content were confirmed by circular dichroism. Structure-guided site-directed mutagenesis experiments indicated that quinacrine makes contact with amino acid residues Y63 in MglA, and K97 in SspA, both located in the “cleft” of the interacting surfaces. In F. tularensis subsp. novicida, quinacrine decreased the transcription of the FPI genes, iglA, iglD, pdpD and pdpA. As a consequence, the intramacrophage survival capabilities of the bacteria were affected. These results support use of the MglA/SspA interacting surface, and quinacrine’s chemical scaffold, for the design of high affinity molecules that will function as therapeutics for the treatment of Tularemia. PMID:23372736

  9. Streptococcus gordonii DL1 adhesin SspB V-region mediates coaggregation via receptor polysaccharide of Actinomyces oris T14V.

    PubMed

    Back, C R; Douglas, S K; Emerson, J E; Nobbs, A H; Jenkinson, H F

    2015-10-01

    Streptococcus gordonii SspA and SspB proteins, members of the antigen I/II (AgI/II) family of Streptococcus adhesins, mediate adherence to cysteine-rich scavenger glycoprotein gp340 and cells of other oral microbial species. In this article we investigated further the mechanism of coaggregation between S. gordonii DL1 and Actinomyces oris T14V. Previous mutational analysis of S. gordonii suggested that SspB was necessary for coaggregation with A. oris T14V. We have confirmed this by showing that Lactococcus lactis surrogate host cells expressing SspB coaggregated with A. oris T14V and PK606 cells, while L. lactis cells expressing SspA did not. Coaggregation occurred independently of expression of A. oris type 1 (FimP) or type 2 (FimA) fimbriae. Polysaccharide was prepared from cells of A. oris T14V and found to contain 1,4-, 4,6- and 3,4-linked glucose, 1,4-linked mannose, and 2,4-linked galactose residues. When immobilized onto plastic wells this polysaccharide supported binding of L. lactis expressing SspB, but not binding of L. lactis expressing other AgI/II family proteins. Purified recombinant NAVP region of SspB, comprising amino acid (aa) residues 41-847, bound A. oris polysaccharide but the C-domain (932-1470 aa residues) did not. A site-directed deletion of 29 aa residues (Δ691-718) close to the predicted binding cleft within the SspB V-region ablated binding of the NAVP region to polysaccharide. These results infer that the V-region head of SspB recognizes an actinomyces polysaccharide ligand, so further characterizing a lectin-like coaggregation mechanism occurring between two important primary colonizers.

  10. meta-Tyrosine in Festuca rubra ssp. commutata (Chewings fescue) is synthesized by hydroxylation of phenylalanine.

    PubMed

    Huang, Tengfang; Rehak, Ludmila; Jander, Georg

    2012-03-01

    m-Tyrosine is a non-protein amino acid that is structurally similar to the common protein amino acids p-tyrosine and phenylalanine. Copious amounts of m-tyrosine can be found in root exudates of the fine fescue cultivar, Festuca rubra L. ssp. commutata (Chewings fescue). The phytotoxicity of m-tyrosine may contribute to the allelopathic potential of F. rubra. m-Tyrosine in Euphorbia myrsinites (donkey-tail spurge), was previously shown to be synthesized via transamination of m-hydroxyphenylpyruvate. Here we show that m-tyrosine biosynthesis in F. rubra occurs through direct hydroxylation of phenylalanine in the root tips, perhaps through the activity of a cytochrome P450 enzyme. Hence, E. myrsinites and F. rubra, the only two plant species known to produce m-tyrosine, use distinct biosynthetic pathways that likely arose independently in evolutionary history.

  11. Embryogenesis and plant regeneration from isolated microspores of Brassica rapa L. ssp. Oleifera.

    PubMed

    Burnett, L; Yarrow, S; Huang, B

    1992-05-01

    Conditions favourable to embryogenesis from isolated microspores of Brassica rapa L. ssp. oleifera (canola quality) were identified. A population with enhanced responsiveness for microspore embryogenesis (C200) was synthesized by crossing individual plants showing microspore embryogenic potential. For optimal microspore embryogenesis, buds (2-3mm in length, containing mid-late uninucieate microspores) were collected from older plants (2 months old) and microspores isolated and washed in iron-free B5 medium. NLN medium with its iron content reduced to half was beneficial for initial microspore culture. An elevated temperature(33-35°C) during the first day of culture, followed by maintenance at 25°C resulted in dozens of embryos from each isolation (about 100 buds). Seeds were obtained from plants regenerated from microsporederived embryos after colchicine treatment.

  12. Lignan formation in hairy root cultures of Edelweiss (Leontopodium nivale ssp. alpinum (Cass.) Greuter)

    PubMed Central

    Wawrosch, Christoph; Schwaiger, Stefan; Stuppner, Hermann; Kopp, Brigitte

    2014-01-01

    A hairy root line of Edelweiss (Leontopodium nivale ssp. alpinum (Cass.) Greuter) was obtained upon transformation with Agrobacterium rhizogenes strain ATCC15834. Elicitation of this line with silver nitrate, sucrose, methyl jasmonate and yeast extract at various concentrations in most cases resulted in a stimulation of lignan biosynthesis. Through elicitation with 6% sucrose the roots accumulated the pharmacologically active lignans leoligin and 5-methoxy-leoligin at levels of 0.0678% and 0.0372%, respectively, without significant growth inhibition. These lignan levels were comparable to those found in intact roots of cultivated Edelweiss. The biotechnological production of leoligin could be an attractive option for the continuous, field culture-independent production of the valuable secondary metabolites leoligin and 5-methoxy-leoligin. PMID:24932777

  13. Herbaspirillum sp. strain GW103 alleviates salt stress in Brassica rapa L. ssp. pekinensis.

    PubMed

    Lee, Gun Woong; Lee, Kui-Jae; Chae, Jong-Chan

    2016-05-01

    Mutual interactions between plant and rhizosphere bacteria facilitate plant growth and reduce risks of biotic and abiotic stresses. The present study demonstrates alleviation of salt stress in Brassica rapa L. ssp. perkinensis (Chinese cabbage) by Herbaspirillum sp. strain GW103 isolated from rhizosphere soil of Phragmites australis. The strain was capable of producing plant beneficial factors, such as auxin, siderophore, and 1-aminocylopropane-1-carboxylic acid deaminase. Treatment of strain GW103 on Chinese cabbage under salt stress increased K(+)/Na(+) ratio in roots generating balance in the ratio of ion homeostasis and consequently contributed to the increase of biomass. In addition, root colonization potential of the strain was observed by green fluorescent protein (GFP)-tagging approach. These results strongly suggest the beneficial impact of strain GW103 by inducing the alleviation of salt stress and development of stress tolerance in Chinese cabbage via plant-microbe interaction.

  14. Antioxidant capacities and polyphenolics of Chinese cabbage (Brassica rapa L. ssp. Pekinensis) leaves.

    PubMed

    Seong, Gi-Un; Hwang, In-Wook; Chung, Shin-Kyo

    2016-05-15

    Chinese cabbage (Brassica rapa L. ssp. Pekinensis) is a green leafy vegetable used mainly in kimchi, salted and fermented dishes. Consumer preference for the leaf portion differs according to the type of dishes. In this study, Chinese cabbage was divided into three parts, and their antioxidant activities were investigated through in vitro assays. The total phenolic contents (TPC), total flavonoid contents (TFC), and vitamin C contents were also determined as indicators of antioxidant contents. The phenolic acids and flavonoids were separated and identified using high performance liquid chromatography (HPLC) and liquid chromatography/mass spectrometry (LC/MS). The outer leaf had the strongest antioxidant activity with the maximum antioxidant contents, followed by the mid- and inner leaves. Principal component analysis (PCA) revealed that outer leaf is positively related to caffeic acid, p-coumaric acid, ferulic acid, and myricetin contents, whereas the mid- and inner leaves are negatively related to sinapic acid contents.

  15. Antioxidant capacities and polyphenolics of Chinese cabbage (Brassica rapa L. ssp. Pekinensis) leaves.

    PubMed

    Seong, Gi-Un; Hwang, In-Wook; Chung, Shin-Kyo

    2016-05-15

    Chinese cabbage (Brassica rapa L. ssp. Pekinensis) is a green leafy vegetable used mainly in kimchi, salted and fermented dishes. Consumer preference for the leaf portion differs according to the type of dishes. In this study, Chinese cabbage was divided into three parts, and their antioxidant activities were investigated through in vitro assays. The total phenolic contents (TPC), total flavonoid contents (TFC), and vitamin C contents were also determined as indicators of antioxidant contents. The phenolic acids and flavonoids were separated and identified using high performance liquid chromatography (HPLC) and liquid chromatography/mass spectrometry (LC/MS). The outer leaf had the strongest antioxidant activity with the maximum antioxidant contents, followed by the mid- and inner leaves. Principal component analysis (PCA) revealed that outer leaf is positively related to caffeic acid, p-coumaric acid, ferulic acid, and myricetin contents, whereas the mid- and inner leaves are negatively related to sinapic acid contents. PMID:26776015

  16. E-cinnamic acid derivatives and phenolics from Chilean strawberry fruits, Fragaria chiloensis ssp. chiloensis.

    PubMed

    Cheel, José; Theoduloz, Cristina; Rodríguez, Jaime; Saud, Guillermo; Caligari, Peter D S; Schmeda-Hirschmann, Guillermo

    2005-11-01

    Three E-cinnamic acid glycosides, tryptophan, and cyanidin-3-O-beta-D-glucopyranoside were isolated from ripe fruits of the Chilean strawberry Fragaria chiloensis ssp. chiloensis. 1-O-E-Cinnamoyl-beta-D-xylopyranoside, 1-O-E-cinnamoyl-beta-D-rhamnopyranoside, and 1-O-E-cinnamoyl-alpha-xylofuranosyl-(1-->6)-beta-D-glucopyranose are reported for the first time. The cinnamic acid glycosides and aromatic compound patterns in F. chiloensis fruits were determined by high-performance liquid chromatography (HPLC). HPLC analyses of extracts showed that cyanidin-3-O-beta-D-glucopyranoside and free ellagic acid are present in achenes while the E-cinnamoyl derivatives and tryptophan were identified only in the thalamus. The free radical scavenging effect of the fruit extract can be associated with the anthocyanin content.

  17. Chemical variability of the needle oil of Juniperus communis ssp. alpina from Corsica.

    PubMed

    Ottavioli, Josephine; Gonny, Marcelle; Casanova, Joseph; Bighelli, Ange

    2009-12-01

    The composition of 109 samples of essential oil isolated from the needles of Juniperus communis ssp. alpina growing wild in Corsica was investigated by GC (in combination with retention indices), GC/MS, and 13C-NMR. Forty-four compounds accounting for 86.7-96.7% of the oil were identified. The oils consisted mainly of monoterpene hydrocarbons, in particular, limonene (9.2-53.9%), beta-phellandrene (3.7-25.2%), alpha-pinene (1.4-33.7%), and sabinene (0.1-33.6%). The 109 oil compositions were submitted to k-means partitioning and principal component analysis, which allowed the distinction of two groups within the oil samples. The composition of the major group (92% of the samples) was dominated by limonene and beta-phellandrene, while the second group contained mainly sabinene beside limonene and beta-phellandrene.

  18. E-cinnamic acid derivatives and phenolics from Chilean strawberry fruits, Fragaria chiloensis ssp. chiloensis.

    PubMed

    Cheel, José; Theoduloz, Cristina; Rodríguez, Jaime; Saud, Guillermo; Caligari, Peter D S; Schmeda-Hirschmann, Guillermo

    2005-11-01

    Three E-cinnamic acid glycosides, tryptophan, and cyanidin-3-O-beta-D-glucopyranoside were isolated from ripe fruits of the Chilean strawberry Fragaria chiloensis ssp. chiloensis. 1-O-E-Cinnamoyl-beta-D-xylopyranoside, 1-O-E-cinnamoyl-beta-D-rhamnopyranoside, and 1-O-E-cinnamoyl-alpha-xylofuranosyl-(1-->6)-beta-D-glucopyranose are reported for the first time. The cinnamic acid glycosides and aromatic compound patterns in F. chiloensis fruits were determined by high-performance liquid chromatography (HPLC). HPLC analyses of extracts showed that cyanidin-3-O-beta-D-glucopyranoside and free ellagic acid are present in achenes while the E-cinnamoyl derivatives and tryptophan were identified only in the thalamus. The free radical scavenging effect of the fruit extract can be associated with the anthocyanin content. PMID:16248546

  19. Herbaspirillum sp. strain GW103 alleviates salt stress in Brassica rapa L. ssp. pekinensis.

    PubMed

    Lee, Gun Woong; Lee, Kui-Jae; Chae, Jong-Chan

    2016-05-01

    Mutual interactions between plant and rhizosphere bacteria facilitate plant growth and reduce risks of biotic and abiotic stresses. The present study demonstrates alleviation of salt stress in Brassica rapa L. ssp. perkinensis (Chinese cabbage) by Herbaspirillum sp. strain GW103 isolated from rhizosphere soil of Phragmites australis. The strain was capable of producing plant beneficial factors, such as auxin, siderophore, and 1-aminocylopropane-1-carboxylic acid deaminase. Treatment of strain GW103 on Chinese cabbage under salt stress increased K(+)/Na(+) ratio in roots generating balance in the ratio of ion homeostasis and consequently contributed to the increase of biomass. In addition, root colonization potential of the strain was observed by green fluorescent protein (GFP)-tagging approach. These results strongly suggest the beneficial impact of strain GW103 by inducing the alleviation of salt stress and development of stress tolerance in Chinese cabbage via plant-microbe interaction. PMID:26358119

  20. Methyl Jasmonate Induces Enhanced Podophyllotoxin Production in Cell Cultures of Thracian Flax (Linum thracicum ssp. thracicum).

    PubMed

    Sasheva, Pavlina; Ionkova, Iliana; Stoilova, Nadezhda

    2015-07-01

    The Linum thracicum ssp. thracicum cell lines developed in this study are a feasible source for the sustainable production of podophyllotoxin, a lignan with an aryltetralin skeleton that is used for the manufacture of the chemotherapeutic drugs etopophos and teniposide. We used mass spectrometry to confirm the presence of the aryltetralin lignan in the thracian flax cell cultures. Next, we explored how changes in the culture medium influenced the podophyllotoxin content. Out of six developed cell lines, four were selected for further experiments and challenged with elicitors. The selected cell lines clustered into two groups: developed in full strength medium (Li) vs developed in half strength medium (HS). While podophyllotoxin production in the Li cell lines was boosted by 80% upon administration of the elicitor methyl jasmonate, the HS lines produced high amounts of the target metabolite triggered by reduced concentration of nutrients and were only slightly influenced by the elicitor.

  1. Space Shuttle Program (SSP) Shock Test and Specification Experience for Reusable Flight Hardware Equipment

    NASA Technical Reports Server (NTRS)

    Larsen, Curtis E.

    2012-01-01

    As commercial companies are nearing a preliminary design review level of design maturity, several companies are identifying the process for qualifying their multi-use electrical and mechanical components for various shock environments, including pyrotechnic, mortar firing, and water impact. The experience in quantifying the environments consists primarily of recommendations from Military Standard-1540, Product Verification Requirement for Launch, Upper Stage, and Space Vehicles. Therefore, the NASA Engineering and Safety Center (NESC) formed a team of NASA shock experts to share the NASA experience with qualifying hardware for the Space Shuttle Program (SSP) and other applicable programs and projects. Several team teleconferences were held to discuss past experience and to share ideas of possible methods for qualifying components for multiple missions. This document contains the information compiled from the discussions

  2. Characterization of a male sterile related gene BcMF15 from Brassica campestris ssp. chinensis.

    PubMed

    Tian, Aimei; Cao, Jiashu; Huang, Li; Yu, Xiaolin; Ye, Wanzhi

    2009-02-01

    Data from cDNA-AFLP analysis based on the genome-wide transcriptional profiling on the flower buds of the male meiotic cytokinesis (mmc) mutant and its wild-type of Brassica campestris L. ssp. chinensis Makino, syn. B. rapa L. ssp. chinensis, indicated that mutation of the MMC gene resulted in changes in expression of a variety of genes. A transcript-derived fragment specifically accumulated in the wild-type flower buds was isolated, and the corresponding full-length cDNA and DNA was subsequently amplified. Bioinformatical analyses of this gene named BcMF15 (GenBank accession number EF600901) showed that it encoded a protein with 103 amino acids. The BcMF15 had a 88% nucleotide similarity to a lipid transfer protein-like gene. Moreover, sequence prediction indicated that BcMF15 might encode a membrane protein with a signal peptide at the N-terminus. Meanwhile, six domains were predicted in the deduced BcMF15 protein, such as the AAI domain existing in some crucial proteins of pollen development-preferential, signal peptide, transmembrane domain, vWF domain, ZnF_C4 domain, and Tryp_alpha_amyl domain. Spatial and temporal expression patterns analysis by RT-PCR indicated that BcMF15 was exclusively expressed in the fertile line, which indicated this gene is male sterile related. Phylogenetic analysis in Cruciferae revealed that the BcMF15 was relative conservative in evolution. We suppose BcMF15 may be a critical molecule in the transmembrane transportation and signal transduction during microspore development.

  3. Assessing the diversity of arbuscular mycorrhizal fungi in semiarid shrublands dominated by Artemisia tridentata ssp. wyomingensis.

    PubMed

    Carter, Keith A; Smith, James F; White, Merlin M; Serpe, Marcelo D

    2014-05-01

    Variation in the abiotic environment and host plant preferences can affect the composition of arbuscular mycorrhizal (AMF) assemblages. This study analyzed the AMF taxa present in soil and seedlings of Artemisia tridentata ssp. wyomingensis collected from sagebrush steppe communities in southwestern Idaho, USA. Our aims were to determine the AMF diversity within and among these communities and the extent to which preferential AMF-plant associations develop during seedling establishment. Mycorrhizae were identified using molecular methods following DNA extraction from field and pot culture samples. The extracted DNA was amplified using Glomeromycota specific primers, and identification of AMF was based on phylogenetic analysis of sequences from the large subunit-D2 rDNA region. The phylogenetic analyses revealed seven phylotypes, two within the Claroideoglomeraceae and five within the Glomeraceae. Four phylotypes clustered with known species including Claroideoglomus claroideum, Rhizophagus irregularis, Glomus microaggregatum, and Funneliformis mosseae. The other three phylotypes were similar to several published sequences not included in the phylogenetic analysis, but all of these were from uncultured and unnamed glomeromycetes. Pairwise distance analysis revealed some phylotypes with high genetic variation. The most diverse was the phylotype that included R. irregularis, which contained sequences showing pairwise differences up to 12 %. Most of the diversity in AMF sequences occurred within sites. The smaller genetic differentiation detected among sites was correlated with differences in soil texture. In addition, multiplication in pot cultures led to differentiation of AMF communities. Comparison of sequences obtained from the soil with those from A. tridentata roots revealed no significant differences between the AMF present in these samples. Overall, the sites sampled were dominated by cosmopolitan AMF taxa, and young seedlings of A. tridentata ssp

  4. Rapid molecular prenatal diagnosis of spondyloepiphyseal dysplasia congenita by PCR-SSP assay.

    PubMed

    Cui, Ying-Xia; Xia, Xin-Yi; Bu, Ying; Zhou, Guo-Hua; Yang, Bin; Lu, Hong-Yong; Shi, Yi-Chao; Pan, Lian-Jun; Huang, Yu-Feng; Li, Xiao-Jun

    2008-12-01

    Heterozygous mutations of COL2A1 gene are responsible for type II collagenopathies. The common skeletal phenotypes include achondrogenesis type II, hypochondrogenesis, Stickler dysplasia, Kniest dysplasia, late onset spondyloepiphyseal dysplasia, and spondyloepiphyseal dysplasia congenita (SEDC). Prevention of SEDC can be achieved by prenatal diagnosis. This study reports the first rapid molecular prenatal diagnosis of SEDC performed in China by polymerase chain reaction sequence-specific primer (PCR-SSP) analysis. The pregnant woman we previously reported with SEDC carried the G to A substitution at nucleotide 1510 in exon 23 of COL2A1 gene, which caused a change from glycine to serine at codon 504 (G504S). By the time the woman got pregnant again, she had terminated two pregnancies and still had no child. In the first pregnancy, the molecular mutation of the family was not yet identified, and therefore prenatal diagnosis was unable to be performed by DNA analysis. In the second pregnancy, G504S mutation was found from fetal DNA. At the time of her third pregnancy, the woman and her husband became extremely worried about the potential SEDC for the fetus. For this reason, a quick and reliable molecular prenatal diagnosis of SEDC was performed by a PCR-SSP on an amniocyte sample collected at the 14th week of pregnancy. No mutation of the fetal DNA was identified. The result was obtained within 24 h after the sample was collected. The technique could be applied in confirmatory diagnosis and prenatal diagnosis for the affected family. PMID:19072565

  5. VizieR Online Data Catalog: SSP in NIR. II. Synthesis models (Meneses-Goytia+, 2015)

    NASA Astrophysics Data System (ADS)

    Meneses-Goytia, S.; Peletier, R. F.; Trager, S. C.; Vazdekis, A.

    2015-08-01

    The present Single Stellar Populations (SSP) models are derived from my Ph.D.'s thesis and this paper. The following nomenclature is used throughout the paper and the website (http://smg.astro-research.net/ssp-models/the-models/) to describe the models, e.g. MarS models use the M08 isochrones (Mar) and the Salpeter (S) IMF. General information about the models is given in table1. For further information, please refer to the paper. Each set of models and their corresponding predictions are available in the website and VIZIER. The spectral energy distributions (SEDs) can be downloaded in a zip-file from those pages in ascii format . The spectral energy distributions (SEDs) can be downloaded in a zip-file from t hose pages in ascii format . The nomenclature of each SED is as follows: isochroneIMFsedXXXXHZX.XXXXXXXXXTgXX.XXXXXXXe+XX - where XXXX tells whether those models contain C-stars or no (COMBO or NOCS respectively) H is the spectral band in which normalization occurred Z_X.XXXXXXXXX is the metallicity in terms of Z Tg_XX.XXXXXXXe+XX is the age in years. Each set of models contains MarS - 96 SEDs GirS - 96 SEDs BaSS - 116 SEDs We have also included in the websites the Integrated colours and line-strength indices from all our models (MarS, GirS and BaSS). The SEDs were convolved to a velocity dispersion of 350km/s before calculating indices. (5 data files).

  6. Intra-specific genetic diversity in wild olives (Olea europaea ssp cuspidata) in Hormozgan Province, Iran.

    PubMed

    Noormohammadi, Z; Samadi-Molayousefi, H; Sheidai, M

    2012-03-19

    Wild olive (O. europaea ssp cuspidata) plants grow in various regions of Iran and are expected to have considerable genetic diversity due to adaptation to the various environmental conditions. We examined the genetic diversity of four populations of wild olive growing in Hormozgan Province located in southern Iran by using 30 RAPDs and 10 ISSR markers. The mean value of polymorphism for RAPD loci was 73.71%, while the value for ISSR loci was 81.74%. The Keshar population had the highest value of intra-population polymorphism for both RAPD and ISSR loci (66.86 and 62.71%, respectively), while the Tudar population had the lowest values (20.35 and 28.81%, respectively). Similarly, the highest and lowest number of effective alleles, Shannon index and Nei's genetic diversity were also found for these two populations. The highest value of H(pop)/H(sp) within population genetic diversity for RAPD and ISSR loci was found for the Keshar population (H(pop) = 0.85 and H(sp) = 0.90). OPA04-750, OPA13-650 and OPA02-350 RAPD bands were specific for Tudar, Bondon and Keshar populations, respectively, while no specific ISSR bands were observed. Analysis of molecular variance as well as the pairwise F(ST) test showed significant differences for RAPD and ISSR markers among the populations. The NJ and UPGMA trees also separated the wild olive populations from each other, indicating their genetic distinctness. UPGMA clustering of the four wild olive populations placed the Tudar population far from the other populations; Keshar and Bokhoon population samples revealed more similarity and were grouped together. We conclude that there is high genetic diversity among O. europaea ssp cuspidata populations located in southern Iran. We also found RAPD and ISSR markers to be useful molecular tools to discriminate and evaluate genetic variations in wild olive trees.

  7. Chemical composition of the essential oils of the berries of Juniperus oxycedrus L. ssp. rufescens (L. K.) and Juniperus oxycedrus L. ssp. macrocarpa (S. & m.) Ball. and their antioxidant activities.

    PubMed

    Hanène, Medini; Ameur, Elaissi; Larbi, Khouja Med; Piras, Alessandra; Porcedda, Silvia; Falconieri, Danilo; Marongiu, Bruno; Farhat, Farhat; Chemli, Rachid

    2012-01-01

    This study is outlined to probe the chemical composition of essential oil and in vitro antioxidant activity of Juniperus oxycedrus ssp. macrocarpa (S. & m.) Ball. and Juniperus oxycedrus L. ssp. rufescens (L. K.) berries, collected from four sites, according to their maturity phase. The chemical composition of the hydrodistilled essential oil was analysed by GC-MS. Forty-eight compounds were identified, accounting for approximately 79.8-98.9% of the oil. The main constituents were α-pinene, germacrene D, myrcene, abietadiene and cis-calamenene, their mean percentage vary according to their phenological stage. The antioxidant activity of the samples was determined by the ABTS and DPPH radical scavenging activities. Hawaria essential oil extracted from mature berries showed the highest antioxidant capacity.

  8. Immunoreactive Proteins of Bifidobacterium longum ssp. longum CCM 7952 and Bifidobacterium longum ssp. longum CCDM 372 Identified by Gnotobiotic Mono-Colonized Mice Sera, Immune Rabbit Sera and Non-immune Human Sera

    PubMed Central

    Górska, Sabina; Dylus, Ewa; Rudawska, Angelika; Brzozowska, Ewa; Srutkova, Dagmar; Schwarzer, Martin; Razim, Agnieszka; Kozakova, Hana; Gamian, Andrzej

    2016-01-01

    The Bifidobacteria show great diversity in the cell surface architecture which may influence the physicochemical properties of the bacterial cell and strain specific properties. The immunomodulatory role of bifidobacteria has been extensively studied, however studies on the immunoreactivity of their protein molecules are very limited. Here, we compared six different methods of protein isolation and purification and we report identification of immunogenic and immunoreactive protein of two human Bifidobacterium longum ssp. longum strains. We evaluated potential immunoreactive properties of proteins employing polyclonal sera obtained from germ free mouse, rabbit and human. The protein yield was isolation method-dependent and the reactivity of proteins detected by SDS-PAGE and Western blotting was heterogeneous and varied between different serum samples. The proteins with the highest immunoreactivity were isolated, purified and have them sequenced. Among the immunoreactive proteins we identified enolase, aspartokinase, pyruvate kinase, DnaK (B. longum ssp. longum CCM 7952) and sugar ABC transporter ATP-binding protein, phosphoglycerate kinase, peptidoglycan synthethase penicillin-binding protein 3, transaldolase, ribosomal proteins and glyceraldehyde 3-phosphate dehydrogenase (B. longum ssp. longum CCDM 372). PMID:27746766

  9. Generation and characterization of Brassica rapa ssp. pekinensis - B. oleracea var. capitata monosomic and disomic alien addition lines.

    PubMed

    Gu, Ai Xia; Shen, Shu Xing; Wang, Yan Hua; Zhao, Jian Jun; Xuan, Shu Xin; Chen, Xue Ping; Li, Xiao Feng; Luo, Shuang Xia; Zhao, Yu Jing

    2015-09-01

    Five monosomic alien addition lines (MAALs) of Brassica rapa ssp. pekinensis - B. oleracea var. capitata were obtained by hybridization and backcrossing between B. rapa ssp. pekinensis (female parent) and B. oleracea var. capitata. The alien linkage groups were identified using 42 B. oleracea var. capitata linkage group-specific markers as B. oleracea linkage groups C2, C3, C6, C7 and C8. Based on the chromosomal karyotype of root tip cells, these five MAALs added individual chromosomes from B. oleracea var. capitata: chr 1 (the longest), chr 2 or 3, chr 5 (small locus of 25S rDNA), chr 7 (satellite-carrying) and chr 9 (the shortest). Five disomic alien addition lines were then generated by selfing their corresponding MAALs.

  10. Embryogenesis and plant regeneration of pakchoi (Brassica rapa L. ssp. chinensis) via in vitro isolated microspore culture.

    PubMed

    Cao, M Q; Li, Y; Liu, F; Doré, C

    1994-05-01

    Isolated microspores of various populations of three varieties of the Chinese cabbage pakchoi (Brassica rapa ssp. chinensis) were cultivated in vitro on NLN82 medium (Lichter 1982) and embryos and plantlets obtained with nine cultivars. The best embryo yield per bud was 57.4. A 33°C one day heat treatment was generally necessary to induce embryogenesis. Analysis of ploidy level through flow cytometry for two cultivars indicated that haploids were present.

  11. Engineering and Safety Partnership Enhances Safety of the Space Shuttle Program (SSP)

    NASA Technical Reports Server (NTRS)

    Duarte, Alberto

    2007-01-01

    Project Management must use the risk assessment documents (RADs) as tools to support their decision making process. Therefore, these documents have to be initiated, developed, and evolved parallel to the life of the project. Technical preparation and safety compliance of these documents require a great deal of resources. Updating these documents after-the-fact not only requires substantial increase in resources - Project Cost -, but this task is also not useful and perhaps an unnecessary expense. Hazard Reports (HRs), Failure Modes and Effects Analysis (FMEAs), Critical Item Lists (CILs), Risk Management process are, among others, within this category. A positive action resulting from a strong partnership between interested parties is one way to get these documents and related processes and requirements, released and updated in useful time. The Space Shuttle Program (SSP) at the Marshall Space Flight Center has implemented a process which is having positive results and gaining acceptance within the Agency. A hybrid Panel, with equal interest and responsibilities for the two larger organizations, Safety and Engineering, is the focal point of this process. Called the Marshall Safety and Engineering Review Panel (MSERP), its charter (Space Shuttle Program Directive 110 F, April 15, 2005), and its Operating Control Plan emphasizes the technical and safety responsibilities over the program risk documents: HRs; FMEA/CILs; Engineering Changes; anomalies/problem resolutions and corrective action implementations, and trend analysis. The MSERP has undertaken its responsibilities with objectivity, assertiveness, dedication, has operated with focus, and has shown significant results and promising perspectives. The MSERP has been deeply involved in propulsion systems and integration, real time technical issues and other relevant reviews, since its conception. These activities have transformed the propulsion MSERP in a truly participative and value added panel, making a

  12. Characterizing the Syphilis-Causing Treponema pallidum ssp. pallidum Proteome Using Complementary Mass Spectrometry

    PubMed Central

    Houston, Simon; Lithgow, Karen V.; Meehan, Conor J.; Strouhal, Michal; Šmajs, David; Cameron, Caroline E.; Van Ostade, Xaveer; Kenyon, Chris R.; Van Raemdonck, Geert A.

    2016-01-01

    Background The spirochete bacterium Treponema pallidum ssp. pallidum is the etiological agent of syphilis, a chronic multistage disease. Little is known about the global T. pallidum proteome, therefore mass spectrometry studies are needed to bring insights into pathogenicity and protein expression profiles during infection. Methodology/Principal Findings To better understand the T. pallidum proteome profile during infection, we studied T. pallidum ssp. pallidum DAL-1 strain bacteria isolated from rabbits using complementary mass spectrometry techniques, including multidimensional peptide separation and protein identification via matrix-assisted laser desorption ionization-time of flight (MALDI-TOF/TOF) and electrospray ionization (ESI-LTQ-Orbitrap) tandem mass spectrometry. A total of 6033 peptides were detected, corresponding to 557 unique T. pallidum proteins at a high level of confidence, representing 54% of the predicted proteome. A previous gel-based T. pallidum MS proteome study detected 58 of these proteins. One hundred fourteen of the detected proteins were previously annotated as hypothetical or uncharacterized proteins; this is the first account of 106 of these proteins at the protein level. Detected proteins were characterized according to their predicted biological function and localization; half were allocated into a wide range of functional categories. Proteins annotated as potential membrane proteins and proteins with unclear functional annotations were subjected to an additional bioinformatics pipeline analysis to facilitate further characterization. A total of 116 potential membrane proteins were identified, of which 16 have evidence supporting outer membrane localization. We found 8/12 proteins related to the paralogous tpr gene family: TprB, TprC/D, TprE, TprG, TprH, TprI and TprJ. Protein abundance was semi-quantified using label-free spectral counting methods. A low correlation (r = 0.26) was found between previous microarray signal data and

  13. Characterization of a novel y-type HMW-GS with eight cysteine residues from Triticum monococcum ssp. monococcum.

    PubMed

    Li, Zenglin; Li, Hongyu; Chen, Gang; Kou, Chunlan; Ning, Shunzong; Yuan, Zhongwei; Jiang, Qi; Zheng, Youliang; Liu, Dengcai; Zhang, Lianquan

    2015-11-15

    The composition and number of high-molecular-weight glutenin subunits (HMW-GSs) play important roles in determining the grain-processing quality of common wheat. The Glu-1Ay allele is silent in common wheat. In this study, an active y-type HMW-GS allele termed 1Ay8.2 (GenBank No. KP137569) was identified from Triticum monococcum L. ssp. monococcum (AmAm, 2n=2x=14), a species with a genome related to the A-genome of common wheat. Compared with previously reported active 1Ay subunits, this novel subunit contained an extra cysteine residue at position 103 of the amino acid sequence in the N-terminal region, in addition to the six cysteines in the N- and C-terminal regions found in most active 1Ay subunits and the one in the repetitive region that appears in only a few 1Ay alleles. This subunit was expressed in an amphiploid (AAAmAmBB, 2n=6x=42) between Triticum turgidum L. ssp. dicoccon and T. monococcum ssp. monococcum. This amphiploid could be used as a bridge to transfer 1Ay8.2 into common wheat cultivars. Replacing the silenced 1Ay in common wheat with the active 1Ay8.2 allele harboring an extra cysteine residue is expected to improve the quality by increasing the number of HMW-GSs and promoting the formation of covalent interactions through disulfide bonds with the extra cysteine residue.

  14. Accumulation of biologically active furanocoumarins in agitated cultures of Ruta graveolens L. and Ruta graveolens ssp. divaricata (Tenore) Gams.

    PubMed

    Ekiert, H; Czygan, F-Ch

    2005-08-01

    This study was designed to investigate the dynamics of accumulation of linear furanocoumarins (psoralen, bergapten, xanthotoxin, isopimpinellin, imperatorin) and their biogenetic precursor, umbelliferone, in agitated cultures of Ruta graveolens L. and Ruta graveolens ssp. divaricata (Tenore) Gams during 6-week growth cycles. The metabolites under study were almost exclusively accumulated in the cultured biomass. The total content of all metabolites increased 4.8- and 2.0-fold, in R. graveolens and R. graveolens ssp. divaricata cultures, respectively. Xanthotoxin and bergapten, which are the most important therapeutic compounds, were the dominating metabolites in cultures of both plants. The maximum content of xanthotoxin (25.0 mg/100 g dry wt.) and bergapten (18.4 mg/100 g dry wt) and the maximum content of all metabolites (64.0 mg/100 g dry wt) in R. graveolens ssp. divaricata callus obtained on the 35th culture day were relatively low. However, maximum contents of xanthotoxin (136.8 mg/100 g dry wt), bergapten (210.4 mg/100 g dry wt.) and isopimpinellin (96.7 mg/100 g dry wt), and total content of all metabolites in R. graveolens shoots (520.8 mg/100 g dry wt) obtained on the 42nd culture day are interesting from a practical point of view.

  15. Bovine PGLYRP1 polymorphisms and their association with resistance to Mycobacterium avium ssp. paratuberculosis.

    PubMed

    Pant, S D; Verschoor, C P; Schenkel, F S; You, Q; Kelton, D F; Karrow, N A

    2011-08-01

    Mycobacterium avium ssp. paratuberculosis (MAP) causes a chronic, granulomatous inflammatory condition of the intestines in ruminants and wild-type species. It causes significant economic losses to the dairy and beef industries owing to reduced productivity, premature culling and mortality. Bovine peptidoglycan recognition protein 1 is an important pattern recognition molecule that is capable of directly killing microorganisms. The goal of this study was to identify single nucleotide polymorphisms (SNPs) in the gene encoding bovine peptidoglycan recognition protein 1 and to assess their association with susceptibility to MAP infection in dairy cattle. Blood and milk samples were collected from Holsteins in Southwestern and Eastern Ontario and tested for MAP infection using blood and milk ELISAs. A resource population consisting of 197 infected (S/P > 0.25) and 242 healthy (S/P < 0.10) cattle was constructed. Sequencing of pooled DNA was used to identify three SNPs (c.102G>C, c.480G>A and c.625C>A) that were genotyped in the resource population. Statistical analysis was performed using a logistic regression model fitting the additive and dominance effects of each SNP in the model. SNP c.480G>A (P = 0.054) was found to be associated with susceptibility to MAP infection. Cows with a copy of the major allele 'G' at this locus had an odds ratio of 1.51 (95% CI: 0.99-2.31) for being infected with MAP.

  16. Identification and preliminary analysis of a new PCP promoter from Brassica rapa ssp. chinensis.

    PubMed

    Zhang, Qiang; Liu, Huizhi; Cao, Jiashu

    2008-12-01

    The promoter of Brassica campestris Male Fertile 5 (BcMF5), a pollen coat protein member, class A (PCP-A) gene family, was isolated from Brassica rapa L. ssp. chinensis Makino (Chinese cabbage-pak-choi) by Thermal Asymmetric Interlaced Polymerase Chain Reaction (TAIL-PCR). Sequence analysis suggested that the 605-bp promoter of BcMF5 appears to be a pollen promoter. In an attempt to confirm the promoter activity of BcMF5 promoter, -609 to +3 bp and -377 to +3 bp fragments of the upstream sequence of BcMF5 were inserted at the site upstream of the coding region of the uidA gene in the sense orientation to construct two deletion expression vectors. Transient expression analysis in onion epidermal cells by particle bombardment showed that both -609 to +3 bp and -377 to +3 bp fragments of BcMF5 promoter were capable of driving beta-glucuronidase gene expression. Furthermore, by Agrobacterium-mediated genetic transformation method, Arabidopsis transgenic Kan(R) plants were obtained. GUS assay analysis revealed that the promoter of BcMF5 induced gene expression at the early stage of anther development and drove high levels of GUS expression in anther walls, upper regions of petals, pollen, and pollen tubes in the middle and late stage of anther development, but did not drive any expression in sepals and pistils.

  17. Genotypic variation of the glucosinolate profile in pak choi (Brassica rapa ssp. chinensis).

    PubMed

    Wiesner, Melanie; Zrenner, Rita; Krumbein, Angelika; Glatt, Hansruedi; Schreiner, Monika

    2013-02-27

    Thirteen different pak choi (Brassica rapa ssp. chinensis) cultivars were characterized regarding their glucosinolate profile analyzed by HPLC-DAD-MS. The identified glucosinolates were subjected to principal component analysis, and three distinct groups of pak choi sprouts were identified. Group differences were marked mainly by variations in the aliphatic glucosinolate profile such as differing levels of 3-butenyl glucosinolate and 2-hydroxy-3-butenyl glucosinolate as well as by their varying proportional ratios. In addition, the three groups of pak choi sprouts varied by the presence or absence of 2-hydroxy-4-pentenyl glucosinolate and in level and composition of butyl glucosinolates. This classification is reflected by relative mRNA expression level of 2-oxoacid-dependent dioxygenase. As in sprouts, the major glucosinolates in mature leaves were found to be the aliphatic glucosinolates. However, unlike in sprouts, an additional aliphatic glucosinolate, 5-methylsulfinylpentyl glucosinolate, was detected as characteristic ontogenetic variation in mature leaves in 12 of the 13 pak choi cultivars analyzed.

  18. Analysis of expressed sequence tags from Brassica rapa L. ssp. pekinensis.

    PubMed

    Lim, J Y; Shin, C S; Chung, E J; Kim, J S; Kim, H U; Oh, S J; Choi, W B; Ryou, C S; Kim, J B; Kwon, M S; Chung, T Y; Song, S I; Kim, J K; Nahm, B H; Hwang, Y S; Eun, M Y; Lee, J S; Cheong, J J; Choi, Y D

    2000-08-31

    Non-redundant expressed sequence tags (ESTs) were generated from six different organs at various developmental stages of Chinese cabbage, Brassica rapa L. ssp. pekinensis. Of the 1,295 ESTs, 915 (71%) showed significantly high homology in nucleotide or deduced amino acid sequences with other sequences deposited in databases, while 380 did not show similarity to any sequences. Briefly, 598 ESTs matched with proteins of identified biological function, 177 with hypothetical proteins or non-annotated Arabidopsis genome sequences, and 140 with other ESTs. About 82% of the top-scored matching sequences were from Arabidopsis or Brassica, but overall 558 (43%) ESTs matched with Arabidopsis ESTs at the nucleotide sequence level. This observation strongly supports the idea that gene-expression profiles of Chinese cabbage differ from that of Arabidopsis, despite their genome structures being similar to each other. Moreover, sequence analyses of 21 Brassica ESTs revealed that their primary structure is different from those of corresponding annotated sequences of Arabidopsis genes. Our data suggest that direct prediction of Brassica gene expression pattern based on the information from Arabidopsis genome research has some limitations. Thus, information obtained from the Brassica EST study is useful not only for understanding of unique developmental processes of the plant, but also for the study of Arabidopsis genome structure.

  19. An intensive understanding of vacuum infiltration transformation of pakchoi (Brassica rapa ssp. chinensis).

    PubMed

    Xu, Hengjian; Wang, Xiufeng; Zhao, Hong; Liu, Fan

    2008-08-01

    Pakchoi (Brassica rapa L. ssp. chinensis), a kind of Chinese cabbage, is an important vegetable in Asian countries. Agrobacterium mediated in planta vacuum infiltration transformation has been performed in pakchoi since 1998, but a detailed study on this technique was lacking. Pakchoi plants 40-50 days old with inflorescences were vacuum infiltrated with Agrobacterium tumefaciens strain C58C1 harboring the binary vector pBBBast-gus-intron. The transformation frequency in the harvested seeds mainly varied from 1 x 10(-4) to 3 x 10(-4) over several years, and it was lower than the frequency in Arabidopsis thaliana. Transformants were obtained from both the upper and the lower parts of the infiltrated plants with or without an elongated inflorescence. Stained ovules and pollen grains were found in the unopened flower 13 days post-infiltration, which was about 0.5-1 mm in diameter at infiltration time with an open ovary as revealed by paraffin sections. Histochemical assays revealed that Agrobacteria were more abundant in the flower tissue than in stem and leaf tissues at all times after infiltration despite the sharp decrease of live Agrobacteria in plant 14 days post infiltration as revealed by the colony forming units on the Agrobacteria culture medium. The results of vacuum infiltration transformation of pakchoi and Arabidopsis thaliana were compared and a strategy to optimize the transformation conditions to increase the transformation frequency in pakchoi was discussed.

  20. Mapping of a QTL for oleic acid concentration in spring turnip rape (Brassica rapa ssp. oleifera).

    PubMed

    Tanhuanpää, P K; Vilkki, J P; Vilkki, H J

    1996-06-01

    Bulk segregant analysis was used to search for RAPD (random amplified polymorphic DNA) markers linked to gene(s) affecting oleic acid concentration in an F2 population from the Brassica rapa ssp. oleifera cross Jo4002 x a high oleic acid individual from line Jo4072. Eight primers (=8 markers) out of 104 discriminated the 'high' and 'low' bulks consisting of extreme individuals from the oleic acid distribution. These markers were analysed throughout the entire F2 population, and their association with oleic acid was studied using both interval mapping and ANOVA analysis. Six of the markers mapped to one linkage group. A quantitative trait locus (QTL) affecting oleic acid concentration was found to reside within this linkage group with a LOD score >15. The most suitable marker for oleic acid content is OPH-17, a codominant marker close (<4cM) to the QTL. The mean seed oleic acid content in the F2 individuals carrying the larger allele of this marker was 80.14±9.76%; in individuals with the smaller allele, 54.53±6.83%; in the heterozygotes, 65.47±8.15%. To increase reproducibility, the RAPD marker was converted into a SCAR (sequence characterized amplied region) marker with specific primers. Marker OPH-17 can be used to select spring turnip rape individuals with the desired oleic acid content.

  1. Second generation bioethanol production from Saccharum spontaneum L. ssp. aegyptiacum (Willd.) Hack.

    PubMed

    Scordia, Danilo; Cosentino, Salvatore L; Jeffries, Thomas W

    2010-07-01

    Saccharum (Saccharum spontaneum L. ssp. aegyptiacum (Willd.) Hack.), is a rapidly growing, wide ranging high-yield perennial, suitable for second generation bioethanol production. This study evaluated oxalic acid as a pretreatment for bioconversion. Overall sugar yields, sugar degradation products, enzymatic glucan hydrolysis and ethanol production were studied as effects of temperature (150-190 degrees C), reaction time (10-40 min) and oxalic acid concentration 2-8% (w/w). Time and temperature were combined into a single parameter, Severity Factor (SF) [Log(R(0))], and related to oxalic acid using a response surface methodology. Maximum total sugar yield was attained at a SF of 2.93 and 6.79% (w/w) oxalic acid, while maximum formation of sugar degradation products was observed at the highest SF (4.05) and 5% (w/w) oxalic acid. These were also the conditions for maximum simultaneous saccharification and fermentation (SSF) of the residual solids. Commercial cellulases and Saccharomyces cerevisiae attained 89.9% glucan conversion and 17.8 g/l ethanol. Pichia stipitis CBS 6054 fermented hemicellulosic hydrolysates from less severe conditions to ethanol with a yield of 0.35 (g(e)/g(s)). Maximal product yields were 69% of theoretical value and 90% of the SSF conversion efficiency for hydrolysate fermentation and SSF, respectively.

  2. Diversity of AMF associated with Ammophila arenaria ssp. arundinacea in Portuguese sand dunes.

    PubMed

    Rodríguez-Echeverría, Susana; Freitas, Helena

    2006-11-01

    Dune vegetation is essential for the formation and preservation of sand dunes and the protection of the coast line. Coastal sand dunes are harsh environments where arbuscular mycorrhizal fungi (AMF) play an important role in promoting plant establishment and growth. We present a study of the diversity of AMF associated with A. arenaria ssp. arundinacea in two locations of the Portuguese coast under a Mediterranean climate. These two locations were selected to compare a well-preserved dune system from a protected area with a degraded dune system from a public beach. AMF diversity was assessed mainly by cloning and sequencing of a fragment of the ribosomal SSU using the primer NS31 and AM1. Most of the 89 AMF clones obtained from the rhizosphere and roots of A. arenaria belonged to the genus Glomus, the largest clade within the Glomeromycota. Higher AMF diversity was found in the least disturbed site, in which spores of Scutellospora persica, Glomus constrictum and Glomus globiferum were found in the rhizosphere of A. arenaria.

  3. The complete chloroplast genome sequence of sugar beet (Beta vulgaris ssp. vulgaris).

    PubMed

    Li, Han; Cao, Hua; Cai, Yan-Fei; Wang, Ji-Hua; Qu, Su-Ping; Huang, Xing-Qi

    2014-06-01

    The complete nucleotide sequence of the sugar beet (Beta vulgaris ssp. vulgaris) chloroplast genome (cpDNA) was determined in this study. The cpDNA was 149,637 bp in length, containing a pair of 24,439 bp inverted repeat regions (IR), which were separated by small and large single copy regions (SSC and LSC) of 17,701 and 83,057 bp, respectively. 53.4% of the sugar beet cpDNA consisted of gene coding regions (protein coding and RNA genes). The gene content and relative positions of 113 individual genes (79 protein encoding genes, 30 tRNA genes, 4 rRNA genes) were almost identical to those of tobacco cpDNA. The overall AT contents of the sugar beet cpDNA were 63.6% and in the LSC, SSC and IR regions were 65.9%, 70.8% and 57.8%, respectively. Fifteen genes contained one intron, while three genes had two introns.

  4. Disentangling the causes of heterogeneity in male fecundity in gynodioecious Beta vulgaris ssp. maritima.

    PubMed

    De Cauwer, I; Arnaud, J-F; Klein, E K; Dufay, M

    2012-08-01

    Variation among individuals in reproductive success is advocated as a major process driving evolution of sexual polymorphisms in plants, such as gynodioecy where females and hermaphrodites coexist. In gynodioecious Beta vulgaris ssp. maritima, sex determination involves cytoplasmic male sterility (CMS) genes and nuclear restorers of male fertility. Both restored CMS and non-CMS hermaphrodites co-occur. Genotype-specific differences in male fitness are theoretically expected to explain the maintenance of cytonuclear polymorphism. Using genotypic information on seedlings and flowering plants within two metapopulations, we investigated whether male fecundity was influenced by ecological, phenotypic and genetic factors, while taking into account the shape and scale of pollen dispersal. Along with spatially restricted pollen flow, we showed that male fecundity was affected by flowering synchrony, investment in reproduction, pollen production and cytoplasmic identity of potential fathers. Siring success of non-CMS hermaphrodites was higher than that of restored CMS hermaphrodites. However, the magnitude of the difference in fecundity depended on the likelihood of carrying restorer alleles for non-CMS hermaphrodites. Our results suggest the occurrence of a cost of silent restorers, a condition supported by scarce empirical evidence, but theoretically required to maintain a stable sexual polymorphism in gynodioecious species.

  5. Comparative Transcriptome Analysis Reveals Heat-Responsive Genes in Chinese Cabbage (Brassica rapa ssp. chinensis)

    PubMed Central

    Wang, Aihua; Hu, Jihong; Huang, Xingxue; Li, Xia; Zhou, Guolin; Yan, Zhixiang

    2016-01-01

    Chinese cabbage (Brassica rapa ssp. chinensis) is an economically and agriculturally significant vegetable crop and is extensively cultivated throughout the world. Heat stress disturbs cellular homeostasis and causes visible growth inhibition of shoots and roots, severe retardation in growth and development, and even death. However, there are few studies on the transcriptome profiling of heat stress in non-heading Chinese cabbage. In this study, we investigated the transcript profiles of non-heading Chinese cabbage from heat-sensitive and heat-tolerant varieties “GHA” and “XK,” respectively, in response to high temperature using RNA sequencing (RNA seq). Approximately 625 genes were differentially expressed between the two varieties. The responsive genes can be divided into three phases along with the time of heat treatment: response to stimulus, programmed cell death and ribosome biogenesis. Differentially expressed genes (DEGs) were identified in the two varieties, including transcription factors (TFs), kinases/phosphatases, genes related to photosynthesis and effectors of homeostasis. Many TFs were involved in the heat stress response of Chinese cabbage, including NAC069 TF which was up-regulated at all the heat treatment stages. And their expression levels were also validated by quantitative real-time-PCR (qRT-PCR). These candidate genes will provide genetic resources for further improving the heat-tolerant characteristics in non-heading Chinese cabbage. PMID:27443222

  6. Isolation and characterization of Lactobacillus delbrueckii ssp. bulgaricus and Streptococcus thermophilus from plants in Bulgaria.

    PubMed

    Michaylova, Michaela; Minkova, Svetlana; Kimura, Katsunori; Sasaki, Takashi; Isawa, Kakuhei

    2007-04-01

    One of the traditional ways of preparation of yogurt starter in Bulgaria is placing a branch of a particular plant species into boiled sheep's milk maintained at about 45 degrees C, which is further incubated until a dense coagulum is obtained. To investigate the possible origin of the yogurt starter bacteria, Lactobacillus delbrueckii ssp. bulgaricus (L. bulgaricus) and Streptococcus thermophilus (S. thermophilus), the traditional way of yogurt-starter preparation was followed. Hundreds of plant samples were collected from four regions in Bulgaria and incubated in sterile skim milk. The two target bacteria at low frequencies from the plant samples collected were successfully isolated. Phenotypic and genotypic characteristics of these bacterial isolates revealed that they were identified as L. bulgaricus and S. thermophilus. Twenty isolates of L. bulgaricus and S. thermophilus, respectively, were selected from the isolated strains and further characterized with regard to their performance in yogurt production. Organoleptic and physical properties of yogurt prepared using the isolated strains from plants were not significantly different from those prepared using commercial yogurt-starter strains. It was therefore suggested that L. bulgaricus and S. thermophilus strains widely used for commercial yogurt production could have originated from plants in Bulgaria. To our knowledge, this is the first report on the isolation and characterization of L. bulgaricus and S. thermophilus strains from plants.

  7. Immunostimulatory oligonucleotide, CpG-like motif exists in Lactobacillus delbrueckii ssp. bulgaricus NIAI B6.

    PubMed

    Kitazawa, Haruki; Watanabe, Hiroshi; Shimosato, Takeshi; Kawai, Yasushi; Itoh, Takatoshi; Saito, Tadao

    2003-08-15

    The present study was conducted to find an immunostimulatory oligonucleotide derived from yogurt starter cultures. The chromosomal DNA was purified from nine strains of Lactobacillus delbrueckii ssp. bulgaricus and six strains of Streptococcus thermophilus. An immunostimulatory ability of the DNA was examined in a proliferation of peyer's patch and splenic B cells. Only the DNA from L. bulgaricus NIAI B6 induced a significant proliferation of both cells. When the DNA was cloned and amplified using PCR, the mitogenic activities to B cells were significantly increased by 13 of 135 DNA clones. Ten homologous nucleotide sequences were found as possible oligonucleotide sequences of mitogens, and were then chemically synthesized (sOL-LB1 to sOL-LB10). One CpG-like motif (sOL-LB7; 5'-CGGCACGCTCACGATTCTTG-3') was identified as an immunostimulatory oligonucleotide, but it did not contain palindromic CpG structure known as a B cell-specific mitogen. The sOL-LB7 substantially bound to B cells and increased the CD69 positive cells in peyer's patch cells. This study demonstrated that L. bulgaricus NIAI B6 was a good candidate of a starter culture for the production of new functional foods, "Bio-Defense Foods".

  8. Evaluating physiological stress in Sumatran tigers (Panthera tigris ssp. sumatrae) managed in Australian zoos.

    PubMed

    Parnell, Tempe; Narayan, Edward J; Magrath, Michael J L; Roe, Sheila; Clark, Giles; Nicolson, Vere; Martin-Vegue, Patrick; Mucci, Al; Hero, Jean-Marc

    2014-01-01

    Glucocorticoid quantification using non-invasive methods provides a powerful tool for assessing the health and welfare of wildlife in zoo-based programmes. In this study, we provide baseline data on faecal-based glucocorticoid (cortisol) monitoring of Sumatran tigers (Panthera tigris ssp. sumatrae) managed at the Melbourne Zoo in Victoria, Australia. We sampled five tigers daily for 60 days. Faecal cortisol metabolites (FCMs) in tiger faecal extracts were quantified using enzyme immunoassays that were successfully validated using parallelism and accuracy recovery checks. Two female tigers had significantly higher mean FCM levels than the two males and another female, suggesting that females may have higher FCM levels. A significant elevation was noted in the FCM levels for one female 2 days after she was darted and anaesthetized; however, the FCM levels returned to baseline levels within 3 days after the event. Comparative analysis of FCM levels of tigers sampled at Melbourne Zoo with tigers sampled earlier at two other Australian Zoos (Dreamworld Themepark and Australia Zoo) showed that FCM levels varied between zoos. Differences in the enclosure characteristics, timing of sampling, size and composition of groupings and training procedures could all contribute to this variation. Overall, we recommend the use of non-invasive sampling for the assessment of adrenocortical activity of felids managed in zoos in Australia and internationally in order to improve the welfare of these charismatic big cats. PMID:27293659

  9. Podophyllotoxin and 6-methoxy podophyllotoxin Production in Hairy Root Cultures of Liunm mucronatum ssp. mucronatum

    PubMed Central

    Samadi, Afsaneh; Jafari, Morad; Nejhad, Nasim Mohammad; Hossenian, Farah

    2014-01-01

    Aim: Two bacterial strains of Agrobacterium rhizogenes, A13 and 9534 were evaluated for induction of transformed hairy roots in Linum mucronatum ssp. mucronatum, a high value medicinal plant. Materials and Methods: The hairy roots were successfully initiated, through infecting the hypocotyl and root explants and the A13 strain performed a high transformation frequency for hairy roots induction. Transgenic status of hairy roots was confirmed by polymerase chain reaction (PCR) analysis of the rol genes. Growth kinetics of transgenic roots induced by two strains indicated a similar pattern of growth, with maximum growth occurring between 42 to 56 days. The lignan contents in hairy roots were analyzed using high-performance liquid chromatography (HPLC) method. Results: Transformed cultures showed significant differences (P < 0.05) in lignan content. The highest amount of Podophyllotoxin (PTOX, 5.78 mg/g DW) and 6-methoxy podophyllotoxin (MPTOX, 49.19 mg/g DW) was found in transformed lines induced by strain A13, which was four times higher than those of non-transformed roots. The results showed that hairy root cultures of L. mucronatum are rich sources of MPTOX. Conclusion: hairy root cultures from L. mucronatum can be used as a useful system for scale-up producing MPTOX and precursors for the production of antitumor agents in substitution with PTOX by considering the appropriate optimizations in future studies. PMID:24914281

  10. The Discovery of Resistant Sources of Spring Barley, Hordeum vulgare ssp. spontaneum, and Unique Greenbug Biotypes.

    PubMed

    Armstrong, J Scott; Mornhinweg, Dolores W; Payton, Mark E; Puterka, Gary J

    2016-02-01

    The genetic sources for host-plant resistance to the greenbug (Schizaphis graminum Rondani) in barley (Hordeum vulgare ssp. spontaneum) are limited in that only two single dominant genes Rsg1 and Rsg2 are available for the complex of greenbug biotypes. We evaluated four new barley lines from the Wild Barley Diversity Collection (WBDC) that previously showed potential for greenbug resistance. Three of those entries, WBDC 53, WBDC 117, WBDC 336, exhibited very dominant sources of resistance to older known biotypes B, C, E, F, H, I, and TX1, which also add to the host-plant differentials used to separate these greenbug biotypes. We also re-evaluated the earlier known set of greenbug biotypes that have been in culture for several years against the known host-plant differentials, and included seven newer greenbug isolates collected from Wyoming to the full complement of small grain differentials. This resulted in the discovery of five new greenbug biotypes, WY10 MC, WY81, WY10 B, WY12 MC, and WY86. Wyoming isolates WY4 A and WY4 B were identical in their phenotypic profile, and should be combined as a single unique greenbug biotype. These barley trials resulted in finding new sources of host-plant resistance, although more research needs to be conducted on what type of resistance was found, and how it can be used. We also document that the Wheatland, Wyoming area serves as a very conducive environment for the development of new greenbug biotypes.

  11. Elemental and nutritional analysis of sea buckthorn (Hippophae rhamnoides ssp. turkestanica) Berries of Pakistani origin.

    PubMed

    Sabir, S M; Maqsood, H; Hayat, Imran; Khan, M Q; Khaliq, A

    2005-01-01

    Sea buckthorn (Hippophae rhamnoides ssp. turkestanica) is a very important medicinal plant in northern areas of Pakistan. The fruit of sea buckthorn is rich in nutrients and medicinal compounds such as vitamins, carotene, flavonoids, essential oil, carbohydrates, organic acids, amino acids, and minerals. In order to compare various populations of sea buckthorn for chemical composition, eight populations from different areas of northern Pakistan were compared using fruit characteristics. Phytochemical analysis of berries showed vitamin C (250-333 mg/100 g), seed oil (7.69-13.7%), oil in softer pulp (19.2-29.1%), phytosterol content of seed oil (3.3-5.5%), and anthocyanin (0.5-25 mg/L), while the mineral element composition analysis revealed high contents of potassium (140-360 ppm), sodium (20-80 ppm), calcium (70-98 ppm), magnesium (150-240 ppm), iron (40-150 ppm), and phosphorus (110-133 ppm). This study established sea buckthorn berries as a good source of biochemical and mineral elements. The high variation between different populations shows the potential of selecting and breeding of the raw material for various defined purposes. PMID:16379565

  12. Characterization of a sagebrush (Artemisia tridentata ssp. wyomingensis) die-off on the Handford Site

    SciTech Connect

    Cardenas, A.; Lewinsohn, J.; Auger, C.; Downs, J.L.; Cadwell, L.L.; Burrows, R.

    1997-09-01

    The Hanford Site contains one of the few remaining contiguous areas of shrub-steppe habitat left in Washington State. This habitat is home to many native plant and wildlife species, some of which are threatened with extinction or are unique to the Site. The importance of the Hanford Site increases as other lands surrounding the Site are developed, and these native species and habitats are lost. Stands of Wyoming big sagebrush (Artemisia tridentata ssp. wyomingensis) on the Site are a particularly important component of shrub-steppe habitat, because a number of wildlife require big sagebrush for food and cover. Since 1993, researchers and field biologists have made anecdotal observations of dying and declining sagebrush in stands of shrubs near the 100 Areas. This study was initiated to delineate and document the general boundary where sagebrush stands appear to be declining. We mapped the areal extent of the die-off using a global positioning system and found that the central portion of the die-off encompasses 280 hectares. Shrub stand defoliation was estimated to be near or greater than 80% in this area. The remainder of the die-off area exhibits varying mixtures of completely defoliated, partially defoliated, and healthy-looking stands. Declining sagebrush stands comprise a total of 1776 hectares.

  13. Improving green roofs and rail road greening systems using Bacillus subtilis and Lactobacillus ssp.

    PubMed

    Grüneberg, H; Oschmann, C; Dunya, S; Ulrichs, C

    2006-01-01

    Aim of the present study was the improvement of existing methods for green roof and rail road greening systems using soil borne bacteria. Bacillus subtilis and Lactobacillus ssp. alone and in combination with vinasse applied to different growing substrates were tested. The substrates were brick chips, textile mats, mineral wool mats, and a commercial available substrate for the Swedish company VegTech. All four substrates were tested along an artificial rail track on the experimental station at Humboldt University Berlin, and partly on an existing rail track in Munich, Germany. Plants selected for the experiments belong to the genus Sedum, which is relatively tolerant to dry conditions. Inoculation of plants with bacteria had no effect on plant growth parameters and on coverage of different mobile bedding systems with Sedum plants. There was no significant difference between the various treatments in Munich. In both experiments, the addition of vinasse alone improved plant growth. Plant growth was significantly different on all substrates, whereas brick chips and the commercial roof soil was the best substrate. Brick chips are a cheap substrate which can be used for rail track greening. The results indicate that the quality of the substrate is the most important factor for remediation and greening of rail tracks and roof tops. The rapid growth of plants can be influenced by the application of vinasse as additional nutrient solution (potash (K) source) or nutrient enriched substrate.

  14. Instability in mitochondrial membranes in Polima cytoplasmic male sterility of Brassica rapa ssp. chinensis.

    PubMed

    Li, Ying; Liu, Tongkun; Duan, Weike; Song, Xiaoming; Shi, Gongjun; Zhang, Jingyi; Deng, Xiaohui; Zhang, Shuning; Hou, Xilin

    2014-06-01

    Cytoplasmic male sterility (CMS) is an important factor to observe heterosis in Brassica rapa. Although several studies have documented the rearrangements of mitochondrial DNA and dysfunction in the mitochondria have been observed in most types of CMS, the basis of the molecular mechanisms involved in these processes and other effects on CMS remain unclear. In this study, suppression subtractive hybridization was performed in the flowers of an alloplasmic Polima CMS system from B. rapa ssp. chinensis to identify genes that are differentially expressed between fertile and sterile plants. A total of 443 clones were isolated (156 were upregulated in fertile buds, and 287 were upregulated in sterile ones). Real-time RT-PCR further demonstrated the credibility of SSH. Among these genes, many membrane protein genes (LTP12, PIP2A, and GRP14) were inhibited in the sterile male line. Mitochondrial membrane potential (MMP) assay was then performed. Results showed that the sterile MMP was unstable and failed to create a potential difference; thus, mitochondrial dysfunction occurred. Moreover, abnormal microtubules and photosynthetic pathways were found in sterile male cells. Unstable MMP, nutritional deficiency, and abnormal microtubules were the causes of Polima CMS in Brassica campestris. H2O2, MDA, and O(2-), accumulated as byproducts of energy metabolism disorder in sterile male cells.

  15. Retrogenes in Rice (Oryza sativa L. ssp. japonica) Exhibit Correlated Expression with Their Source Genes

    PubMed Central

    Sakai, Hiroaki; Mizuno, Hiroshi; Kawahara, Yoshihiro; Wakimoto, Hironobu; Ikawa, Hiroshi; Kawahigashi, Hiroyuki; Kanamori, Hiroyuki; Matsumoto, Takashi; Itoh, Takeshi; Gaut, Brandon S.

    2011-01-01

    Gene duplication occurs by either DNA- or RNA-based processes; the latter duplicates single genes via retroposition of messenger RNA. The expression of a retroposed gene copy (retrocopy) is expected to be uncorrelated with its source gene because upstream promoter regions are usually not part of the retroposition process. In contrast, DNA-based duplication often encompasses both the coding and the intergenic (promoter) regions; hence, expression is often correlated, at least initially, between DNA-based duplicates. In this study, we identified 150 retrocopies in rice (Oryza sativa L. ssp japonica), most of which represent ancient retroposition events. We measured their expression from high-throughput RNA sequencing (RNAseq) data generated from seven tissues. At least 66% of the retrocopies were expressed but at lower levels than their source genes. However, the tissue specificity of retrogenes was similar to their source genes, and expression between retrocopies and source genes was correlated across tissues. The level of correlation was similar between RNA- and DNA-based duplicates, and they decreased over time at statistically indistinguishable rates. We extended these observations to previously identified retrocopies in Arabidopsis thaliana, suggesting they may be general features of the process of retention of plant retrogenes. PMID:22042334

  16. FurA contributes to the oxidative stress response regulation of Mycobacterium avium ssp. paratuberculosis

    PubMed Central

    Eckelt, Elke; Meißner, Thorsten; Meens, Jochen; Laarmann, Kristin; Nerlich, Andreas; Jarek, Michael; Weiss, Siegfried; Gerlach, Gerald-F.; Goethe, Ralph

    2015-01-01

    The ferric uptake regulator A (FurA) is known to be involved in iron homeostasis and stress response in many bacteria. In mycobacteria the precise role of FurA is still unclear. In the presented study, we addressed the functional role of FurA in the ruminant pathogen Mycobacterium avium ssp. paratuberculosis (MAP) by construction of a furA deletion strain (MAPΔfurA). RNA deep sequencing revealed that the FurA regulon consists of repressed and activated genes associated to stress response or intracellular survival. Not a single gene related to metal homeostasis was affected by furA deletion. A decisive role of FurA during intracellular survival in macrophages was shown by significantly enhanced survival of MAPΔfurA compared to the wildtype, indicating that a principal task of mycobacterial FurA is oxidative stress response regulation in macrophages. This resistance was not associated with altered survival of mice after long term infection with MAP. Our results demonstrate for the first time, that mycobacterial FurA is not involved in the regulation of iron homeostasis. However, they provide strong evidence that FurA contributes to intracellular survival as an oxidative stress sensing regulator. PMID:25705205

  17. Selenium alleviates chromium toxicity by preventing oxidative stress in cabbage (Brassica campestris L. ssp. Pekinensis) leaves.

    PubMed

    Qing, Xuejiao; Zhao, Xiaohu; Hu, Chengxiao; Wang, Peng; Zhang, Ying; Zhang, Xuan; Wang, Pengcheng; Shi, Hanzhi; Jia, Fen; Qu, Chanjuan

    2015-04-01

    The beneficial role of selenium (Se) in alleviation of chromium (Cr)-induced oxidative stress is well established. However, little is known about the underlying mechanism. The impacts of exogenous Se (0.1mg/L) on Cr(1mg/L)-induced oxidative stress and antioxidant systems in leaves of cabbage (Brassica campestris L. ssp. Pekinensis) were investigated by using cellular and biochemical approaches. The results showed that supplementation of the medium with Se was effective in reducing Cr-induced increased levels of lipid peroxides and superoxide free radicals (O(-)2(·)), as well as increasing activities of superoxide dismutase (SOD) and peroxidase (POD). Meanwhile, 1mg/L Cr induced loss of plasma membrane integrity, growth inhibition, as well as ultrastructural changes of leaves were significantly reversed due to Se supplementation in the medium. In addition, Se application significantly altered the subcellular distribution of Cr which transported from mitochondria, nucleus and the cell-wall material to the soluble fraction and chloroplasts. However, Se application did no significant alteration of Cr effects on osmotic adjustment accumulating products. The study suggested that Se is able to protect leaves of cabbage against Cr toxicity by alleviation of Cr induced oxidative stress, and re-distribution of Cr in the subcellular of the leaf. Furthermore, free radicals, lipid peroxides, activity of SOD and POD, and subcellular distribution of Cr can be considered the efficient biomarkers to indicate the efficiency of Se to detoxification Cr.

  18. Comparative Transcriptome Analysis Reveals Heat-Responsive Genes in Chinese Cabbage (Brassica rapa ssp. chinensis).

    PubMed

    Wang, Aihua; Hu, Jihong; Huang, Xingxue; Li, Xia; Zhou, Guolin; Yan, Zhixiang

    2016-01-01

    Chinese cabbage (Brassica rapa ssp. chinensis) is an economically and agriculturally significant vegetable crop and is extensively cultivated throughout the world. Heat stress disturbs cellular homeostasis and causes visible growth inhibition of shoots and roots, severe retardation in growth and development, and even death. However, there are few studies on the transcriptome profiling of heat stress in non-heading Chinese cabbage. In this study, we investigated the transcript profiles of non-heading Chinese cabbage from heat-sensitive and heat-tolerant varieties "GHA" and "XK," respectively, in response to high temperature using RNA sequencing (RNA seq). Approximately 625 genes were differentially expressed between the two varieties. The responsive genes can be divided into three phases along with the time of heat treatment: response to stimulus, programmed cell death and ribosome biogenesis. Differentially expressed genes (DEGs) were identified in the two varieties, including transcription factors (TFs), kinases/phosphatases, genes related to photosynthesis and effectors of homeostasis. Many TFs were involved in the heat stress response of Chinese cabbage, including NAC069 TF which was up-regulated at all the heat treatment stages. And their expression levels were also validated by quantitative real-time-PCR (qRT-PCR). These candidate genes will provide genetic resources for further improving the heat-tolerant characteristics in non-heading Chinese cabbage.

  19. Antioxidant activity of Nepeta nuda L. ssp. nuda essential oil rich in nepetalactones from Greece.

    PubMed

    Gkinis, George; Bozin, Biljana; Mimica-Dukic, Neda; Tzakou, Olga

    2010-10-01

    Essential oils from air-dried leaves and verticillasters of Nepeta nuda ssp. nuda from Greece were analyzed by means of gas chromatography-flame ionization detection and gas chromatography-mass spectrometry. The dominant constituent in the verticillaster oils was 4aα,7α,7aß-nepetalactone (75.7%). The main metabolites of the leaf oil were 1,8-cineole (16.7%), 4aα,7α,7aß-nepetalactone (24.7%), and caryophyllene oxide (16.3%). The oils were examined for their antioxidant activity. Neutralization of stable 2,2-diphenyl-1-picrylhydrazyl radical ranged from 10.83% (2.50 μg/mL) to 58.64% (50.00 μg/mL) for verticillaster oil and from 6.25% (2.50 μg/mL) to 57.79% (50.00 μg/mL) for leaf oil. The essential oil from verticillasters had significant effects on lipid peroxidation (in the range of 41.18-59.23%), compared to tert-butylated hydroxytoluene (37.04%). In contrast, the essential oil from leaves exhibited pro-oxidant activity at the highest concentration applied.

  20. Chemical composition and nutritional evaluation of the seeds of Acacia tortilis (Forssk.) Hayne ssp. raddiana.

    PubMed

    Embaby, Hassan E; Rayan, Ahmed M

    2016-06-01

    Chemical composition and nutritional evaluation as well as physicochemical and functional properties of seed flour of Acacia tortilis (Forssk.) Hayne ssp. raddiana were studied. The results indicated that seeds contained 5.30% moisture, 3.99% ash, 9.19% fat, 14.31% fiber, 27.21% protein and 45.30% carbohydrates. Potassium was the predominant element followed by calcium and then phosphorous. Phytic acid, tannins and trypsin inhibitor as antinutrients were detected. The amino acid profile compared well with FAO/WHO recommended pattern except for cystine/methionine, isoleucine, tyrosine/phenylalanine, lysine and threonine. Also, the first limiting amino acid was lysine. Fatty acid composition showed that linoleic acid was the major fatty acid, followed by palmitic, stearic, oleic and arachidic acids. The seed oil showed absorbance in the ultraviolet ranges, thus it can be used as a broad spectrum UV protectant. For physicochemical and functional properties, acacia seeds flour had excellent water holding index, swelling index, foaming capacity and foam stability.

  1. FurA contributes to the oxidative stress response regulation of Mycobacterium avium ssp. paratuberculosis.

    PubMed

    Eckelt, Elke; Meißner, Thorsten; Meens, Jochen; Laarmann, Kristin; Nerlich, Andreas; Jarek, Michael; Weiss, Siegfried; Gerlach, Gerald-F; Goethe, Ralph

    2015-01-01

    The ferric uptake regulator A (FurA) is known to be involved in iron homeostasis and stress response in many bacteria. In mycobacteria the precise role of FurA is still unclear. In the presented study, we addressed the functional role of FurA in the ruminant pathogen Mycobacterium avium ssp. paratuberculosis (MAP) by construction of a furA deletion strain (MAPΔfurA). RNA deep sequencing revealed that the FurA regulon consists of repressed and activated genes associated to stress response or intracellular survival. Not a single gene related to metal homeostasis was affected by furA deletion. A decisive role of FurA during intracellular survival in macrophages was shown by significantly enhanced survival of MAPΔfurA compared to the wildtype, indicating that a principal task of mycobacterial FurA is oxidative stress response regulation in macrophages. This resistance was not associated with altered survival of mice after long term infection with MAP. Our results demonstrate for the first time, that mycobacterial FurA is not involved in the regulation of iron homeostasis. However, they provide strong evidence that FurA contributes to intracellular survival as an oxidative stress sensing regulator.

  2. Detection of Mycobacterium avium subspecies paratuberculosis in several herds of Arctic Caribou (Rangifer tarandus ssp.).

    PubMed

    Forde, Taya; Orsel, Karin; De Buck, Jeroen; Côté, Steeve D; Cuyler, Christine; Davison, Tracy; Elkin, Brett; Kelly, Allicia; Kienzler, Martin; Popko, Richard; Taillon, Joëlle; Veitch, Alasdair; Kutz, Susan

    2012-10-01

    Mycobacterium avium subspecies paratuberculosis (MAP) is a common pathogen in domestic ruminants that causes granulomatous inflammation of the small intestine leading to emaciation and wasting. Clinical disease (Johne's disease) is also reported for several wild ruminant species. Between 2007 and 2009 we collected 561 fecal samples from caribou (Rangifer tarandus ssp.) representing 10 herds of migratory caribou, two herds of caribou from Greenland, and three populations of boreal woodland caribou. Feces were tested for MAP by bacterial culture and PCR targeting the IS900 insertion sequence. In total, 31 samples from eight different populations representing all three ecotypes were found positive for MAP by PCR, with one sample from the Rivière-aux-Feuilles herd also being culture positive for the type II (cattle) strain. The proportion of positive animals was particularly high in the Akia-Maniitsoq herd in Greenland, and Rivière-aux-Feuilles and Riviè re-George herds in northeastern Canada (23.4, 11.5, and 10.0%, respectively). Our results indicate that MAP is present in several caribou herds of different ecotypes in northern Canada and Greenland and that MAP circulates within wildlife populations that do not have ongoing contact with domestic livestock. The epidemiology, pathogenicity, and effects on the health of caribou in northern ecosystems remain unknown.

  3. Comparative Transcriptome Analysis Reveals Heat-Responsive Genes in Chinese Cabbage (Brassica rapa ssp. chinensis).

    PubMed

    Wang, Aihua; Hu, Jihong; Huang, Xingxue; Li, Xia; Zhou, Guolin; Yan, Zhixiang

    2016-01-01

    Chinese cabbage (Brassica rapa ssp. chinensis) is an economically and agriculturally significant vegetable crop and is extensively cultivated throughout the world. Heat stress disturbs cellular homeostasis and causes visible growth inhibition of shoots and roots, severe retardation in growth and development, and even death. However, there are few studies on the transcriptome profiling of heat stress in non-heading Chinese cabbage. In this study, we investigated the transcript profiles of non-heading Chinese cabbage from heat-sensitive and heat-tolerant varieties "GHA" and "XK," respectively, in response to high temperature using RNA sequencing (RNA seq). Approximately 625 genes were differentially expressed between the two varieties. The responsive genes can be divided into three phases along with the time of heat treatment: response to stimulus, programmed cell death and ribosome biogenesis. Differentially expressed genes (DEGs) were identified in the two varieties, including transcription factors (TFs), kinases/phosphatases, genes related to photosynthesis and effectors of homeostasis. Many TFs were involved in the heat stress response of Chinese cabbage, including NAC069 TF which was up-regulated at all the heat treatment stages. And their expression levels were also validated by quantitative real-time-PCR (qRT-PCR). These candidate genes will provide genetic resources for further improving the heat-tolerant characteristics in non-heading Chinese cabbage. PMID:27443222

  4. Evaluating physiological stress in Sumatran tigers (Panthera tigris ssp. sumatrae) managed in Australian zoos.

    PubMed

    Parnell, Tempe; Narayan, Edward J; Magrath, Michael J L; Roe, Sheila; Clark, Giles; Nicolson, Vere; Martin-Vegue, Patrick; Mucci, Al; Hero, Jean-Marc

    2014-01-01

    Glucocorticoid quantification using non-invasive methods provides a powerful tool for assessing the health and welfare of wildlife in zoo-based programmes. In this study, we provide baseline data on faecal-based glucocorticoid (cortisol) monitoring of Sumatran tigers (Panthera tigris ssp. sumatrae) managed at the Melbourne Zoo in Victoria, Australia. We sampled five tigers daily for 60 days. Faecal cortisol metabolites (FCMs) in tiger faecal extracts were quantified using enzyme immunoassays that were successfully validated using parallelism and accuracy recovery checks. Two female tigers had significantly higher mean FCM levels than the two males and another female, suggesting that females may have higher FCM levels. A significant elevation was noted in the FCM levels for one female 2 days after she was darted and anaesthetized; however, the FCM levels returned to baseline levels within 3 days after the event. Comparative analysis of FCM levels of tigers sampled at Melbourne Zoo with tigers sampled earlier at two other Australian Zoos (Dreamworld Themepark and Australia Zoo) showed that FCM levels varied between zoos. Differences in the enclosure characteristics, timing of sampling, size and composition of groupings and training procedures could all contribute to this variation. Overall, we recommend the use of non-invasive sampling for the assessment of adrenocortical activity of felids managed in zoos in Australia and internationally in order to improve the welfare of these charismatic big cats.

  5. Evaluating physiological stress in Sumatran tigers (Panthera tigris ssp. sumatrae) managed in Australian zoos

    PubMed Central

    Parnell, Tempe; Narayan, Edward J.; Magrath, Michael J. L.; Roe, Sheila; Clark, Giles; Nicolson, Vere; Martin-Vegue, Patrick; Mucci, Al; Hero, Jean-Marc

    2014-01-01

    Glucocorticoid quantification using non-invasive methods provides a powerful tool for assessing the health and welfare of wildlife in zoo-based programmes. In this study, we provide baseline data on faecal-based glucocorticoid (cortisol) monitoring of Sumatran tigers (Panthera tigris ssp. sumatrae) managed at the Melbourne Zoo in Victoria, Australia. We sampled five tigers daily for 60 days. Faecal cortisol metabolites (FCMs) in tiger faecal extracts were quantified using enzyme immunoassays that were successfully validated using parallelism and accuracy recovery checks. Two female tigers had significantly higher mean FCM levels than the two males and another female, suggesting that females may have higher FCM levels. A significant elevation was noted in the FCM levels for one female 2 days after she was darted and anaesthetized; however, the FCM levels returned to baseline levels within 3 days after the event. Comparative analysis of FCM levels of tigers sampled at Melbourne Zoo with tigers sampled earlier at two other Australian Zoos (Dreamworld Themepark and Australia Zoo) showed that FCM levels varied between zoos. Differences in the enclosure characteristics, timing of sampling, size and composition of groupings and training procedures could all contribute to this variation. Overall, we recommend the use of non-invasive sampling for the assessment of adrenocortical activity of felids managed in zoos in Australia and internationally in order to improve the welfare of these charismatic big cats. PMID:27293659

  6. Effects of amendments on the uptake and distribution of DDT in Cucurbita pepo ssp pepo plants.

    PubMed

    Whitfield Aslund, Melissa L; Lunney, Alissa I; Rutter, Allison; Zeeb, Barbara A

    2010-02-01

    The effects of soil amendments on the phytoextraction of summation operatorDDT (DDT + DDD + DDE) from soil ([ summation operatorDDT] approximately 1500 ng/g) by a pumpkin variety of Cucurbita pepo ssp pepo were tested and the patterns of summation operatorDDT storage throughout the plant shoot were examined. The soil amendments did not increase the total amount of summation operatorDDT extracted into plant shoots, but new information about summation operatorDDT distribution in the plants was obtained. As observed previously, the summation operatorDDT concentration in plant leaves (mean 290 ng/g) was significantly lower than in plant stems (mean 2600 ng/g). Further analysis revealed that summation operatorDDT composition was consistent throughout the plant shoot and that summation operatorDDT concentration in leaves and stems decreased exponentially as distance from the root increased, which was previously unknown. This new information about the patterns of summation operatorDDT uptake and translocation within pumpkin plants highlights the need for appropriate plant sampling strategies in future POPs phytoextraction research.

  7. Chemical composition and nutritional evaluation of the seeds of Acacia tortilis (Forssk.) Hayne ssp. raddiana.

    PubMed

    Embaby, Hassan E; Rayan, Ahmed M

    2016-06-01

    Chemical composition and nutritional evaluation as well as physicochemical and functional properties of seed flour of Acacia tortilis (Forssk.) Hayne ssp. raddiana were studied. The results indicated that seeds contained 5.30% moisture, 3.99% ash, 9.19% fat, 14.31% fiber, 27.21% protein and 45.30% carbohydrates. Potassium was the predominant element followed by calcium and then phosphorous. Phytic acid, tannins and trypsin inhibitor as antinutrients were detected. The amino acid profile compared well with FAO/WHO recommended pattern except for cystine/methionine, isoleucine, tyrosine/phenylalanine, lysine and threonine. Also, the first limiting amino acid was lysine. Fatty acid composition showed that linoleic acid was the major fatty acid, followed by palmitic, stearic, oleic and arachidic acids. The seed oil showed absorbance in the ultraviolet ranges, thus it can be used as a broad spectrum UV protectant. For physicochemical and functional properties, acacia seeds flour had excellent water holding index, swelling index, foaming capacity and foam stability. PMID:26830561

  8. Patterns of diversity and recombination along chromosome 1 of maize (Zea mays ssp. mays L.).

    PubMed Central

    Tenaillon, Maud I; Sawkins, Mark C; Anderson, Lorinda K; Stack, Stephen M; Doebley, John; Gaut, Brandon S

    2002-01-01

    We investigate the interplay between genetic diversity and recombination in maize (Zea mays ssp. mays). Genetic diversity was measured in three types of markers: single-nucleotide polymorphisms, indels, and microsatellites. All three were examined in a sample of previously published DNA sequences from 21 loci on maize chromosome 1. Small indels (1-5 bp) were numerous and far more common than large indels. Furthermore, large indels (>100 bp) were infrequent in the population sample, suggesting they are slightly deleterious. The 21 loci also contained 47 microsatellites, of which 33 were polymorphic. Diversity in SNPs, indels, and microsatellites was compared to two measures of recombination: C (=4Nc) estimated from DNA sequence data and R based on a quantitative recombination nodule map of maize synaptonemal complex 1. SNP diversity was correlated with C (r = 0.65; P = 0.007) but not with R (r = -0.10; P = 0.69). Given the lack of correlation between R and SNP diversity, the correlation between SNP diversity and C may be driven by demography. In contrast to SNP diversity, microsatellite diversity was correlated with R (r = 0.45; P = 0.004) but not C (r = -0.025; P = 0.55). The correlation could arise if recombination is mutagenic for microsatellites, or it may be consistent with background selection that is apparent only in this class of rapidly evolving markers. PMID:12454083

  9. Influence of mowing Artemisia tridentata ssp. wyomingensis on winter habitat for wildlife.

    PubMed

    Davies, Kirk W; Bates, Jonathan D; Johnson, Dustin D; Nafus, Aleta M

    2009-07-01

    Mowing is commonly implemented to Artemisia tridentata ssp. wyomingensis (Beetle & A. Young) S.L. Welsh (Wyoming big sagebrush) plant communities to improve wildlife habitat, increase forage production for livestock, and create fuel breaks for fire suppression. However, information detailing the influence of mowing on winter habitat for wildlife is lacking. This information is crucial because many wildlife species depended on A. tridentata spp. wyomingensis plant communities for winter habitat and consume significant quantities of Artemisia during this time. Furthermore, information is generally limited describing the recovery of A. tridentata spp. wyomingensis to mowing and the impacts of mowing on stand structure. Stand characteristics and Artemisia leaf tissue crude protein (CP), acid detergent fiber (ADF), and neutral detergent fiber (NDF) concentrations were measured in midwinter on 0-, 2-, 4-, and 6-year-old fall-applied mechanical (mowed at 20 cm height) treatments and compared to adjacent untreated (control) areas. Mowing compared to the control decreased Artemisia cover, density, canopy volume, canopy elliptical area, and height (P < 0.05), but all characteristics were recovering (P < 0.05). Mowing A. tridentata spp. wyomingensis plant communities slightly increases the nutritional quality of Artemisia leaves (P < 0.05), but it simultaneously results in up to 20 years of decrease in Artemisia structural characteristics. Because of the large reduction in A. tridentata spp. wyomingensis for potentially 20 years following mowing, mowing should not be applied in Artemisia facultative and obligate wildlife winter habitat. Considering the decline in A. tridentata spp. wyomingensis-dominated landscapes, we caution against mowing these communities.

  10. Short communication: Evaluation of sampling socks for detection of Mycobacterium avium ssp. paratuberculosis on dairy farms.

    PubMed

    Wolf, R; Orsel, K; De Buck, J; Kanevets, U; Barkema, H W

    2016-04-01

    Mycobacterium avium ssp. paratuberculosis (MAP) causes Johne's disease, a production-limiting disease in cattle. Detection of infected herds is often done using environmental samples (ES) of manure, which are collected in cattle pens and manure storage areas. Disadvantages of the method are that sample accuracy is affected by cattle housing and type of manure storage area. Furthermore, some sampling locations (e.g., manure lagoons) are frequently not readily accessible. However, sampling socks (SO), as used for Salmonella spp. testing in chicken flocks, might be an easy to use and accurate alternative to ES. The objective of the study was to assess accuracy of SO for detection of MAP in dairy herds. At each of 102 participating herds, 6 ES and 2 SO were collected. In total, 45 herds had only negative samples in both methods and 29 herds had ≥1 positive ES and ≥1 positive SO. Furthermore, 27 herds with ≥1 positive ES had no positive SO, and 1 herd with no positive ES had 1 positive SO. Bayesian simulation with informative priors on sensitivity of ES and MAP herd prevalence provided a posterior sensitivity for SO of 43.5% (95% probability interval=33-58), and 78.5% (95% probability interval=62-93) for ES. Although SO were easy to use, accuracy was lower than for ES. Therefore, with improvements in the sampling protocol (e.g., more SO per farm and more frequent herd visits), as well as improvements in the laboratory protocol, perhaps SO would be a useful alternative for ES. PMID:26851860

  11. Coding sequence divergence between two closely related plant species: Arabidopsis thaliana and Brassica rapa ssp. pekinensis.

    PubMed

    Tiffin, Peter; Hahn, Matthew W

    2002-06-01

    To characterize the coding-sequence divergence of closely related genomes, we compared DNA sequence divergence between sequences from a Brassica rapa ssp. pekinensis EST library isolated from flower buds and genomic sequences from Arabidopsis thaliana. The specific objectives were (i) to determine the distribution of and relationship between K(a) and K(s), (ii) to identify genes with the lowest and highest K(a): K(s) values, and (iii) to evaluate how codon usage has diverged between two closely related species. We found that the distribution of K(a): K(s) was unimodal, and that substitution rates were more variable at nonsynonymous than synonymous sites, and detected no evidence that K(a) and K(s) were positively correlated. Several genes had K(a): K(s) values equal to or near zero, as expected for genes that have evolved under strong selective constraint. In contrast, there were no genes with K(a): K(s) >1 and thus we found no strong evidence that any of the 218 sequences we analyzed have evolved in response to positive selection. We detected a stronger codon bias but a lower frequency of GC at synonymous sites in A. thaliana than B. rapa. Moreover, there has been a shift in the profile of most commonly used synonymous codons since these two species diverged from one another. This shift in codon usage may have been caused by stronger selection acting on codon usage or by a shift in the direction of mutational bias in the B. rapa phylogenetic lineage.

  12. Influence of mowing Artemisia tridentata ssp. wyomingensis on winter habitat for wildlife.

    PubMed

    Davies, Kirk W; Bates, Jonathan D; Johnson, Dustin D; Nafus, Aleta M

    2009-07-01

    Mowing is commonly implemented to Artemisia tridentata ssp. wyomingensis (Beetle & A. Young) S.L. Welsh (Wyoming big sagebrush) plant communities to improve wildlife habitat, increase forage production for livestock, and create fuel breaks for fire suppression. However, information detailing the influence of mowing on winter habitat for wildlife is lacking. This information is crucial because many wildlife species depended on A. tridentata spp. wyomingensis plant communities for winter habitat and consume significant quantities of Artemisia during this time. Furthermore, information is generally limited describing the recovery of A. tridentata spp. wyomingensis to mowing and the impacts of mowing on stand structure. Stand characteristics and Artemisia leaf tissue crude protein (CP), acid detergent fiber (ADF), and neutral detergent fiber (NDF) concentrations were measured in midwinter on 0-, 2-, 4-, and 6-year-old fall-applied mechanical (mowed at 20 cm height) treatments and compared to adjacent untreated (control) areas. Mowing compared to the control decreased Artemisia cover, density, canopy volume, canopy elliptical area, and height (P < 0.05), but all characteristics were recovering (P < 0.05). Mowing A. tridentata spp. wyomingensis plant communities slightly increases the nutritional quality of Artemisia leaves (P < 0.05), but it simultaneously results in up to 20 years of decrease in Artemisia structural characteristics. Because of the large reduction in A. tridentata spp. wyomingensis for potentially 20 years following mowing, mowing should not be applied in Artemisia facultative and obligate wildlife winter habitat. Considering the decline in A. tridentata spp. wyomingensis-dominated landscapes, we caution against mowing these communities. PMID:19159967

  13. [Effects of ruta SSP on the activity of the smooth gastrointestinal muscle isolated of rat].

    PubMed

    Grigorjev, Carlota A; Brizuela, Nilda Y

    2010-01-01

    Ruta graveolens L. and Ruta chalepensis L. are plants used in folkloric medicine as antispasmodics, digestive and for intestinal gases. Animals used as experimental model were rats of the Wistar line, adult females, clinically healthy and with a weight average of 250 grams. We used strips of stomach and duodenum. Each one of the segments mounted on two stirrups in a bath of organ isolated with Ringer-lactate solution, at 37° C , pH: 7.3-7.4, and bubbled with 95% O2, 5% CO2. One of the stirrups was connected vertically to the bottom of the bath and the other to a transducer of tension connected to a Beckman polygraph. We applied 500 mgs of basal tension. After the stabilization, the ethanolic extract of Ruta ssp was added in increasing doses. At 50μl/ml the tone lower 23% in small intestine and lower 27% in stomach. However at 100 μl/ml the tone lower 32% and 35% respectively. In the other parameters the amplitude decrease 50% in the stomach at dose of 5 μl/ml while in the small intestine the amplitude lower 60%. With 10 μl/ml the amplitude change in both organs ( 96% in small intestine, and 75% in stomach). The frequency changes in both organs ( 32% in small intestine, and 50% in stomach) at 10 μl/ml Rue showed decreased effects on isolated small intestine and stomach were is dose dependent, maybe we were demonstrated the effects digestive of Ruta. PMID:21450144

  14. Ontogenetic differences in mesophyll structure and chlorophyll distribution in Eucalyptus globulus ssp. globulus.

    PubMed

    James, S A; Smith, W K; Vogelmann, T C

    1999-02-01

    Mesophyll structure has been associated with the photosynthetic performance of leaves via the regulation of internal light and CO(2) profiles. Differences in mesophyll structure and chlorophyll distribution within three ontogenetically different leaf types of Eucalyptus globulus ssp. globulus were investigated. Juvenile leaves are blue-grey in color, dorsiventral (adaxial palisade layer only), hypostomatous, and approximately horizontal in orientation. In contrast, adult leaves are dark green in color, isobilateral (adaxial and abaxial palisade), amphistomatous, and nearly vertical in orientation. The transitional leaf type has structural features that appear intermediate between the juvenile and adult leaves. The ratio of mesophyll cell surface area per unit leaf surface area (A(mes)/A) of juvenile leaves was maximum at the base of a single, adaxial palisade layer and declined through the spongy mesophyll. Chlorophyll a + b content showed a coincident pattern, while the chlorophyll a:b ratio declined linearly from the adaxial to abaxial epidermis. In comparison, the mesophyll of adult leaves had a bimodal distribution of A(mes)/A, with maxima occurring beneath both the adaxial and abaxial surfaces within the first layer of multiple palisade layers. The distribution of chlorophyll a + b content had a similar pattern, although the maximum ratio of chlorophyll a:b occurred immediately beneath the adaxial and abaxial epidermis. The matching distributions of A(mes)/A and chlorophyll provide further evidence that mesophyll structure may act to influence photosynthetic performance. These changes in internal leaf structure at different life stages of E. globulus may be an adaptation for increased xeromorphy under increasing light exposure experienced from the seedling to adult tree, similar to the characteristics reported for different species according to sunlight exposure and water availability within their native habitats.

  15. Influence of Mowing Artemisia tridentata ssp. wyomingensis on Winter Habitat for Wildlife

    NASA Astrophysics Data System (ADS)

    Davies, Kirk W.; Bates, Jonathan D.; Johnson, Dustin D.; Nafus, Aleta M.

    2009-07-01

    Mowing is commonly implemented to Artemisia tridentata ssp. wyomingensis (Beetle & A. Young) S.L. Welsh (Wyoming big sagebrush) plant communities to improve wildlife habitat, increase forage production for livestock, and create fuel breaks for fire suppression. However, information detailing the influence of mowing on winter habitat for wildlife is lacking. This information is crucial because many wildlife species depended on A. tridentata spp. wyomingensis plant communities for winter habitat and consume significant quantities of Artemisia during this time . Furthermore, information is generally limited describing the recovery of A. tridentata spp. wyomingensis to mowing and the impacts of mowing on stand structure. Stand characteristics and Artemisia leaf tissue crude protein (CP), acid detergent fiber (ADF), and neutral detergent fiber (NDF) concentrations were measured in midwinter on 0-, 2-, 4-, and 6-year-old fall-applied mechanical (mowed at 20 cm height) treatments and compared to adjacent untreated (control) areas. Mowing compared to the control decreased Artemisia cover, density, canopy volume, canopy elliptical area, and height ( P < 0.05), but all characteristics were recovering ( P < 0.05). Mowing A. tridentata spp. wyomingensis plant communities slightly increases the nutritional quality of Artemisia leaves ( P < 0.05), but it simultaneously results in up to 20 years of decrease in Artemisia structural characteristics. Because of the large reduction in A. tridentata spp. wyomingensis for potentially 20 years following mowing, mowing should not be applied in Artemisia facultative and obligate wildlife winter habitat. Considering the decline in A. tridentata spp. wyomingensis-dominated landscapes, we caution against mowing these communities.

  16. [Effects of ruta SSP on the activity of the smooth gastrointestinal muscle isolated of rat].

    PubMed

    Grigorjev, Carlota A; Brizuela, Nilda Y

    2010-01-01

    Ruta graveolens L. and Ruta chalepensis L. are plants used in folkloric medicine as antispasmodics, digestive and for intestinal gases. Animals used as experimental model were rats of the Wistar line, adult females, clinically healthy and with a weight average of 250 grams. We used strips of stomach and duodenum. Each one of the segments mounted on two stirrups in a bath of organ isolated with Ringer-lactate solution, at 37° C , pH: 7.3-7.4, and bubbled with 95% O2, 5% CO2. One of the stirrups was connected vertically to the bottom of the bath and the other to a transducer of tension connected to a Beckman polygraph. We applied 500 mgs of basal tension. After the stabilization, the ethanolic extract of Ruta ssp was added in increasing doses. At 50μl/ml the tone lower 23% in small intestine and lower 27% in stomach. However at 100 μl/ml the tone lower 32% and 35% respectively. In the other parameters the amplitude decrease 50% in the stomach at dose of 5 μl/ml while in the small intestine the amplitude lower 60%. With 10 μl/ml the amplitude change in both organs ( 96% in small intestine, and 75% in stomach). The frequency changes in both organs ( 32% in small intestine, and 50% in stomach) at 10 μl/ml Rue showed decreased effects on isolated small intestine and stomach were is dose dependent, maybe we were demonstrated the effects digestive of Ruta.

  17. Flowering time in wild beet ( Beta vulgaris ssp. maritima) along a latitudinal cline

    NASA Astrophysics Data System (ADS)

    Dijk, Henk Van; Boudry, Pierre; McCombre, Helen; Vernet, Philippe

    The wild beet ( Beta vulgaris ssp. maritima, a perennial species from the Mediterranean and the European Atlantic coasts) shows marked variation in flowering time in terms of both the year of first flowering and flowering date in a given year. Much of this variability is related to latitude. Beta vulgaris plants flower either in the same year as they germinate or in their second year. This is mainly due to differences in their requirement for vernalization, which is determined by a single gene B/b and by quantitative trait loci. The more southern the origin of the plants, the less vernalization is required. Also the B allele, which cancels vernalization requirement completely, has a high frequency in the Mediterranean region, but is completely absent in the northern part of the distribution of this species. We found that flowering date variation in relation to the latitude of origin is maintained under greenhouse conditions but does not follow a simple clinal relationship. From the Mediterranean northwards to the west coast of Brittany, flowering occurs progressively earlier, but from Brittany northwards to south-east England and The Netherlands it is progressively later. A possible explanation for this difference is that in the southern part of the range sensitivity to daylength and warmth control flowering time, whereas further north vernalization requirement is also a key factor. A substantial part of all differences in flowering time was heritable: heritability within populations was measured as 0.33 under greenhouse conditions. The high heritability implies evolutionary change may occur in this character.

  18. Systemic and mucosal immune reactivity upon Mycobacterium avium ssp. paratuberculosis infection in mice.

    PubMed

    Koc, Arzu; Bargen, Imke; Suwandi, Abdulhadi; Roderfeld, Martin; Tschuschner, Annette; Rath, Timo; Gerlach, Gerald F; Hornef, Mathias; Goethe, Ralph; Weiss, Siegfried; Roeb, Elke

    2014-01-01

    Mycobacterium avium ssp. paratuberculosis (MAP) is the cause of Johne's disease, an inflammatory bowel disorder of ruminants. Due to the similar pathology, MAP was also suggested to cause Crohn's disease (CD). Despite of intensive research, this question is still not settled, possibly due to the lack of versatile mouse models. The aim of this study was to identify basic immunologic mechanisms in response to MAP infection. Immune compromised C57BL/6 Rag2-/- mice were infected with MAP intraperitoneally. Such chronically infected mice were then reconstituted with CD4+ and CD8+ T cells 28 days after infection. A systemic inflammatory response, detected as enlargement of the spleen and granuloma formation in the liver, was observed in mice infected and reconstituted with CD4+ T cells. Whereby inflammation in infected and CD4+CD45RB(hi) T cell reconstituted animals was always higher than in the other groups. Reconstitution of infected animals with CD8+ T cells did not result in any inflammatory signs. Interestingly, various markers of inflammation were strongly up-regulated in the colon of infected mice reconstituted with CD4+CD45RB(lo/int) T cells. We propose, the usual non-colitogenic CD4+CD45RB(lo/int) T cells were converted into inflammatory T cells by the interaction with MAP. However, the power of such cells might be not sufficient for a fully established inflammatory response in the colon. Nevertheless, our model system appears to mirror aspects of an inflammatory bowel disease (IBD) like CD and Johne's diseases. Thus, it will provide an experimental platform on which further knowledge on IBD and the involvement of MAP in the induction of CD could be acquired.

  19. The development-specific protein (Ssp1) from Sclerotinia sclerotiorum is encoded by a novel gene expressed exclusively in sclerotium tissues.

    PubMed

    Li, Moyi; Rollins, Jeffrey A

    2009-01-01

    The gene encoding a development-specific protein (Ssp1) was identified; it previously was described as the major protein present in mature sclerotia of Sclerotinia sclerotiorum. To determine the developmental specificity of ssp1 gene expression in relation to protein accumulation we examined transcript and protein accumulation during various growth and development stages of the lifecycle. We found that ssp1 transcript accumulated exclusively within developing sclerotium tissue and not in any other examined stage of growth or development. In contrast high levels of Sspl protein were detectable by western blot and tandem mass spectrometry analyses in all stages of sclerotium as well as apothecium development. Immunolocalization further indicated that Ssp1 protein bodies were depleted from the sclerotium tissue surrounding the site of apothecium germination, but by this method Sspl was not detected in the apothecium. Together these findings suggest that Sspl is not metabolized during carpogenic germination, instead it is translocated from the sclerotium to the apothecium in an antigenically novel form. Outside the Sclerotiniaceae ssp1 homologs were found only from the sclerotium-forming Aspergillus species A. flavus and A. oryzae. Further studies concerning the regulation and function of this gene and its occurrence in other species have the potential to inform our understanding of sclerotium development and the evolution of sclerotia and other forms of fungal stroma. PMID:19271669

  20. Bifidobacterium animalis ssp. lactis CNCM-I2494 Restores Gut Barrier Permeability in Chronically Low-Grade Inflamed Mice

    PubMed Central

    Martín, Rebeca; Laval, Laure; Chain, Florian; Miquel, Sylvie; Natividad, Jane; Cherbuy, Claire; Sokol, Harry; Verdu, Elena F.; van Hylckama Vlieg, Johan; Bermudez-Humaran, Luis G.; Smokvina, Tamara; Langella, Philippe

    2016-01-01

    Growing evidence supports the efficacy of many probiotic strains in the management of gastrointestinal disorders associated with deregulated intestinal barrier function and/or structure. In particular, bifidobacteria have been studied for their efficacy to both prevent and treat a broad spectrum of animal and/or human gut disorders. The aim of the current work was thus to evaluate effects on intestinal barrier function of Bifidobacterium animalis ssp. lactis CNCM-I2494, a strain used in fermented dairy products. A chronic dinitrobenzene sulfonic acid (DNBS)-induced low-grade inflammation model causing gut dysfunction in mice was used in order to study markers of inflammation, intestinal permeability, and immune function in the presence of the bacterial strain. In this chronic low-grade inflammation mice model several parameters pointed out the absence of an over active inflammation process. However, gut permeability, lymphocyte populations, and colonic cytokines were found to be altered. B. animalis ssp. lactis CNCM-I2494 was able to protect barrier functions by restoring intestinal permeability, colonic goblet cell populations, and cytokine levels. Furthermore, tight junction (TJ) proteins levels were also measured by qRT-PCR showing the ability of this strain to specifically normalize the level of several TJ proteins, in particular for claudin-4. Finally, B. lactis strain counterbalanced CD4+ lymphocyte alterations in both spleen and mesenteric lymphoid nodes. It restores the Th1/Th2 ratio altered by the DNBS challenge (which locally augments CD4+ Th1 cells) by increasing the Th2 response as measured by the increase in the production of major representative Th2 cytokines (IL-4, IL-5, and IL-10). Altogether, these data suggest that B. animalis ssp. lactis CNCM-I2494 may efficiently prevent disorders associated with increased barrier permeability. PMID:27199937

  1. BrpSPL9 (Brassica rapa ssp. pekinensis SPL9) controls the earliness of heading time in Chinese cabbage.

    PubMed

    Wang, Yali; Wu, Feijie; Bai, Jinjuan; He, Yuke

    2014-04-01

    The leafy heads of cabbage (Brassica oleracea), Chinese cabbage (Brassica rapa ssp. pekinensis), Brussels sprouts (B. oleracea ssp. gemmifera) and lettuce (Lactuca sativa) comprise extremely incurved leaves that are edible vegetable products. The heading time is important for high quality and yield of these crops. Here, we report that BrpSPL9-2 (B. rapa ssp. pekinensis SQUAMOSA PROMOTER BINDING-LIKE 9-2), a target gene of microRNA brp-miR156, controls the heading time of Chinese cabbage. Quantitative measurements of leaf shapes, sizes, colour and curvature indicated that heading is a late adult phase of vegetative growth. During the vegetative period, miR156 levels gradually decreased from the seedling stage to the heading one, whereas BrpSPL9-2 and BrpSPL15-1 mRNAs increased progressively and reached the highest levels at the heading stage. Overexpression of a mutated miR156-resistant form of BrpSPL9-2 caused the significant earliness of heading, concurrent with shortening of the seedling and rosette stages. By contrast, overexpression of miR156 delayed the folding time, concomitant with prolongation of the seedling and rosette stages. Morphological analysis reveals that the significant earliness of heading in the transgenic plants overexpressing BrpSPL9-2 gene was produced because the juvenile phase was absent and the early adult phase shortened, whereas the significant delay of folding in the transgenic plants overexpressing Brp-MIR156a was due to prolongation of the juvenile and early adult phases. Thus, miR156 and BrpSPL9 genes are potentially important for genetic improvement of earliness of Chinese cabbage and other crops.

  2. Bifidobacterium animalis ssp. lactis CNCM-I2494 Restores Gut Barrier Permeability in Chronically Low-Grade Inflamed Mice.

    PubMed

    Martín, Rebeca; Laval, Laure; Chain, Florian; Miquel, Sylvie; Natividad, Jane; Cherbuy, Claire; Sokol, Harry; Verdu, Elena F; van Hylckama Vlieg, Johan; Bermudez-Humaran, Luis G; Smokvina, Tamara; Langella, Philippe

    2016-01-01

    Growing evidence supports the efficacy of many probiotic strains in the management of gastrointestinal disorders associated with deregulated intestinal barrier function and/or structure. In particular, bifidobacteria have been studied for their efficacy to both prevent and treat a broad spectrum of animal and/or human gut disorders. The aim of the current work was thus to evaluate effects on intestinal barrier function of Bifidobacterium animalis ssp. lactis CNCM-I2494, a strain used in fermented dairy products. A chronic dinitrobenzene sulfonic acid (DNBS)-induced low-grade inflammation model causing gut dysfunction in mice was used in order to study markers of inflammation, intestinal permeability, and immune function in the presence of the bacterial strain. In this chronic low-grade inflammation mice model several parameters pointed out the absence of an over active inflammation process. However, gut permeability, lymphocyte populations, and colonic cytokines were found to be altered. B. animalis ssp. lactis CNCM-I2494 was able to protect barrier functions by restoring intestinal permeability, colonic goblet cell populations, and cytokine levels. Furthermore, tight junction (TJ) proteins levels were also measured by qRT-PCR showing the ability of this strain to specifically normalize the level of several TJ proteins, in particular for claudin-4. Finally, B. lactis strain counterbalanced CD4(+) lymphocyte alterations in both spleen and mesenteric lymphoid nodes. It restores the Th1/Th2 ratio altered by the DNBS challenge (which locally augments CD4(+) Th1 cells) by increasing the Th2 response as measured by the increase in the production of major representative Th2 cytokines (IL-4, IL-5, and IL-10). Altogether, these data suggest that B. animalis ssp. lactis CNCM-I2494 may efficiently prevent disorders associated with increased barrier permeability. PMID:27199937

  3. A study of System Interface Sets (SIS) for the host, target and integration environments of the Space Station Program (SSP)

    NASA Technical Reports Server (NTRS)

    Mckay, Charles; Auty, David; Rogers, Kathy

    1987-01-01

    System interface sets (SIS) for large, complex, non-stop, distributed systems are examined. The SIS of the Space Station Program (SSP) was selected as the focus of this study because an appropriate virtual interface specification of the SIS is believed to have the most potential to free the project from four life cycle tyrannies which are rooted in a dependance on either a proprietary or particular instance of: operating systems, data management systems, communications systems, and instruction set architectures. The static perspective of the common Ada programming support environment interface set (CAIS) and the portable common execution environment (PCEE) activities are discussed. Also, the dynamic perspective of the PCEE is addressed.

  4. Refractory Toxic Shock-Like Syndrome from Streptococcus dysgalactiae ssp. equisimilis and Intravenous Immunoglobulin as Salvage Therapy: A Case Series.

    PubMed

    Islam, Marjan; Karter, Dennis; Altshuler, Jerry; Altshuler, Diana; Schwartz, David; Torregrossa, Gianluca

    2016-01-01

    Infections from Streptococcus dysgalactiae ssp. equisimilis (SDSE) can cause a wide variety of infections, ranging from mild cellulitis to invasive disease, such as endocarditis and streptococcal toxic shock-like syndrome (TSLS). Despite prompt and appropriate antibiotics, mortality rates associated with shock have remained exceedingly high, prompting the need for adjunctive therapy. IVIG has been proposed as a possible adjunct, given its ability to neutralize a wide variety of superantigens and modulate a dysregulated inflammatory response. We present the first reported cases of successful IVIG therapy for reversing shock in the treatment of SDSE TSLS. PMID:27597908

  5. Refractory Toxic Shock-Like Syndrome from Streptococcus dysgalactiae ssp. equisimilis and Intravenous Immunoglobulin as Salvage Therapy: A Case Series

    PubMed Central

    Karter, Dennis; Altshuler, Jerry; Altshuler, Diana; Schwartz, David; Torregrossa, Gianluca

    2016-01-01

    Infections from Streptococcus dysgalactiae ssp. equisimilis (SDSE) can cause a wide variety of infections, ranging from mild cellulitis to invasive disease, such as endocarditis and streptococcal toxic shock-like syndrome (TSLS). Despite prompt and appropriate antibiotics, mortality rates associated with shock have remained exceedingly high, prompting the need for adjunctive therapy. IVIG has been proposed as a possible adjunct, given its ability to neutralize a wide variety of superantigens and modulate a dysregulated inflammatory response. We present the first reported cases of successful IVIG therapy for reversing shock in the treatment of SDSE TSLS. PMID:27597908

  6. Effector MiSSP7 of the mutualistic fungus Laccaria bicolor stabilizes the Populus JAZ6 protein and represses jasmonic acid (JA) responsive genes.

    PubMed

    Plett, Jonathan M; Daguerre, Yohann; Wittulsky, Sebastian; Vayssières, Alice; Deveau, Aurelie; Melton, Sarah J; Kohler, Annegret; Morrell-Falvey, Jennifer L; Brun, Annick; Veneault-Fourrey, Claire; Martin, Francis

    2014-06-01

    Ectomycorrhizal fungi, such as Laccaria bicolor, support forest growth and sustainability by providing growth-limiting nutrients to their plant host through a mutualistic symbiotic relationship with host roots. We have previously shown that the effector protein MiSSP7 (Mycorrhiza-induced Small Secreted Protein 7) encoded by L. bicolor is necessary for the establishment of symbiosis with host trees, although the mechanistic reasoning behind this role was unknown. We demonstrate here that MiSSP7 interacts with the host protein PtJAZ6, a negative regulator of jasmonic acid (JA)-induced gene regulation in Populus. As with other characterized JASMONATE ZIM-DOMAIN (JAZ) proteins, PtJAZ6 interacts with PtCOI1 in the presence of the JA mimic coronatine, and PtJAZ6 is degraded in plant tissues after JA treatment. The association between MiSSP7 and PtJAZ6 is able to protect PtJAZ6 from this JA-induced degradation. Furthermore, MiSSP7 is able to block--or mitigate--the impact of JA on L. bicolor colonization of host roots. We show that the loss of MiSSP7 production by L. bicolor can be complemented by transgenically varying the transcription of PtJAZ6 or through inhibition of JA-induced gene regulation. We conclude that L. bicolor, in contrast to arbuscular mycorrhizal fungi and biotrophic pathogens, promotes mutualism by blocking JA action through the interaction of MiSSP7 with PtJAZ6. PMID:24847068

  7. Effector MiSSP7 of the mutualistic fungus Laccaria bicolor stabilizes the Populus JAZ6 protein and represses jasmonic acid (JA) responsive genes

    PubMed Central

    Plett, Jonathan M.; Daguerre, Yohann; Wittulsky, Sebastian; Vayssières, Alice; Deveau, Aurelie; Melton, Sarah J.; Kohler, Annegret; Morrell-Falvey, Jennifer L.; Brun, Annick; Veneault-Fourrey, Claire; Martin, Francis

    2014-01-01

    Ectomycorrhizal fungi, such as Laccaria bicolor, support forest growth and sustainability by providing growth-limiting nutrients to their plant host through a mutualistic symbiotic relationship with host roots. We have previously shown that the effector protein MiSSP7 (Mycorrhiza-induced Small Secreted Protein 7) encoded by L. bicolor is necessary for the establishment of symbiosis with host trees, although the mechanistic reasoning behind this role was unknown. We demonstrate here that MiSSP7 interacts with the host protein PtJAZ6, a negative regulator of jasmonic acid (JA)-induced gene regulation in Populus. As with other characterized JASMONATE ZIM-DOMAIN (JAZ) proteins, PtJAZ6 interacts with PtCOI1 in the presence of the JA mimic coronatine, and PtJAZ6 is degraded in plant tissues after JA treatment. The association between MiSSP7 and PtJAZ6 is able to protect PtJAZ6 from this JA-induced degradation. Furthermore, MiSSP7 is able to block—or mitigate—the impact of JA on L. bicolor colonization of host roots. We show that the loss of MiSSP7 production by L. bicolor can be complemented by transgenically varying the transcription of PtJAZ6 or through inhibition of JA-induced gene regulation. We conclude that L. bicolor, in contrast to arbuscular mycorrhizal fungi and biotrophic pathogens, promotes mutualism by blocking JA action through the interaction of MiSSP7 with PtJAZ6. PMID:24847068

  8. Subsurface irrigation of potato crop (Solanum tuberosum ssp. Andigena) in Suka Kollus with different drainage systems

    NASA Astrophysics Data System (ADS)

    Serrano-Coronel, Genaro; Chipana-Rivera, René; Fátima Moreno-Pérez, María; Roldán-Cañas, José

    2016-04-01

    Among the most important hydraulic structures of pre-Hispanic ancestral technology developed in the Andean region, we find the suka kollus, aymara word, called also waru waru, en quechua or raised fields, in English. They are raised platforms surrounded by water canals that irrigate subsurface, but also have the function of draining, to deal with floods because they are surrounding Lake Titicaca. They also have the property of generating a thermoregulatory effect to crops, depending on the configuration of the channels and platforms. Such agro-ecosystems are being abandoned, however, if properly addressed crop management and some drainage canals are replaced by underground drains for increased crop area could be very useful in enabling marginal soils affected by salts and / or excess water. For these reasons, the objective of this study was to evaluate the subsurface irrigation in the potato crop in suka kollus under a system of surface drainage, and mixed drainage (surface and subsurface). The study was conducted in marginal soils of Kallutaca area, located 30 km from the city of La Paz, Bolivia, at a height of 3892 m.a.s.l. The cultivation of the potato (Solanum tuberosum ssp. Andigena) was used. Four treatments were tested with different widths of the platforms: T1 (Control) with drainage through channels; T2 (replacing a channel by a drain); T3 (replacing two channels by two drains); T4 (replacing three channels by three drains). The flow of water into the soil from the water table was predominantly upward, except during periods of high rainfall. In terms of treatments, the flow in T1 was higher, mainly at weeks 8 to 11 after seedling emergence, coinciding with the phenological phases of flowering and at the beginning of the tuber ripening. It was followed by T3, T2 and T4 treatments, respectively. Tuber yield, if one considers that the channels detract arable land, was higher in the T3 treatment,16.4 Mg / ha, followed by T2 treatment, 15.2 Mg / ha, T1

  9. BcMF13, a new reproductive organ-specific gene from Brassica rapa. ssp. chinensis, affects pollen development.

    PubMed

    Li, Yanyan; Cao, Jiashu; Huang, Li; Yu, Xiaolin; Xiang, Xun

    2008-06-01

    A transcript-derived fragment (GenBank accession number DN237920.1) accumulated in the wild-type flower buds of Chinese cabbage (Brassica campestris L. ssp. chinensis Makino, syn. B. rapa ssp. chinensis) was isolated and further investigated. The full length DNA and cDNA of the fragment were cloned by rapid amplification of cDNA ends. The gene, BcMF13, encodes a protein of 73 amino acids and is interrupted by an intron of 106 bp (GenBank accession number EF158459). Southern blot analysis revealed that BcMF13 could be a single-copy gene in the Chinese cabbage genome. Sequence blast analysis showed that BcMF13 was a new gene. In EST database, those sequences share 96-98% identity with BcMF13 cDNA all came from flower buds, microspores, anthers of Brassica, which proved BcMF13 homologs closely related to the development of male gametogenesis in Brassica. RT-PCR discovered that it is exclusively expressed in stage four and five flower buds of fertile line, strongly expressed in stamens. Successful suppression of BcMF13 gene expression by RNA antisense strategy greatly reduced the normal pollen grains, suggesting that BcMF13 was essential in pollen development in Brassica.

  10. Characterization of a putative pollen-specific arabinogalactan protein gene, BcMF8, from Brassica campestris ssp. chinensis.

    PubMed

    Huang, Li; Cao, Jia-Shu; Zhang, Ai-Hong; Ye, Yi-Qun

    2008-12-01

    The BcMF8 (Brassica campestris male fertility 8) gene, possessing the features of 'classical' arabinogalactan protein (AGP) was isolated from Brassica campestris L. ssp. chinensis, Makino syn. B. rapa L. ssp. chinensis. This gene was highly abundant in the fertile flower buds but silenced in the sterile ones of genic male sterile A/B line ('ZUBajh97-01A/B') in B. campestris. Expression patterns analysis suggested BcMF8 was a pollen-specific gene, whose transcript started to be expressed at the uninucleate stage and maintained throughout to the pollen at pollination stage. BcMF8 is highly homologous to the known pollen-specific AGP genes Sta 39-4 and Sta 39-3 from B. napus. Isolation and multiple alignment of the homologs of BcMF8 gene in the family Cruciferae indicated that BcMF8 was highly conserved in this family, which reflect the conservation in biological function and importance of this putative AGP gene in plant development. Similarity analysis also demonstrated Sta 39-4 and Sta 39-3 may originate from different genomes.

  11. Functional analysis of a novel male fertility CYP86MF gene in Chinese cabbage (Brassica campestris L. ssp. chinensis makino).

    PubMed

    Cao, J S; Yu, X L; Ye, W Z; Lu, G; Xiang, X

    2006-01-01

    In our earlier work, a cytochrome P450 CYP86MF gene was isolated from floral bud of Chinese cabbage (Brassica campestris L. ssp. chinensis Makino, syn. B. rapa L.) by mRNA differential display PCR (DD-PCR) and rapid amplification of cDNA ends (RACE). To unravel the biological function of CYP86MF gene, the antisense fragment from the CYP86MF gene was transferred into Chinese cabbage pak-choi (B. campestris ssp. chinensis var. communis Tsen et Lee). Out of 22 plants transformed with the antisense gene constructed from the CYP86MF, 20 reached to flowering stage. Morphological investigations showed that the transgenic plants developed the normal floral organ. However, they remained self-infertile, even when artificial self-pollination was performed in the bud stage. Pollen germination test indicated that the pollen from the transgenic line TB-2 could not germinate normally. Further physiological, biochemical and cytological analyses showed that only significant difference was detectable in contents of the endogenous hormones, and a layer of unknown material adhered to the surface of microspore. The present studies thus provided valuable clues for understanding the biological function of the CYP86C subfamily genes. Furthermore, our studies also demonstrate a novel method for obtaining artificial male sterility line of Chinese cabbage.

  12. Essential oil composition and antifungal activity of aerial parts of Ballota nigra ssp foetida collected at flowering and fruiting times.

    PubMed

    Fraternale, Daniele; Ricci, Donata

    2014-07-01

    The present study reports the results of gas chromatographic-mass spectrometric (GC/MS) analyses of the essential oils from the aerial parts of Ballota nigra L. ssp foetida (Lamiaceae) collected at flowering and fruiting times, as well as their in vitro antifungal activity against nine plant pathogenic fungi. Moreover, the essential oils were evaluated for their antifungal activity using the agar dilution method, and also MICs (minimum inhibitory concentrations) and MFCs (minimum fungicidal concentrations) were determined. The major compounds identified in the flowering and fruiting aerial parts oils respectively were beta-caryophyllene (22.6% and 21.8%), caryophyllene oxide (18.0% and 20.5%) and germacrene-D (16.5 and 13.1%). The oils showed in vitro antifungal activity against some species of Fusarium, Botrytis cinerea, and Alternaria solani. Our study indicates that the oil of B. nigra ssp foetida could be used as a control agent for plant pathogenic fungi in natural formulations. PMID:25230517

  13. Effect of organic matter additions on uptake of weathered DDT by Cucurbita pepo ssp. pepo cv. Howden.

    PubMed

    Lunney, Alissa I; Rutter, Allison; Zeeb, Barbara A

    2010-01-01

    Greenhouse studies were conducted to assess the impact of organic matter additions on plant uptake of DDT [2,2-bis(chlorophenyl)-1,1,1-trichloroethane] from weathered soil. Cucurbita pepo ssp. pepo cv. Howden pumpkins were grown in 100 g of DDT contaminated soil ([DDT] - 1100 ng/g) mixed with equal volumes of either clean soil, perlite, vermiculite, peat, potting soil, or granular activated carbon (GAC) to give total organic carbon contents of 2.4%, 2.5%, 2.6%, 11.5%, 12.2%, and 27.3%, respectively. As in other studies, root DDT concentrations were significantly lower in soils with high organic matter. Root bioaccumulation factors (BAF = [DDT]root/[DDT]soil) approximated this trend. Root concentrations correlated with organic matter concentrations and not with soil DDT concentrations. Conversely, shoot DDT concentrations, shoot BAFs and translocation factors (TLF = BAF(shoot)/BAF(root)) were not significantly different between treatment groups, except for plants grown in GAC/DDT soil. This suggests that amendments with a range of organic matter contents may be added to improve soil conditions at industrial sites without significant adverse effects on phytoextraction potential of C. pepo ssp. pepo. PMID:20734916

  14. A SNP and SSR based genetic map of asparagus bean (Vigna. unguiculata ssp. sesquipedialis) and comparison with the broader species.

    PubMed

    Xu, Pei; Wu, Xiaohua; Wang, Baogen; Liu, Yonghua; Ehlers, Jeffery D; Close, Timothy J; Roberts, Philip A; Diop, Ndeye-Ndack; Qin, Dehui; Hu, Tingting; Lu, Zhongfu; Li, Guojing

    2011-01-06

    Asparagus bean (Vigna. unguiculata ssp. sesquipedialis) is a distinctive subspecies of cowpea [Vigna. unguiculata (L.) Walp.] that apparently originated in East Asia and is characterized by extremely long and thin pods and an aggressive climbing growth habit. The crop is widely cultivated throughout Asia for the production of immature pods known as 'long beans' or 'asparagus beans'. While the genome of cowpea ssp. unguiculata has been characterized recently by high-density genetic mapping and partial sequencing, little is known about the genome of asparagus bean. We report here the first genetic map of asparagus bean based on SNP and SSR markers. The current map consists of 375 loci mapped onto 11 linkage groups (LGs), with 191 loci detected by SNP markers and 184 loci by SSR markers. The overall map length is 745 cM, with an average marker distance of 1.98 cM. There are four high marker-density blocks distributed on three LGs and three regions of segregation distortion (SDRs) identified on two other LGs, two of which co-locate in chromosomal regions syntenic to SDRs in soybean. Synteny between asparagus bean and the model legume Lotus. japonica was also established. This work provides the basis for mapping and functional analysis of genes/QTLs of particular interest in asparagus bean, as well as for comparative genomics study of cowpea at the subspecies level.

  15. Complement sensitivity and factor H binding of European Francisella tularensis ssp. holarctica strains in selected animal species.

    PubMed

    Kreizinger, Zsuzsa; Bhide, Mangesh; Bencurova, Elena; Dolinska, Saskia; Gyuranecz, Miklós

    2015-09-01

    Francisella tularensis is a Gram-negative bacterium, the causative agent of the zoonotic disease tularaemia. The bacterium has developed several extracellular and intracellular strategies to evade the hosts' innate and adaptive immune responses. The aims of the study were to examine complement sensitivity of wild and attenuated F. tularensis ssp. holarctica strains in animal hosts of distinct sensitivity to the bacterium, to compare the complement-evading ability of wild strains of different phylogeographic background, and to examine the role of factor H in the host-pathogen interactions. Complement sensitivity assays were carried out on various F. tularensis ssp. holarctica wild strains and on the attenuated live vaccine strain (LVS) with sera of the highly sensitive house mouse (Mus musculus), the moderately sensitive European brown hare (Lepus europaeus) and the relatively resistant cattle (Bos taurus). Specific binding of complement regulator factor H to bacterial membrane proteins was examined by Western blot assays. All wild strains interacted with the hosts' complement system and showed no significant differences in their survivability. The attenuated LVS was resistant to serum killing in mouse, but was lysed in the sera of hare and cattle. Direct binding of factor H to F. tularensis membrane proteins was not detected.

  16. Genetic Transformation of Hordeum vulgare ssp. spontaneum for the Development of a Transposon-Based Insertional Mutagenesis System.

    PubMed

    Cardinal, Marie-Josée; Kaur, Rajvinder; Singh, Jaswinder

    2016-10-01

    Domestication and intensive selective breeding of plants has triggered erosion of genetic diversity of important stress-related alleles. Researchers highlight the potential of using wild accessions as a gene source for improvement of cereals such as barley, which has major economic and social importance worldwide. Previously, we have successfully introduced the maize Ac/Ds transposon system for gene identification in cultivated barley. The objective of current research was to investigate the response of Hordeum vulgare ssp. spontaneum wild barley accessions in tissue culture to standardize parameters for introduction of Ac/Ds transposons through genetic transformation. We investigated the response of ten wild barley genotypes for callus induction, regenerative green callus induction and regeneration of fertile plants. The activity of exogenous Ac/Ds elements was observed through a transient assay on immature wild barley embryos/callus whereby transformed embryos/calli were identified by the expression of GUS. Transient Ds expression bombardment experiments were performed on 352 pieces of callus (3-5 mm each) or immature embryos in 4 genotypes of wild barley. The transformation frequency of putative transgenic callus lines based on transient GUS expression ranged between 72 and100 % in wild barley genotypes. This is the first report of a transformation system in H. vulgare ssp. spontaneum. PMID:27480175

  17. [Analgesic activity of different nonvolatile extracts of Nepeta atlantica Ball and Nepeta Tuberosa L. ssp. reticulata (Desf.) Maire].

    PubMed

    Bouidida, El Houcine; Alaoui, Katim; Cherrah, Yahia; Chammache, Malika; Il Idrissi, Abdelkader

    2008-01-01

    Different extracts of Nepeta atlantica Ball and Nepeta tuberosa L. ssp. reticulata (Desf.) Maire contain mainly secondary metabolites with iridoïd lactonic and glucosidic type, also with triterpine lupan type.The aerial part of each species is crushed, then extracted in methanol by cold maceration, called global extracts. The global extracts will be extracted through various solvents: initially by hexane, then by dichloromethane, after that by ethyl acetate and at the end by buthanol. Each one of the obtained extracts will be used for the following trials: i) Tail flick trial on the rat for central morphine-like analgesic activity; ii) Koster trial on the mouse for peripheral analgesic activity. The evaluation of the central and peripheral analgesic activities for the pre-cited extracts was realized after optimal doses determination of the global extracts activities for both species.The peripheral analgesic activity test on the mouse showed that, for 60 mg/kg intra peritoneum (IP), the hexanic, dichloromethanic, ethyl acetate and butanic extracts have a protection power against abdominal cramp respectively around 89.78%, 81.73%, 70.9% et 69.05% for Nepeta atlantica Ball, and around 89.16%, 82.98%, 71.52% et 70.27% for Nepeta tuberosa L. ssp. reticulata.Central morphine-like analgesic activity on the rat showed that, for both spices under 60 mg/kg IP, the central analgesic activity effect is significantly for two extracts only: dichloromethane and ethyl acetate.

  18. A comparison of isozyme and quantitative genetic variation in Pinus contorta ssp. latifolia by F{sub ST}

    SciTech Connect

    Yang, Rong-Cai; Yeh, F.C.; Yanchuk, A.D.

    1996-03-01

    We employed F-statistics to analyze quantitative and isozyme variation among five populations of Pinus contorta ssp. latifolia, a wind-pollinated outcrossing conifer with wide and continuous distribution in west North America. Estimates of population differentiation (F{sub ST}) for six quantitative traits were compared with the overall estimate of the differentiation (F*{sub ST}) from 19 isozymes that tested neutral to examine whether similar evolutionary processes were involved in morphological and isozyme differentiation. While the F{sub ST} estimates for specific gravity, stem diameter, stem height and branch length were significantly greater than the F*{sub ST} estimate, as judged from the 95% confidence intervals by bootstrapping, the F{sub ST} estimates for branch angle and branch diameter were indistinguishable from the F*{sub ST} estimate. Differentiation in stem height and stem diameter might reflect the inherent adaptation of the populations for rapid growth to escape suppression by neighboring plants during establishment and to regional differences in photoperiod, precipitation and temperature. In contrast, divergences in wood specific gravity and branch length might be correlated responses to population differentiation in stem growth. Possible bias in the estimation of F{sub ST} due to Hardy-Weinberg disequilibrium (F{sub IS} {ne} 0), linkage disequilibrium, maternal effects and nonadditive genetic effects was discussed with special reference to P. contorta ssp. latifolia. 48 refs., 1 fig., 3 tabs.

  19. The genetic basis of zinc tolerance in the metallophyte Arabidopsis halleri ssp. halleri (Brassicaceae): an analysis of quantitative trait loci.

    PubMed

    Willems, Glenda; Dräger, Dörthe B; Courbot, Mikael; Godé, Cécile; Verbruggen, Nathalie; Saumitou-Laprade, Pierre

    2007-05-01

    The species Arabidopsis halleri, an emerging model for the study of heavy metal tolerance and accumulation in plants, has evolved a high level of constitutive zinc tolerance. Mapping of quantitative trait loci (QTL) was used to investigate the genetic architecture of zinc tolerance in this species. A first-generation backcross progeny of A. halleri ssp. halleri from a highly contaminated industrial site and its nontolerant relative A. lyrata ssp. petraea was produced and used for QTL mapping of zinc tolerance. A genetic map covering most of the A. halleri genome was constructed using 85 markers. Among these markers, 65 were anchored in A. thaliana and revealed high synteny with other Arabidopsis genomes. Three QTL of comparable magnitude on three different linkage groups were identified. At all QTL positions zinc tolerance was enhanced by A. halleri alleles, indicating directional selection for higher zinc tolerance in this species. The two-LOD support intervals associated with these QTL cover 24, 4, and 13 cM. The importance of each of these three regions is emphasized by their colocalization with HMA4, MTP1-A, and MTP1-B, respectively, three genes well known to be involved in metal homeostasis and tolerance in plants.

  20. GA4 and IAA were involved in the morphogenesis and development of flowers in Agapanthus praecox ssp. orientalis.

    PubMed

    Zhang, Di; Ren, Li; Yue, Jian-hua; Wang, Ling; Zhuo, Li-huan; Shen, Xiao-hui

    2014-07-01

    The transition from vegetative to reproductive growth represents a major phase change in angiosperms. Hormones play important roles in this process. In this study, gibberellic acid (GA), cytokinins (CKs), indoleacetic acid (IAA), and abscisic acid (ABA) were analyzed during the flowering in Agapanthus praecox ssp. orientalis. Eleven types of endogenous gibberellins in addition to GA1 were detected in various organs. GA9 was detected with the highest concentrations, followed by GA5, GA8, and GA19. However, GA4 was the main bioactive GA that was involved in the regulation of flowering. Eight types of endogenous cytokinins were detected in A. praecox ssp. orientalis, and zeatin, zeatin riboside, zeatin-O-glucoside, and N(6)-isopentenyladenosine-5-monophosphate were present at higher levels throughout the study, of which zeatin plays an important role in the development of various organs. IAA increased by 581% in the shoot tips from the vegetative to inflorescence bud stages and had the most significant changes during flowering. Phytohormone immunolocalization analysis suggested that IAA involved in differentiation and development of each floral organs, GA and zeatin play important roles in floret primordia differentiation and ovule development. Using exogenous plant growth regulators proved that GA signaling regulate the scape elongation and stimulate early-flowering, and IAA signaling is involved in the pedicel and corolla elongation and delay flowering slightly. PMID:24913054

  1. First report of 'Candidatus Liberibacter solanacearum' on carrot in Africa

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In March of 2014, carrot plants (Daucus carota L. var. Mascot) exhibiting symptoms of yellowing, purpling, and curling of leaves, proliferation of shoots, formation of hairy secondary roots, general stunting and plant decline were observed in commercial fields in the Gharb region of Morocco. The sym...

  2. The carrot genome sequence brings colors out of the dark.

    PubMed

    Garcia-Mas, Jordi; Rodriguez-Concepcion, Manuel

    2016-05-27

    The genome sequence of carrot (Daucus carota L.) is the first completed for an Apiaceae species, furthering knowledge of the evolution of the important euasterid II clade. Analyzing the whole-genome sequence allowed for the identification of a gene that may regulate the accumulation of carotenoids in the root. PMID:27230684

  3. Meloidogyne incognita nematode resistance QTL in carrot

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Root-knot nematodes (Meloidogyne spp.) are major pests attacking carrots (Daucus carota) worldwide, causing galling and forking of the storage roots, rendering them unacceptable for market. Genetic resistance could significantly reduce the need for broad-spectrum soil fumigants in carrot production....

  4. 21 CFR 73.300 - Carrot oil.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 1 2013-04-01 2013-04-01 false Carrot oil. 73.300 Section 73.300 Food and Drugs... ADDITIVES EXEMPT FROM CERTIFICATION Foods § 73.300 Carrot oil. (a) Identity. (1) The color additive carrot... extraction of edible carrots (Daucus carota L.) with subsequent removal of the hexane by vacuum...

  5. 21 CFR 73.300 - Carrot oil.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 1 2012-04-01 2012-04-01 false Carrot oil. 73.300 Section 73.300 Food and Drugs... ADDITIVES EXEMPT FROM CERTIFICATION Foods § 73.300 Carrot oil. (a) Identity. (1) The color additive carrot... extraction of edible carrots (Daucus carota L.) with subsequent removal of the hexane by vacuum...

  6. 21 CFR 73.300 - Carrot oil.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 1 2014-04-01 2014-04-01 false Carrot oil. 73.300 Section 73.300 Food and Drugs... ADDITIVES EXEMPT FROM CERTIFICATION Foods § 73.300 Carrot oil. (a) Identity. (1) The color additive carrot... extraction of edible carrots (Daucus carota L.) with subsequent removal of the hexane by vacuum...

  7. 21 CFR 73.300 - Carrot oil.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 1 2010-04-01 2010-04-01 false Carrot oil. 73.300 Section 73.300 Food and Drugs... ADDITIVES EXEMPT FROM CERTIFICATION Foods § 73.300 Carrot oil. (a) Identity. (1) The color additive carrot... extraction of edible carrots (Daucus carota L.) with subsequent removal of the hexane by vacuum...

  8. 21 CFR 73.300 - Carrot oil.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 1 2011-04-01 2011-04-01 false Carrot oil. 73.300 Section 73.300 Food and Drugs... ADDITIVES EXEMPT FROM CERTIFICATION Foods § 73.300 Carrot oil. (a) Identity. (1) The color additive carrot... extraction of edible carrots (Daucus carota L.) with subsequent removal of the hexane by vacuum...

  9. First report of ‘Candidatus Liberibacter solanacearum’ associated with the Psyllid Bactericera trigonica Hodkinson on carrots in northern Africa

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Carrot plants (Daucus carota L.) exhibiting symptoms of yellowing, purpling, and curling of leaves, proliferation of shoots, formation of hairy secondary roots, and plant decline were observed in March 2014 and February 2015 in commercial fields in the Gharb region of Morocco. The symptoms resembled...

  10. Promoter Strength Comparisons of Maize Shrunken 1 and Alcohol Dehydrogenase 1 and 2 Promoters in Mono- and Dicotyledonous Species 1

    PubMed Central

    Hauptmann, R. M.; Ashraf, M.; Vasil, V.; Hannah, L. C.; Vasil, I. K.; Ferl, R.

    1988-01-01

    Promoter strengths of two maize alcohol dehydrogenase genes, Adh1 and Adh2, and the maize shrunken-1 gene, Sh1, were evaluated by transient expression in cultured protoplasts of Panicum maximum, Triticum monococcum, and Daucus carota. Promoter elements were ligated in correct and opposite orientations as transcriptional gene fusions to the chloramphenicol acetyl transferase gene containing the nopaline synthase 3′ polyadenylation signal. The relative levels of gene expression were compared to the cauliflower mosaic virus 35S promoter. The full length Adh1 promoter (−1100 to +15) functioned in all species, but at a reduced level in D. carota. An Adh1 promoter deletion from −304 to −1100 did not express at detectable levels in any species nor did the Sh1 promoter construction. The Adh2 promoter (−860 to +90) only expressed in D. carota. The full length Adh1 promoter gave the highest level of CAT expression in the monocot cells but at levels which were approximately 30% compared to the CaMV 35S promoter. This was reduced further in D. carota to approximately 4%. These data suggest that at least some of the regulatory factors responsible for promoter function are somewhat species specific and that these differences should be considered in gene expression studies. Images Fig. 2 PMID:16666422

  11. First Report of 'Candidatus Liberibacter solanacearum' in Carrots in Europe

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Carrot (Daucus carota) plants exhibiting symptoms that resembled those of carrot psyllid (Trioza apicalis) damage were observed in commercial fields in southern Finland in August 2008. Carrot psyllid is a serious pest of carrots in northern and central Europe, where it can cause up to 100% yield los...

  12. Molecular detection of aster yellows phytoplasma and 'Candidatus Liberibacter solanacearum' in carrots affected by the psyllid Trioza apicalis (Hemiptera: triozidae) in Finland

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Carrot psyllid (Trioza apicalis Förster) causes considerable damage to carrot (Daucus carota L.) in many parts of Europe. It was recently established that the new bacterium “Candidatus Liberibacter solanacearum” is associated with carrot psyllid and plants affected by this insect pest. No other path...

  13. Association of 'Candidatus Liberibacter solanacearum' with the Psyllid, Trioza apicalis (Hemiptera: Triozidae) in Europe

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Carrot psyllid (Trioza apicalis Förster) is a serious pest of carrots (Daucus carota L.) in northern and central Europe. Carrots exhibiting symptoms of psyllid damage were observed in commercial fields in southern Finland in 2008. Symptoms in affected plants included leaf curling, yellow and purple ...

  14. A high-quality carrot genome assembly provides new insights into carotenoid accumulation and asterid genome evolution

    Technology Transfer Automated Retrieval System (TEKTRAN)

    We report a chromosome-scale assembly and analysis of the Daucus carota genome, an important source of provitamin A in the human diet and the first sequenced genome among members of the Euasterid II clade. We characterized two new polyploidization events, both occurring after the divergence of carro...

  15. First report of 'Candidatus Liberibacter solanacearum' associated with psyllid-affected carrots in Sweden

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Carrot (Daucus carota) plants with symptoms resembling those of the carrot psyllid Trioza apicalis and “Candidatus Liberibacter solanacearum” damage were observed in 70% of commercial fields in southern Sweden in August 2011; all cultivars grown were affected, at about 1 to 45% symptomatic plants pe...

  16. Optimization of flat plate drying of carrot pomace (abstract)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Carrot (Daucus carota var. sativus) pomace is a co-product of the carrot juice and cut-carrot industry; it has high nutritional value but is currently underutilized. Drum drying is one method that could be used to dry and stabilize carrot pomace. However, optimum conditions for the dryer surface tem...

  17. Survey of 'Candidatus Liberibacter solanacearum' in carrot crops affected by the psyllid Trioza apicalis (Hemiptera: Triozidae) in Norway

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The carrot psyllid Trioza apicalis Förster (Hemiptera: Triozidae) is a serious insect pest of carrot (Daucus carota L.) in northern Europe, where it can cause up to 100% crop loss. Although it was long believed that T. apicalis causes damage to carrot by injection of toxins into the plant, it was re...

  18. Crop candidates for the bioregenerative life support systems in China

    NASA Astrophysics Data System (ADS)

    Chunxiao, Xu; Hong, Liu

    The use of plants for life support applications in space is appealing because of the multiple life support functions by the plants. Research on crops that were grown in the life support system to provide food and oxygen, remove carbon dioxide was begun from 1960. To select possible crops for research on the bioregenerative life support systems in China, criteria for the selection of potential crops were made, and selection of crops was carried out based on these criteria. The results showed that 14 crops including 4 food crops (wheat, rice, soybean and peanut) and 7 vegetables (Chinese cabbage, lettuce, radish, carrot, tomato, squash and pepper) won higher scores. Wheat ( Triticum aestivum L.), rice ( Oryza sativa L.), soybean ( Glycine max L.) and peanut ( Arachis hypogaea L.) are main food crops in China. Chinese cabbage ( Brassica campestris L. ssp. chinensis var. communis), lettuce ( Lactuca sativa L. var. longifolia Lam.), radish ( Raphanus sativus L.), carrot ( Daucus carota L. var. sativa DC.), tomato ( Lycopersicon escalentum L.), squash ( Cucurbita moschata Duch.) and pepper ( Capsicum frutescens L. var. longum Bailey) are 7 vegetables preferred by Chinese. Furthermore, coriander ( Coriandum sativum L.), welsh onion ( Allium fistulosum L. var. giganteum Makino) and garlic ( Allium sativum L.) were selected as condiments to improve the taste of space crew. To each crop species, several cultivars were selected for further research according to their agronomic characteristics.

  19. Enhanced natural killer cell activation by exopolysaccharides derived from yogurt fermented with Lactobacillus delbrueckii ssp. bulgaricus OLL1073R-1.

    PubMed

    Makino, Seiya; Sato, Asako; Goto, Ayako; Nakamura, Marie; Ogawa, Miho; Chiba, Yoshika; Hemmi, Jun; Kano, Hiroshi; Takeda, Kazuyoshi; Okumura, Ko; Asami, Yukio

    2016-02-01

    Yogurt is generally recognized as a beneficial food for our health, but research into its physiological effects has focused mainly on intestinal dysfunctions such as constipation and diarrhea. We previously found yogurt fermented with Lactobacillus delbrueckii ssp. bulgaricus OLL1073R-1 (hereafter OLL1073R-1) could reduce risks of catching the common cold and flu in human trials. It was assumed that immunostimulatory exopolysaccharide (EPS) produced from OLL1073R-1 play an important role in this context. However, few studies have examined the immunostimulatory effects of traditional Bulgarian yogurts fermented with different strains of lactobacilli and their metabolites. Therefore, we screened 139 L. delbrueckii ssp. bulgaricus strains and identified OLL1073R-1 as the most robust producer of EPS. This strain was also the only strain that induced the production of IFN-γ in vitro. Oral administration of the EPS or yogurt fermented with OLL1073R-1 and Streptococcus thermophilus OLS3059 (OLL1073R-1 yogurt) augmented natural killer (NK) cell activity and induced IFN-γ production in spleen cells in mice, whereas 2 other yogurts fermented with other strains had no effect on NK cell activity. Cellular preparations of the OLL1073R-1 strain also slightly augmented NK cell activity, but were less effective than EPS itself. The EPS-dependent stimulation of NK cell activity was abrogated in IFN-γ knockout mice and in myeloid differentiation factor 88 knockout mice. Furthermore, IFN-γ production from spleen cells stimulated with EPS was completely blocked with both anti-IL-12 and anti-IL-18 antibodies in vitro. These findings suggest that NK cell activation by OLL1073R-1 yogurt is EPS-dependent, occurs via IL-12- and IL-18-mediated IFN-γ production, and requires myeloid differentiation factor 88. We showed that traditional Bulgarian yogurt could exert immunostimulatory effects by selecting starter strains and part of the mechanisms depend on IFN-γ inducible EPS produced

  20. Enhanced natural killer cell activation by exopolysaccharides derived from yogurt fermented with Lactobacillus delbrueckii ssp. bulgaricus OLL1073R-1.

    PubMed

    Makino, Seiya; Sato, Asako; Goto, Ayako; Nakamura, Marie; Ogawa, Miho; Chiba, Yoshika; Hemmi, Jun; Kano, Hiroshi; Takeda, Kazuyoshi; Okumura, Ko; Asami, Yukio

    2016-02-01

    Yogurt is generally recognized as a beneficial food for our health, but research into its physiological effects has focused mainly on intestinal dysfunctions such as constipation and diarrhea. We previously found yogurt fermented with Lactobacillus delbrueckii ssp. bulgaricus OLL1073R-1 (hereafter OLL1073R-1) could reduce risks of catching the common cold and flu in human trials. It was assumed that immunostimulatory exopolysaccharide (EPS) produced from OLL1073R-1 play an important role in this context. However, few studies have examined the immunostimulatory effects of traditional Bulgarian yogurts fermented with different strains of lactobacilli and their metabolites. Therefore, we screened 139 L. delbrueckii ssp. bulgaricus strains and identified OLL1073R-1 as the most robust producer of EPS. This strain was also the only strain that induced the production of IFN-γ in vitro. Oral administration of the EPS or yogurt fermented with OLL1073R-1 and Streptococcus thermophilus OLS3059 (OLL1073R-1 yogurt) augmented natural killer (NK) cell activity and induced IFN-γ production in spleen cells in mice, whereas 2 other yogurts fermented with other strains had no effect on NK cell activity. Cellular preparations of the OLL1073R-1 strain also slightly augmented NK cell activity, but were less effective than EPS itself. The EPS-dependent stimulation of NK cell activity was abrogated in IFN-γ knockout mice and in myeloid differentiation factor 88 knockout mice. Furthermore, IFN-γ production from spleen cells stimulated with EPS was completely blocked with both anti-IL-12 and anti-IL-18 antibodies in vitro. These findings suggest that NK cell activation by OLL1073R-1 yogurt is EPS-dependent, occurs via IL-12- and IL-18-mediated IFN-γ production, and requires myeloid differentiation factor 88. We showed that traditional Bulgarian yogurt could exert immunostimulatory effects by selecting starter strains and part of the mechanisms depend on IFN-γ inducible EPS produced

  1. Survival, acid and bile tolerance, and surface hydrophobicity of microencapsulated B. animalis ssp. lactis Bb12 during storage at room temperature.

    PubMed

    Dianawati, Dianawati; Shah, Nagendra P

    2011-01-01

    Survival, acid and bile tolerance, and surface hydrophobicity of microencapsulated Bifidobacterium animalis ssp. lactis Bb12 were studied during storage at room temperature (25 °C) at low water activity (0.07, 0.1, and 0.2). Two types of alginate-based systems were prepared with and without mannitol as microencapsulant of B. animalis ssp. lactis Bb12. Formation of gel beads containing cells was achieved by dropping each emulsion into CaCl(2) solution; then, the beads were freeze dried. Survival, acid tolerance during 2-h exposure in de Man, Rogosa, Sharpe (MRS) broth at pH 2.0, bile tolerance during 8-h exposure in MRS broth containing taurocholic acid at pH 5.8, and retention of surface hydrophobicity were determined after freeze drying and during storage. The result showed that neither alginate nor alginate-mannitol formulation was effective in protecting B. animalis ssp. lactis Bb12 during freezing and freeze drying. The viability in alginate-mannitol and alginate formulations after freeze drying was 6.61 and 6.34 log CFU/g, respectively. Storage at low a(w) improved survival, acid tolerance, bile tolerance, and surface hydrophobicity retention of microencapsulated B. animalis ssp. lactis Bb12 when compared with controlled storage in an aluminum foil (with a(w) of 0.38 and 0.40 for alginate-mannitol and alginate formulations, respectively). Alginate mannitol was more effective than the alginate system during a short period of storage, but its effectiveness decreased during a long period of storage (80% survival at 10 wk). Nevertheless, storage of microencapsulated B. animalis ssp. lactis Bb12 in an aluminum foil without a(w) adjustment during 10 wk at room temperature was not effective (survival was 64% to 65%). PMID:22416710

  2. Comparative essential oil composition and antifungal effect of bitter fennel (Foeniculum vulgare ssp. piperitum) fruit oils obtained during different vegetation.

    PubMed

    Ozcan, Mehmet Musa; Chalchat, Jean-Claude; Arslan, Derya; Ateş, Ayşe; Unver, Ahmet

    2006-01-01

    The chemical composition of the flower and unripe and ripe fruits from fennel (bitter) (Foeniculum vulgare ssp. piperitum) has been examined by gas chromatography and gas chromatography-mass spectrometry. The main identified components of the flower and unripe and ripe fruit oils were estragole (53.08%, 56.11%, and 61.08%), fenchone (13.53%, 19.18%, and 23.46%), and alpha-phellandrene (5.77%, 3.30%, and 0.72%), respectively. Minor qualitative and major quantitative variations for some compounds of essential oils were determined with respect to the different parts of F. vulgare. The oils exerted varying levels of antifungal effects on the experimental mycelial growth of Alternaria alternata, Fusarium oxysporum, and Rhizoctonia solani. The 40 ppm concentrations of fennel oils showed inhibitory effect against mycelial growth of A. alternaria, whereas 10 ppm levels were ineffective. The analyses show that fennel oils exhibited different degrees of fungistatic activity depending on the doses.

  3. Enhanced glucosinolates in root exudates of Brassica rapa ssp. rapa mediated by salicylic acid and methyl jasmonate.

    PubMed

    Schreiner, Monika; Krumbein, Angelika; Knorr, Dietrich; Smetanska, Iryna

    2011-02-23

    Elicitation studies with salicylic acid (SA) and methyl jasmonate (MJ) inducing a targeted rhizosecretion of high levels of anticarcinogenic glucosinolates in Brassica rapa ssp. rapa plants were conducted. Elicitor applications not only led to an accumulation of individual indole glucosinolates and the aromatic 2-phenylethyl glucosinolate in the turnip organs but also in turnip root exudates. This indicates an extended systemic response, which comprises the phyllosphere with all aboveground plant organs and the rhizosphere including the belowground root system and also root exudates. Both elicitor applications induced a doubling in 2-phenylethyl glucosinolate in root exudates, whereas application of MJ enhanced rhizosecreted indole glucosinolates up to 4-fold. In addition, the time course study revealed that maximal elicitation was observed on the 10th day of SA and MJ treatment. This study may provide an essential contribution using these glucosinolates as bioactive additives in functional foods and nutraceuticals.

  4. Genetic variability of the low-molecular-weight glutenin subunits in spelt wheat (Triticum aestivum ssp. spelta L. em Thell.).

    PubMed

    Caballero, L; Martín, L M; Alvarez, J B

    2004-03-01

    The low-molecular-weight glutenin subunit composition of a collection of 403 accessions of spelt wheat ( Triticum aestivum ssp. spelta L. em. Thell) was analyzed by SDS-PAGE. Extensive variation was found, including 46 different patterns for zone B and 16 for zone C. Patterns within zone B exhibited from two to six bands and patterns in zone C had between four and six bands in SDS-PAGE gels. A higher number of bands was observed when urea was added to the gels. Zone B exhibited between six and 11 bands, and we identified 14 new patterns in this zone. For zone C, up to ten new patterns that comprised between five and nine bands were detected. For both zones, 86 patterns were found. The variability detected in this material is greater than that detected in other hulled wheats.

  5. Cloning and expression of the gene encoding alpha-acetolactate decarboxylase from Acetobacter aceti ssp. xylinum in brewer's yeast.

    PubMed

    Yamano, S; Tanaka, J; Inoue, T

    1994-02-14

    Acetobacter aceti ssp. xylinum genomic library was constructed using cosmid pJB8 in Escherichia coli. The gene encoding alpha-acetolactate decarboxylase (ALDC) was isolated from the library by direct measurement of ALDC activity. The ALDC gene was expressed by its own promoter in E. coli. The nucleotide sequence was determined, and an open reading frame which may encode a protein composed of 304 amino acids with a molecular weight of 33,747 was found. A brewer's yeast was transformed with the YEp-type plasmid containing the ALDC gene placed under the control of the glyceraldehyde-3-phosphate dehydrogenase promoter. The laboratory-scale growth test confirmed that the total diacetyl concentration was considerably reduced by the transformant. The analysis of the wort indicates that the Acetobacter ALDC reduces the concentration of diacetyl more effectively than that of 2,3-pentanedione.

  6. Construction of a brewer's yeast having alpha-acetolactate decarboxylase gene from Acetobacter aceti ssp. xylinum integrated in the genome.

    PubMed

    Yamano, S; Kondo, K; Tanaka, J; Inoue, T

    1994-02-14

    alpha-Acetolactate decarboxylase (ALDC) gene from Acetobacter aceti ssp. xylinum has several possible initiation codons in the N-terminus. To determine the initiation codon of the ALDC giving the highest expression levels, glyceraldehyde-3-phosphate dehydrogenase (GPD) promoter was linked just upstream of each possible initiation codon. The ALDC whose translation starts 130 bp downstream from the first ATG codon had the highest activity in yeast cells. When expression levels of the ALDC gene were compared using three strong yeast promoters of glycolytic genes, alcohol dehydrogenase I (ADC1), phosphoglycerate kinase (PGK) and GPD, the GPD promoter was the strongest. The ALDC gene was integrated in a ribosomal RNA gene of a brewer's yeast by co-transformation with an expression plasmid of G418-resistance gene. The laboratory-scale growth test confirmed that the total diacetyl concentration was reduced in wort.

  7. Effects of gentiopicroside, sweroside and swertiamarine, secoiridoids from gentian (Gentiana lutea ssp. symphyandra), on cultured chicken embryonic fibroblasts.

    PubMed

    Oztürk, Nilgün; Korkmaz, Seval; Oztürk, Yusuf; Başer, K Hüsnü Can

    2006-03-01

    Wound healing properties of Gentian (Gentiana lutea ssp. symphyandra) extract and its main constituents, gentiopicroside, sweroside and swertiamarine (compounds 1-3, respectively) were evaluated by comparison with dexpanthenol on cultured chicken embryonic fibroblasts. The extract was also analyzed by HPLC to quantify its constituents. Chicken embryonic fibroblasts from fertilized eggs were incubated with the plant extract and its constituents, compounds 1-3. Using microscopy, mitotic ability, morphological changes and collagen production in the cultured fibroblasts were evaluated as parameters. Wound healing activity of Gentian seems to be mainly due to the increase in the stimulation of collagen production and the mitotic activity by compounds 2 and 3, respectively (p < 0.005 in all cases). All three compounds also exhibited cytoprotective effects, which may cause a synergism in terms of wound healing activity of Gentian. The findings demonstrated the wound healing activity of Gentian, which has previously been based only on ethnomedical data.

  8. Vitamin B12 production from crude glycerol by Propionibacterium freudenreichii ssp. shermanii: optimization of medium composition through statistical experimental designs.

    PubMed

    Kośmider, Alicja; Białas, Wojciech; Kubiak, Piotr; Drożdżyńska, Agnieszka; Czaczyk, Katarzyna

    2012-02-01

    A two-step statistical experimental design was employed to optimize the medium for vitamin B(12) production from crude glycerol by Propionibacterium freudenreichii ssp. shermanii. In the first step, using Plackett-Burman design, five of 13 tested medium components (calcium pantothenate, NaH(2)PO(4)·2H(2)O, casein hydrolysate, glycerol and FeSO(4)·7H(2)O) were identified as factors having significant influence on vitamin production. In the second step, a central composite design was used to optimize levels of medium components selected in the first step. Valid statistical models describing the influence of significant factors on vitamin B(12) production were established for each optimization phase. The optimized medium provided a 93% increase in final vitamin concentration compared to the original medium. PMID:22178491

  9. Twist-induced defects of the P-SSP7 genome revealed by modeling the cryo-EM density.

    PubMed

    Wang, Qian; Myers, Christopher G; Pettitt, B Montgomery

    2015-04-16

    We consider the consequences of assuming that DNA inside of phages can be approximated as a strongly nonlinear persistence length polymer. Recent cryo-EM experiments find a hole in the density map of P-SSP7 phage, located in the DNA segment filling the portal channel of the phage. We use experimentally derived structural constraints with coarse-grained simulation techniques to consider contrasting model interpretations of reconstructed density in the portal channel. The coarse-grained DNA models used are designed to capture the effects of torsional strain and electrostatic environment. Our simulation results are consistent with the interpretation that the vacancy or hole in the experimental density map is due to DNA strain leading to strand separation. We further demonstrate that a moderate negative twisting strain is able to account for the strand separation. This effect of nonlinear persistence length may be important in other aspects of phage DNA packing. PMID:25793549

  10. A high-resolution karyotype of Brassica rapa ssp. pekinensis revealed by pachytene analysis and multicolor fluorescence in situ hybridization.

    PubMed

    Koo, Dal-Hoe; Plaha, Prikshit; Lim, Yong Pyo; Hur, Yoonkang; Bang, Jae-Wook

    2004-11-01

    A molecular cytogenetic map of Chinese cabbage (Brassica rapa ssp. pekinensis, 2 n=20) was constructed based on the 4'-6-diamino-2-phenylindole dihydrochloride-stained mitotic metaphase and pachytene chromosomes and multicolor fluorescence in situ hybridization (McFISH), using three repetitive DNA sequences, 5S rDNA, 45S rDNA, and C11-350H. The lengths of mitotic metaphase chromosomes ranged from 1.46 microm to 3.30 microm. Five 45S and three 5S rDNA loci identified were assigned to different chromosomes. The C11-350H loci were located on all the mitotic metaphase chromosomes, except chromosomes 2 and 4. The pachytene karyotype consisted of two metacentric (chromosomes 1 and 6), five submetacentric (chromosomes 3, 4, 5, 9 and 10), two subtelocentric (chromosomes 7 and 8), and one acrocentric (chromosome 2) chromosome(s). The mean lengths of ten pachytene chromosomes ranged from 23.7 microm to 51.3 microm, with a total of 385.3 microm, which is 17.5-fold longer than that of the mitotic metaphase chromosomes. In the proposed pachytene karyotype, all the chromosomes of B. rapa ssp. pekinensis can be identified on the basis of chromosome length, centromere position, heterochromatin pattern, and the location of the three repetitive sequences. Moreover, the precise locations of the earlier reported loci of 5S rDNA, 45S rDNA, and Chinese cabbage tandem DNA repeat C11-350H were established using McFISH analysis. We also identified a 5S rDNA locus on the long arm of pachytene bivalent 7, which could not be detected in the mitotic metaphase chromosomes in the present and earlier studies. The deduced karyotype will be useful for structural and functional genomic studies in B. rapa.

  11. Emergence of gynodioecy in wild beet (Beta vulgaris ssp. maritima L.): a genealogical approach using chloroplastic nucleotide sequences.

    PubMed

    Fénart, Stéphane; Touzet, Pascal; Arnaud, Jean-François; Cuguen, Joël

    2006-06-01

    Gynodioecy is a breeding system where both hermaphroditic and female individuals coexist within plant populations. This dimorphism is the result of a genomic interaction between maternally inherited cytoplasmic male sterility (CMS) genes and bi-parentally inherited nuclear male fertility restorers. As opposed to other gynodioecious species, where every cytoplasm seems to be associated with male sterility, wild beet Beta vulgaris ssp. maritima exhibits a minority of sterilizing cytoplasms among numerous non-sterilizing ones. Many studies on population genetics have explored the molecular diversity of different CMS cytoplasms, but questions remain concerning their evolutionary dynamics. In this paper we report one of the first investigations on phylogenetic relationships between CMS and non-CMS lineages. We investigated the phylogenetic relationships between 35 individuals exhibiting different mitochondrial haplotypes. Relying on the high linkage disequilibrium between chloroplastic and mitochondrial genomes, we chose to analyse the nucleotide sequence diversity of three chloroplastic fragments (trnK intron, trnD-trnT and trnL-trnF intergenic spacers). Nucleotide diversity appeared to be low, suggesting a recent bottleneck during the evolutionary history of B. vulgaris ssp. maritima. Statistical parsimony analyses revealed a star-like genealogy and showed that sterilizing haplotypes all belong to different lineages derived from an ancestral non-sterilizing cytoplasm. These results suggest a rapid evolution of male sterility in this taxon. The emergence of gynodioecy in wild beet is confronted with theoretical expectations, describing either gynodioecy dynamics as the maintenance of CMS factors through balancing selection or as a constant turnover of new CMSs.

  12. Comparative cDNA-AFLP analysis of Cd-tolerant and -sensitive genotypes derived from crosses between the Cd hyperaccumulator Arabidopsis halleri and Arabidopsis lyrata ssp. petraea.

    PubMed

    Craciun, Adrian Radu; Courbot, Mikael; Bourgis, Fabienne; Salis, Pietrino; Saumitou-Laprade, Pierre; Verbruggen, Nathalie

    2006-01-01

    Cadmium (Cd) tolerance seems to be a constitutive species-level trait in Arabidopsis halleri. In order to identify genes potentially implicated in Cd tolerance, a backcross (BC1) segregating population was produced from crosses between A. halleri ssp. halleri and its closest non-tolerant relative A. lyrata ssp. petraea. The most sensitive and tolerant genotypes of the BC1 were analysed on a transcriptome-wide scale by cDNA-amplified fragment length polymorphism (AFLP). A hundred and thirty-four genes expressed more in the root of tolerant genotypes than in sensitive genotypes were identified. Most of the identified genes showed no regulation in their expression when exposed to Cd in a hydroponic culture medium and belonged to diverse functional classes, including reactive oxygen species (ROS) detoxification, cellular repair, metal sequestration, water transport, signal transduction, transcription regulation, and protein degradation, which are discussed. PMID:16916885

  13. Inactivation of Mycobacterium bovis ssp. caprae in high-temperature, short-term pasteurized pilot-plant milk.

    PubMed

    Hammer, P; Richter, E; Rüsch-Gerdes, S; Walte, H-G C; Matzen, S; Kiesner, C

    2015-03-01

    Experiments to determine the efficacy of high temperature, short time (HTST) pasteurization of milk in terms of inactivation of pathogenic microorganisms were mainly performed between 1930 and 1960. Among the target organisms were Mycobacterium bovis and Mycobacterium tuberculosis. As a result, the Codex Alimentarius prescribes that HTST treatment of milk should lead to a significant reduction of pathogenic microorganisms during milk pasteurization. Due to the development of improved methods for the detection of survivors and of more advanced heating technology, verification of this requirement seemed to be necessary. To address recent outbreaks of tuberculosis in cattle caused by M. bovis ssp. caprae (M. caprae) in the southern regions of Germany, this organism was tested and compared with M. bovis ssp. bovis (M. bovis). Experiments were performed in a pilot plant for HTST pasteurization of milk with 3 strains of M. caprae and 1 strain of M. bovis. In preliminary trials at a fixed holding time of 25 s, the temperature at which significant inactivation occurred was 62.5°C for all strains. To determine D-values (decimal reduction times) for the inactivation kinetics, the strains were tested at 65, 62.5, and 60°C at holding times of 16.5, 25, and 35 s. At 65°C, the D-values of all strains ranged from 6.8 to 7.8 s, and at 62.5°C, D-values ranged from 14.5 to 18.1 s. Low inactivation was observed at 60°C. When the low slope of the inactivation curve allowed calculation of a D-value, these ranged from 40.8 to 129.9 s. In terms of log10 reductions, the highest values for all strains were 4.1 to 4.9 log at 65°C, with a holding time of 35 s. The tested strains of M. caprae and M. bovis showed similar low resistance to heat. Standard HTST treatment should result in a high reduction of these organisms and thus the requirements of the Codex Alimentarius for inactivation of pathogens by this process are far exceeded.

  14. Genome-wide analysis of the AP2/ERF transcription factor superfamily in Chinese cabbage (Brassica rapa ssp. pekinensis)

    PubMed Central

    2013-01-01

    Background Chinese cabbage (Brassica rapa ssp. pekinensis) is a member of one of the most important leaf vegetables grown worldwide, which has experienced thousands of years in cultivation and artificial selection. The entire Chinese cabbage genome sequence, and more than forty thousand proteins have been obtained to date. The genome has undergone triplication events since its divergence from Arabidopsis thaliana (13 to 17 Mya), however a high degree of sequence similarity and conserved genome structure remain between the two species. Arabidopsis is therefore a viable reference species for comparative genomics studies. Variation in the number of members in gene families due to genome triplication may contribute to the broad range of phenotypic plasticity, and increased tolerance to environmental extremes observed in Brassica species. Transcription factors are important regulators involved in plant developmental and physiological processes. The AP2/ERF proteins, one of the most important families of transcriptional regulators, play a crucial role in plant growth, and in response to biotic and abiotic stressors. Our analysis will provide resources for understanding the tolerance mechanisms in Brassica rapa ssp. pekinensis. Results In the present study, 291 putative AP2/ERF transcription factor proteins were identified from the Chinese cabbage genome database, and compared with proteins from 15 additional species. The Chinese cabbage AP2/ERF superfamily was classified into four families, including AP2, ERF, RAV, and Soloist. The ERF family was further divided into DREB and ERF subfamilies. The AP2/ERF superfamily was subsequently divided into 15 groups. The identification, classification, phylogenetic reconstruction, conserved motifs, chromosome distribution, functional annotation, expression patterns, and interaction networks of the AP2/ERF transcription factor superfamily were predicted and analyzed. Distribution mapping results showed AP2/ERF superfamily genes were

  15. A simplified PCR-SSP method for HLA-A2 subtype in a population of Wuhan, China.

    PubMed

    Liang, Bing; Zhu, Lijun; Liang, Zhihui; Weng, Xiufang; Lu, Xiaoling; Zhang, Cai'e; Li, Hui; Wu, Xiongwen

    2006-12-01

    HLA-A2 is the most frequent HLA-A allele in all ethnic populations, and an important restriction element for peptide presentation to T cells in infectious disease and cancer. However, the HLA-A2 supertype consisting of up to 75 subtypes, mutation studies and analyses using cytotoxic T lymphocytes suggest the functional relevance of subtype-specific differences in HLA-A2 molecules for peptide binding and T-cell recognition. Therefore, it is necessary for T-cell response study to discriminate the HLA-A2 subtypes and to understand the profile of HLA-A2 allelic distribution in a given population. In this study, we developed a simple, robust approach based on the nested polymerase chain reaction using sequence-specific primers (PCR-SSP) to discriminate 17 HLA-A2 subtypes which cover the most HLA-A2 alleles (> 99% allele frequency) reported in Chinese, using 15 combinations of 19 allelic specific primers. In the first round of PCR, 3 combinations of 5 primers were used to determine whether the tested sample was HLA-A2 positive, meanwhile the subtypes of HLA-A*0209 and HLA-A*0215N were determined for the variant position of these two subtypes is in exon 4 instead of exon 2, 3. Samples of HLA-A2 positive were subtyped in the second round of PCR, using PCR products of the first round as templates. This strategy was applied to test the samples of 78 random HLA-A2 positive individuals for their HLA-A2 subtypes. Those samples were screened for HLA-A2 positive by the first round PCR-SSP from 154 healthy blood donors in Wuhan, China. The subtyping results were verified by using flow cytometric analysis (FCM) with HLA-A2 specific monoclonal antibody BB7.2 and DNA sequencing. The typing results of the samples show 50.7% random individuals in the population carry HLA-A2, HLA-A*0201 ranks the first (allele frequency = 15.5%), followed by A*0207 (5.8%), A*0206 (4.7%), A*0203 (2.6%), A*0210 (0.7%), and these 5 alleles account for 99.0% HLA-A2 subtypes of allele frequency. Our study

  16. Chemical Composition of Ballota macedonica Vandas and Ballota nigra L. ssp. foetida (Vis.) Hayek Essential Oils - The Chemotaxonomic Approach.

    PubMed

    Đorđević, Aleksandra S; Jovanović, Olga P; Zlatković, Bojan K; Stojanović, Gordana S

    2016-06-01

    The essential oils isolated from fresh aerial parts of Ballota macedonica (two populations) and Ballota nigra ssp. foetida were analyzed by GC and GC/MS. Eighty five components were identified in total; 60 components in B. macedonica oil (population from the Former Yugoslav Republic of Macedonia), 34 components in B. macedonica oil (population from the Republic of Serbia), and 33 components in the oil of B. nigra ssp. foetida accounting for 93.9%, 98.4%, and 95.8% of the total oils, respectively. The most abundant components in B. macedonica oils were carotol (13.7 - 52.1%), germacrene D (8.6 - 24.6%), and (E)-caryophyllene (6.5 - 16.5%), while B. nigra ssp. foetida oil was dominated by (E)-phytol (56.9%), germacrene D (10.0%), and (E)-caryophyllene (4.7%). Multivariate statistical analyses (agglomerative hierarchical cluster analysis and principal component analysis) were used to compare and discuss relationships among Ballota species examined so far based on their volatile profiles. The chemical compositions of B. macedonica essential oils are reported for the first time. PMID:27144646

  17. A clinical study on SSP (silver spike point) electro-therapy combined with splint therapy for temporo-mandibular joint dysfunction.

    PubMed

    Sugimoto, K; Konda, T; Shimahara, M; Hyodo, M; Kitade, T

    1995-01-01

    When the functional limits of the muscles related to the temporo-mandibular joint and adjacent tissue exceed their anatomical capability, pain, crepitation, and functional abnormality appear as the main complaints. Although the precise nature of the condition is unknown, pain at the temporo-mandibular joint sometimes in combination with muscular tension is assumed to be due to compression of the myoneural mechanism. It is reported that occlusal lifting using a splint enables the alleviation of this muscular tension. On the other hand, there are only a few reports on the usefulness of SSP therapy for Temporo-Mandibular Joint Dysfunction. We studied the efficacy of SSP therapy combined with splint therapy in 33 patients diagnosed as having Temporo-Mandibular Joint Dysfunction who consulted our department primarily due to pain, and report our findings below. Evaluation of the results was conducted 2 weeks later. Very beneficial results were seen in 6 cases. Beneficial results were seen in 7 cases. Slightly beneficial results were seen in 18 cases, while there were no changes found in 2 cases. When combined SSP and splint therapies were conducted for Temporo-Mandibular Joint Dysfunction, favorable results were seen in about 90% of the cases.

  18. Mapping of isolate-specific QTLs for clubroot resistance in Chinese cabbage (Brassica rapa L. ssp. pekinensis).

    PubMed

    Sakamoto, Koji; Saito, Atsuo; Hayashida, Nobuaki; Taguchi, Goro; Matsumoto, Etsuo

    2008-09-01

    A number of clubroot resistant (CR) Chinese cabbage cultivars have been developed in Japan using resistant genes from CR European fodder turnips (B. rapa ssp. rapifera). Clubroot resistance in European fodder turnips are known to be controlled by the combined action of several dominant resistance genes. We have developed three Chinese cabbage clubroot-resistant doubled haploid (DH) lines--T136-8, K10, and C9--which express resistance in different manners against two isolates of Plasmodiophora brassicae, M85 and K04. Depending on the isolates, we identified two CR loci, CRk and CRc. CRk was identified by quantitative trait loci (QTL) analysis of an F(2) population derived from a cross between K10 and Q5. This locus showed resistance to both isolates and is located close to Crr3 in linkage group R3. The other locus, CRc was identified by QTL analysis of an F(2) population derived from a cross between C9 and susceptible DH line, 6R. This locus was mapped to linkage group R2 and is independent from any published CR loci. We developed sequence-tagged site markers linked to this locus.

  19. Identification and mapping of resistance gene analogs and a white rust resistance locus in Brassica rapa ssp. oleifera.

    PubMed

    Tanhuanpää, P

    2004-04-01

    The objective of this investigation was to tag a locus for white rust resistance in a Brassica rapa ssp. oleifera F(2) population segregating for this trait, using bulked segregant analysis with random amplified polymorphic DNA (RAPD) markers, linkage mapping and a candidate gene approach based on resistance gene analogs (RGAs). The resistance source was the Finnish line Bor4109. The reaction against white rust races 7a and 7v was scored in 20 seedlings from each self-pollinated F(2 )individual. The proportion of resistant plants among these F(3) families varied from 0 to 67%. Bulked segregant analysis did not reveal any markers linked with resistance and, therefore, a linkage map with 81 markers was created. A locus that accounted for 18.4% of the variation in resistance to white rust was mapped to linkage group (LG) 2 near the RAPD marker Z19a. During the study, a bacterial resistance gene homologous to Arabidopsis RPS2 and six different RGAs were sequenced. RPS2 and five of the RGAs were mapped to linkage groups LG1, LG4 and LG9. Unfortunately, none of the RGAs could be shown to be associated with white rust resistance.

  20. The dynamic growth exhibition and accumulation of cadmium of Pak choi (Brassica campestris L. ssp. chinensis) grown in contaminated soils.

    PubMed

    Lai, Hung-Yu; Chen, Bo-Ching

    2013-10-25

    The accumulation of heavy metals, especially cadmium (Cd), in leafy vegetables was compared with other vegetables. Pak choi (Brassica campestris L. ssp. chinensis) is a leafy vegetable consumed in Taiwan and its safety for consumption after growing in contaminated soils is a public concern. A pot experiment (50 days) was conducted to understand the dynamic accumulation of Cd by pak choi grown in artificially contaminated soils. The edible parts of pak choi were sampled and analyzed every 2-3 days. The dry weight (DW) of pak choi was an exponential function of leaf length, leaf width, and chlorophyll content. The accumulation of Cd increased when the soil Cd concentration was raised, but was kept at a constant level during different growth stages. Pak choi had a high bioconcentration factor (BCF = ratio of the concentration in the edible parts to that in the soils), at values of 3.5-4.0. The consumption of pak choi grown in soils contaminated at levels used in this study would result in the ingestion of impermissible amounts of Cd and could possibly have harmful effects on health.

  1. HrpM is involved in glucan biosynthesis, biofilm formation and pathogenicity in Xanthomonas citri ssp. citri.

    PubMed

    Malamud, Florencia; Conforte, Valeria P; Rigano, Luciano A; Castagnaro, Atilio P; Marano, María Rosa; Morais do Amaral, Alexandre; Vojnov, Adrian A

    2012-12-01

    Xanthomonas citri ssp. citri (Xcc) is the causal agent of citrus canker. This bacterium develops a characteristic biofilm on both biotic and abiotic surfaces. A biofilm-deficient mutant was identified in a screening of a transposon mutagenesis library of the Xcc 306 strain constructed using the commercial Tn5 transposon EZ-Tn5 Tnp Transposome (Epicentre). Sequence analysis of a mutant obtained in the screening revealed that a single copy of the EZ-Tn5 was inserted at position 446 of hrpM, a gene encoding a putative enzyme involved in glucan synthesis. We demonstrate for the first time that the product encoded by the hrpM gene is involved in β-1,2-glucan synthesis in Xcc. A mutation in hrpM resulted in no disease symptoms after 4 weeks of inoculation in lemon and grapefruit plants. The mutant also showed reduced ability to swim in soft agar and decreased resistance to H(2)O(2) in comparison with the wild-type strain. All defective phenotypes were restored to wild-type levels by complementation with the plasmid pBBR1-MCS containing an intact copy of the hrpM gene and its promoter. These results indicate that the hrpM gene contributes to Xcc growth and adaptation in its host plant. PMID:22738424

  2. Sea Buckthorn (Hippophaë rhamnoides ssp. rhamnoides) Berries in Nordic Environment: Compositional Response to Latitude and Weather Conditions.

    PubMed

    Zheng, Jie; Kallio, Heikki; Yang, Baoru

    2016-06-22

    Flavonol glycosides (FGs) in sea buckthorn (Hippophaë rhamnoides ssp. rhamnoides) berries of varieties 'Tytti' and 'Terhi', cultivated in northern Finland (68°02' N) for six years and southern Finland (60°23' N) for seven years, were investigated and compared by HPLC-DAD-ESI-MS/MS. The average total content of 23 identified glycosides of isorhamnetin and quercetin was 103 ± 23 and 110 ± 21 mg/100 g fresh berries in 'Terhi' and 'Tytti', respectively. The total contents of FGs, flavonol diglycosides, and triglycosides in both varieties were higher in the north than in the south, whereas total flavonol monoglycoside content behaved vice versa (p < 0.05). Among the 89 weather variables studied, the sum of the daily mean temperatures that are 5 °C or higher from the start of growth season until the day of harvest was the most important variable which associated negatively with the accumulation of FGs in berries. Such influence was much stronger in berries from the north than from the south.

  3. Evolutionary change in flowering phenology in the iteroparous herb Beta vulgaris ssp. maritima: a search for the underlying mechanisms.

    PubMed

    Van Dijk, Henk

    2009-01-01

    The potential for evolutionary change in flowering time has gained considerable attention in view of the current global climate change. To explore this potential and its underlying mechanisms in the iteroparous perennial Beta vulgaris ssp. maritima (sea beet), artificial selection for earlier and later flowering date was applied under semi-natural greenhouse conditions. Mean flowering date occurred more than 30 d earlier in 13 generations in the early selection line, but response was weaker in the late selection line. Taking advantage of the growing knowledge on the genetics and the physiology of flowering induction, particularly in Arabidopsis thaliana, the results obtained here were analysed in terms of the four different pathways of flowering induction known in this species. A first significant correlated response was stem elongation (bolting) in the vegetative stage, suggesting that plants were thus able to flower earlier as long as other requirements were satisfied. Vernalization had a clear influence on flowering date and its influence increased during the selection process, together with sensitivity to photoperiod. Vernalization and photoperiod could compensate for each other: each additional week of vernalization at 5 degrees C decreased the necessary daylength for flowering by about 15 min during the later selection stages, while in unselected plants, it was about 7 min. Devernalizing effects were observed at short days combined with higher temperatures. Special attention was given to the role of the B (bolting) gene that cancels the vernalization requirement. The results here obtained suggest that all four known pathways may simultaneously participate in evolutionary change.

  4. Transcriptomic profiling of the salt stress response in excised leaves of the halophyte Beta vulgaris ssp. maritima.

    PubMed

    Skorupa, Monika; Gołębiewski, Marcin; Domagalski, Krzysztof; Kurnik, Katarzyna; Abu Nahia, Karim; Złoch, Michał; Tretyn, Andrzej; Tyburski, Jarosław

    2016-02-01

    Beta vulgaris ssp. maritima is a halophytic relative of cultivated beets. In the present work a transcriptome response to acute salt stress imposed to excised leaves of sea beet was investigated. Salt treatments consisted of adding NaCl directly to the transpiration stream by immersing the petioles of excised leaves into the salt solutions. Sequencing libraries were generated from leaves subjected to either moderate or strong salt stress. Control libraries were constructed from untreated leaves. Sequencing was performed using the Illumina MiSeq platform. We obtained 32970 unigenes by assembling the pooled reads from all the libraries with Trinity software. Screening the nr database returned 18,362 sequences with functional annotation. Using the reference transcriptome we identified 1,246 genes that were differentially expressed after 48 h of NaCl stress. Genes related to several cellular functions such as membrane transport, osmoprotection, molecular chaperoning, redox metabolism or protein synthesis were differentially expressed in response to salt stress. The response of sea beet leaves to salt treatments was marked out by transcriptomic up-regulation of genes related to photosynthetic carbon fixation, ribosome biogenesis, cell wall-building and cell wall expansion. Furthermore, several novel and undescribed transcripts were responsive to salinity in leaves of sea beet.

  5. HrpM is involved in glucan biosynthesis, biofilm formation and pathogenicity in Xanthomonas citri ssp. citri.

    PubMed

    Malamud, Florencia; Conforte, Valeria P; Rigano, Luciano A; Castagnaro, Atilio P; Marano, María Rosa; Morais do Amaral, Alexandre; Vojnov, Adrian A

    2012-12-01

    Xanthomonas citri ssp. citri (Xcc) is the causal agent of citrus canker. This bacterium develops a characteristic biofilm on both biotic and abiotic surfaces. A biofilm-deficient mutant was identified in a screening of a transposon mutagenesis library of the Xcc 306 strain constructed using the commercial Tn5 transposon EZ-Tn5 Tnp Transposome (Epicentre). Sequence analysis of a mutant obtained in the screening revealed that a single copy of the EZ-Tn5 was inserted at position 446 of hrpM, a gene encoding a putative enzyme involved in glucan synthesis. We demonstrate for the first time that the product encoded by the hrpM gene is involved in β-1,2-glucan synthesis in Xcc. A mutation in hrpM resulted in no disease symptoms after 4 weeks of inoculation in lemon and grapefruit plants. The mutant also showed reduced ability to swim in soft agar and decreased resistance to H(2)O(2) in comparison with the wild-type strain. All defective phenotypes were restored to wild-type levels by complementation with the plasmid pBBR1-MCS containing an intact copy of the hrpM gene and its promoter. These results indicate that the hrpM gene contributes to Xcc growth and adaptation in its host plant.

  6. TP0262 is a modulator of promoter activity of tpr Subfamily II genes of Treponema pallidum ssp. pallidum.

    PubMed

    Giacani, Lorenzo; Godornes, Charmie; Puray-Chavez, Maritza; Guerra-Giraldez, Cristina; Tompa, Martin; Lukehart, Sheila A; Centurion-Lara, Arturo

    2009-06-01

    Transcriptional regulation in Treponema pallidum ssp. pallidum is poorly understood, primarily because this organism cannot be cultivated in vitro or genetically manipulated. We have recently shown a phase variation mechanism controlling transcription initiation of Subfamily II tpr (T. pallidumrepeat) genes (tprE, tprG and tprJ), a group of virulence factor candidates. Furthermore, the same study suggested that additional mechanisms might influence the level of transcription of these tprs. The T. pallidum genome sequence has revealed a few open reading frames with similarity to known bacterial transcription factors, including four catabolite activator protein homologues. In this work, sequences matching the Escherichia coli cAMP receptor protein (CRP) binding motif were identified in silico upstream of tprE, tprG and tprJ. Using elecrophoretic mobility shift assay and DNaseI footprinting assay, recombinant TP0262, a T. pallidum CRP homologue, was shown to bind specifically to amplicons obtained from the tpr promoters containing putative CRP binding motifs. Using a heterologous reporter system, binding of TP0262 to these promoters was shown to either increase (tprE and tprJ) or decrease (tprG) tpr promoter activity. This is the first characterization of a T. pallidum transcriptional modulator that influences tpr promoter activity.

  7. [Outbreak of subclinical mastitis due to beta hemolytic group L streptococci (S. dysgalactiae ssp. equisimilis) in an Austrian dairy herd].

    PubMed

    Baumgartner, Martina; Giffinger, Friederike; Hoppe, Jan Christoph; Spergser, Joachim

    2011-01-01

    This study is reporting an outbreak of subclinical mastitis due to beta-hemolytic group L streptococci in an Austrian dairy herd with a history of high somatic cell count. At the first survey 16 of 33 lactating cows (28 quarters of 132) were cultured positive for beta-hemolytic, CAMP and esculin negative cocci that grew on Columbia blood agar with small grey catalase negative colonies. With the commercial API 20 Strep system (bioMerieux, F) isolates were classified as members of streptococci group L. All tested strains (eight of 28) produced acid from ribose, lactose, trehalose, amidon and glycogen; they hydrolysed hippurate and showed beta-glucuronidase, beta-galactosidase, alkaline phosphatase, leucinaminopeptidase and arginindehydrolase activity. Isolates were sensitive to bacitracin but resistant to tetracycline. Using phenotypic characterisation as well as sequence analysis of the 16S-23S intergenic spacer region of a representative strain, recovered isolates were identified as Streptococcus (S.) dysgalactiae ssp. equisimilis. Mastitis was characterized by normal milk secretions and absence of clinical abnormalities but high elevations of somatic cell count. Based on the characteristics of the strains and on the observations during the first herd survey, contagious transmission during milking as a result of poor milking hygiene was assumed. The mastitis was controlled through implementation of a strict hygiene protocol including use of single-use udder towels, post milking teat desinfection and cluster disinfection between milking cows in combination with antibiotic treatment of infected udders. PMID:22059292

  8. The maize (Zea mays ssp. mays var. B73) genome encodes 33 members of the purple acid phosphatase family

    PubMed Central

    González-Muñoz, Eliécer; Avendaño-Vázquez, Aida-Odette; Montes, Ricardo A. Chávez; de Folter, Stefan; Andrés-Hernández, Liliana; Abreu-Goodger, Cei; Sawers, Ruairidh J. H.

    2015-01-01

    Purple acid phosphatases (PAPs) play an important role in plant phosphorus nutrition, both by liberating phosphorus from organic sources in the soil and by modulating distribution within the plant throughout growth and development. Furthermore, members of the PAP protein family have been implicated in a broader role in plant mineral homeostasis, stress responses and development. We have identified 33 candidate PAP encoding gene models in the maize (Zea mays ssp. mays var. B73) reference genome. The maize Pap family includes a clear single-copy ortholog of the Arabidopsis gene AtPAP26, shown previously to encode both major intracellular and secreted acid phosphatase activities. Certain groups of PAPs present in Arabidopsis, however, are absent in maize, while the maize family contains a number of expansions, including a distinct radiation not present in Arabidopsis. Analysis of RNA-sequencing based transcriptome data revealed accumulation of maize Pap transcripts in multiple plant tissues at multiple stages of development, and increased accumulation of specific transcripts under low phosphorus availability. These data suggest the maize PAP family as a whole to have broad significance throughout the plant life cycle, while highlighting potential functional specialization of individual family members. PMID:26042133

  9. Identification of surface-associated proteins of Bifidobacterium animalis ssp. lactis KLDS 2.0603 by enzymatic shaving.

    PubMed

    Zhu, Dequan; Sun, Yu; Liu, Fei; Li, Aili; Yang, Limei; Meng, Xiang-Chen

    2016-07-01

    Bifidobacteria are commensal microorganisms of the human and animal intestinal tract, and their surface proteins can mediate bacterial communication and chemical sensing in the environment, as well as facilitate interactions between bacteria and the host. However, a systematic study of the outer surface-associated proteome of bifidobacteria has not been undertaken. In the present study, the proteins located on the surface of Bifidobacterium animalis ssp. lactis KLDS 2.0603 were systematically identified by a nongel proteomic approach, which consisted of the shaving of the bacterial surface with trypsin and an analysis of the released peptides by liquid chromatography-tandem mass spectrometry. A total of 105 surface-associated proteins were found, of which 15 proteins could potentially be involved in adhesion and interactions between bifidobacteria and the host. The proteins related to adhesion and interaction between bacteria and the host include pilus structure proteins (Fim A, Fim B), 10 moonlighting proteins, an NLP/P60 family protein, an immunogenic secreted protein, and a putative sugar-binding secreted protein. The results provide the basis for future studies on the molecular mechanisms of the interactions between bifidobacteria and the host. PMID:27132091

  10. A suite of genetic markers useful in assessing wildcat (Felis silvestris ssp.)-domestic cat (Felis silvestris catus) admixture.

    PubMed

    Driscoll, Carlos; Yamaguchi, Nobuyuki; O'Brien, Stephen J; Macdonald, David W

    2011-01-01

    The wildcat (Felis silvestris ssp.) is a conservation concern largely due to introgressive hybridization with its congener F. s. catus, the common domestic cat. Because of a recent divergence and entirely overlapping ranges, hybridization is common and pervasive between these taxa threatening the genetic integrity of remaining wildcat populations. Identifying pure wildcats for inclusion in conservation programs using current morphological discriminants is difficult because of gross similarity between them and the domestic, critically hampering conservation efforts. Here, we present a vetted panel of microsatellite loci and mitochondrial polymorphisms informative for each of the 5 naturally evolved wildcat subspecies and the derived domestic cat. We also present reference genotypes for each assignment class. Together, these marker sets and corresponding reference genotypes allow for the development of a genetic rational for defining "units of conservation" within a phylogenetically based taxonomy of the entire F. silvestris species complex. We anticipate this marker panel will allow conservators to assess genetic integrity and quantify admixture in managed wildcat populations and to be a starting point for more in-depth analysis of hybridization.

  11. Gene expression profiling of PBMCs from Holstein and Jersey cows sub-clinically infected with Mycobacterium avium ssp. paratuberculosis.

    PubMed

    Verschoor, Chris P; Pant, Sameer D; You, Qiumei; Kelton, David F; Karrow, Niel A

    2010-09-15

    Infection of calves with intracellular Mycobacterium avium ssp. paratuberculosis (MAP) commonly results in a granulomatous, chronic inflammatory bowel disease known as Johne's disease. The asymptomatic stage of this infection can persist for the entire production life of an adult cow, resulting in reduced performance and premature culling, as well as transmission of MAP to progeny and herd-mates. It has been previously shown that the gene expression profiles of peripheral blood mononuclear cells (PBMCs) of healthy cows, and those chronically infected with MAP are inherently different, and that these changes may be indicative of disease progression. Since resistance to MAP infection is a heritable trait, and has been proposed to differ amongst domestic dairy cattle breeds, the objective of the present study was to compare gene expression profiles of PBMCs from healthy adult Holstein and Jersey cows to those considered to be sub-clinically infected with MAP, as indicated by serum ELISA. Microarray analysis using a platform containing more than 10,000 probes and ontological analysis identified differences in gene expression between a) healthy and infected cows, including genes involved in the inflammatory response, and calcium binding, and b) infected Holsteins and Jerseys, including genes involved in the immune response, and antigen processing and presentation. These results suggest a mixed pro- and anti-inflammatory phenotype of PBMCs from MAP-infected as compared to healthy control animals, and inherently different levels of immune and inflammatory-related gene expression between MAP-infected Holsteins and Jerseys.

  12. Genome-wide identification and characterization of aquaporin genes (AQPs) in Chinese cabbage (Brassica rapa ssp. pekinensis).

    PubMed

    Tao, Peng; Zhong, Xinmin; Li, Biyuan; Wang, Wuhong; Yue, Zhichen; Lei, Juanli; Guo, Weiling; Huang, Xiaoyun

    2014-12-01

    Aquaporins (AQPs) are members of a superfamily of integral membrane proteins and play a significant role in the transportation of small molecules across membranes. However, currently little is known about the AQP genes in Chinese cabbage (Brassica rapa ssp. pekinensis). In this study, a genome-wide analysis was carried out to identify the AQP genes in Chinese cabbage. In total, 53 non-redundant AQP genes were identified that were located on all of the 10 chromosomes. The number of AQP genes in Chinese cabbage was greater than in Arabidopsis. They were classified into four subfamilies, including PIP, TIP, NIP, and SIP. Thirty-three groups of AQP orthologous genes were identified between Chinese cabbage and Arabidopsis, but orthologs corresponding to AtNIP1;1 and AtPIP2;8 were not detected. Seventeen groups of paralogous genes were identified in Chinese cabbage. Three-dimensional models of the AQPs of Chinese cabbage were constructed using Phyre2, and ar/R selectivity filters were analyzed comparatively between Chinese cabbage and Arabidopsis. Generally, gene structure was conserved within each subfamily, especially in the SIP subfamily. Intron loss events have occurred during the evolution of the PIP, TIP, and NIP subfamilies. The expression of AQP genes in Chinese cabbage was analyzed in different organs. Most AQP genes were downregulated in response to salt stress. This work shows that the AQP genes of Chinese cabbage have undergone triplication and subsequent biased gene loss.

  13. Genome-wide survey and expression analysis of the PUB family in Chinese cabbage (Brassica rapa ssp. pekinesis).

    PubMed

    Wang, Cheng; Duan, Weike; Riquicho, Ali RamuliMaquina; Jing, Zange; Liu, Tongkun; Hou, Xilin; Li, Ying

    2015-12-01

    U-box proteins are widely distributed among eukaryotic organisms and show a higher prevalence in plants than in other organisms. Plant U-box (PUB) proteins play crucial regulatory roles in various developmental and physiological processes. Previously, 64 and 77 PUB genes have been identified in Arabidopsis (Arabidopsis thaliana) and rice (Oryza sativa), respectively. In this study, 101 putative PUB genes were identified in the Chinese cabbage (Brassica rapa ssp. pekinensis line Chiifu-401-42) genome and compared with other 15 representative plants. By specific protein domains and a phylogenetic analysis, the B. rapa PUB (BrPUB) gene family was subdivided into 10 groups. Localization of BrPUB genes showed an uneven distribution on the ten chromosomes of B. rapa. The orthologous and co-orthologous PUB gene pairs were identified between B. rapa and A. thaliana. RNA-seq transcriptome data of different tissues revealed tissue-specific and differential expression profiles of the BrPUBs, and quantitative real-time PCR analysis showed inverse gene expression patterns of the BrPUB-ARMs in response to cold and heat stresses. Altogether, the identification, classification, phylogenetic analysis, chromosome distribution, conserved motifs, and expression patterns of BrPUBs were predicted and analysed. Importantly, this study of BrPUBs provides a rich resource that will aid in the determination of PUB functions in plant development.

  14. Ogura-CMS in Chinese cabbage (Brassica rapa ssp. pekinensis) causes delayed expression of many nuclear genes.

    PubMed

    Dong, Xiangshu; Kim, Wan Kyu; Lim, Yong-Pyo; Kim, Yeon-Ki; Hur, Yoonkang

    2013-02-01

    We investigated the mechanism regulating cytoplasmic male sterility (CMS) in Brassica rapa ssp. pekinensis using floral bud transcriptome analyses of Ogura-CMS Chinese cabbage and its maintainer line in B. rapa 300-K oligomeric probe (Br300K) microarrays. Ogura-CMS Chinese cabbage produced few and infertile pollen grains on indehiscent anthers. Compared to the maintainer line, CMS plants had shorter filaments and plant growth, and delayed flowering and pollen development. In microarray analysis, 4646 genes showed different expression, depending on floral bud size, between Ogura-CMS and its maintainer line. We found 108 and 62 genes specifically expressed in Ogura-CMS and its maintainer line, respectively. Ogura-CMS line-specific genes included stress-related, redox-related, and B. rapa novel genes. In the maintainer line, genes related to pollen coat and germination were specifically expressed in floral buds longer than 3mm, suggesting insufficient expression of these genes in Ogura-CMS is directly related to dysfunctional pollen. In addition, many nuclear genes associated with auxin response, ATP synthesis, pollen development and stress response had delayed expression in Ogura-CMS plants compared to the maintainer line, which is consistent with the delay in growth and development of Ogura-CMS plants. Delayed expression may reduce pollen grain production and/or cause sterility, implying that mitochondrial, retrograde signaling delays nuclear gene expression.

  15. Codon optimisation to improve expression of a Mycobacterium avium ssp. paratuberculosis-specific membrane-associated antigen by Lactobacillus salivarius.

    PubMed

    Johnston, Christopher; Douarre, Pierre E; Soulimane, Tewfik; Pletzer, Daniel; Weingart, Helge; MacSharry, John; Coffey, Aidan; Sleator, Roy D; O'Mahony, Jim

    2013-06-01

    Subunit and DNA-based vaccines against Mycobacterium avium ssp. paratuberculosis (MAP) attempt to overcome inherent issues associated with whole-cell formulations. However, these vaccines can be hampered by poor expression of recombinant antigens from a number of disparate hosts. The high G+C content of MAP invariably leads to a codon bias throughout gene expression. To investigate if the codon bias affects recombinant MAP antigen expression, the open reading frame of a MAP-specific antigen MptD (MAP3733c) was codon optimised for expression against a Lactobacillus salivarius host. Of the total 209 codons which constitute MAP3733c, 172 were modified resulting in a reduced G+C content from 61% for the native gene to 32.7% for the modified form. Both genes were placed under the transcriptional control of the PnisA promoter; allowing controlled heterologous expression in L. salivarius. Expression was monitored using fluorescence microscopy and microplate fluorometry via GFP tags translationally fused to the C-termini of the two MptD genes. A > 37-fold increase in expression was observed for the codon-optimised MAP3733synth variant over the native gene. Due to the low cost and improved expression achieved, codon optimisation significantly improves the potential of L. salivarius as an oral vaccine stratagem against Johne's disease. PMID:23620276

  16. The maize (Zea mays ssp. mays var. B73) genome encodes 33 members of the purple acid phosphatase family.

    PubMed

    González-Muñoz, Eliécer; Avendaño-Vázquez, Aida-Odette; Montes, Ricardo A Chávez; de Folter, Stefan; Andrés-Hernández, Liliana; Abreu-Goodger, Cei; Sawers, Ruairidh J H

    2015-01-01

    Purple acid phosphatases (PAPs) play an important role in plant phosphorus nutrition, both by liberating phosphorus from organic sources in the soil and by modulating distribution within the plant throughout growth and development. Furthermore, members of the PAP protein family have been implicated in a broader role in plant mineral homeostasis, stress responses and development. We have identified 33 candidate PAP encoding gene models in the maize (Zea mays ssp. mays var. B73) reference genome. The maize Pap family includes a clear single-copy ortholog of the Arabidopsis gene AtPAP26, shown previously to encode both major intracellular and secreted acid phosphatase activities. Certain groups of PAPs present in Arabidopsis, however, are absent in maize, while the maize family contains a number of expansions, including a distinct radiation not present in Arabidopsis. Analysis of RNA-sequencing based transcriptome data revealed accumulation of maize Pap transcripts in multiple plant tissues at multiple stages of development, and increased accumulation of specific transcripts under low phosphorus availability. These data suggest the maize PAP family as a whole to have broad significance throughout the plant life cycle, while highlighting potential functional specialization of individual family members. PMID:26042133

  17. Phenotype definition is a main point in genome-wide association studies for bovine Mycobacterium avium ssp. paratuberculosis infection status.

    PubMed

    Küpper, J; Brandt, H; Donat, K; Erhardt, G

    2014-10-01

    Paratuberculosis caused by Mycobacterium avium ssp. paratuberculosis (MAP) causes economic losses and is present in dairy herds worldwide. Different studies used different diagnostic tests to detect infection status and are the basis of genome-wide association (GWA) studies with inconsistent results. Therefore, the aim of this study was to identify and compare genomic regions associated with MAP susceptibility in the same cohort of cattle using different diagnostic tests. The GWA study was performed in German Holsteins within a case-control assay using 305 cows tested for MAP by fecal culture and additional with four different commercial ELISA-tests. Genotyping was performed with the Illumina Bovine SNP50 BeadChip. The results using fecal culture or ELISA test led to the identification of different genetic loci. Two single-nucleotide polymorphisms showed significant association with the ELISA-status. However, no significant association for MAP infection could be confirmed. Our results show that the definition of the MAP-phenotype has an important impact on the outcome of GWA studies for paratuberculosis.

  18. Transcriptomic profiling of the salt stress response in excised leaves of the halophyte Beta vulgaris ssp. maritima.

    PubMed

    Skorupa, Monika; Gołębiewski, Marcin; Domagalski, Krzysztof; Kurnik, Katarzyna; Abu Nahia, Karim; Złoch, Michał; Tretyn, Andrzej; Tyburski, Jarosław

    2016-02-01

    Beta vulgaris ssp. maritima is a halophytic relative of cultivated beets. In the present work a transcriptome response to acute salt stress imposed to excised leaves of sea beet was investigated. Salt treatments consisted of adding NaCl directly to the transpiration stream by immersing the petioles of excised leaves into the salt solutions. Sequencing libraries were generated from leaves subjected to either moderate or strong salt stress. Control libraries were constructed from untreated leaves. Sequencing was performed using the Illumina MiSeq platform. We obtained 32970 unigenes by assembling the pooled reads from all the libraries with Trinity software. Screening the nr database returned 18,362 sequences with functional annotation. Using the reference transcriptome we identified 1,246 genes that were differentially expressed after 48 h of NaCl stress. Genes related to several cellular functions such as membrane transport, osmoprotection, molecular chaperoning, redox metabolism or protein synthesis were differentially expressed in response to salt stress. The response of sea beet leaves to salt treatments was marked out by transcriptomic up-regulation of genes related to photosynthetic carbon fixation, ribosome biogenesis, cell wall-building and cell wall expansion. Furthermore, several novel and undescribed transcripts were responsive to salinity in leaves of sea beet. PMID:26795151

  19. A novel suicide shuttle plasmid for Streptococcus suis serotype 2 and Streptococcus equi ssp. zooepidemicus gene mutation

    PubMed Central

    Liu, Rui; Zhang, Ping; Su, Yiqi; Lin, Huixing; Zhang, Hui; Yu, Lei; Ma, Zhe; Fan, Hongjie

    2016-01-01

    The mariner-based Himar1 system has been utilized for creating mutant libraries of many Gram-positive bacteria. Streptococcus suis serotype 2 (SS2) and Streptococcus equi ssp. zooepidemicus (SEZ) are primary pathogens of swine that threaten the swine industry in China. To provide a forward-genetics technology for finding virulent phenotype-related genes in these two pathogens, we constructed a novel temperature-sensitive suicide shuttle plasmid, pMar4s, which contains the Himar1 system transposon, TnYLB-1, and the Himar1 C9 transposase from pMarA and the repTAs temperature-sensitive fragment from pSET4s. The kanamycin (Kan) resistance gene was in the TnYLB-1 transposon. Temperature sensitivity and Kan resistance allowed the selection of mutant strains and construction of the mutant library. The SS2 and SEZ mutant libraries were successfully constructed using the pMar4s plasmid. Inverse-Polymerase Chain Reaction (Inverse-PCR) results revealed large variability in transposon insertion sites and that the library could be used for phenotype alteration screening. The thiamine biosynthesis gene apbE was screened for its influence on SS2 anti-phagocytosis; likewise, the sagF gene was identified to be a hemolytic activity-related gene in SEZ. pMar4s was suitable for mutant library construction, providing more information regarding SS2 and SEZ virulence factors and illustrating the pathogenesis of swine streptococcosis. PMID:27256117

  20. Sea Buckthorn (Hippophaë rhamnoides ssp. rhamnoides) Berries in Nordic Environment: Compositional Response to Latitude and Weather Conditions.

    PubMed

    Zheng, Jie; Kallio, Heikki; Yang, Baoru

    2016-06-22

    Flavonol glycosides (FGs) in sea buckthorn (Hippophaë rhamnoides ssp. rhamnoides) berries of varieties 'Tytti' and 'Terhi', cultivated in northern Finland (68°02' N) for six years and southern Finland (60°23' N) for seven years, were investigated and compared by HPLC-DAD-ESI-MS/MS. The average total content of 23 identified glycosides of isorhamnetin and quercetin was 103 ± 23 and 110 ± 21 mg/100 g fresh berries in 'Terhi' and 'Tytti', respectively. The total contents of FGs, flavonol diglycosides, and triglycosides in both varieties were higher in the north than in the south, whereas total flavonol monoglycoside content behaved vice versa (p < 0.05). Among the 89 weather variables studied, the sum of the daily mean temperatures that are 5 °C or higher from the start of growth season until the day of harvest was the most important variable which associated negatively with the accumulation of FGs in berries. Such influence was much stronger in berries from the north than from the south. PMID:27215398

  1. The Dynamic Growth Exhibition and Accumulation of Cadmium of Pak Choi (Brassica campestris L. ssp. chinensis) Grown in Contaminated Soils

    PubMed Central

    Lai, Hung-Yu; Chen, Bo-Ching

    2013-01-01

    The accumulation of heavy metals, especially cadmium (Cd), in leafy vegetables was compared with other vegetables. Pak choi (Brassica campestris L. ssp. chinensis) is a leafy vegetable consumed in Taiwan and its safety for consumption after growing in contaminated soils is a public concern. A pot experiment (50 days) was conducted to understand the dynamic accumulation of Cd by pak choi grown in artificially contaminated soils. The edible parts of pak choi were sampled and analyzed every 2–3 days. The dry weight (DW) of pak choi was an exponential function of leaf length, leaf width, and chlorophyll content. The accumulation of Cd increased when the soil Cd concentration was raised, but was kept at a constant level during different growth stages. Pak choi had a high bioconcentration factor (BCF = ratio of the concentration in the edible parts to that in the soils), at values of 3.5–4.0. The consumption of pak choi grown in soils contaminated at levels used in this study would result in the ingestion of impermissible amounts of Cd and could possibly have harmful effects on health. PMID:24284350

  2. Esterase SeE of Streptococcus equi ssp. equi is a Novel Non-specific Carboxylic Ester Hydrolase

    PubMed Central

    Xie, Gang; Liu, Mengyao; Zhu, Hui; Lei, Benfang

    2009-01-01

    Extracellular carboxylic ester hydrolases are produced by many bacterial pathogens and have been shown recently to be important for virulence of some pathogens. However, these hydrolases are poorly characterized in enzymatic activity. This study prepared and characterized the secreted ester hydrolase of Streptococcus equi ssp. equi (designated SeE for S. equi esterase). SeE hydrolyzes ethyl acetate, acetylsalicylic acid, and tributyrin but not ethyl butyrate. This substrate specificity pattern does not match those of the three conventional types of non-specific carboxylic ester hydrolases (carboxylesterases, arylesterases, and acetylesterases). To determine whether SeE has lipase activity, a number of triglycerides and vinyl esters were tested in SeE-catalyzed hydrolysis. SeE does not hydrolyze triglycerides and vinyl esters of long chain carboxylic acids nor display interfacial activation, indicating that SeE is not a lipase. Like the conventional carboxylesterases, SeE is inhibited by diisopropylfluorophosphate. These findings indicate that SeE is a novel non-specific carboxylic ester hydrolase that has broader substrate specificity than the conventional carboxylesterases. PMID:19054107

  3. A novel suicide shuttle plasmid for Streptococcus suis serotype 2 and Streptococcus equi ssp. zooepidemicus gene mutation.

    PubMed

    Liu, Rui; Zhang, Ping; Su, Yiqi; Lin, Huixing; Zhang, Hui; Yu, Lei; Ma, Zhe; Fan, Hongjie

    2016-01-01

    The mariner-based Himar1 system has been utilized for creating mutant libraries of many Gram-positive bacteria. Streptococcus suis serotype 2 (SS2) and Streptococcus equi ssp. zooepidemicus (SEZ) are primary pathogens of swine that threaten the swine industry in China. To provide a forward-genetics technology for finding virulent phenotype-related genes in these two pathogens, we constructed a novel temperature-sensitive suicide shuttle plasmid, pMar4s, which contains the Himar1 system transposon, TnYLB-1, and the Himar1 C9 transposase from pMarA and the repTAs temperature-sensitive fragment from pSET4s. The kanamycin (Kan) resistance gene was in the TnYLB-1 transposon. Temperature sensitivity and Kan resistance allowed the selection of mutant strains and construction of the mutant library. The SS2 and SEZ mutant libraries were successfully constructed using the pMar4s plasmid. Inverse-Polymerase Chain Reaction (Inverse-PCR) results revealed large variability in transposon insertion sites and that the library could be used for phenotype alteration screening. The thiamine biosynthesis gene apbE was screened for its influence on SS2 anti-phagocytosis; likewise, the sagF gene was identified to be a hemolytic activity-related gene in SEZ. pMar4s was suitable for mutant library construction, providing more information regarding SS2 and SEZ virulence factors and illustrating the pathogenesis of swine streptococcosis. PMID:27256117

  4. Genome-wide survey and expression analysis of the PUB family in Chinese cabbage (Brassica rapa ssp. pekinesis).

    PubMed

    Wang, Cheng; Duan, Weike; Riquicho, Ali RamuliMaquina; Jing, Zange; Liu, Tongkun; Hou, Xilin; Li, Ying

    2015-12-01

    U-box proteins are widely distributed among eukaryotic organisms and show a higher prevalence in plants than in other organisms. Plant U-box (PUB) proteins play crucial regulatory roles in various developmental and physiological processes. Previously, 64 and 77 PUB genes have been identified in Arabidopsis (Arabidopsis thaliana) and rice (Oryza sativa), respectively. In this study, 101 putative PUB genes were identified in the Chinese cabbage (Brassica rapa ssp. pekinensis line Chiifu-401-42) genome and compared with other 15 representative plants. By specific protein domains and a phylogenetic analysis, the B. rapa PUB (BrPUB) gene family was subdivided into 10 groups. Localization of BrPUB genes showed an uneven distribution on the ten chromosomes of B. rapa. The orthologous and co-orthologous PUB gene pairs were identified between B. rapa and A. thaliana. RNA-seq transcriptome data of different tissues revealed tissue-specific and differential expression profiles of the BrPUBs, and quantitative real-time PCR analysis showed inverse gene expression patterns of the BrPUB-ARMs in response to cold and heat stresses. Altogether, the identification, classification, phylogenetic analysis, chromosome distribution, conserved motifs, and expression patterns of BrPUBs were predicted and analysed. Importantly, this study of BrPUBs provides a rich resource that will aid in the determination of PUB functions in plant development. PMID:26054324

  5. Genome-wide identification and characterization of aquaporin genes (AQPs) in Chinese cabbage (Brassica rapa ssp. pekinensis).

    PubMed

    Tao, Peng; Zhong, Xinmin; Li, Biyuan; Wang, Wuhong; Yue, Zhichen; Lei, Juanli; Guo, Weiling; Huang, Xiaoyun

    2014-12-01

    Aquaporins (AQPs) are members of a superfamily of integral membrane proteins and play a significant role in the transportation of small molecules across membranes. However, currently little is known about the AQP genes in Chinese cabbage (Brassica rapa ssp. pekinensis). In this study, a genome-wide analysis was carried out to identify the AQP genes in Chinese cabbage. In total, 53 non-redundant AQP genes were identified that were located on all of the 10 chromosomes. The number of AQP genes in Chinese cabbage was greater than in Arabidopsis. They were classified into four subfamilies, including PIP, TIP, NIP, and SIP. Thirty-three groups of AQP orthologous genes were identified between Chinese cabbage and Arabidopsis, but orthologs corresponding to AtNIP1;1 and AtPIP2;8 were not detected. Seventeen groups of paralogous genes were identified in Chinese cabbage. Three-dimensional models of the AQPs of Chinese cabbage were constructed using Phyre2, and ar/R selectivity filters were analyzed comparatively between Chinese cabbage and Arabidopsis. Generally, gene structure was conserved within each subfamily, especially in the SIP subfamily. Intron loss events have occurred during the evolution of the PIP, TIP, and NIP subfamilies. The expression of AQP genes in Chinese cabbage was analyzed in different organs. Most AQP genes were downregulated in response to salt stress. This work shows that the AQP genes of Chinese cabbage have undergone triplication and subsequent biased gene loss. PMID:24972664

  6. Antioxidant and antimicrobial activities of essential oils obtained from oregano (Origanum vulgare ssp. hirtum) by using different extraction methods.

    PubMed

    Karakaya, Sibel; El, Sedef Nehir; Karagözlü, Nural; Sahin, Serpil

    2011-06-01

    In this study, antioxidant and antimicrobial activities of essential oils obtained from oregano (Origanum vulgare ssp. hirtum) were determined by using solvent-free microwave extraction (SFME), supercritical fluid extraction, and conventional hydrodistillation (CH) methods. The inhibitory effects on the 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) radical of essential oils obtained from oregano by using SFME and CH were similar. However, essential oil extracted by CH showed greater (2.69 μmol/μL of oil) Trolox equivalent antioxidant capacity (TEAC) than oregano oils obtained by SFME (P < .05). The difference between percentage inhibition and TEAC values most probably is due to the fact that undiluted and diluted samples are used in the percentage inhibition assay and the TEAC assay, respectively. TEAC values of oregano essential oils obtained by SFME at different microwave power levels were found to be similar and ranged from 0.72 to 0.84 μmol/μL of oil. Essential oils obtained by CH and SFME at different microwave powers inhibited the survival of Listeria monocytogenes, Salmonella typhimurium, and Escherichia coli O157:H7, whereas survival of Staphylococcus aureus was not influenced. In addition, oregano oil obtained by SFME at 40% power level did not show any inhibitory effect on E. coli O157:H7.

  7. A comprehensive analysis of flowering transition in Agapanthus praecox ssp. orientalis (Leighton) Leighton by using transcriptomic and proteomic techniques.

    PubMed

    Zhang, Di; Ren, Li; Yue, Jian-Hua; Wang, Ling; Zhuo, Li-Huan; Shen, Xiao-Hui

    2013-03-27

    Comprehensive transcriptomic and proteomic analyses were performed to gain further understanding of the molecular mechanisms of floral initiation in Agapanthus praecox ssp. orientalis. Samples of stem apexes were collected at three different time points including the vegetative, induced, and reproductive period. A total of 374 transcript-derived fragments and 72 proteins showed significant differential expression between the samples. The largest proportion of the identified genes and proteins are involved in metabolism, followed by signal transduction, protein fate, cellular transport, and biogenesis of cellular components. A large number of these genes and proteins were upregulated during the induced and reproductive stages. Their expression profiles demonstrate that carbohydrate metabolism provides nutrients foundation for floral initiation in Agapanthus. Furthermore, a transcription factors GAI (GA insensitive protein) that negatively regulates gibberellin signaling, auxin receptor protein TIR1 (Transport inhibitor response 1), a key enzyme of ethylene biosynthesis SAMS (S-adenosylmethionine synthase), and ethylene receptor protein ETR were isolated and identified. Expression patterns of these proteins, in combination with the results of quantitative phytohormone and immunolocalization analyses, indicated that GA, indole-acetic acid (IAA), and ethylene regulate floral morphogenesis and flowering. In conclusion, these data provide novel insight into the early regulatory steps of flowering in Agapanthus. PMID:23333928

  8. [Outbreak of subclinical mastitis due to beta hemolytic group L streptococci (S. dysgalactiae ssp. equisimilis) in an Austrian dairy herd].

    PubMed

    Baumgartner, Martina; Giffinger, Friederike; Hoppe, Jan Christoph; Spergser, Joachim

    2011-01-01

    This study is reporting an outbreak of subclinical mastitis due to beta-hemolytic group L streptococci in an Austrian dairy herd with a history of high somatic cell count. At the first survey 16 of 33 lactating cows (28 quarters of 132) were cultured positive for beta-hemolytic, CAMP and esculin negative cocci that grew on Columbia blood agar with small grey catalase negative colonies. With the commercial API 20 Strep system (bioMerieux, F) isolates were classified as members of streptococci group L. All tested strains (eight of 28) produced acid from ribose, lactose, trehalose, amidon and glycogen; they hydrolysed hippurate and showed beta-glucuronidase, beta-galactosidase, alkaline phosphatase, leucinaminopeptidase and arginindehydrolase activity. Isolates were sensitive to bacitracin but resistant to tetracycline. Using phenotypic characterisation as well as sequence analysis of the 16S-23S intergenic spacer region of a representative strain, recovered isolates were identified as Streptococcus (S.) dysgalactiae ssp. equisimilis. Mastitis was characterized by normal milk secretions and absence of clinical abnormalities but high elevations of somatic cell count. Based on the characteristics of the strains and on the observations during the first herd survey, contagious transmission during milking as a result of poor milking hygiene was assumed. The mastitis was controlled through implementation of a strict hygiene protocol including use of single-use udder towels, post milking teat desinfection and cluster disinfection between milking cows in combination with antibiotic treatment of infected udders.

  9. Regulation of bolting and identification of the α-tubulin gene family in Brassica rapa L. ssp pekinensis.

    PubMed

    Zhang, Y W; Jin, D; Xu, C; Zhang, L; Guo, M H; Fang, Z Y

    2016-01-01

    Microtubules are important components of eukaryotic cells, and they play vital roles in cell morphogenesis, carrying of signaling molecules, transport of materials, and establishing the cell polarity. During bolting of biennial plants, cell division and elongation are involved, and cell elongation inevitably involves the microtubules arrangement and expression of related genes. So we deduce that it is of great significance to figure out the mechanism of bolting and flowering in which TUA genes are involved. In the present study, bioinformatic methods were used to predict and identify the α-tubulin gene family (BrTUAs) in Brassica rapa L. ssp pekinensis (Chinese cabbage) through the alignment of AtTUA gene sequence from Arabidopsis thaliana with the B. rapa genome database (http://brassicadb.org/brad/) using the basic local alignment search tool. The change in the structure and functions of BrTUAs during the process of evolution, cis-acting elements in the promoter sequences of BrTUAs, and the expression of the identified genes was also analyzed. Twelve members of the α-tubulin gene family were identified from Chinese cabbage. The gene length, intron, exon, and promoter regions were determined to have changed significantly during the genome evolution. Only five of the 12 members were encoded completely and were observed to differ in their spatial and temporal expression. The five BrTUA promoter sequences contained different numbers of cis-elements responsive to light and low-temperature response, cis-elements responsive among which hormonal responses were significantly different. We also report that the BrTUAs were involved in the regulation of the bolting in Chinese cabbage, and propose that this process could be controlled by regulating the expression of BrTUAs.

  10. Molecular cloning and stress-dependent regulation of potassium channel gene in Chinese cabbage (Brassica rapa ssp. Pekinensis).

    PubMed

    Zhang, Yidong; Wang, Zeyun; Zhang, Lida; Cao, Youfang; Huang, Danfeng; Tang, Kexuan

    2006-09-01

    Potassium channels are important for many physiological functions in plants, one of which is to regulate plant adaption to stress conditions. In this study, KCT2, the gene encoding a membrane-bound protein potassium channel (GenBank accession number: ), was isolated from Chinese cabbage (Brassica rapa ssp. Pekinensis) by RACE-PCR technique. Bioinformatics methods were performed for the gene structure and molecular similarity analysis. The KCT2 expression patterns under various stress conditions were studied by semi-quantitative RT-PCR. DNA gel blot was used to analyze genomic organization. The putative KCT2 was found to contain five membrane-spanning segments, a pore-forming domain (P-domain) between the last two transmembrane spans, a TxxTxGYGD motif in the P-domain and a putative cyclic nucleotide-binding-like domain within a long C-terminal region. KCT2 is closest to KAT2 in Arabidopsis. KCT2 could be a one-copy gene with different isoforms or belong to a small gene family with four or five members. KCT2 was expressed more strongly in leaves than in shoots and roots. KCT2 transcription products were up-regulated by a 4-h-incubation in abscisic acid (ABA) and various stress treatment including cold stress (4 degrees C) for 24 h, drought stress for 1h, and salt stress for 12 h. KCT2 transcription was not affected by anoxia stress for 8h and was down-regulated with cold stress for 48 h. KCT2 was cloned for the first time from the genus Brassica. Expression analysis indicated that in the early stage of plant adaption to stress conditions KCT2 is up-regulated, which results in a stimulation of potassium transport.

  11. The construction of a genetic linkage map of non-heading Chinese cabbage (Brassica campestris ssp. chinensis Makino).

    PubMed

    Cheng, Yan; Geng, Jianfeng; Zhang, Jingyi; Wang, Qian; Ban, Qingyu; Hou, Xilin

    2009-08-01

    Non-heading Chinese cabbage (Brassica campestris ssp. chinensis Makino) is one of the most important vegetables in eastern China. A genetic linkage map was constructed using 127 doubled haploid (DH) lines, and the DH population was derived from a commercial hybrid "Hanxiao" (lines SW-13 x L-118). Out of the 614 polymorphic markers, 43.49% were not assigned to any of the linkage groups(LGs). Chi-square tests showed that 42.67% markers were distorted from expected Mendelian segregation ratios, and the direction of distorted segregation was mainly toward the paternal parent L-118. After sequentially removing the markers that had an interval distance smaller than 1 cM from the upper marker, the overall quality of the linkage map was increased. Two hundred and sixty-eight molecular markers were mapped into 10 LGs, which were anchored to the corresponding chromosome of the B. rapa reference map based on common simple sequence repeat (SSR) markers. The map covers 973.38 cM of the genome and the average interval distance between markers was 3.63 cM. The number of markers on each LG ranged from 18 (R08) to 64 (R07), with an average interval distance within a single LG from 1.70 cM (R07) to 6.71 cM (R06). Among these mapped markers, 169 were sequence-related amplified polymorphism (SRAP) molecular markers, 50 were SSR markers and 49 were random amplification polymorphic DNA (RAPD) markers. With further saturation to the LG, the current map offers a genetic tool for loci analysis for important agronomic traits.

  12. Enhancement of tolerance to soft rot disease in the transgenic Chinese cabbage (Brassica rapa L. ssp. pekinensis) inbred line, Kenshin.

    PubMed

    Vanjildorj, Enkhchimeg; Song, Seo Young; Yang, Zhi Hong; Choi, Jae Eul; Noh, Yoo Sun; Park, Suhyoung; Lim, Woo Jin; Cho, Kye Man; Yun, Han Dae; Lim, Yong Pyo

    2009-10-01

    We developed a transgenic Chinese cabbage (Brassica rapa L. ssp. pekinensis) inbred line, Kenshin, with high tolerance to soft rot disease. Tolerance was conferred by expression of N-acyl-homoserine lactonase (AHL-lactonase) in Chinese cabbage through an efficient Agrobacterium-mediated transformation method. To synthesize and express the AHL-lactonase in Chinese cabbage, the plant was transformed with the aii gene (AHL-lactonase gene from Bacillus sp. GH02) fused to the PinII signal peptide (protease inhibitor II from potato). Five transgenic lines were selected by growth on hygromycin-containing medium (3.7% transformation efficiency). Southern blot analysis showed that the transgene was stably integrated into the genome. Among these five transgenic lines, single copy number integrations were observed in four lines and a double copy number integration was observed in one transgenic line. Northern blot analysis confirmed that pinIISP-aii fusion gene was expressed in all the transgenic lines. Soft rot disease tolerance was evaluated at tissue and seedling stage. Transgenic plants showed a significantly enhanced tolerance (2-3-fold) to soft rot disease compared to wild-type plants. Thus, expression of the fusion gene pinIISP-aii reduces susceptibility to soft rot disease in Chinese cabbage. We conclude that the recombinant AHL-lactonase, encoded by aii, can effectively quench bacterial quorum-sensing and prevent bacterial population density-dependent infections. To the best of our knowledge, the present study is the first to demonstrate the transformation of Chinese cabbage inbred line Kenshin, and the first to describe the effect of the fusion gene pinIISP-aii on enhancement of soft rot disease tolerance.

  13. Ethyl Acetate Extract of Origanum vulgare L. ssp. hirtum Prevents Streptozotocin-Induced Diabetes in C57BL/6 Mice.

    PubMed

    Vujicic, Milica; Nikolic, Ivana; Kontogianni, Vassiliki G; Saksida, Tamara; Charisiadis, Pantelis; Vasic, Bobana; Stosic-Grujicic, Stanislava; Gerothanassis, Ioannis P; Tzakos, Andreas G; Stojanovic, Ivana

    2016-07-01

    Type 1 diabetes (T1D) is an autoimmune disease that develops as a consequence of pancreatic β-cell death induced by proinflammatory mediators. Because Origanum vulgare L. ssp. hirtum (Greek oregano) contains antiinflammatory molecules, we hypothesized that it might be beneficial for the treatment of T1D. An ethyl acetate extract of oregano (EAO) was prepared from the leaves by a polar extraction method. Phytochemical composition was determined by liquid chromatography-UV diode array coupled to ion-trap mass spectrometry with electrospray ionization interface (LC/DAD/ESI-MS(n) ). In vitro immunomodulatory effect of EAO was estimated by measuring proliferation (MTT) or cytokine secretion (ELISA) from immune cells. Diabetes was induced by multiple low doses of streptozotocin (MLDS) in male C57BL/6 mice and EAO was administered intraperitoneally for 10 d. Determination of cellular composition (flow cytometry) and cytokine production (ELISA) was performed on 12th d after diabetes induction. EAO suppressed the function of both macrophages and lymphocytes in vitro. In vivo, EAO treatment significantly preserved pancreatic islets and reduced diabetes incidence in MLDS-challenged mice. Besides down-modulatory effect on macrophages, EAO reduced the number of total CD4(+) and activated CD4(+) CD25(+) T cells. Furthermore, EAO affected the number of T helper 1 (Th1) and T helper 17 (Th17) cells through downregulation of their key transcription factors T-bet and RORγT. Because EAO treatment protects mice from development of hyperglycemia by reducing proinflammatory macrophage/Th1/Th17 response, this plant extract could represent a basis for future diabetes therapy. PMID:27219840

  14. The effects of progressing and nonprogressing Mycobacterium avium ssp. paratuberculosis infection on milk production in dairy cows.

    PubMed

    Smith, Rebecca L; Gröhn, Y T; Pradhan, A K; Whitlock, R H; Van Kessel, J S; Smith, J M; Wolfgang, D R; Schukken, Y H

    2016-02-01

    Longitudinal data from 3 commercial dairy herds in the northeast United States, collected from 2004 to 2011, were analyzed to determine the effect of Mycobacterium avium ssp. paratuberculosis (MAP) infection status and progression path on milk production. Disease status, as indicated by MAP test results, was determined through quarterly ELISA serum testing, biannual fecal culture, and culture of tissues and feces at slaughter. Milk production data were collected from the Dairy Herd Information Association. Animals with positive MAP test results were categorized, based on test results over the full course of the study, as high path (at least one high-positive culture) or low path (at least one positive culture or ELISA). The cumulative numbers of positive ELISA and culture results were recorded. The effects of both MAP infection path, status, and number of positive tests on milk production were analyzed using a mixed linear model with an autocorrelation random effect structure. Low- and high-path animals produced more milk before their first positive test than always-negative animals, especially high-path animals. Although mean production decreased after a first positive test, low-path animals were shown to recover some productivity. High-path animals continued to exhibit a decrease in milk production, especially after their first high-positive fecal culture. These results show that not all animals that test positive for MAP will have long-term production losses. Milk production decreased significantly with each additional positive test. Ultimately, production loss appeared to be a function of MAP infection progression. PMID:26686721

  15. Laboratory and field evaluation of Teknar HP-D, a biolarvicidal formulation of Bacillus thuringiensis ssp. israelensis, against mosquito vectors.

    PubMed

    Gunasekaran, K; Doss, P S Boopathi; Vaidyanathan, K

    2004-10-01

    Larvicidal efficacy of Teknar HP-D, an improved biolarvicidal formulation of Bacillus thuringiensis ssp. israelensis (Bti), against Anopheles stephensi, Culex quinquefasciatus and Aedes aegypti was determined in the laboratory, and in field the efficacy of the formulation was tested against Cx. quinquefasciatus breeding in cesspits, unused wells and drains. The toxicity of the formulation to Gambusia affinis (larvivorous fish), Notonecta sp. and Diplonychus indicus (water bugs) was also evaluated in the laboratory. Teknar HP-D was field tested at three recommended dosages, 1, 1.5 and 2l/ha, selecting five habitats for each dosage. Another five habitats were kept untreated as controls. Ae. aegypti showed greatest susceptibility to the Bti toxin in the laboratory. In cesspits, all the three dosages caused >80% reduction of pupal recruitment up to day 6 post-treatment, indicating that a weekly application at the lowest would be necessary for sustained control. The residual activity of the formulation was longer in unused wells, causing >80% reduction of pupal recruitment for 17 days from the day of treatment. In controlling pupal recruitment the three dosages produced equal effect. Application of Teknar HP-D at 1 l/ha once in three weeks is therefore recommended to control Cx. quinquefasciatus in unused wells. However, in drains, >80% reduction of pupal recruitment was observed for only 3 days and hence, application of Teknar HP-D at 2 l/ha that caused significantly higher level of reduction twice in a week at 3-day interval is necessary. At dosages from 0.032 to 3.2 mg/l (ppm), Teknar HP-D was non-toxic to Gambusia fish. The two predatory water bugs, Notonecta sp. and Diplonychus indicus that fed on the surviving larvae of Cx. quinquefasciatus exposed to the sub-lethal doses (LC(50) and LC(80)) of Teknar HP-D were safe with out having any mortality.

  16. Plant phenolic acids affect the virulence of Pectobacterium aroidearum and P. carotovorum ssp. brasiliense via quorum sensing regulation.

    PubMed

    Joshi, Janak Raj; Burdman, Saul; Lipsky, Alexander; Yariv, Shaked; Yedidia, Iris

    2016-05-01

    Several studies have reported effects of the plant phenolic acids cinnamic acid (CA) and salicylic acid (SA) on the virulence of soft rot enterobacteria. However, the mechanisms involved in these processes are not yet fully understood. Here, we investigated whether CA and SA interfere with the quorum sensing (QS) system of two Pectobacterium species, P. aroidearum and P. carotovorum ssp. brasiliense, which are known to produce N-acyl-homoserine lactone (AHL) QS signals. Our results clearly indicate that both phenolic compounds affect the QS machinery of the two species, consequently altering the expression of bacterial virulence factors. Although, in control treatments, the expression of QS-related genes increased over time, the exposure of bacteria to non-lethal concentrations of CA or SA inhibited the expression of QS genes, including expI, expR, PC1_1442 (luxR transcriptional regulator) and luxS (a component of the AI-2 system). Other virulence genes known to be regulated by the QS system, such as pecS, pel, peh and yheO, were also down-regulated relative to the control. In agreement with the low levels of expression of expI and expR, CA and SA also reduced the level of the AHL signal. The effects of CA and SA on AHL signalling were confirmed in compensation assays, in which exogenous application of N-(β-ketocaproyl)-l-homoserine lactone (eAHL) led to the recovery of the reduction in virulence caused by the two phenolic acids. Collectively, the results of gene expression studies, bioluminescence assays, virulence assays and compensation assays with eAHL clearly support a mechanism by which CA and SA interfere with Pectobacterium virulence via the QS machinery.

  17. Effects of biological and methodological factors on volatile organic compound patterns during cultural growth of Mycobacterium avium ssp. paratuberculosis.

    PubMed

    Küntzel, Anne; Fischer, Sina; Bergmann, Andreas; Oertel, Peter; Steffens, Markus; Trefz, Phillip; Miekisch, Wolfram; Schubert, Jochen K; Reinhold, Petra; Köhler, Heike

    2016-01-01

    Mycobacterium avium ssp. paratuberculosis (MAP) causes chronic granulomatous enteritis in ruminants. Bacterial growth is still the diagnostic 'gold standard', but is very time consuming. MAP-specific volatile organic compounds (VOCs) above media could accelerate cultural diagnosis. The aim of this project was to assess the kinetics of a VOC profile linked to the growth of MAP in vitro. The following sources of variability were taken into account: five different culture media, three different MAP strains, inoculation with different bacterial counts, and different periods of incubation. Needle-trap microextraction was employed for pre-concentration of VOCs, and gas chromatography-mass spectrometry for subsequent analysis. All volatiles were identified and calibrated by analysing pure references at different concentration levels. More than 100 VOCs were measured in headspaces above MAP-inoculated and control slants. Results confirmed different VOC profiles above different culture media. Emissions could be assigned to either egg-containing media or synthetic ingredients. 43 VOCs were identified as potential biomarkers of MAP growth on Herrold's Egg Yolk Medium without significant differences between the tree MAP strains. Substances belonged to the classes of alcohols, aldehydes, esters, ketones, aliphatic and aromatic hydrocarbons. With increasing bacterial density the VOC concentrations above MAP expressed different patterns: the majority of substances increased (although a few decreased after reaching a peak), but nine VOCs clearly decreased. Data support the hypotheses that (i) bacteria emit different metabolites on different culture media; (ii) different MAP strains show uniform VOC patterns; and (iii) cultural diagnosis can be accelerated by taking specific VOC profiles into account. PMID:27604146

  18. Impact of biologically synthesized silver nanoparticles on the growth and physiological responses in Brassica rapa ssp. pekinensis.

    PubMed

    Baskar, Venkidasamy; Venkatesh, Jelli; Park, Se Won

    2015-11-01

    Silver nanoparticles (AgNPs) were extensively used in various fields, particularly in medicine as an antimicrobial agent. The unavoidable and extensive usage of AgNPs in turn accumulates in the environment. Plants are the essential base of ecosystem and are ready to disturb by environmental pollutants. Therefore, in the present study, we have planned to evaluate the impact of biologically synthesized AgNPs on the essential food crop Chinese cabbage (Brassica rapa ssp. pekinensis). The effects of AgNP-induced plant morphological and physiological changes were investigated in different concentrations (100, 250, and 500 mg/L). The results of morphological features showed that AgNPs at lower concentrations (100 mg/L) exhibit growth-stimulating activity, whereas at higher concentrations (250 and 500 mg/L), particularly, 500 mg/L exhibited growth-suppressing activities which are in terms of reduced root, shoot growth, and fresh biomass. The increased reactive oxygen species (ROS) generation, malondialdehyde production, anthocyanin biosynthesis, and decreased chlorophyll content were also more obviously present at higher concentrations of AgNPs. The concentration-dependent DNA damage was observed in the AgNP-treated plants. The molecular responses of AgNPs indicate that most of the genes related to secondary metabolism (glucosinolates, anthocyanin) and antioxidant activities were induced at higher concentrations of AgNP treatment. The dose-dependent phytotoxicity effects of AgNPs were also observed. Taken together, the highest concentration of AgNPs (500 mg/L) could induce growth-suppressing activities via the induction of ROS generation and other molecular changes in B. rapa seedlings.

  19. Genetic mapping of stem rust resistance gene Sr13 in tetraploid wheat (Triticum turgidum ssp. durum L.)

    PubMed Central

    Simons, Kristin; Abate, Zewdie; Chao, Shiaoman; Zhang, Wenjun; Rouse, Matt; Jin, Yue; Elias, Elias; Dubcovsky, Jorge

    2016-01-01

    Wheat stem rust caused by Puccinia graminis f. sp. tritici, can cause significant yield losses. To combat the disease breeders have deployed resistance genes both individually and in combinations to increase resistance durability. A new race, TTKSK (Ug99), identified in Uganda in 1999, is virulent on most of the resistance genes currently deployed, and is rapidly spreading to other regions of the world. It is therefore important to identify, map, and deploy resistance genes that are still effective against TTKSK. One of these resistance genes, Sr13, was previously assigned to the long arm of chromosome 6A, but its precise map location was not known. In this study, the genome location of Sr13 was determined in four tetraploid wheat (T. turgidum ssp. durum) mapping populations involving the TTKSK resistant varieties Kronos, Kofa, Medora and Sceptre. Our results showed that resistance was linked to common molecular markers in all four populations, suggesting that these durum lines carry the same resistance gene. Based on its chromosome location and infection types against different races of stem rust, this gene is postulated to be Sr13. Sr13 was mapped within a 1.2 to 2.8 cM interval (depending on the mapping population) between EST markers CD926040 and BE471213, which corresponds to a 285-kb region in rice chromosome 2, and a 3.1-Mb region in Brachypodium chromosome 3. These maps will be the foundation for developing high-density maps, identifying diagnostic markers, and positional cloning of Sr13. PMID:20857083

  20. Genome-wide identification, classification, and expression analysis of sHSP genes in Chinese cabbage (Brassica rapa ssp pekinensis).

    PubMed

    Tao, P; Guo, W L; Li, B Y; Wang, W H; Yue, Z C; Lei, J L; Zhong, X M

    2015-10-05

    Small heat shock proteins (sHSPs) are essential for the plant's normal development and stress responses, especially the heat stress response. The information regarding sHSP genes in Chinese cabbage (Brassica rapa ssp pekinensis) is sparse, hence we performed a genome-wide analysis to identify sHSP genes in this species. We identified 26 non-redundant sHSP genes distributed on all chromosomes, except chromosome A7, with one additional sHSP gene identified from an expressed sequence tag library. Chinese cabbage was found to contain more sHSP genes than Arabidopsis. The 27 sHSP genes were classified into 11 subfamilies. We identified 22 groups of sHSP syntenic orthologous genes between Chinese cabbage and Arabidopsis. In addition, eight groups of paralogous genes were uncovered in Chinese cabbage. Protein structures of the 27 Chinese cabbage sHSPs were modeled using Phyre2, which revealed that all of them contain several conserved β strands across different subfamilies. In general, gene structure was conserved within each subfamily between Chinese cabbage and Arabidopsis, except for peroxisome sHSP. Analysis of promoter motifs showed that most sHSP genes contain heat shock elements or variants. We also found that biased gene loss has occurred during the evolution of the sHSP subfamily in Chinese cabbage. Expression analysis indicated that the greatest transcript abundance of most Chinese cabbage sHSP genes was found in siliques and early cotyledon embryos. Thus, genome-wide identification and characterization of sHSP genes is a first and important step in the investigation of sHSPs in Chinese cabbage.

  1. Genotypic differences in nitrate uptake, translocation and assimilation of two Chinese cabbage cultivars [Brassica campestris L. ssp. Chinensis (L.)].

    PubMed

    Tang, Yafang; Sun, Xuecheng; Hu, Chengxiao; Tan, Qiling; Zhao, Xiaohu

    2013-09-01

    A hydroponic trial was conducted to investigate genotypic differences in nitrate uptake, translocation and assimilation between low nitrate accumulator L18 and high accumulator H96 of Chinese cabbage [Brassica campestris L. ssp. Chinensis (L.)]. The results suggested that H96 could uptake more nitrate than L18 in the root but lower transport inside leaf cells and assimilation in the leaf. It was showed that root morphology parameters - length, surface area and volume of H96 were 18.0%, 31.6% and 46.5% higher than L18. Nitrate transporters NRT1.1 and NRT2.1 transcription levels were 41.6% and 269.6% higher than those of L18 respectively. NRT1.1 and NRT2.1 expression amount in leaf blade of two cultivars were opposite to in the root, L18 NRT1.1 and NRT2.1 expressions were 279.2% and 80.0% higher than H96. In addition, nitrate assimilation capacity of L18 was significantly higher than H96 in leaf. It was showed that Nitrate Reductase (NR; EC 1.7.1.1) activity and NIA expression level of L18 leaf were 234 0.4% and 105.4% higher than those of H96, Glutamine Synthetase (GS; EC 6.3.1.2) activity, Gln1 and Gln2 expression levels in the leaf blade of L18 were 43.9%, 331.5% and 124.8% higher than those of H96 respectively. Nitrate assimilation products-Glu, total amino acid, soluble protein content in the leaf of L18 were all significantly higher than those of H96. The results above suggested that nitrate accumulation differences were due to differential capacities to uptake, mechanisms for nitrate transport in leaves and assimilate nitrate. Comparing contribution of three aspects in nitrate accumulation, translocation and assimilation were contributed more in low nitrate concentration in the leaf blade.

  2. High-throughput sequencing discovery of conserved and novel microRNAs in Chinese cabbage (Brassica rapa L. ssp. pekinensis).

    PubMed

    Wang, Fengde; Li, Libin; Liu, Lifeng; Li, Huayin; Zhang, Yihui; Yao, Yingyin; Ni, Zhongfu; Gao, Jianwei

    2012-07-01

    MicroRNAs (miRNAs) are a class of 21-24 nucleotide non-coding RNAs that down-regulate gene expression by cleaving or inhibiting the translation of target gene transcripts. miRNAs have been extensively analyzed in a few model plant species such as Arabidopsis, rice and Populus, and partially investigated in other non-model plant species. However, only a few conserved miRNAs have been identified in Chinese cabbage, a common and economically important crop in Asia. To identify novel and conserved miRNAs in Chinese cabbage (Brassica rapa L. ssp. pekinensis) we constructed a small RNA library. Using high-throughput Solexa sequencing to identify microRNAs we found 11,210 unique sequences belonging to 321 conserved miRNA families and 228 novel miRNAs. We ran a Blast search with these sequences against the Chinese cabbage mRNA database and found 2,308 and 736 potential target genes for 221 conserved and 125 novel miRNAs, respectively. The BlastX search against the Arabidopsis genome and GO analysis suggested most of the targets were involved in plant growth, metabolism, development and stress response. This study provides the first large scale-cloning and characterization of Chinese cabbage miRNAs and their potential targets. These miRNAs add to the growing database of new miRNAs, prompt further study on Chinese cabbage miRNA regulation mechanisms, and help toward a greater understanding of the important roles of miRNAs in Chinese cabbage.

  3. ANALYSIS OF GENOMIC DNA METHYLATION AND GENE EXPRESSION IN CHINESE CABBAGE (Brassica rapa L. ssp. pekinensis) AFTER CONTINUOUS SEEDLING BREEDING.

    PubMed

    Tao, L; Wang, X L; Guo, M H; Zhang, Y W

    2015-08-01

    Vernalization plays a key role in the bolting and flowering of Chinese cabbage (Brassica rapa L. ssp. pekinensis). Plants can switch from vegetative to reproductive growth and then bolt and flower under low temperature induction. The economic benefits of Chinese cabbage will decline significantly when the bolting happens before the vegetative body fully grows due to a lack of the edible value. It was found that continuous seedling breeding reduced the heading of Chinese cabbage and led to bolt and flower more easily. In the present study, two inbred lines, termed A161 and A105, were used as experiment materials. These two lines were subjected to vernalization and formed four types: seeds-seedling breeding once, seedling breeding twice, seedling breeding thrice and normal type. Differences in plant phenotype were compared. DNA methylation analysis was performed based on MSAP method. The differential fragments were cloned and analyzed by qPCR. Results showed that plants after seedling breeding thrice had a loosen heading leaves, elongated center axis and were easier to bolt and flower. It is suggested that continuous seedling breeding had a weaker winterness. It was observed that genome methylation level decreased with increasing generation. Four differential genes were identified, short for BraAPC1, BraEMP3, BraUBC26, and BraAL5. Fluorescent qPCR analysis showed that expression of four genes varied at different reproduction modes and different vernalization time. It is indicated that these genes might be involve in the development and regulation of bolting and flowering of plants. Herein, the molecular mechanism that continuous seedling breeding caused weaker winterness was analyzed preliminarily. It plays an important guiding significance for Chinese cabbage breeding.

  4. Regulation of bolting and identification of the α-tubulin gene family in Brassica rapa L. ssp pekinensis.

    PubMed

    Zhang, Y W; Jin, D; Xu, C; Zhang, L; Guo, M H; Fang, Z Y

    2016-01-01

    Microtubules are important components of eukaryotic cells, and they play vital roles in cell morphogenesis, carrying of signaling molecules, transport of materials, and establishing the cell polarity. During bolting of biennial plants, cell division and elongation are involved, and cell elongation inevitably involves the microtubules arrangement and expression of related genes. So we deduce that it is of great significance to figure out the mechanism of bolting and flowering in which TUA genes are involved. In the present study, bioinformatic methods were used to predict and identify the α-tubulin gene family (BrTUAs) in Brassica rapa L. ssp pekinensis (Chinese cabbage) through the alignment of AtTUA gene sequence from Arabidopsis thaliana with the B. rapa genome database (http://brassicadb.org/brad/) using the basic local alignment search tool. The change in the structure and functions of BrTUAs during the process of evolution, cis-acting elements in the promoter sequences of BrTUAs, and the expression of the identified genes was also analyzed. Twelve members of the α-tubulin gene family were identified from Chinese cabbage. The gene length, intron, exon, and promoter regions were determined to have changed significantly during the genome evolution. Only five of the 12 members were encoded completely and were observed to differ in their spatial and temporal expression. The five BrTUA promoter sequences contained different numbers of cis-elements responsive to light and low-temperature response, cis-elements responsive among which hormonal responses were significantly different. We also report that the BrTUAs were involved in the regulation of the bolting in Chinese cabbage, and propose that this process could be controlled by regulating the expression of BrTUAs. PMID:26909938

  5. Impact of biologically synthesized silver nanoparticles on the growth and physiological responses in Brassica rapa ssp. pekinensis.

    PubMed

    Baskar, Venkidasamy; Venkatesh, Jelli; Park, Se Won

    2015-11-01

    Silver nanoparticles (AgNPs) were extensively used in various fields, particularly in medicine as an antimicrobial agent. The unavoidable and extensive usage of AgNPs in turn accumulates in the environment. Plants are the essential base of ecosystem and are ready to disturb by environmental pollutants. Therefore, in the present study, we have planned to evaluate the impact of biologically synthesized AgNPs on the essential food crop Chinese cabbage (Brassica rapa ssp. pekinensis). The effects of AgNP-induced plant morphological and physiological changes were investigated in different concentrations (100, 250, and 500 mg/L). The results of morphological features showed that AgNPs at lower concentrations (100 mg/L) exhibit growth-stimulating activity, whereas at higher concentrations (250 and 500 mg/L), particularly, 500 mg/L exhibited growth-suppressing activities which are in terms of reduced root, shoot growth, and fresh biomass. The increased reactive oxygen species (ROS) generation, malondialdehyde production, anthocyanin biosynthesis, and decreased chlorophyll content were also more obviously present at higher concentrations of AgNPs. The concentration-dependent DNA damage was observed in the AgNP-treated plants. The molecular responses of AgNPs indicate that most of the genes related to secondary metabolism (glucosinolates, anthocyanin) and antioxidant activities were induced at higher concentrations of AgNP treatment. The dose-dependent phytotoxicity effects of AgNPs were also observed. Taken together, the highest concentration of AgNPs (500 mg/L) could induce growth-suppressing activities via the induction of ROS generation and other molecular changes in B. rapa seedlings. PMID:26154034

  6. Genome-wide identification, classification, and expression analysis of sHSP genes in Chinese cabbage (Brassica rapa ssp pekinensis).

    PubMed

    Tao, P; Guo, W L; Li, B Y; Wang, W H; Yue, Z C; Lei, J L; Zhong, X M

    2015-01-01

    Small heat shock proteins (sHSPs) are essential for the plant's normal development and stress responses, especially the heat stress response. The information regarding sHSP genes in Chinese cabbage (Brassica rapa ssp pekinensis) is sparse, hence we performed a genome-wide analysis to identify sHSP genes in this species. We identified 26 non-redundant sHSP genes distributed on all chromosomes, except chromosome A7, with one additional sHSP gene identified from an expressed sequence tag library. Chinese cabbage was found to contain more sHSP genes than Arabidopsis. The 27 sHSP genes were classified into 11 subfamilies. We identified 22 groups of sHSP syntenic orthologous genes between Chinese cabbage and Arabidopsis. In addition, eight groups of paralogous genes were uncovered in Chinese cabbage. Protein structures of the 27 Chinese cabbage sHSPs were modeled using Phyre2, which revealed that all of them contain several conserved β strands across different subfamilies. In general, gene structure was conserved within each subfamily between Chinese cabbage and Arabidopsis, except for peroxisome sHSP. Analysis of promoter motifs showed that most sHSP genes contain heat shock elements or variants. We also found that biased gene loss has occurred during the evolution of the sHSP subfamily in Chinese cabbage. Expression analysis indicated that the greatest transcript abundance of most Chinese cabbage sHSP genes was found in siliques and early cotyledon embryos. Thus, genome-wide identification and characterization of sHSP genes is a first and important step in the investigation of sHSPs in Chinese cabbage. PMID:26505345

  7. Using Small-Scale Studies to Prioritize Threats and Guide Recovery of a Rare Hemiparasitic Plant: Cordylanthus rigidus ssp. littoralis

    PubMed Central

    Watts, Sean M.; Uhl, Melissa M.; Maurano, Stephen P.; Nuccio, Erin E.

    2010-01-01

    Background Recovering endangered species would benefit from identifying and ranking of the factors that threaten them. Simply managing for multiple positive influences will often aid in recovery; however, the relative impacts of multiple threats and/or interactions among them are not always predictable. We used a series of experiments and quantitative observational studies to examine the importance of five potential limiting factors to the abundance of a state-listed endangered hemiparasitic annual forb, Cordylanthus rigidus ssp. littoralis (C.r.l., California, USA): host availability, mammalian herbivores, insect seed predators, fire suppression, and exotic species. While this initial assessment is certainly not a complete list, these factors stem from direct observation and can inform provisional recommendations for management and further research. Methodology and Principal Findings Studies were conducted at five sites and included assessments of the influence of host availability, exotic species, exclusion of mammalian herbivores and insect seed predators on C.r.l. productivity, and simulated effects of fire on seed germination. C.r.l. was limited by multiple threats: individuals with access to host species produced up to three times more inflorescences than those lacking hosts, mammalian herbivory reduced C.r.l. size and fecundity by more than 50% and moth larvae reduced seed production by up to 40%. Litter deposition and competition from exotic plant species also appears to work in conjunction with other factors to limit C.r.l. throughout its life cycle. Conclusions and Significance The work reported here highlights the contribution that a series of small-scale studies can make to conservation and restoration. Taken as a whole, the results can be used immediately to inform current management and species recovery strategies. Recovery of C.r.l. will require management that addresses competition with exotic plant species, herbivore pressure, and availability of

  8. Seed germination ecology of the annual grass Leptochloa panicea ssp. mucronata and a comparison with L. panicoides and L. fusca

    NASA Astrophysics Data System (ADS)

    Baskin, Carol C.; Baskin, Jerry M.; Chester, Edward W.

    1999-09-01

    Leptochloa panicea ssp. mucronata is an annual grass that grows in relatively dry habitats. Requirements for dormancy loss and germination were determined for seeds of this species and compared to those of two species from wet habitats. Seeds of L. panicea were dormant at maturity in autumn, but when exposed to actual or simulated autumn temperatures (e.g. 20/10, 15/6 °C), they entered conditional dormancy and thus germinated to high percentages in light at 35/20 °C. Seeds buried in non-flooded soil exposed to natural seasonal temperature changes in Kentucky (USA) were non-dormant by the following summer and germinated to 80-100 % in light at 25/15, 30/15 and 35/20 °C. Seeds buried in non-flooded soil exhibited an annual conditional dormancy/non-dormancy cycle, with seeds mostly germinating to 80-100 % in light at 30/15 and 35/20 °C throughout the year but to 80-100 % in light at 25/15 °C only in summer. Results for L. panicea were compared to published data for L. panicoides and L. fusca. Whereas seeds of L. panicea buried in flooded soil failed to come out of dormancy, those of L. panicoides, an annual of moist habitats such as mudflats, exhibited an annual conditional dormancy/non-dormancy cycle, and those of L. fusca, a semi-aquatic, required flooding for both dormancy loss and germination. Differences in dormancy breaking and germination responses of seeds of Leptochloa species may help to explain why this genus occupies a wide range of habitats with regard to soil moisture conditions.

  9. Ethyl Acetate Extract of Origanum vulgare L. ssp. hirtum Prevents Streptozotocin-Induced Diabetes in C57BL/6 Mice.

    PubMed

    Vujicic, Milica; Nikolic, Ivana; Kontogianni, Vassiliki G; Saksida, Tamara; Charisiadis, Pantelis; Vasic, Bobana; Stosic-Grujicic, Stanislava; Gerothanassis, Ioannis P; Tzakos, Andreas G; Stojanovic, Ivana

    2016-07-01

    Type 1 diabetes (T1D) is an autoimmune disease that develops as a consequence of pancreatic β-cell death induced by proinflammatory mediators. Because Origanum vulgare L. ssp. hirtum (Greek oregano) contains antiinflammatory molecules, we hypothesized that it might be beneficial for the treatment of T1D. An ethyl acetate extract of oregano (EAO) was prepared from the leaves by a polar extraction method. Phytochemical composition was determined by liquid chromatography-UV diode array coupled to ion-trap mass spectrometry with electrospray ionization interface (LC/DAD/ESI-MS(n) ). In vitro immunomodulatory effect of EAO was estimated by measuring proliferation (MTT) or cytokine secretion (ELISA) from immune cells. Diabetes was induced by multiple low doses of streptozotocin (MLDS) in male C57BL/6 mice and EAO was administered intraperitoneally for 10 d. Determination of cellular composition (flow cytometry) and cytokine production (ELISA) was performed on 12th d after diabetes induction. EAO suppressed the function of both macrophages and lymphocytes in vitro. In vivo, EAO treatment significantly preserved pancreatic islets and reduced diabetes incidence in MLDS-challenged mice. Besides down-modulatory effect on macrophages, EAO reduced the number of total CD4(+) and activated CD4(+) CD25(+) T cells. Furthermore, EAO affected the number of T helper 1 (Th1) and T helper 17 (Th17) cells through downregulation of their key transcription factors T-bet and RORγT. Because EAO treatment protects mice from development of hyperglycemia by reducing proinflammatory macrophage/Th1/Th17 response, this plant extract could represent a basis for future diabetes therapy.

  10. UV-induced changes in pigment content and light penetration in the fruticose lichen Cladonia arbuscula ssp. mitis.

    PubMed

    Buffoni Hall, Roberta S; Bornman, Janet F; Björn, Lars Olof

    2002-02-01

    The response of the lichen, Cladonia arbuscula (Wallr.) Flot. ssp. mitis (Sandst.) Ruoss to enhanced UV-B (280-315 nm) radiation was investigated with respect to: (a) changes in phenolic content; (b) differential pigment accumulation under visible and UV radiation with increasing distance from thallus apices; and (c) the internal distribution of UV-B radiation within the thallus measured with quartz optical fibres. In a short-term experiment, lichens were exposed for 7 days in a growth chamber to visible light with or without additional UV-B radiation. For a longer term experiment, lichens were grown outdoors under both natural UV radiation, and supplemental UV-A (315-400 nm)+UV-B provided by lamps. Controls were placed under filters that removed the radiation below 290 nm from the natural sunlight. The concentration of total phenolic compounds was measured spectrophotometrically at the termination of the experiments, in different parts of the lichen podetia. UV-exposed lichens showed increased accumulation of phenolics compared to those not grown under UV. At the termination of the long-term experiment, fibre optic measurements of the penetration of radiation into lichen thallus reflected the influence of growth under UV radiation, whereby UV was more strongly attenuated as compared to that in lichens not exposed to enhanced levels of UV-B radiation. Results indicated that in Cladonia, UV-B radiation induces the accumulation of phenolic compounds that may have a protective role. In addition, the morphological distribution of phenolic compounds was different under visible and supplemental UV-B radiation. Internal radiation measurements served to visualise the attenuation of radiation with thallus depth for different wavelengths in the UV-B waveband.

  11. [Acute toxicity and analgesic activity of the global extracts of Nepeta atlantica Ball and Nepeta tuberosa L. ssp. reticulata (Desf.) Maire].

    PubMed

    Bouidida, El Houcine; Alaoui, Katim; Cherrah, Yahia; Fkih-Tetouani, Souad; Idrissi, Abdelkader Il

    2006-01-01

    The global extracts of Nepeta atlantica Ball and Nepeta tuberosa L. ssp. reticulata (Desf.) Maire are especially rich in secondary metabolites of the type iridoid lactonique and glucosidique and of type lupane triterpine. The aerial part of each species is crushed, and then extracted by cold maceration in methanol. These total extracts are in the form of suspension in Arabic gum with 5%, they are tested on the mice for the tests of acute toxicity like for the peripheral analgesic activity according to the test of Koster; and also on the rats for the central analgesic activity of the morphine type based on the test "Tail Flick". The acute toxicity evaluation of these extracts follows upon the determination of the lethal amounts 50% of essential oils from these two species, already given it is specified here by the lethal dose 50% (DL50) of 1672 +/- 232 mg/kg with confidence limits [1030 - 2320] mg/kg for Nepeta atlantica and 1401 +/- 97.29 mg/kg with confidence limits [1130 - 1670] mg/kg for Nepeta tuberosa L. ssp. reticulata. The tests of Koster in the mouse and the "Tail Flik" in the rat showed that the global extracts of the studied species have all two greatly peripheral analgesic activity with an important protection against abdominal cramp 67.91% and 75.53% for 60 mg/kg IP respectively for Nepeta atlantica and Nepeta tuberosa L. ssp. Reticulata, which rise up to 90.10% and 92.89% for 120 mg/kg IP. A central morphine like analgesic activity is record with 120 mg/kg IP for the two species.

  12. Reduced-gravity Testing of The Huygens Probe Ssp Tiltmeter and Hasi Accelerometer Sensors and Their Role In Reconstruction of The Probe Descent Dynamics

    NASA Astrophysics Data System (ADS)

    Ghafoor, N.; Zarnecki, J.

    When the ESA Huygens Probe arrives at Titan in 2005, measurements taken during and after the descent through the atmosphere are likely to revolutionise our under- standing of SaturnSs most enigmatic moon. The accurate atmospheric profiling of Titan from these measurements will require knowledge of the probe descent trajectory and in some cases attitude history, whilst certain atmospheric information (e.g. wind speeds) may be inferred directly from the probe dynamics during descent. Two of the instruments identified as contributing valuable information for the reconstruction of the probeSs parachute descent dynamics are the Surface Science Package Tilt sensor (SSP-TIL) and the Huygens Atmospheric Structure Instrument servo accelerometer (HASI-ACC). This presentation provides an overview of these sensors and their static calibration before describing an investigation into their real-life dynamic performance under simulated Titan-gravity conditions via a low-cost parabolic flight opportunity. The combined use of SSP-TIL and HASI-ACC in characterising the aircraft dynam- ics is also demonstrated and some important challenges are highlighted. Results from some simple spin tests are also presented. Finally, having validated the performance of the sensors under simulated Titan conditions, estimates are made as to the output of SSP-TIL and HASI-ACC under a variety of probe dynamics, ranging from verti- cal descent with spin to a simple 3 degree-of-freedom parachute descent model with horizontal gusting. It is shown how careful consideration must be given to the instru- mentsS principles of operation in each case, and also the impact of the sampling rates and resolutions as selected for the Huygens mission. The presentation concludes with a discussion of ongoing work on more advanced descent modelling and surface dy- namics modelling, and also of a proposal for the testing of the sensors on a sea-surface.

  13. Application of the ligase chain reaction to the detection of nisinA and nisinZ genes in Lactococcus lactis ssp. lactis.

    PubMed

    Ward, L J; Brown, J C; Davey, G P

    1994-03-15

    This paper reports on the application of the ligase chain reaction (LCR) to the specific detection of variants of the nisin structural gene (nisinA and nisinZ) in nisin producing strains of Lactococcus lactis ssp lactis. The LCR assay was used to screen nisin producing strains to determine which form of the nisin structural gene they contained. This method of differentiating the nisin structural gene variants provides a useful alternative to the only other available genetic differentiation, that of sequencing the gene.

  14. Implications of PCR and ELISA results on the routes of bulk-tank contamination with Mycobacterium avium ssp. paratuberculosis.

    PubMed

    Beaver, A; Cazer, C L; Ruegg, P L; Gröhn, Y T; Schukken, Y H

    2016-02-01

    Mycobacterium avium ssp. paratuberculosis (MAP), the etiologic agent of Johne's disease in dairy cattle, may enter the bulk tank via environmental contamination or direct excretion into milk. Traditionally, diagnostics to identify MAP in milk target either MAP antibodies (by ELISA) or the organism itself (by culture or PCR). High ELISA titers may be directly associated with excretion of MAP into milk but only indirectly linked to environmental contamination of the bulk tank. Patterns of bulk-milk ELISA and bulk-milk PCR results could therefore provide insight into the routes of contamination and level of infection or environmental burden. Coupled with questionnaire responses pertaining to management, the results of these diagnostic tests could reveal correlations with herd characteristics or on-farm practices that distinguish herds with high and low environmental bulk-tank MAP contamination. A questionnaire on hygiene, management, and Johne's specific parameters was administered to 292 dairy farms in New York, Oregon, and Wisconsin. Bulk-tank samples were collected from each farm for evaluation by real-time PCR and ELISA. Before DNA extraction and testing of the unknown samples, bulk-milk template preparation was optimized with respect to parameters such as MAP fractionation patterns and lysis. Two regression models were developed to explore the relationships among bulk-tank PCR, ELISA, environmental predictors, and herd characteristics. First, ELISA optical density (OD) was designated as the outcome in a linear regression model. Second, the log odds of being PCR positive in the bulk tank were modeled using binary logistic regression with penalized maximum likelihood. The proportion of PCR-positive bulk tanks was highest for New York and for organic farms, providing a clue as to the geographical patterns of MAP-positive bulk-tank samples and relationship to production type. Bulk-milk PCR positivity was also higher for large relative to small herds. The models

  15. A new phenanthrene derivative and two diarylheptanoids from the roots of Brassica rapa ssp. campestris inhibit the growth of cancer cell lines and LDL-oxidation.

    PubMed

    Wu, Qian; Cho, Jin-Gyeong; Yoo, Ki-Hyun; Jeong, Tae-Sook; Park, Ji-Hae; Kim, Su-Yeon; Kang, Ji-Hyun; Chung, In-Sik; Choi, Myung-Sook; Lee, Kyung-Tae; Chung, Hae-Gon; Bang, Myun-Ho; Baek, Nam-In

    2013-04-01

    Brassica rapa ssp. campestris (Brassicaceae) is a conical, deep purple, edible root vegetable commonly known as a turnip. We initiated phytochemical and pharmacological studies to search for biological active compounds from the roots of B. rapa ssp. campestris. We isolated a novel phenanthrene derivative, 6-methoxy-1-[10-methoxy-7-(3-methylbut-2-enyl)phenanthren-3-yl]undecane-2,4-dione, named brassicaphenanthrene A (3) along with two known diarylheptanoid compounds, 6-paradol (1) and trans-6-shogaol (2), through the repeated silica gel (SiO2), octadecyl silica gel, and Sephadex LH-20 column chromatography. The chemical structures of the compounds were determined by spectroscopic data analyses including nuclear magnetic resonance, mass spectrometry, ultraviolet spectroscopy, and infra-red spectroscopy. All compounds exhibited high inhibitory activity against the growth of human cancer lines, HCT-116, MCF-7, and HeLa, with IC50 values ranging from 15.0 to 35.0 μM and against LDL-oxidation with IC50 values ranging from 2.9 to 7.1 μM.

  16. Transcriptional differences between the male-sterile mutant bcms and wild-type Brassica campestris ssp. chinensis reveal genes related to pollen development.

    PubMed

    Huang, L; Cao, J; Ye, W; Liu, T; Jiang, L; Ye, Y

    2008-05-01

    A novel male-sterile mutant which lacks mature pollen, Brassisa campestris male sterile (bcms), was identified in Brassica campestris L. ssp. chinensis Makino (syn. B. rapa ssp. chinensis). Genetic analysis revealed that bcms was controlled by a single recessive mutation locus. Genome-wide transcriptional profiling was performed on the flower buds of both the bcms mutant and the wild-type from which it originated, and profiling analysis indicated that there were numerous changes in gene expression attributable to the gene mutation. This mutation resulted in down-regulation of a variety of genes and up-regulated expression of a few other genes. A total of 51 transcript-derived fragments (TDFs) were isolated: 32 specifically and 12 predominantly accumulated in wild-type flower buds, and two specifically and five predominantly accumulated in bcms flower buds. Sequence analysis showed that some of these TDFs share significant similarities with genes involved in different aspects of cellular development, such as signal transduction, cell wall biosynthesis and regulation. Most other TDFs showed no or very poor sequence similarities to entries in any database and might represent new candidate proteins involved in pollen development. Furthermore, spatial and temporal expression pattern analysis of 20 genes derived from cDNA-amplified fragment length polymorphism in different tissues of both the bcms and wild-type plants revealed their complex and dynamic expression patterns. The bcms mutant and the genes isolated in this paper provide excellent material for future studies on the molecular mechanism of male sterility.

  17. Morphological and functional characterization of BcMF13 in the antisense-silenced plants of Brassica campestris ssp. chinensis var. parachinensis.

    PubMed

    Li, Yanyan; Cao, Jiashu

    2009-05-01

    The gene Brassica campestris male fertility 13 (BcMF13, GenBank accession number EF158459) was isolated as a reproductive organ-specific gene from Chinese cabbage (Brassica campestris L. ssp. chinensis Makino, syn. B. rapa ssp. chinensis). It is exclusively expressed in stage four and five flower buds of fertile lines and is most strongly expressed in stamens. Here, we report a functional characterization of this BcMF13 gene in the antisense-silenced plants. The inflorescence of the BcMF13 mutant was compacted with anthers curved outside. The fertility of this mutant was greatly reduced with less than 5 seeds per silique. Under scanning electron microscopy, the mutant demonstrated numerous shriveled pollen grains with deep invaginations. The frequency of normal pollen grains was just 45.34%. The pollen mother cell, the tetrad, and the mature pollen of the BcMF13 mutant were abnormal resulting in the poor pollen vitality. Germination test in vivo suggested BcMF13 delayed the pollen tubes' extension in the style. All these indicated BcMF13 had a vital role in pollen development of Chinese cabbage.

  18. Genetic variability in anthocyanin composition and nutritional properties of blue, purple, and red bread (Triticum aestivum L.) and durum (Triticum turgidum L. ssp. turgidum convar. durum) wheats.

    PubMed

    Ficco, Donatella B M; De Simone, Vanessa; Colecchia, Salvatore A; Pecorella, Ivano; Platani, Cristiano; Nigro, Franca; Finocchiaro, Franca; Papa, Roberto; De Vita, Pasquale

    2014-08-27

    Renewed interest in breeding for high anthocyanins in wheat (Triticum ssp.) is due to their antioxidant potential. A collection of different pigmented wheats was used to investigate the stability of anthocyanins over three crop years. The data show higher anthocyanins in blue-aleurone bread wheat (Triticum aestivum L.), followed by purple- and red-pericarp durum wheat (Triticum turgidum L. ssp. turgidum convar. durum), using cyanidin 3-O-glucoside as standard. HPLC of the anthocyanin components shows five to eight major anthocyanins for blue wheat extracts, compared to three anthocyanins for purple and red wheats. Delphinidin 3-O-rutinoside, delphinidin 3-O-glucoside, and malvidin 3-O-glucoside are predominant in blue wheat, with cyanidin 3-O-glucoside, peonidin 3-O-galactoside, and malvidin 3-O-glucoside in purple wheat. Of the total anthocyanins, 40-70% remain to be structurally identified. The findings confirm the high heritability for anthocyanins, with small genotype × year effects, which will be useful for breeding purposes, to improve the antioxidant potential of cereal-based foods.

  19. Growth and exopolysaccharide yield of Lactobacillus delbrueckii ssp. bulgaricus DSM 20081 in batch and continuous bioreactor experiments at constant pH.

    PubMed

    Mende, Susann; Krzyzanowski, Leona; Weber, Jost; Jaros, Doris; Rohm, Harald

    2012-02-01

    Some Lactobacillus delbrueckii ssp. bulgaricus strains are able to synthesize exopolysaccharides (EPS) and are therefore highly important for the dairy industry as starter cultures. The aim of this study was to investigate the nutritional requirements for growth and EPS production of Lactobacillus delbrueckii ssp. bulgaricus DSM 20081. A medium was developed from a semi-defined medium (SDM) in which glucose was replaced by lactose and different combinations of supplements (nucleobases, vitamins, salts, sodium formate and orotic acid) were added. Constant pH batch fermentation with the modified medium resulted in an EPS yield of approximately 210 mg glucose equivalents per liter medium. This was a 10-fold increase over flask cultivation of this strain in SDM. Although not affecting cell growth, the mixture of salts enhanced the EPS synthesis. Whereas EPS production was approximately 12 mg/g dry biomass without salt supplementation, a significantly higher yield (approximately 20 mg/g dry biomass) was observed after adding the salt mixture. In continuous fermentation, a maximal EPS concentration was obtained at a dilution rate of 0.31/h (80 mg EPS/L), which corresponded to a specific EPS production of 49 mg/g dry biomass.

  20. Analysis of chromosomal polymorphism in barley (Hordeum vulgare L. ssp. vulgare) and between H. vulgare and H. chilense using three-color fluorescence in situ hybridization (FISH).

    PubMed

    Szakács, É; Kruppa, K; Molnár-Láng, M

    2013-11-01

    The aim of the present work was to study chromosomal polymorphism within cultivated barley (Hordeum vulgare ssp. vulgare) using three-color fluorescence in situ hybridization (FISH). The physical distribution of the most frequently used, highly repetitive DNA sequences (GAA)7 specific for pericentromeric heterochromatic regions, the ribosomal DNA clone pTa71, specific for the 45S rDNA, and the barley-specific telomere-associated sequence HvT01, was investigated to reveal genetic diversity in metaphase spreads of ten barley genotypes with diverse geographical origin, growth habit and row number. A wild relative of barley, Hordeum chilense was also studied in order to compare the polymorphism between and within Hordeum species. Significant differences in the hybridization patterns of all three DNA probes could be detected between the two related species, but only probes pTa71 and HvT01 showed variation in the intensity and/or position of hybridization sites among genotypes of H. vulgare ssp. vulgare. The extent of polymorphism was less than that earlier reported for molecular markers and was restricted to the long chromosome arms, with differences between the chromosomes. 1H and 3H proved to be the most variable chromosomes and 4H and 6H the most conserved.

  1. Comparative sequence analysis of the potato cyst nematode resistance locus H1 reveals a major lack of co-linearity between three haplotypes in potato (Solanum tuberosum ssp.).

    PubMed

    Finkers-Tomczak, Anna; Bakker, Erin; de Boer, Jan; van der Vossen, Edwin; Achenbach, Ute; Golas, Tomasz; Suryaningrat, Suwardi; Smant, Geert; Bakker, Jaap; Goverse, Aska

    2011-02-01

    The H1 locus confers resistance to the potato cyst nematode Globodera rostochiensis pathotypes 1 and 4. It is positioned at the distal end of chromosome V of the diploid Solanum tuberosum genotype SH83-92-488 (SH) on an introgression segment derived from S. tuberosum ssp. andigena. Markers from a high-resolution genetic map of the H1 locus (Bakker et al. in Theor Appl Genet 109:146-152, 2004) were used to screen a BAC library to construct a physical map covering a 341-kb region of the resistant haplotype coming from SH. For comparison, physical maps were also generated of the two haplotypes from the diploid susceptible genotype RH89-039-16 (S. tuberosum ssp. tuberosum/S. phureja), spanning syntenic regions of 700 and 319 kb. Gene predictions on the genomic segments resulted in the identification of a large cluster consisting of variable numbers of the CC-NB-LRR type of R genes for each haplotype. Furthermore, the regions were interspersed with numerous transposable elements and genes coding for an extensin-like protein and an amino acid transporter. Comparative analysis revealed a major lack of gene order conservation in the sequences of the three closely related haplotypes. Our data provide insight in the evolutionary mechanisms shaping the H1 locus and will facilitate the map-based cloning of the H1 resistance gene.

  2. Physical mapping and microsynteny of Brassica rapa ssp. pekinensis genome corresponding to a 222 kbp gene-rich region of Arabidopsis chromosome 4 and partially duplicated on chromosome 5.

    PubMed

    Park, J Y; Koo, D H; Hong, C P; Lee, S J; Jeon, J W; Lee, S H; Yun, P Y; Park, B S; Kim, H R; Bang, J W; Plaha, P; Bancroft, I; Lim, Y P

    2005-12-01

    We constructed a bacterial artificial chromosome (BAC) library, designated as KBrH, from high molecular weight genomic DNA of Brassica rapa ssp. pekinensis (Chinese cabbage). This library, which was constructed using HindIII-cleaved genomic DNA, consists of 56,592 clones with average insert size of 115 kbp. Using a partially duplicated DNA sequence of Arabidopsis, represented by 19 and 9 predicted genes on chromosome 4 and 5, respectively, and BAC clones from the KBrH library, we studied conservation and microsynteny corresponding to the Arabidopsis regions in B. rapa ssp. pekinensis. The BAC contigs assembled according to the Arabidopsis homoeologues revealed triplication and rearrangements in the Chinese cabbage. In general, collinearity of genes in the paralogous segments was maintained, but gene contents were highly variable with interstitial losses. We also used representative BAC clones, from the assembled contigs, as probes and hybridized them on mitotic (metaphase) and/or meiotic (leptotene/pachytene/metaphase I) chromosomes of Chinese cabbage using bicolor fluorescence in situ hybridization. The hybridization pattern physically identified the paralogous segments of the Arabidopsis homoeologues on B. rapa ssp. pekinensis chromosomes. The homoeologous segments corresponding to chromosome 4 of Arabidopsis were located on chromosomes 2, 8 and 7, whereas those of chromosome 5 were present on chromosomes 6, 1 and 4 of B. rapa ssp. pekinensis.

  3. Characterization of a novel RNA regulator of Erwinia carotovora ssp. carotovora that controls production of extracellular enzymes and secondary metabolites.

    PubMed

    Liu, Y; Cui, Y; Mukherjee, A; Chatterjee, A K

    1998-07-01

    The enterobacterium Erwinia carotovora ssp. carotovora strain 71 (hereafter Ecc71) produces extracellular enzymes such as pectate lyase isozymes (Pels), cellulase (Cel), polygalacturonase (Peh) and protease (Prt). These enzymes degrade plant cell wall components and are largely responsible for the elicitation of soft-rot diseases in plants and plant products. Ecc71 also produces HarpinEcc, the elicitor of hypersensitive reaction (HR) and the quorum-sensing signal, N-(3-oxohexanoyl)-L-homoserine lactone (OHL). OHL controls extracellular enzyme and HarpinEcc production. The levels of these enzymes, as well as the expression of hrpNEcc, the structural gene for HarpinEcc, and ohll, the gene specifying OHL synthesis, are negatively regulated by RsmaA. rsmB, formerly aepH, on the other hand, positively regulates extracellular enzyme production. 6His-RsmA recombinant protein purified from E. coli binds rsmB RNA as indicated by gel mobility shift assays. rsmB comprises 547 bp DNA, which is transcribed from a single start site immediately after a sigma70-like promoter. In Ecc71, two rsmB RNA species are detected: a full-length 479 base rsmB RNA and a 259 base rsmB' RNA. rsmB' DNA hybridizes with the 259 base and the 479 base transcripts. A 3' RNase protection assay revealed that the 259 base and the 479 base RNA species end at the same position immediately after the putative rho-independent terminator. The expression of rsmB-lacZ transcriptional fusions established that the rsmB' RNA is not produced because of the activation of an internal promoter. These data strongly suggest that the 259 base rsmB' RNA is derived by processing of the primary rsmB RNA. In Ecc71, rsmB' expression driven by the lac promoter causes overproduction of Pel, Peh, Cel and Prt, and accumulation of pel-1, peh-1, hrpNEcc and ohll transcripts. By contrast, a plasmid with the rsmB' DNA sequence deleted fails to cause overproduction of the extracellular enzymes in Ecc71. The rsmB' effect also occurs in

  4. Genome-wide quantitative trait locus mapping identifies multiple major loci for brittle rachis and threshability in Tibetan semi-wild wheat (Triticum aestivum ssp. tibetanum Shao).

    PubMed

    Jiang, Yun-Feng; Lan, Xiu-Jin; Luo, Wei; Kong, Xing-Chen; Qi, Peng-Fei; Wang, Ji-Rui; Wei, Yu-Ming; Jiang, Qian-Tao; Liu, Ya-Xi; Peng, Yuan-Ying; Chen, Guo-Yue; Dai, Shou-Fen; Zheng, You-Liang

    2014-01-01

    Tibetan semi-wild wheat (Triticum aestivum ssp. tibetanum Shao) is a semi-wild hexaploid wheat resource that is only naturally distributed in the Qinghai-Tibet Plateau. Brittle rachis and hard threshing are two important characters of Tibetan semi-wild wheat. A whole-genome linkage map of T. aestivum ssp. tibetanum was constructed using a recombinant inbred line population (Q1028×ZM9023) with 186 lines, 564 diversity array technology markers, and 117 simple sequence repeat markers. Phenotypic data on brittle rachis and threshability, as two quantitative traits, were evaluated on the basis of the number of average spike rachis fragments per spike and percent threshability in 2012 and 2013, respectively. Quantitative trait locus (QTL) mapping performed using inclusive composite interval mapping analysis clearly identified four QTLs for brittle rachis and three QTLs for threshability. However, three loci on 2DS, 2DL, and 5AL showed pleiotropism for brittle rachis and threshability; they respectively explained 5.3%, 18.6%, and 18.6% of phenotypic variation for brittle rachis and 17.4%, 13.2%, and 35.2% of phenotypic variation for threshability. A locus on 3DS showed an independent effect on brittle rachis, which explained 38.7% of the phenotypic variation. The loci on 2DS and 3DS probably represented the effect of Tg and Br1, respectively. The locus on 5AL was in very close proximity to the Q gene, but was different from the predicted q in Tibetan semi-wild wheat. To our knowledge, the locus on 2DL has never been reported in common wheat but was prominent in T. aestivum ssp. tibetanum accession Q1028. It remarkably interacted with the locus on 5AL to affect brittle rachis. Several major loci for brittle rachis and threshability were identified in Tibetan semi-wild wheat, improving the understanding of these two characters and suggesting the occurrence of special evolution in Tibetan semi-wild wheat.

  5. Detection of Campylobacter antibodies in sheep sera by a Dot-ELISA using acid extracts from c. fetus ssp. fetus and c. jejuni strains and comparison with a complement fixation test.

    PubMed

    Gürtürk, K; Ekin, I H; Aksakal, A; Solmaz, H

    2002-04-01

    In this study, a dot-enzyme-linked immunosorbent assay (Dot-ELISA) was evaluated in comparison with a complement fixation test (CFT) for the detection of Campylobacter antibodies in sheep sera. Acid glycine extracts (AGE) of both Campylobacter fetus ssp. fetus and Campylobacter jejuni strains that had been isolated from the gall-bladder of slaughtered sheep was used as antigen in both tests. A total of 153 sheep sera from aborted (74) and slaughtered (79) sheep were examined by both Dot-ELISA and CFT. Twenty-two sera showed anti-complementary activity were not suitable for CFT. Of the 22 sera showing anti-complementary activity, two sera were found to be positive in Dot-ELISA. Eighty-eight (67.2%) of the remaining 131 sera were negative by both Dot-ELISA and CFT using AGE of both Campylobacter strains whereas 43 sera (32.8%) gave different reaction patterns in Dot-ELISA and CFT with the extracts of both Campylobacter strains. Twelve sera were positive by both tests using AGE of C. fetus ssp. fetus but CFT failed to detect antibodies in nine of these sera when AGE of C. jejuni was used. Twelve sera were positive by both tests only when AGE of C. fetus ssp. fetus was used. Eleven sera were positive only by CFT. Seven of these reacted only with the AGE of C. fetus ssp. fetus and four sera were positive by using AGE of both Campylobacter strains. The remaining eight sera were found to be positive only by dot-immunobinding assay either with the AGE of both Campylobacter strains or with the AGE of one of the Campylobacter strains. It is concluded that Dot-ELISA using AGE from C. fetus ssp. fetus could be employed for the detection of Campylobacter antibodies in sheep sera and the additional use of AGE from C. jejuni as antigen appeared not to be profitable for this purpose.

  6. Host-specific races in the holoparasitic angiosperm Orobanche minor: implications for speciation in parasitic plants

    PubMed Central

    Thorogood, C. J.; Rumsey, F. J.; Hiscock, S. J.

    2009-01-01

    Background and Aims Orobanche minor is a root-holoparasitic angiosperm that attacks a wide range of host species, including a number of commonly cultivated crops. The extent to which genetic divergence among natural populations of O. minor is influenced by host specificity has not been determined previously. Here, the host specificity of natural populations of O. minor is quantified for the first time, and evidence that this species may comprise distinct physiological races is provided. Methods A tripartite approach was used to examine the physiological basis for the divergence of populations occurring on different hosts: (1) host–parasite interactions were cultivated in rhizotron bioassays in order to quantify the early stages of the infection and establishment processes; (2) using reciprocal-infection experiments, parasite races were cultivated on their natural and alien hosts, and their fitness determined in terms of biomass; and (3) the anatomy of the host–parasite interface was investigated using histochemical techniques, with a view to comparing the infection process on different hosts. Key Results Races occurring naturally on red clover (Trifolium pratense) and sea carrot (Daucus carota ssp. gummifer) showed distinct patterns of host specificity: parasites cultivated in cross-infection studies showed a higher fitness on their natural hosts, suggesting that races show local adaptation to specific hosts. In addition, histological evidence suggests that clover and carrot roots vary in their responses to infection. Different root anatomy and responses to infection may underpin a physiological basis for host specificity. Conclusions It is speculated that host specificity may isolate races of Orobanche on different hosts, accelerating divergence and ultimately speciation in this genus. The rapid life cycle and broad host range of O. minor make this species an ideal model with which to study the interactions of parasitic plants with their host associates. PMID

  7. Search path of a fossorial herbivore, Geomys bursarius, foraging in structurally complex plant communities

    USGS Publications Warehouse

    Andersen, Douglas C.

    1990-01-01

    The influence of habitat patchiness and unpalatable plants on the search path of the plains pocket gopher (Geomys bursarius) was examined in outdoor enclosures. Separate experiments were used to evaluate how individual animals explored (by tunnel excavation) enclosures free of plants except for one or more dense patches of a palatable plant (Daucus carota), a dense patch of an unpalatable species (Pastinaca sativa) containing a few palatable plants (D. carota), or a relatively sparse mixture of palatable (D. carota) and unpalatable (Raphanus sativus) species. Only two of eight individuals tested showed the predicted pattern of concentrating search effort in patches of palatable plants. The maintenance of relatively high levels of effort in less profitable sites may reflect the security afforded food resources by the solitary social system and fossorial lifestyle of G. bursarius. Unpalatable plants repelled animals under some conditions, but search paths in the sparsely planted mixed-species treatment suggest animals can use visual or other cues to orient excavations. Evidence supporting area-restricted search was weak. More information about the use of visual cues by G. bursarius and the influence of experience on individual search mode is needed for refining current models of foraging behavior in this species.

  8. Characterization of flavonol conjugates in immature leaves of pak choi [Brassica rapa L. Ssp. chinensis L. (Hanelt.)] by HPLC-DAD and LC-MS/MS.

    PubMed

    Rochfort, Simone J; Imsic, Michael; Jones, Rod; Trenerry, V Craige; Tomkins, Bruce

    2006-06-28

    The flavonoid composition of immature leaves of pak choi [Brassica rapa L. ssp. chinensis L. (Hanelt.)] was investigated. Flavonol aglycone content was measured in 11 pak choi varieties, indicating significant differences (P < 0.05) in content between varieties and relatively high contents of kaempferol and isorhamnetin. Levels of quercetin ranged from 3.2 to 6.1 mg/100 g of dry weight (DW), whereas levels of isorhamnetin and kaempferol were significantly higher (8.1-35.1 and 36.0-102.6 mg/100 g of DW, respectively). A large number of glycoside and hydroxycinnamic acid derivatives of quercetin, kaempferol, and isorhamnetin were identified in cv. 'Shanghai' by LC/UV-DAD/ESI-MS/MS. The UV-DAD data allowed identification of hydroxycinnamic acid derivatives, but detailed MS/MS fragmentations were required for the structure elucidation. Pak choi could be a potentially important source of dietary flavonols, in particular, kaempferol and isorhamnetin.

  9. Pollen limitation of female reproductive success at fine spatial scale in a gynodioecious and wind-pollinated species, Beta vulgaris ssp. maritima.

    PubMed

    De Cauwer, I; Arnaud, J-F; Schmitt, E; Dufay, M

    2010-12-01

    In sexually polymorphic plants, the spatial distribution of sexes is usually not random. Local variation in phenotype frequencies is expected to affect individual fitness of the different phenotypes. For gynodioecious species, with co-occurrence of hermaphrodites and females, if sexual phenotypes are structured in space and pollen flow is spatially restricted, local pollen availability should vary among patches. Female fitness may thus be low when hermaphrodites are locally rare. To test this hypothesis, we analysed how the reproductive output of females varied among patches within two natural study sites of the gynodioecious wind-pollinated Beta vulgaris ssp. maritima. Plants growing in female-biased areas and experiencing pollen limitation were found to have low fruit and seed sets but did not reallocate resources towards better offspring. Our results show that fine-scale processes influence individual fitness and the evolution of sex ratio in sexually polymorphic plants.

  10. Potential for evolutionary change in the seasonal timing of germination in sea beet (Beta vulgaris ssp. maritima) mediated by seed dormancy.

    PubMed

    Wagmann, Kristen; Hautekèete, Nina-Coralie; Piquot, Yves; Van Dijk, Henk

    2010-07-01

    In sea beet (Beta vulgaris ssp. maritima), germination occurs in autumn or spring and is mediated by dormancy which can be released by cold or dry periods. Environmental change such as current climate change may require evolutionary response in seasonal timing. Here, we explore the potential for such evolutionary change. Seed dormancy was studied in a composite population based on seeds from all over the species range in France together with several generations of reciprocal crosses. We found high, repeatable variability for dormancy rate among individuals under greenhouse conditions and confirmed its relevance for germination phenology in the field. Our data fitted best with an exclusively maternal determination of the dormancy phenotype. Narrow-sense heritability, h(2) approximately 0.5 in the composite population and approximately 0.4 in the original local populations, was such that rapid evolutionary change in the relative proportions of autumn and spring germination may be possible.

  11. Determination of antimutagenic properties of apigenin-7-O-rutinoside, a flavonoid isolated from Mentha longifolia (L.) Huds. ssp. longifolia with yeast DEL assay.

    PubMed

    Gulluce, Medine; Orhan, Furkan; Adiguzel, Ahmet; Bal, Tugba; Guvenalp, Zuhal; Dermirezer, Lutfiye Omur

    2013-07-01

    Lamiaceae is an important plant family that has been investigated for its medicinal properties due to its large amounts of phenolic acids and flavonoids. Flavonoids have been shown to have antioxidant and antimutagenic activities in different test systems, but their certain mechanisms are still unclear. This study was designed to evaluate the mutagenic and antimutagenic activities of apigenin 7-O-rutinoside, a flavonoid isolated from Mentha longifolia (L.) Huds. ssp. longifolia. The possible antimutagenic potential of apigenin 7-O-rutinoside (A7R) was examined against mutagens ethyl methanesulfonate (EMS) and acridine (AC) in a eukaryotic cell system Saccharomyces cerevisiae RS112. The results showed that A7R has different inhibition rates against EMS and AC-induced mutagenicity. Thus, the properties of A7R are of great pharmacological importance and might be beneficial for reducing the risk of reactive oxygen species-related diseases.

  12. Influence of harvest time on the quality of oil-based compounds in sea buckthorn (Hippophae rhamnoides L. ssp. sinensis) seed and fruit.

    PubMed

    St George, Susan D; Cenkowski, Stefan

    2007-10-01

    The effect of the harvest time on oil-based bioactive compounds in sea buckthorn berries ( Hippophae rhamnoides L. ssp. sinensis) was investigated. Sea buckthorn berries were collected at early maturity (September), maturity (November), and postmaturity (January) during the 2003-2004 harvest year. Whole berries were analyzed for physical characteristics, and fruit and seed fractions were analyzed for bioactive content. November-harvested berries yielded the highest values for berry sizes, CIELab factor a*, and total carotenoid content in the fruit fraction ( p < 0.05). September yielded significantly higher ( p < 0.05) levels of major compounds, alpha-tocopherol and beta-sitosterol, in the fruit fraction. Seed characteristics and bioactive compounds did not vary significantly with respect to the harvest time ( p > 0.05). These results have identified the most suitable level of maturity for the optimization of certain compounds and the losses that may occur with winter harvest, commonly practiced in cold climates.

  13. Chemical composition and biological activity of the essential oil of Origanum vulgare ssp. hirtum from different areas in the Southern Apennines (Italy).

    PubMed

    Mancini, Emilia; Camele, Ippolito; Elshafie, Hazem S; De Martino, Laura; Pellegrino, Carlo; Grulova, Daniela; De Feo, Vincenzo

    2014-04-01

    The chemical composition of the essential oils of Origanum vulgare ssp. hirtum, growing wild in three different localities in the Southern Apennines, was studied by GC-FID and GC/MS analyses. In total, 103 compounds were identified. The oils were mainly composed of phenolic compounds and all oils belonged to the chemotype carvacrol/thymol. The three essential oils were evaluated for their in vitro phytotoxic activity by determining their influence on the germination and initial radicle elongation of Sinapis arvensis L., Phalaris canariensis L., Lepidium sativum L., and Raphanus sativus L. The seed germination and radicle growth were affected in various degrees. Moreover, the antifungal activity of the three essential oils was assayed against three species causing pre- and postharvest fruit decay (Monilinia laxa, M. fructigena, and M. fructicola). At 1000 ppm, the three oils completely inhibited fungal growth. The hemolytic activity of the oils was assayed and showed no effect on the cell membranes of bovine erythrocytes.

  14. [Effect of Genotype and Medium Culture Content on Microspore-Derived Embryo Formation in Chinese Cabbage (Brassica rapa ssp. chinensis) Cv. Lastochka].

    PubMed

    Shumilina, D V; Shmykova, N A; Bondareva, L L; Suprunova, T P

    2015-01-01

    The influence of different factors on microspore embryogenesis in Chinese cabbage (B. rapa ssp. chinensis) was studied. A genotype dependence for embryo formation was observed. The majority of embryos and plants were obtained from microspores isolated from flower buds (2-2.9 mm in length) and cultured in the NLN liquid medium with 13% sucrose (w/v) supplemented with 24-epibrassinolide and 1% activated charcoal. Embryos cultured on the 1/2 Murashige-Skoog culture medium with 2% sucrose (w/v), 0.1 mg/L benzylaminopurine, and 3g/L Phytagel stimulated the formation of secondary embryos that resulted in development of large number of doubled haploid plants.

  15. Characterization of a novel gene, BcMF7, that is expressed preferentially in pollen of Brassica campestris L. ssp. chinensis Makino.

    PubMed

    Huang, Li; Cao, JiaShu; Zhang, YuChao; Ye, YiQun

    2007-08-01

    Pollen formation is important for plant sexual reproduction. To identify the genes that are involved in pollen formation, we performed the genome-wide transcriptional profiling in the flower buds of both male meiotic cytokinesis (mmc) mutant and its wild-type plants of Brassica campestris L. ssp. chinensis, syn. B. rapa L. ssp. chinensis. cDNA-amplified fragment length polymorphism (cDNA-AFLP) analysis showed that the mmc mutation resulted in changes in expression of a variety of genes. BcMF7, a transcript-derived fragment (TDF) accumulated in the wild-type flower buds was further characterized. The BcMF7 gene has 1161 bp in length with two introns. The full-length BcMF7 cDNA has 609 bp in length and encodes a protein of 129 amino acids. The deduced amino acid sequence of BcMF7 protein shares no similarity to any function-known protein in Swiss-Prot database, but has 8 protein kinase C phosphorylation sites, 2 caselin kinase II phosphorylation sites, 2 tyrosine kinase phosphorylation sites, 2 N-glycosylation sites and 2 N-myristolyation sites. Spatial and temporal expression patterns analysis showed that BcMF7 was expressed exclusively in pollen. The expression signal of BcMF7 was first detected at the tetrad stage of microspore development, reached a peak level at the uninucleate stage, and decreased to a slightly low level at the mature pollen stage. All these results show that BcMF7 may play a certain role in the signal transduction during pollen development.

  16. The Non-universality of the Low-mass End of the IMF is Robust against the Choice of SSP Model

    NASA Astrophysics Data System (ADS)

    Spiniello, C.; Trager, S. C.; Koopmans, L. V. E.

    2015-04-01

    We perform a direct comparison of two state-of-the art single stellar population (SSP) models that have been used to demonstrate the non-universality of the low-mass end of the initial mass function (IMF) slope. The two public versions of the SSP models are restricted to either solar abundance patterns or solar metallicity, too restrictive if one aims to disentangle elemental enhancements, metallicity changes, and IMF variations in massive early-type galaxies (ETGs) with star formation histories different from those in the solar neighborhood. We define response functions (to metallicity and α-abundance) to extend the parameter space for each set of models. We compare these extended models with a sample of Sloan Digital Sky Survey (SDSS) ETG spectra with varying velocity dispersions. We measure equivalent widths of optical IMF-sensitive stellar features to examine the effect of the underlying model assumptions and ingredients, such as stellar libraries or isochrones, on the inference of the IMF slope down to ∼0.1 M⊙. We demonstrate that the steepening of the low-mass end of the IMF based on a non-degenerate set of spectroscopic optical indicators is robust against the choice of the stellar population model. Although the models agree in a relative sense (i.e., both imply more bottom-heavy IMFs for more massive systems), we find non-negligible differences in the absolute values of the IMF slope inferred at each velocity dispersion by using the two different models. In particular, we find large inconsistencies in the quantitative predictions of the IMF slope variations and abundance patterns when sodium lines are used. We investigate the possible reasons for these inconsistencies.

  17. Spelt (Triticum aestivum ssp. spelta) as a source of breadmaking flours and bran naturally enriched in oleic acid and minerals but not phytic acid.

    PubMed

    Ruibal-Mendieta, Nike L; Delacroix, Dominique L; Mignolet, Eric; Pycke, Jean-Marie; Marques, Carole; Rozenberg, Raoul; Petitjean, Géraldine; Habib-Jiwan, Jean-Louis; Meurens, Marc; Quetin-Leclercq, Joëlle; Delzenne, Nathalie M; Larondelle, Yvan

    2005-04-01

    The nutritional value of breadmaking cereal spelt (Triticum aestivum ssp. spelta) is said to be higher than that of common wheat (Triticum aestivum ssp. vulgare), but this traditional view is not substantiated by scientific evidence. In an attempt to clarify this issue, wholemeal and milling fractions (sieved flour, fine bran, and coarse bran) from nine dehulled spelt and five soft winter wheat samples were compared with regard to their lipid, fatty acid, and mineral contents. In addition, tocopherol (a biochemical marker of germ) was measured in all wholemeals, whereas phytic acid and phosphorus levels were determined in fine bran and coarse bran samples after 1 month of storage. Results showed that, on average, spelt wholemeals and milling fractions were higher in lipids and unsaturated fatty acids as compared to wheat, whereas tocopherol content was lower in spelt, suggesting that the higher lipid content of spelt may not be related to a higher germ proportion. Although milling fractionation produced similar proportions of flour and brans in spelt and wheat, it was found that ash, copper, iron, zinc, magnesium, and phosphorus contents were higher in spelt samples, especially in aleurone-rich fine bran and in coarse bran. Even though phosphorus content was higher in spelt than in wheat brans, phytic acid content showed the opposite trend and was 40% lower in spelt versus wheat fine bran, which may suggest that spelt has either a higher endogenous phytase activity or a lower phytic acid content than wheat. The results of this study give important indications on the real nutritional value of spelt compared to wheat. Moreover, they show that the Ca/Fe ratio, combined with that of oleate/palmitate, provides a highly discriminating tool to authenticate spelt from wheat flours and to face the growing issue of spelt flour adulteration. Finally, they suggest that aleurone differences, the nature of which still needs to be investigated, may account for the differential

  18. Differences between the rhizosphere microbiome of Beta vulgaris ssp. maritima—ancestor of all beet crops—and modern sugar beets

    PubMed Central

    Zachow, Christin; Müller, Henry; Tilcher, Ralf; Berg, Gabriele

    2014-01-01

    The structure and function of the plant microbiome is driven by plant species and prevailing environmental conditions. Effectuated by breeding efforts, modern crops diverge genetically and phenotypically from their wild relatives but little is known about consequences for the associated microbiota. Therefore, we studied bacterial rhizosphere communities associated with the wild beet B. vulgaris ssp. maritima grown in their natural habitat soil from coastal drift lines (CS) and modern sugar beets (Beta vulgaris ssp. vulgaris) cultivated in CS and potting soil (PS) under greenhouse conditions. Analysis of 16S rRNA gene fingerprints and pyrosequencing-based amplicon libraries revealed plant genotype- and soil-specific microbiomes. Wild beet plants harbor distinct operational taxonomic units (OTUs) and a more diverse bacterial community than the domesticated sugar beet plants. Although the rhizospheres of both plant genotypes were dominated by Proteobacteria and Planctomycetes, 37.5% of dominant OTUs were additionally detected in the wild beet rhizosphere. Analysis of the cultivable fraction confirmed these plant genotype-specific differences at functional level. The proportion of isolates displayed in vitro activity against phytopathogens was lower for wild beet (≤45.8%) than for sugar beet (≤57.5%). Conversely, active isolates from the wild beet exhibited stronger ability to cope with abiotic stresses. From all samples, active isolates of Stenotrophomonas rhizophila were frequently identified. In addition, soil type-specific impacts on the composition of bacterial communities were found: Acidobacteria, Chloroflexi, and Planctomycetes were only detected in plants cultivated in CS; whereas Bacteroidetes and Proteobacteria dominated in PS. Overall, in comparison to modern sugar beets, wild beets were associated with taxonomically and functionally distinct microbiomes. PMID:25206350

  19. Differences between the rhizosphere microbiome of Beta vulgaris ssp. maritima-ancestor of all beet crops-and modern sugar beets.

    PubMed

    Zachow, Christin; Müller, Henry; Tilcher, Ralf; Berg, Gabriele

    2014-01-01

    The structure and function of the plant microbiome is driven by plant species and prevailing environmental conditions. Effectuated by breeding efforts, modern crops diverge genetically and phenotypically from their wild relatives but little is known about consequences for the associated microbiota. Therefore, we studied bacterial rhizosphere communities associated with the wild beet B. vulgaris ssp. maritima grown in their natural habitat soil from coastal drift lines (CS) and modern sugar beets (Beta vulgaris ssp. vulgaris) cultivated in CS and potting soil (PS) under greenhouse conditions. Analysis of 16S rRNA gene fingerprints and pyrosequencing-based amplicon libraries revealed plant genotype- and soil-specific microbiomes. Wild beet plants harbor distinct operational taxonomic units (OTUs) and a more diverse bacterial community than the domesticated sugar beet plants. Although the rhizospheres of both plant genotypes were dominated by Proteobacteria and Planctomycetes, 37.5% of dominant OTUs were additionally detected in the wild beet rhizosphere. Analysis of the cultivable fraction confirmed these plant genotype-specific differences at functional level. The proportion of isolates displayed in vitro activity against phytopathogens was lower for wild beet (≤45.8%) than for sugar beet (≤57.5%). Conversely, active isolates from the wild beet exhibited stronger ability to cope with abiotic stresses. From all samples, active isolates of Stenotrophomonas rhizophila were frequently identified. In addition, soil type-specific impacts on the composition of bacterial communities were found: Acidobacteria, Chloroflexi, and Planctomycetes were only detected in plants cultivated in CS; whereas Bacteroidetes and Proteobacteria dominated in PS. Overall, in comparison to modern sugar beets, wild beets were associated with taxonomically and functionally distinct microbiomes.

  20. Targeted sequence capture provides insight into genome structure and genetics of male sterility in a gynodioecious diploid strawberry, Fragaria vesca ssp. bracteata (Rosaceae).

    PubMed

    Tennessen, Jacob A; Govindarajulu, Rajanikanth; Liston, Aaron; Ashman, Tia-Lynn

    2013-08-07

    Gynodioecy is a sexual system wherein females coexist with hermaphrodites. It is of interest not only because male-sterile plants are advantageous in plant breeding but also because it can be a crucial step in the evolutionary transition to entirely separate sexes (dioecy) from a hermaphroditic ancestor. The gynodioecious diploid wild strawberry, Fragaria vesca ssp. bracteata (Rosaceae), is a member of a clade with both dioecious and cultivated species, making it an ideal model in which to study the genetics of male sterility. To create a genetic map of F. v. ssp. bracteata, we identified informative polymorphisms from genomic sequencing (3-5x coverage) of two outbred plants from the same population. Using targeted enrichment, we sequenced 200 bp surrounding each of 6575 polymorphisms in 48 F1 offspring, yielding genotypes at 98% of targeted sites with mean coverage >100x, plus more than 600-kb high-coverage nontargeted sequence. With the resulting linkage map of 7802 stringently filtered markers (5417 targeted), we assessed recombination rates and genomic incongruities. Consistent with past work in strawberries, male sterility is dominant, segregates 1:1, and maps to a single location in the female. Further mapping an additional 55 offspring places male sterility in a gene-dense, 338-kb region of chromosome 4. The region is not syntenic with the sex-determining regions in the closely related octoploids, F. chiloensis and F. virginiana, suggesting either independent origins or translocation. The 57 genes in this region do not include protein families known to control male sterility and thus suggest alternate mechanisms for the suppression of male function.

  1. Functional identification of genes responsible for the biosynthesis of 1-methoxy-indol-3-ylmethyl-glucosinolate in Brassica rapa ssp. chinensis

    PubMed Central

    2014-01-01

    Background Brassica vegetables contain a class of secondary metabolites, the glucosinolates (GS), whose specific degradation products determine the characteristic flavor and smell. While some of the respective degradation products of particular GS are recognized as health promoting substances for humans, recent studies also show evidence that namely the 1-methoxy-indol-3-ylmethyl GS might be deleterious by forming characteristic DNA adducts. Therefore, a deeper knowledge of aspects involved in the biosynthesis of indole GS is crucial to design vegetables with an improved secondary metabolite profile. Results Initially the leafy Brassica vegetable pak choi (Brassica rapa ssp. chinensis) was established as suitable tool to elicit very high concentrations of 1-methoxy-indol-3-ylmethyl GS by application of methyl jasmonate. Differentially expressed candidate genes were discovered in a comparative microarray analysis using the 2 × 104 K format Brassica Array and compared to available gene expression data from the Arabidopsis AtGenExpress effort. Arabidopsis knock out mutants of the respective candidate gene homologs were subjected to a comprehensive examination of their GS profiles and confirmed the exclusive involvement of polypeptide 4 of the cytochrome P450 monooxygenase subfamily CYP81F in 1-methoxy-indol-3-ylmethyl GS biosynthesis. Functional characterization of the two identified isoforms coding for CYP81F4 in the Brassica rapa genome was performed using expression analysis and heterologous complementation of the respective Arabidopsis mutant. Conclusions Specific differences discovered in a comparative microarray and glucosinolate profiling analysis enables the functional attribution of Brassica rapa ssp. chinensis genes coding for polypeptide 4 of the cytochrome P450 monooxygenase subfamily CYP81F to their metabolic role in indole glucosinolate biosynthesis. These new identified Brassica genes will enable the development of genetic tools for breeding

  2. Molecular characterization of BrMYB28 and BrMYB29 paralogous transcription factors involved in the regulation of aliphatic glucosinolate profiles in Brassica rapa ssp. pekinensis.

    PubMed

    Baskar, Venkidasamy; Park, Se Won

    2015-07-01

    Glucosinolates (GSL) are one of the major secondary metabolites of the Brassicaceae family. In the present study, we aim at characterizing the multiple paralogs of aliphatic GSL regulators, such as BrMYB28 and BrMYB29 genes in Brassica rapa ssp. pekinensis, by quantitative real-time PCR (qRT-PCR) analysis in different tissues and at various developmental stages. An overlapping gene expression pattern between the BrMYBs as well as their downstream genes (DSGs) was found at different developmental stages. Among the BrMYB28 and BrMYB29 paralogous genes, the BrMYB28.3 and BrMYB29.1 genes were dominantly expressed in most of the developmental stages, compared to the other paralogs of the BrMYB genes. Furthermore, the differential expression pattern of the BrMYBs was observed under various stress treatments. Interestingly, BrMYB28.2 showed the least expression in most developmental stages, while its expression was remarkably high in different stress conditions. More specifically, the BrMYB28.2, BrMYB28.3, and BrMYB29.1 genes were highly responsive to various abiotic and biotic stresses, further indicating their possible role in stress tolerance. Moreover, the in silico cis motif analysis in the upstream regulatory regions of BrMYBs showed the presence of various putative stress-specific motifs, which further indicated their responsiveness to biotic and abiotic stresses. These observations suggest that the dominantly expressed BrMYBs, both in different developmental stages and under various stress treatments (BrMYB28.3 and BrMYB29.1), may be potential candidate genes for altering the GSL level through genetic modification studies in B. rapa ssp. pekinensis.

  3. Molecular cloning and expression of the male sterility-related CtYABBY1 gene in flowering Chinese cabbage (Brassica campestris L. ssp chinensis var. parachinensis).

    PubMed

    Zhang, X L; Zhang, L G

    2014-06-10

    Expression of the YABBY gene family in the abaxial surface of lateral plant organs determines abaxial destiny of cells, enhances growth and expansion of lateral organs, and plays an important role in polar establishment of lateral organs. However, the YABBY gene has not been studied in male sterility and fertility restoration. We homologously cloned the CtYABBY1 gene of male-sterile TC1 in Brassica campestris L. ssp chinensis var. parachinensis; its expression was analyzed by real-time PCR. A 937-bp sequence was cloned from TC1 and named CtYABBY1. The ORF of this gene has 702 bp, contains a "C2C2 zinc finger" motif at the N-terminal end, and a "YABBY" structural domain at the C-terminal end. This gene had the highest homology with DBC43-3-2 gene in B. campetris ssp pekinensis. Expression of CtYABBY1 gene has a wide range of functions. It is involved in growth and development of lateral organs, such as leaves and flowers, enhancing expansion of the area and volume of young organs. CtYABBY1 is a gene that promotes thermo-sensitive fertility restoration. At room temperature, expression level of this gene was found to be lower in the stamens of sterile flowers. After treating TC1 at a low temperature of 2°-6°C for 20 days, expression of this gene was upregulated in the stamen of fertile flowers. We conclude that male sterility in TC1 is negatively regulated by this gene, which facilitates transition from male sterility to fertility.

  4. Differences between the rhizosphere microbiome of Beta vulgaris ssp. maritima-ancestor of all beet crops-and modern sugar beets.

    PubMed

    Zachow, Christin; Müller, Henry; Tilcher, Ralf; Berg, Gabriele

    2014-01-01

    The structure and function of the plant microbiome is driven by plant species and prevailing environmental conditions. Effectuated by breeding efforts, modern crops diverge genetically and phenotypically from their wild relatives but little is known about consequences for the associated microbiota. Therefore, we studied bacterial rhizosphere communities associated with the wild beet B. vulgaris ssp. maritima grown in their natural habitat soil from coastal drift lines (CS) and modern sugar beets (Beta vulgaris ssp. vulgaris) cultivated in CS and potting soil (PS) under greenhouse conditions. Analysis of 16S rRNA gene fingerprints and pyrosequencing-based amplicon libraries revealed plant genotype- and soil-specific microbiomes. Wild beet plants harbor distinct operational taxonomic units (OTUs) and a more diverse bacterial community than the domesticated sugar beet plants. Although the rhizospheres of both plant genotypes were dominated by Proteobacteria and Planctomycetes, 37.5% of dominant OTUs were additionally detected in the wild beet rhizosphere. Analysis of the cultivable fraction confirmed these plant genotype-specific differences at functional level. The proportion of isolates displayed in vitro activity against phytopathogens was lower for wild beet (≤45.8%) than for sugar beet (≤57.5%). Conversely, active isolates from the wild beet exhibited stronger ability to cope with abiotic stresses. From all samples, active isolates of Stenotrophomonas rhizophila were frequently identified. In addition, soil type-specific impacts on the composition of bacterial communities were found: Acidobacteria, Chloroflexi, and Planctomycetes were only detected in plants cultivated in CS; whereas Bacteroidetes and Proteobacteria dominated in PS. Overall, in comparison to modern sugar beets, wild beets were associated with taxonomically and functionally distinct microbiomes. PMID:25206350

  5. THE NON-UNIVERSALITY OF THE LOW-MASS END OF THE IMF IS ROBUST AGAINST THE CHOICE OF SSP MODEL

    SciTech Connect

    Spiniello, C.; Trager, S. C.; Koopmans, L. V. E.

    2015-04-20

    We perform a direct comparison of two state-of-the art single stellar population (SSP) models that have been used to demonstrate the non-universality of the low-mass end of the initial mass function (IMF) slope. The two public versions of the SSP models are restricted to either solar abundance patterns or solar metallicity, too restrictive if one aims to disentangle elemental enhancements, metallicity changes, and IMF variations in massive early-type galaxies (ETGs) with star formation histories different from those in the solar neighborhood. We define response functions (to metallicity and α-abundance) to extend the parameter space for each set of models. We compare these extended models with a sample of Sloan Digital Sky Survey (SDSS) ETG spectra with varying velocity dispersions. We measure equivalent widths of optical IMF-sensitive stellar features to examine the effect of the underlying model assumptions and ingredients, such as stellar libraries or isochrones, on the inference of the IMF slope down to ∼0.1 M{sub ⊙}. We demonstrate that the steepening of the low-mass end of the IMF based on a non-degenerate set of spectroscopic optical indicators is robust against the choice of the stellar population model. Although the models agree in a relative sense (i.e., both imply more bottom-heavy IMFs for more massive systems), we find non-negligible differences in the absolute values of the IMF slope inferred at each velocity dispersion by using the two different models. In particular, we find large inconsistencies in the quantitative predictions of the IMF slope variations and abundance patterns when sodium lines are used. We investigate the possible reasons for these inconsistencies.

  6. Comparative sequence analysis of the potato cyst nematode resistance locus H1 reveals a major lack of co-linearity between three haplotypes in potato (Solanum tuberosum ssp.)

    PubMed Central

    Bakker, Erin; de Boer, Jan; van der Vossen, Edwin; Achenbach, Ute; Golas, Tomasz; Suryaningrat, Suwardi; Smant, Geert; Bakker, Jaap; Goverse, Aska

    2010-01-01

    The H1 locus confers resistance to the potato cyst nematode Globodera rostochiensis pathotypes 1 and 4. It is positioned at the distal end of chromosome V of the diploid Solanum tuberosum genotype SH83-92-488 (SH) on an introgression segment derived from S. tuberosum ssp. andigena. Markers from a high-resolution genetic map of the H1 locus (Bakker et al. in Theor Appl Genet 109:146–152, 2004) were used to screen a BAC library to construct a physical map covering a 341-kb region of the resistant haplotype coming from SH. For comparison, physical maps were also generated of the two haplotypes from the diploid susceptible genotype RH89-039-16 (S. tuberosum ssp. tuberosum/S. phureja), spanning syntenic regions of 700 and 319 kb. Gene predictions on the genomic segments resulted in the identification of a large cluster consisting of variable numbers of the CC-NB-LRR type of R genes for each haplotype. Furthermore, the regions were interspersed with numerous transposable elements and genes coding for an extensin-like protein and an amino acid transporter. Comparative analysis revealed a major lack of gene order conservation in the sequences of the three closely related haplotypes. Our data provide insight in the evolutionary mechanisms shaping the H1 locus and will facilitate the map-based cloning of the H1 resistance gene. Electronic supplementary material The online version of this article (doi:10.1007/s00122-010-1472-9) contains supplementary material, which is available to authorized users. PMID:21049265

  7. Molecular cloning and expression of the male sterility-related CtYABBY1 gene in flowering Chinese cabbage (Brassica campestris L. ssp chinensis var. parachinensis).

    PubMed

    Zhang, X L; Zhang, L G

    2014-01-01

    Expression of the YABBY gene family in the abaxial surface of lateral plant organs determines abaxial destiny of cells, enhances growth and expansion of lateral organs, and plays an important role in polar establishment of lateral organs. However, the YABBY gene has not been studied in male sterility and fertility restoration. We homologously cloned the CtYABBY1 gene of male-sterile TC1 in Brassica campestris L. ssp chinensis var. parachinensis; its expression was analyzed by real-time PCR. A 937-bp sequence was cloned from TC1 and named CtYABBY1. The ORF of this gene has 702 bp, contains a "C2C2 zinc finger" motif at the N-terminal end, and a "YABBY" structural domain at the C-terminal end. This gene had the highest homology with DBC43-3-2 gene in B. campetris ssp pekinensis. Expression of CtYABBY1 gene has a wide range of functions. It is involved in growth and development of lateral organs, such as leaves and flowers, enhancing expansion of the area and volume of young organs. CtYABBY1 is a gene that promotes thermo-sensitive fertility restoration. At room temperature, expression level of this gene was found to be lower in the stamens of sterile flowers. After treating TC1 at a low temperature of 2°-6°C for 20 days, expression of this gene was upregulated in the stamen of fertile flowers. We conclude that male sterility in TC1 is negatively regulated by this gene, which facilitates transition from male sterility to fertility. PMID:25036178

  8. Molecular characterization of BrMYB28 and BrMYB29 paralogous transcription factors involved in the regulation of aliphatic glucosinolate profiles in Brassica rapa ssp. pekinensis.

    PubMed

    Baskar, Venkidasamy; Park, Se Won

    2015-07-01

    Glucosinolates (GSL) are one of the major secondary metabolites of the Brassicaceae family. In the present study, we aim at characterizing the multiple paralogs of aliphatic GSL regulators, such as BrMYB28 and BrMYB29 genes in Brassica rapa ssp. pekinensis, by quantitative real-time PCR (qRT-PCR) analysis in different tissues and at various developmental stages. An overlapping gene expression pattern between the BrMYBs as well as their downstream genes (DSGs) was found at different developmental stages. Among the BrMYB28 and BrMYB29 paralogous genes, the BrMYB28.3 and BrMYB29.1 genes were dominantly expressed in most of the developmental stages, compared to the other paralogs of the BrMYB genes. Furthermore, the differential expression pattern of the BrMYBs was observed under various stress treatments. Interestingly, BrMYB28.2 showed the least expression in most developmental stages, while its expression was remarkably high in different stress conditions. More specifically, the BrMYB28.2, BrMYB28.3, and BrMYB29.1 genes were highly responsive to various abiotic and biotic stresses, further indicating their possible role in stress tolerance. Moreover, the in silico cis motif analysis in the upstream regulatory regions of BrMYBs showed the presence of various putative stress-specific motifs, which further indicated their responsiveness to biotic and abiotic stresses. These observations suggest that the dominantly expressed BrMYBs, both in different developmental stages and under various stress treatments (BrMYB28.3 and BrMYB29.1), may be potential candidate genes for altering the GSL level through genetic modification studies in B. rapa ssp. pekinensis. PMID:26043798

  9. A novel cold-inducible gene from Pak-choi (Brassica campestris ssp. chinensis), BcWRKY46, enhances the cold, salt and dehydration stress tolerance in transgenic tobacco.

    PubMed

    Wang, Feng; Hou, Xilin; Tang, Jun; Wang, Zhen; Wang, Shuming; Jiang, Fangling; Li, Ying

    2012-04-01

    WRKY TFs belong to one of the largest families of transcriptional regulators in plants and form integral parts of signaling webs that modulate many plant processes. BcWRKY46, a cDNA clone encoding a polypeptide of 284 amino acids and exhibited the structural features of group III of WRKY protein family, was isolated from the cold-treated leaves of Pak-choi (Brassica campestris ssp. chinensis Makino, syn. B. rapa ssp. chinensis) using the cDNA-AFLP technique. Expression of this gene was induced quickly and strongly in response to various environmental stresses, including low temperatures, ABA, salt and dehydration. Constitutive expression of BcWRKY46 in tobacco under the control of the CaMV35S promoter reduced the susceptibility of transgenic tobacco to freezing, ABA, salt and dehydration stresses. Our studies suggest that BcWRKY46 plays an important role in responding to ABA and abiotic stress.

  10. Space Shuttle Program (SSP) Orbiter Main Propulsion System (MPS) Gaseous Hydrogen (GH2) Flow Control Valve (FCV) Poppet Eddy Current (EC) Inspection Probability of Detection (POD) Study. Volume 1

    NASA Technical Reports Server (NTRS)

    Piascik, Robert S.; Prosser, William H.

    2011-01-01

    The Director of the NASA Engineering and Safety Center (NESC), requested an independent assessment of the anomalous gaseous hydrogen (GH2) flow incident on the Space Shuttle Program (SSP) Orbiter Vehicle (OV)-105 during the Space Transportation System (STS)-126 mission. The main propulsion system (MPS) engine #2 GH2 flow control valve (FCV) LV-57 transition from low towards high flow position without being commanded. Post-flight examination revealed that the FCV LV-57 poppet had experienced a fatigue failure that liberated a section of the poppet flange. The NESC assessment provided a peer review of the computational fluid dynamics (CFD), stress analysis, and impact testing. A probability of detection (POD) study was requested by the SSP Orbiter Project for the eddy current (EC) nondestructive evaluation (NDE) techniques that were developed to inspect the flight FCV poppets. This report contains the findings and recommendations from the NESC assessment.

  11. Space Shuttle Program (SSP) Orbiter Main Propulsion System (MPS) Gaseous Hydrogen (GH2) Flow Control Valve (FCV) Poppet Eddy Current (EC) Inspection Probability of Detection (POD) Study. Volume 2; Appendices

    NASA Technical Reports Server (NTRS)

    Piascik, Robert S.; Prosser, William H.

    2011-01-01

    The Director of the NASA Engineering and Safety Center (NESC), requested an independent assessment of the anomalous gaseous hydrogen (GH2) flow incident on the Space Shuttle Program (SSP) Orbiter Vehicle (OV)-105 during the Space Transportation System (STS)-126 mission. The main propulsion system (MPS) engine #2 GH2 flow control valve (FCV) LV-57 transition from low towards high flow position without being commanded. Post-flight examination revealed that the FCV LV-57 poppet had experienced a fatigue failure that liberated a section of the poppet flange. The NESC assessment provided a peer review of the computational fluid dynamics (CFD), stress analysis, and impact testing. A probability of detection (POD) study was requested by the SSP Orbiter Project for the eddy current (EC) nondestructive evaluation (NDE) techniques that were developed to inspect the flight FCV poppets. This report contains the Appendices to the main report.

  12. Rapid DNA typing for HLA-C using sequence-specific primers (PCR-SSP): identification of serological and non-serologically defined HLA-C alleles including several new alleles.

    PubMed

    Bunce, M; Welsh, K I

    1994-01-01

    Detection of HLA-C antigens by complement mediated cytotoxicity using human alloantisera is often difficult. Between 20 to 40% of individuals in every race have undetectable HLA-C locus antigens and 9 out of the 29 sequenced HLA-C alleles so far published encode serologically undetected antigens. In addition, HLA-C molecules are expressed at the cell surface at about 10% of the levels of HLA-A and HLA-B. Recently, amplification of DNA using sequence-specific primers (PCR-SSP) has proved a reliable and rapid method for typing HLA-DR, HLA-DQA and HLA-DQB genes. PCR-SSP takes two hours to perform and is therefore suitable for the genotyping of cadaveric donors. We have designed a set of primers which will positively identify the HLA-C alleles corresponding to the serologically defined series HLA-Cw1, Cw2, Cw3, Cw4, Cw5, Cw6, Cw7 and Cw8. The serologically undetectable alleles have also been detected in groups according to sequence homology. In addition, three new unsequenced variants have been identified. DNA samples from 56 International Histocompatibility Workshop reference cell lines and 103 control individuals have been typed by the HLA-C PCR-SSP technique. 4/56 cell line types and 11/103 normal control individuals types were discrepant with the reported serological types. All combinations of serologically detectable and most of the serologically blank HLA-C antigens can be readily identified. DNA typing for HLA-Cw by PCR-SSP can take as little as 130 minutes from start to finish, including DNA preparation.

  13. Insecticidal efficacy of silica gel with Juniperus oxycedrus ssp. oxycedrus (Pinales: Cupressaceae) essential oil against Sitophilus oryzae (Coleoptera: Curculionidae) and Tribolium confusum (Coleoptera: Tenebrionidae).

    PubMed

    Athanassiou, Christos G; Kavallieratos, Nickolas C; Evergetis, Epameinondas; Katsoula, Anna-Maria; Haroutounian, Serkos A

    2013-08-01

    Laboratory bioassays were carried out to evaluate the effect of silica gel enhanced with the essential oil (EO) of Juniperus oxycedrus L. ssp. oxycedrus (Pinales: Cupressaceae) (derived from berry specimens from Greece) against adults of Sitophilus oryzae (L.) (Coleoptera: Curculionidae) and Tribolium confusum Jacquelin du Val (Coleoptera: Tenebrionidae). For that purpose, a dry mixture consisting of 500 mg of silica gel that had absorbed 2.18 mg of EO (total weight: 502.18 mg) was tested at three doses; 0.125, 0.250, and 0.5 g/kg of wheat, corresponding to 125, 250, and 500 ppm, respectively, and silica gel alone at 0.5 g/kg of wheat corresponding to 500 ppm, at different exposure intervals (24 and 48 h and 7 and 14 d for S. oryzae; 24 and 48 h and 7, 14, and 21 d for T. confusum). The chemical content of the specific EO was determined by gas chromatography (GC)-mass spectrometry (MS) analyses indicating the presence of 31 constituents with myrcene and germacrene-D being the predominant compounds. The bioactivity results for S. oryzae indicated that 48 h of exposure in wheat resulted in an 82% mortality for treatment with 500 ppm of the enhanced silica gel. For 7 d of exposure, 100 and 98% of S. oryzae adults died when they were treated with 500 and 250 ppm of enhanced silica gel, respectively. At 14 d of exposure, all adults died both at 250 and 500 ppm of enhanced silica gel. At 48 h, 7 and 14 d of exposure significantly less S. oryzae adults died in wheat treated with silica gel alone than at 250 or 500 ppm of enhanced silica gel. In the case of T. confusum, at 7 d of exposure, mortality in wheat treated with silica gel only was significantly higher in comparison to the other treatments. At the 14 d of exposure mortality in wheat treated with 500 ppm of silica gel alone was significantly higher than 125 and 250 ppm of the enhanced silica gel. Similar trends were also noted at 21 d of exposure, indicating that there is no enhancement effect from the addition of

  14. Effect of feeding heat-treated colostrum on risk for infection with Mycobacterium avium ssp. paratuberculosis, milk production, and longevity in Holstein dairy cows.

    PubMed

    Godden, S M; Wells, S; Donahue, M; Stabel, J; Oakes, J M; Sreevatsan, S; Fetrow, J

    2015-08-01

    In summer 2007, a randomized controlled field trial was initiated on 6 large Midwest commercial dairy farms to investigate the effect of feeding heat-treated (HT) colostrum on transmission of Mycobacterium avium ssp. paratuberculosis (MAP) and on future milk production and longevity within the herd. On each farm, colostrum was collected daily from fresh cows, pooled, divided into 2 aliquots, and then 1 aliquot was heat-treated in a commercial batch pasteurizer at 60°C for 60min. A sample from each batch of colostrum was collected for PCR testing (MAP-positive vs. MAP-negative). Newborn heifer calves were removed from the dam within 30 to 60min of birth and systematically assigned to be fed 3.8 L of either fresh (FR; n=434) or heat-treated (HT; n=490) colostrum within 2h of birth. After reaching adulthood (>2 yr old), study animals were tested once annually for 3 yr (2010, 2011, 2012) for infection with MAP using serum ELISA and fecal culture. Lactation records describing milk production data and death or culling events were collected during the 3-yr testing period. Multivariable model logistic and linear regression was used to investigate the effect of feeding HT colostrum on risk for testing positive to MAP during the 3-yr testing period (positive/negative; logistic regression) and on first and second lactation milk yield (kg/cow; linear regression), respectively. Cox proportional hazards regression was used to investigate the effect of feeding HT colostrum on risk and time to removal from the herd. Fifteen percent of all study animals were fed PCR-positive colostrum. By the end of the 3-yr testing period, no difference was noted in the proportion of animals testing positive for MAP, with either serum ELISA or fecal culture, when comparing the HT group (10.5%) versus the FR group (8.1%). There was no effect of treatment on first- (HT=11.797kg; FR=11,671kg) or second-lactation (HT=11,013kg; FR=11,235kg) milk production. The proportion of cows leaving the herd by

  15. Cucumis melo ssp. Agrestis var. Agrestis Ameliorates High Fat Diet Induced Dyslipidemia in Syrian Golden Hamsters and Inhibits Adipogenesis in 3T3-L1 Adipocytes

    PubMed Central

    Shankar, Kripa; Singh, Sumit K.; Kumar, Durgesh; Varshney, Salil; Gupta, Abhishek; Rajan, Sujith; Srivastava, Ankita; Beg, Muheeb; Srivastava, Anurag Kumar; Kanojiya, Sanjeev; Mishra, Dipak K.; Gaikwad, Anil N.

    2015-01-01

    Background: Cucumis melo ssp. agrestis var. agrestis (CMA) is a wild variety of C. melo. This study aimed to explore anti-dyslipidemic and anti-adipogenic potential of CMA. Materials and Methods: For initial anti-dyslipidemic and antihyperglycemic potential of CMA fruit extract (CMFE), male Syrian golden hamsters were fed a chow or high-fat diet with or without CMFE (100 mg/kg). Further, we did fractionation of this CMFE into two fractions namely; CMA water fraction (CMWF) and CMA hexane fraction (CMHF). Phytochemical screening was done with liquid chromatography-mass spectrometry LC- (MS)/MS and direct analysis in real time-MS to detect active compounds in the fractions. Further, high-fat diet fed dyslipidemic hamsters were treated with CMWF and CMHF at 50 mg/kg for 7 days. Results: Oral administration of CMFE and both fractions (CMWF and CMHF) reduced the total cholesterol, triglycerides, low‐density lipoprotein cholesterol, and very low‐density lipoprotein-cholesterol levels in high fat diet-fed dyslipidemic hamsters. CMHF also modulated expression of genes involved in lipogenesis, lipid metabolism, and reverse cholesterol transport. Standard biochemical diagnostic tests suggested that neither of fractions causes any toxicity to hamster liver or kidneys. CMFE and CMHF also decreased oil-red-O accumulation in 3T3-L1 adipocytes. Conclusion: Based on these results, it is concluded that CMA possesses anti-dyslipidemic and anti-hyperglycemic activity along with the anti-adipogenic activity. SUMMARY The oral administration of Cucumis melo agrestis fruit extract (CMFE) and its fractions (CMWF and CMHF) improved serum lipid profile in HFD fed dyslipidemic hamsters.CMFE, CMWF and CMHF significantly attenuated body weight gain and eWAT hypertrophy.The CMHF decreased lipogenesis in both liver and adipose tissue.CMFE and CMHF also inhibited adipogenesis in 3T3-L1 adipocytes. Abbreviation used: CMA: Cucumis melo ssp. agrestis var. agrestis, CMFE: CMA fruit extract, CMWF

  16. Identification of a RAPD marker for palmitic-acid concentration in the seed oil of spring turnip rape (Brassica rapa ssp. oleifera).

    PubMed

    Tanhuanpää, P K; Vilkki, J P; Vilkki, H J

    1995-08-01

    F2 progeny (105 individuals) from the cross Jo4002 x Sv3402 were used to identify DNA markers associated with palmitic-acid content in spring turnip rape (Brassica rapa ssp. oleifera). QTL mapping and ANOVA analysis of 140 markers exposed one linkage group with a locus controlling palmitic-acid content (LOD score 27), and one RAPD (random amplified polymorphic DNA) marker, OPB-11a, closely linked (1.4 cM) to this locus. Palmitic-acid content in the 62 F2 plants with the visible allele of marker OPB-11a was 8.45 ±3.15%, while that in the 24 plants without it was 4.59 ±0.97%. As oleic-acid concentration is affected by a locus on the same linkage group as the palmitic-acid locus, this locus probably controls the chain elongation from palmitic acid to oleic acid (through stearic acid). Marker OPB-11a may be used in future breeding programs of spring turnip rape to simplify and hasten the selection for palmitic-acid content.

  17. Carbohydrate derivatives from the roots of Brassica rapa ssp. campestris and their effects on ROS production and glutamate-induced cell death in HT-22 cells.

    PubMed

    Wu, Qian; Cho, Jin-Gyeong; Lee, Dong-Sung; Lee, Dae-Young; Song, Na-Young; Kim, Youn-Chul; Lee, Kyung-Tae; Chung, Hae-Gon; Choi, Myung-Sook; Jeong, Tae-Sook; Ahn, Eun-Mi; Kim, Geum-Soog; Baek, Nam-In

    2013-05-01

    Phytochemical investigation of the roots of Brassica rapa ssp. campestris led to the isolation of three new carbohydrate derivatives, namely sucrose 3,3',4'-triisovalerate (2), sucrose 6,3',4'-triisovalerate (3), and ethanone-1-C-β-d-glucopyranoside (3,7-anhydro-1-deoxy-d-glycero-d-gulo-2-octulose, 6), along with four known carbohydrate derivatives, 2,6,3',4'-tetraisovalerate (1), ethyl β-d-glucopyranoside (4), n-butyl β-d-fructofuranoside (5), and n-pentyl β-d-fructofuranoside (7), which were initially isolated from plants of the Brassica genus. Structures of the isolated compounds were established by spectroscopic analyses, including UV, IR, MS, and NMR. All of the isolated carbohydrate derivatives were evaluated to determine their effect on ROS production and glutamate-induced cell death in HT-22 cells. Compound 6 showed the most significant ROS reduction and a protective effect with IC50 values of 69.4 ± 3.8 μM and 4.96 ± 0.32 μM, respectively, which were equivalent to those of the positive control, Trolox.

  18. Induced production of 1-methoxy-indol-3-ylmethyl glucosinolate by jasmonic acid and methyl jasmonate in sprouts and leaves of pak choi (Brassica rapa ssp. chinensis).

    PubMed

    Wiesner, Melanie; Hanschen, Franziska S; Schreiner, Monika; Glatt, Hansruedi; Zrenner, Rita

    2013-01-01

    Pak choi plants (Brassica rapa ssp. chinensis) were treated with different signaling molecules methyl jasmonate, jasmonic acid, linolenic acid, and methyl salicylate and were analyzed for specific changes in their glucosinolate profile. Glucosinolate levels were quantified using HPLC-DAD-UV, with focus on induction of indole glucosinolates and special emphasis on 1-methoxy-indol-3-ylmethyl glucosinolate. Furthermore, the effects of the different signaling molecules on indole glucosinolate accumulation were analyzed on the level of gene expression using semi-quantitative realtime RT-PCR of selected genes. The treatments with signaling molecules were performed on sprouts and mature leaves to determine ontogenetic differences in glucosinolate accumulation and related gene expression. The highest increase of indole glucosinolate levels, with considerable enhancement of the 1-methoxy-indol-3-ylmethyl glucosinolate content, was achieved with treatments of sprouts and mature leaves with methyl jasmonate and jasmonic acid. This increase was accompanied by increased expression of genes putatively involved in the indole glucosinolate biosynthetic pathway. The high levels of indole glucosinolates enabled the plant to preferentially produce the respective breakdown products after tissue damage. Thus, pak choi plants treated with methyl jasmonate or jasmonic acid, are a valuable tool to analyze the specific protection functions of 1-methoxy-indole-3-carbinole in the plants defense strategy in the future.

  19. Identification of the BrRHP1 locus that confers resistance to downy mildew in Chinese cabbage (Brassica rapa ssp. pekinensis) and development of linked molecular markers.

    PubMed

    Kim, Sunggil; Song, Young Ha; Lee, Ji-Yeon; Choi, Su Ryun; Dhandapani, Vignesh; Jang, Chang Soon; Lim, Yong Pyo; Han, Taeho

    2011-11-01

    Inheritance of resistance to downy mildew (Hyaloperonospora parasitica) in Chinese cabbage (Brassica rapa ssp. pekinensis) was studied using inbred parental lines RS1 and SS1 that display strong resistance and severe susceptibility, respectively. F(1), F(2), and BC(1)F(1) populations were evaluated for their responses to downy mildew infection. Resistance to downy mildew was conditioned by a single dominant locus designated BrRHP1. A random amplified polymorphic DNA (RAPD) marker linked to BrRHP1 was identified using bulked segregant analysis and two molecular markers designated BrPERK15A and BrPERK15B were developed. BrPERK15B was polymorphic between the parental lines used to construct the reference linkage map of B. rapa, allowing the mapping of the BrRHP1 locus to the A1 linkage group. Using bacterial artificial chromosome clone sequences anchored to the A1 linkage group, six simple polymerase chain reaction (PCR) markers were developed for use in marker-assisted breeding of downy mildew resistance in Chinese cabbage. Four simple PCR markers flanking the BrRHP1 locus were shown to be collinear with the long-arm region of Arabidopsis chromosome 3. The two closely linked flanking markers delimit the BrRHP1 locus within a 2.2-Mb interval of this Arabidopsis syntenic region.

  20. Isolation of fertility-related genes of multiple-allele-inherited male sterility in Brassica rapa ssp pekinensis by cDNA-AFLP.

    PubMed

    Ji, R Q; Song, Q; Xin, X F; Zhou, X; Feng, H

    2011-01-01

    To better understand the molecular mechanisms of multiple-allele-inherited male sterility in Chinese cabbage (Brassica rapa ssp pekinensis), differentially expressed genes in fertile and sterile plants must be isolated. We used cDNA-AFLP analysis to isolate differentially expressed genes in fertile and sterile buds of the two-type line, AB01. Sixteen high-quality sequences were generated, 11 of which were up-regulated in fertile buds, and five of which were up-regulated in sterile buds. Based on BLAST screening and functional annotation, these genes have homology with genes encoding known flower- or bud-specific proteins, metabolism-related proteins and cell-structure proteins. In addition, the full-length cDNA sequences of the actin gene were cloned from the cabbage plants by RACE and used as an internal standard for semi-quantitative reverse transcription-PCR. Expression of three flower- or bud-specific differentially expressed transcript-derived fragments in fertile and sterile buds was examined using RT-PCR; the expression patterns of these genes were similar to the patterns observed in the cDNA-AFLP analysis.

  1. Chemical composition and biological activity of the essential oil of Origanum vulgare ssp. hirtum from different areas in the Southern Apennines (Italy).

    PubMed

    Mancini, Emilia; Camele, Ippolito; Elshafie, Hazem S; De Martino, Laura; Pellegrino, Carlo; Grulova, Daniela; De Feo, Vincenzo

    2014-04-01

    The chemical composition of the essential oils of Origanum vulgare ssp. hirtum, growing wild in three different localities in the Southern Apennines, was studied by GC-FID and GC/MS analyses. In total, 103 compounds were identified. The oils were mainly composed of phenolic compounds and all oils belonged to the chemotype carvacrol/thymol. The three essential oils were evaluated for their in vitro phytotoxic activity by determining their influence on the germination and initial radicle elongation of Sinapis arvensis L., Phalaris canariensis L., Lepidium sativum L., and Raphanus sativus L. The seed germination and radicle growth were affected in various degrees. Moreover, the antifungal activity of the three essential oils was assayed against three species causing pre- and postharvest fruit decay (Monilinia laxa, M. fructigena, and M. fructicola). At 1000 ppm, the three oils completely inhibited fungal growth. The hemolytic activity of the oils was assayed and showed no effect on the cell membranes of bovine erythrocytes. PMID:24706631

  2. Microsatellite analysis of a population crash and bottleneck in the Mauna Kea silversword, Argyroxiphium sandwicense ssp. sandwicense (Asteraceae), and its implications for reintroduction.

    PubMed

    Friar, E A; Ladoux, T; Roalson, E H; Robichaux, R H

    2000-12-01

    The Mauna Kea silversword, Argyroxiphium sandwicense ssp. sandwicense, has experienced both a severe population crash associated with an increase in alien ungulate populations on Mauna Kea, and a population bottleneck associated with reintroduction. In this paper, we address the genetic consequences of both demographic events using eight microsatellite loci. The population crash was not accompanied by a significant reduction in number of alleles or heterozygosity. However, the population bottleneck was accompanied by significant reductions in observed number of alleles, effective number of alleles, and expected heterozygosity, though not in observed heterozygosity. The effective size of the population bottleneck was calculated using both observed heterozygosities and allele frequency variances. Both methods corroborated the historical census size of the population bottleneck of at most three individuals. The results suggest that: (i) small populations, even those that result from severe reductions in historical population size and extent, are not necessarily genetically depauperate; and (ii) species reintroduction plans need to be conceived and implemented carefully, with due consideration to the genetic impact of sampling for reintroduction.

  3. Microsatellite analysis of a population crash and bottleneck in the Mauna Kea silversword, Argyroxiphium sandwicense ssp. sandwicense (Asteraceae), and its implications for reintroduction.

    PubMed

    Friar, E A; Ladoux, T; Roalson, E H; Robichaux, R H

    2000-12-01

    The Mauna Kea silversword, Argyroxiphium sandwicense ssp. sandwicense, has experienced both a severe population crash associated with an increase in alien ungulate populations on Mauna Kea, and a population bottleneck associated with reintroduction. In this paper, we address the genetic consequences of both demographic events using eight microsatellite loci. The population crash was not accompanied by a significant reduction in number of alleles or heterozygosity. However, the population bottleneck was accompanied by significant reductions in observed number of alleles, effective number of alleles, and expected heterozygosity, though not in observed heterozygosity. The effective size of the population bottleneck was calculated using both observed heterozygosities and allele frequency variances. Both methods corroborated the historical census size of the population bottleneck of at most three individuals. The results suggest that: (i) small populations, even those that result from severe reductions in historical population size and extent, are not necessarily genetically depauperate; and (ii) species reintroduction plans need to be conceived and implemented carefully, with due consideration to the genetic impact of sampling for reintroduction. PMID:11123615

  4. Anthocyanin Accumulation, Antioxidant Ability and Stability, and a Transcriptional Analysis of Anthocyanin Biosynthesis in Purple Heading Chinese Cabbage (Brassica rapa L. ssp. pekinensis).

    PubMed

    He, Qiong; Zhang, Zhanfeng; Zhang, Lugang

    2016-01-13

    Heading Chinese cabbage (Brassica rapa L. ssp. pekinensis) is a significant dietary vegetable for its edible heading leaves in Asia countries. The new purple anthocyanin-rich pure line (11S91) was successfully bred, and the anthocyanins were mainly distributed in 2-3 cell layers beneath the leaf epidermis, whereas siliques and stems accumulated only a cell layer of anthocyanins. The anthocyanins of 11S91 were more stable at pHs below 3.0 and temperatures below 45 °C. The total antioxidant ability was highly positive correlated with the anthocyanin content in 11S91. Thirty-two anthocyanins were separated and identified, and 70% of them were glycosylated and acylated cyanidins. The four major anthocyanins present were cyanidin-3-sophoroside(p-coumaroyl)-5-glucoside(malonyl), cyanidin-3-sophoroside(ferulyl)-5-glucoside(malonyl), cyanidin-3-sophoroside(sinapyl-p-coumaroyl)-5-glucoside(malonyl), and cyanidin-3-sophoroside-(sinapyl-ferulyl)-5-glucoside(malonyl). According to the expression of biosynthetic genes and the component profile of anthocyanins in 11S91 and its parents, regulatory genes BrMYB2 and BrTT8 probably activate the anthocyanin biosynthesis but other factors may govern the primary anthocyanins and the distribution. PMID:26709726

  5. Relationships between functional genes in Lactobacillus delbrueckii ssp. bulgaricus isolates and phenotypic characteristics associated with fermentation time and flavor production in yogurt elucidated using multilocus sequence typing.

    PubMed

    Liu, Wenjun; Yu, Jie; Sun, Zhihong; Song, Yuqin; Wang, Xueni; Wang, Hongmei; Wuren, Tuoya; Zha, Musu; Menghe, Bilige; Heping, Zhang

    2016-01-01

    Lactobacillus delbrueckii ssp. bulgaricus (L. bulgaricus) is well known for its worldwide application in yogurt production. Flavor production and acid producing are considered as the most important characteristics for starter culture screening. To our knowledge this is the first study applying functional gene sequence multilocus sequence typing technology to predict the fermentation and flavor-producing characteristics of yogurt-producing bacteria. In the present study, phenotypic characteristics of 35 L. bulgaricus strains were quantified during the fermentation of milk to yogurt and during its subsequent storage; these included fermentation time, acidification rate, pH, titratable acidity, and flavor characteristics (acetaldehyde concentration). Furthermore, multilocus sequence typing analysis of 7 functional genes associated with fermentation time, acid production, and flavor formation was done to elucidate the phylogeny and genetic evolution of the same L. bulgaricus isolates. The results showed that strains significantly differed in fermentation time, acidification rate, and acetaldehyde production. Combining functional gene sequence analysis with phenotypic characteristics demonstrated that groups of strains established using genotype data were consistent with groups identified based on their phenotypic traits. This study has established an efficient and rapid molecular genotyping method to identify strains with good fermentation traits; this has the potential to replace time-consuming conventional methods based on direct measurement of phenotypic traits. PMID:26547656

  6. Sensitivity of solid culture, broth culture, and real-time PCR assays for milk and colostrum samples from Mycobacterium avium ssp. paratuberculosis-infectious dairy cows.

    PubMed

    Laurin, Emilie; McKenna, Shawn; Chaffer, Marcelo; Keefe, Greg

    2015-12-01

    Mycobacterium avium ssp. paratuberculosis (MAP) can be shed in feces, milk, and colostrum. The goal of this study was to assess assays that detect MAP in these sample types, including effects of lactation stage or season. Understanding the performance of these assays could improve how they are used, limiting the risk of infection to calves. Forty-six previously confirmed MAP-positive cows from 7 Atlantic Canadian dairy farms were identified for colostrum sampling and monthly sampling of milk and feces over a 12-mo period. Samples were assayed for MAP using solid culture, broth culture, and direct real-time PCR (qPCR). Across assay types, test sensitivity when applied to milk samples averaged 25% of that when applied to fecal samples. For colostrum samples, sensitivity depended on assay type, with sensitivity of qPCR being approximately 46% of that in feces. Across sample types, sensitivity of qPCR was higher than that of the other assays. Sensitivity of qPCR, when applied to milk samples, was significantly higher in summer than in other seasons. Summer was also the season with highest agreement between milk and fecal samples collected within the same month. Our results suggest that qPCR would detect more cows shedding MAP in their milk and colostrum than solid or broth culture assays, particularly during the summer, thus providing better management information to limit exposure of calves to this infectious organism. PMID:26476944

  7. Geosmin (2β,6α-dimethylbicyclo[4.4.0]decan-1β-ol) production associated with Beta vulgaris ssp. vulgaris is cultivar specific.

    PubMed

    Freidig, Amy K; Goldman, Irwin L

    2014-03-01

    The characteristic earthy flavor and aroma of table beet [Beta vulgaris ssp. vulgaris (garden beet group)] is due to the presence of geosmin, C₁₂H₂₂O, a volatile terpenoid compound commonly produced by many soil microorganisms. This study screened beet and related subspecies cultivars grown in three different environments (field, greenhouse in nonautoclaved soil, greenhouse in autoclaved soil) to evaluate the effect of cultivar and environment on geosmin level in table beet. There was no significant difference between years or between cultivars grown in autoclaved and nonautoclaved soil, indicating geosmin content may not be primarily attributable to microbial associations. A significant interaction between cultivar and environment was found, but generalizations could be made for high- or low-producing cultivars, demonstrating that geosmin levels were cultivar specific. 'Bull's Blood', 'Chioggia', and sugar beet exhibited the highest geosmin levels. Cultivars grown in the field had the smallest range of geosmin production, from 4.84 to 20.82 μg geosmin (kg root tissue)⁻¹. The high degree of consistency in cultivar performance across years and in ranking for geosmin levels across environments as well as the lack of a significant difference between plants grown in autoclaved and nonautoclaved soil suggests characteristic levels of geosmin may be present in and produced endogenously by cultivars of table beet. It may be possible to establish breeding populations with defined geosmin levels and to identify variety-specific aroma and flavor intensities that would be durable across environments.

  8. Molecular cloning and characterization of a novel pollen predominantly membrane protein gene BcMF12 from Brassica campestris ssp. chinensis.

    PubMed

    Song, Jianghua; Zhang, Lixin; Cao, Jiashu

    2009-11-01

    A novel membrane protein gene, BcMF12, was isolated from Chinese cabbage (Brassica campestris L. ssp. chinensis Makino) using rapid amplification of the cDNA ends based on a pollen-specific cDNA fragment (DN237936). The cDNA was 1,155 bp in length with an open reading frame of 894 bp capable of encoding a putative polypeptide of 297 amino acids with an estimated molecular mass of 34.6 kDa and a predicted isoelectric point of 9.6. Comparative and bioinformatics analyses revealed that BcMF12 showed high similarities with some membrane protein sequences previously published in the public database and contained six highly conserved transmembrane domains corresponding to six highly hydrophobic regions. This indicates that BcMF12 may be a putative membrane protein. RNA gel blot analysis indicated that the transcripts of BcMF12 were abundant in the flower bud, flower and anther, but not detected in the root, stem, leaf and pistil. Moreover, the BcMF12 transcripts were detectable at the late stages of pollen development. Morphological investigations of pollen from the BcMF12 antisense transgenic plants showed that most of pollen grains of transgenic plants were abnormal. These results strongly suggest that BcMF12 is a novel pollen-preferentially membrane protein which play an important role during the pollen development in Chinese cabbage.

  9. Degradation kinetics of peroxidase enzyme, phenolic content, and physical and sensorial characteristics in broccoli (Brassica oleracea L. ssp. Italica) during blanching.

    PubMed

    Gonçalves, Elsa M; Pinheiro, Joaquina; Alegria, Carla; Abreu, Marta; Brandão, Teresa R S; Silva, Cristina L M

    2009-06-24

    The effects of water blanching treatment on peroxidase inactivation, total phenolic content, color parameters [-a*/b* and hue (h degrees*)], texture (maximum shear force), and sensory attributes (color and texture, evaluated by a trained panel) of broccoli (Brassica oleracea L. ssp. Italica) were studied at five temperatures (70, 75, 80, 85, and 90 degrees C). Experimental results showed that all studied broccoli quality parameters suffered significative changes due to blanching treatments. The vegetal total phenolic content showed a marked decline. Degradation on objective color and texture measurements and alterations in sensorial attributes were detected. Correlations between sensory and instrumental measurements have been found. Under the conditions 70 degrees C and 6.5 min or 90 degrees C and 0.4 min, 90% of the initial peroxidase activity was reduced. At these conditions, no significant alterations were detected by panelists, and a small amount of phenolic content was lost (ca. 16 and 10%, respectively). The peroxidase inactivation and phenolic content degradation were found to follow first-order reaction models. The zero-order reaction model showed a good fit to the broccoli color (-a*/b* and h degrees*), texture, and sensory parameters changes. The temperature effect was well-described by the Arrhenius law.

  10. Comparative assessment of feeding damage by pod-sucking bugs (Heteroptera: Coreoidea) associated with cowpea, Vigna unguiculata ssp. unguiculata in Nigeria.

    PubMed

    Soyelu, O L; Akingbohungbe, A E

    2007-02-01

    Feeding trials were conducted on three (young, mid-fill and mature) developmental stages of cowpea Vigna unguiculata ssp. unguiculata pods in the screenhouse using fourth instar nymphs and adults of Anoplocnemis curvipes (Fabricius), Riptortus dentipes (Fabricius), Mirperus jaculus (Thunberg), Clavigralla tomentosicollis Stål and C. shadabi Dolling. Anoplocnemis curvipes was observed to be the most damaging coreoid species causing a yield reduction of 26.4-51.7% followed by R. dentipes (24.4-29.4%), M. jaculus (21.9-26.9%), C. tomentosicollis (17.9-22.4%) and C. shadabi (15.9-20.4%). The fourth instar nymphs of each pod-sucking bug species caused a significantly higher cowpea yield reduction than their respective adults. Similarly, infestation on young pods compared to mid-fill and mature stages resulted in significantly higher yield reduction. The results suggest that infestation levels of two fourth instar nymphs of A. curvipes or three fourth instar nymphs of the other four pod-sucking bug species per young pod should be adequate for screening of cowpea varieties for resistance to the coreoid bugs.

  11. Esterification of bio-oil from mallee (Eucalyptus loxophleba ssp. gratiae) leaves with a solid acid catalyst: Conversion of the cyclic ether and terpenoids into hydrocarbons.

    PubMed

    Hu, Xun; Gunawan, Richard; Mourant, Daniel; Wang, Yi; Lievens, Caroline; Chaiwat, Weerawut; Wu, Liping; Li, Chun-Zhu

    2012-11-01

    Bio-oil from pyrolysis of mallee (Eucalyptus loxophleba ssp. gratiae) leaves differs from that obtained with wood by its content of cyclic ethers, terpenoids and N-containing organic compounds. Upgrading of the leaf bio-oil in methanol with a solid acid catalyst was investigated and it was found that the N-containing organics in the bio-oil lead to deactivation of the catalyst in the initial stage of exposure and have to be removed via employing high catalyst loading to allow the occurrence of other acid-catalysed reactions. Eucalyptol, the main cyclic ether in the bio-oil, could be converted into the aromatic hydrocarbon, p-cymene, through a series of intermediates including α-terpineol, terpinolene, and α-terpinene. Various steps such as ring-opening, dehydration, isomerisation, and aromatization were involved in the conversion of eucalyptol. The terpenoids in bio-oil could also be converted into aromatic hydrocarbons that can serve as starting materials for the synthesis of fine chemicals, via the similar processes.

  12. Spatial analysis of nuclear and cytoplasmic DNA diversity in wild sea beet (Beta vulgaris ssp. maritima) populations: do marine currents shape the genetic structure?

    PubMed

    Fievet, Virgil; Touzet, Pascal; Arnaud, Jean-François; Cuguen, Joël

    2007-05-01

    Patterns of seed dispersal in the wild sea beet (Beta vulgaris ssp. maritima) are predicted to be influenced by marine currents because populations are widely distributed along the European Atlantic coast. We investigated the potential influence of marine currents on the pattern of spatial genetic structuring in natural populations of sea beet. Populations were located along the French coasts of the Anglo-Norman gulf that features peculiar marine currents in the Channel. Thirty-three populations were sampled, among which 23 were continental and 10 were insular populations located in Jersey, Guernsey and Chausey, for a total of 1224 plants genotyped. To validate the coastal topography influence and the possibility of marine current orientated gene flow on the genetic features of sea beet populations, we assessed patterns of genetic structuring of cytoplasmic and nuclear diversity by: (i) searching for an isolation-by-distance (IBD) pattern using spatial autocorrelation tools; (ii) using the Monmonier algorithm to identify genetic boundaries in the area studied; and (iii) performing assignment tests that are based on multilocus genotype information to ascertain population membership of individuals. Our results showed a highly contrasted cytoplasmic and nuclear genetic differentiation and highlighted the peculiar situation of island populations. Beyond a classical isolation-by-distance due to short-range dispersal, genetic barriers fitting the orientation of marine currents were clearly identified. This suggests the occurrence of long-distance seed dispersal events and an asymmetrical gene flow separating the eastern and western part of the Anglo-Norman gulf.

  13. Effect of the absence of the CcpA gene on growth, metabolic production, and stress tolerance in Lactobacillus delbrueckii ssp. bulgaricus.

    PubMed

    Li, C; Sun, J W; Zhang, G F; Liu, L B

    2016-01-01

    The catabolite control protein A (CcpA) is a kind of multi-effect regulatory protein. In the study, the effect of the inactivation of CcpA and aerobic conditions on the growth, metabolic production, and stress tolerance to heat, oxidative, and cold stresses in Lactobacillus delbrueckii ssp. bulgaricus was investigated. Results showed that inactivation of CcpA distinctly hindered growth. Total lactic acid concentration was significantly lower in aerobiosis for both strains and was lower for the mutant strain than L. bulgaricus. Acetic acid production from the mutant strain was higher than L. bulgaricus in aerobiosis compared with anaerobiosis. Enzyme activities, lactate dehydrogenase (LDH), phosphate fructose kinase (PFK), pyruvate kinase (PK), and pyruvic dehydrogenase (PDH), were significantly lower in the mutant strain than L. bulgaricus. The diameters of inhibition zone were 13.59 ± 0.02 mm and 9.76 ± 0.02 mm for L. bulgaricus in anaerobiosis and aerobiosis, respectively; and 8.12 ± 0.02 mm and 7.38 ± 0.02 mm for the mutant in anaerobiosis and aerobiosis, respectively. For both strains, cells grown under aerobic environment possess more stress tolerance. This is the first study in which the CcpA-negative mutant of L. bulgaricus is constructed and the effect of aerobic growth on stress tolerance of L. bulgaricus is evaluated. Although aerobic cultivation does not significantly improve growth, it does improve stress tolerance.

  14. Interchangeable effects of gibberellic acid and temperature on embryo growth, seed germination and epicotyl emergence in Ribes multiflorum ssp. sandalioticum (Grossulariaceae).

    PubMed

    Mattana, E; Pritchard, H W; Porceddu, M; Stuppy, W H; Bacchetta, G

    2012-01-01

    Morphophysiological dormancy was investigated in seeds of Ribes multiflorum Kit ex Roem et Schult. ssp. sandalioticum Arrigoni, a rare mountain species endemic to Sardinia (Italy). There were no differences in imbibition rates between intact and scarified seeds, suggesting a lack of physical dormancy, while methylene blue solution (0.5%) highlighted a preferential pathway for solution entrance through the raphe. Embryos were small at seed dispersal, with an initial embryo:seed ratio (E:S) of ca. 0.2 (embryo length, ca. 0.5 mm), whereas the critical E:S ratio for germination was three times longer (ca. 0.6). Gibberellic acid (GA(3), 250 mg · l(-1)) and warm stratification (25 °C for 3 months) followed by low temperature (<15 °C) enhanced embryo growth rate (maximum of ca. 0.04 mm · day(-1) at 10 °C) and subsequent seed germination (radicle emergence; ca. 80% at 10 °C). Low germination occurred at warmer temperatures, and cold stratification (5 °C for 3 months) induced secondary dormancy. After radicle emergence, epicotyl emergence was delayed for ca. 2 months for seeds from three different populations. Mean time of epicotyl emergence was affected by GA(3) . Seeds of this species showed non-deep simple (root) - non-deep simple (epicotyl) morphophysiological dormancy, highlighting a high synchronisation with Mediterranean seasonality in all the investigated populations.

  15. Induced Production of 1-Methoxy-indol-3-ylmethyl Glucosinolate by Jasmonic Acid and Methyl Jasmonate in Sprouts and Leaves of Pak Choi (Brassica rapa ssp. chinensis)

    PubMed Central

    Wiesner, Melanie; Hanschen, Franziska S.; Schreiner, Monika; Glatt, Hansruedi; Zrenner, Rita

    2013-01-01

    Pak choi plants (Brassica rapa ssp. chinensis) were treated with different signaling molecules methyl jasmonate, jasmonic acid, linolenic acid, and methyl salicylate and were analyzed for specific changes in their glucosinolate profile. Glucosinolate levels were quantified using HPLC-DAD-UV, with focus on induction of indole glucosinolates and special emphasis on 1-methoxy-indol-3-ylmethyl glucosinolate. Furthermore, the effects of the different signaling molecules on indole glucosinolate accumulation were analyzed on the level of gene expression using semi-quantitative realtime RT-PCR of selected genes. The treatments with signaling molecules were performed on sprouts and mature leaves to determine ontogenetic differences in glucosinolate accumulation and related gene expression. The highest increase of indole glucosinolate levels, with considerable enhancement of the 1-methoxy-indol-3-ylmethyl glucosinolate content, was achieved with treatments of sprouts and mature leaves with methyl jasmonate and jasmonic acid. This increase was accompanied by increased expression of genes putatively involved in the indole glucosinolate biosynthetic pathway. The high levels of indole glucosinolates enabled the plant to preferentially produce the respective breakdown products after tissue damage. Thus, pak choi plants treated with methyl jasmonate or jasmonic acid, are a valuable tool to analyze the specific protection functions of 1-methoxy-indole-3-carbinole in the plants defense strategy in the future. PMID:23873294

  16. Short communication: Herd-level prevalence of Mycobacterium avium ssp. paratuberculosis is not associated with participation in a voluntary Alberta Johne's disease control program.

    PubMed

    Ritter, C; Wolf, R; Adams, C L; Kelton, D F; Pickel, C; Mason, S; Orsel, K; De Buck, J; Barkema, H W

    2016-03-01

    Johne's disease (JD) control programs for dairy farms have the general objective of reducing both cow- and herd-level prevalence of Mycobacterium avium ssp. paratuberculosis (MAP). An important aspect of many programs is herd testing for MAP to determine the infection status of participating farms. However, it is uncertain whether MAP herd-level prevalence on farms voluntarily participating in a JD control program is different from that on nonparticipating farms. Therefore, the aim was to compare MAP infection status of participants and nonparticipants in the Alberta Johne's Disease Initiative (AJDI), a voluntary JD control program initiated in 2010 in Alberta, Canada. Between September 2012 and August 2013, environmental fecal samples were collected from 93 randomly selected farms not enrolled in the AJDI. Additionally, 81 farms that initially enrolled in the AJDI during the same time interval were also sampled. Samples were collected from 6 defined locations on each farm and cultured for MAP. Results were confirmed using conventional IS900 PCR and F 0285 quantitative PCR. Overall, 51% of participating and 51% of nonparticipating farms were identified as being MAP-infected. Furthermore, based on multivariable logistic regression, the number of MAP-positive samples was not associated with AJDI participation (taking herd size into account as a potentially modifying or confounding variable). In conclusion, there was no indication that voluntary participation in the AJDI was associated with herd-level MAP prevalence.

  17. Glucosinolate Accumulation and Related Gene Expression in Pak Choi (Brassica rapa L. ssp. chinensis var. communis [N. Tsen & S.H. Lee] Hanelt) in Response to Insecticide Application.

    PubMed

    Zhu, Biao; Yang, Jing; He, Yong; Zang, Yunxiang; Zhu, Zhujun

    2015-11-11

    Glucosinolates and their breakdown products are well-known for their cancer-chemoprotective functions and biocidal activities against pathogens and generalist herbivores. Insecticides are commonly used in the production of pak choi (Brassica rapa L. ssp. chinensis var. communis [N. Tsen & S.H. Lee] Hanelt). We studied the effects of four commonly used insecticides, namely, β-cypermethrin, acephate, pymetrozine, and imidacloprid, on glucosinolate metabolism in pak choi. All insecticides significantly increased both the transcription of glucosinolate biosynthetic genes and the aliphatic and total glucosinolate accumulations in pak choi. β-Cypermethrin and acephate caused gradual and continuous up-regulation of gene expression from 0.5 to 24 h after treatment, whereas pymetrozine and imidacloprid did so more rapidly, reaching a peak at 1 h and returning to normal at 3 h. Our findings indicate that the four insecticides affect glucosinolate metabolism in pak choi plants to various degrees and suggest that glucosinolates may be involved in plant insecticide metabolism.

  18. Short communication: Herd-level prevalence of Mycobacterium avium ssp. paratuberculosis is not associated with participation in a voluntary Alberta Johne's disease control program.

    PubMed

    Ritter, C; Wolf, R; Adams, C L; Kelton, D F; Pickel, C; Mason, S; Orsel, K; De Buck, J; Barkema, H W

    2016-03-01

    Johne's disease (JD) control programs for dairy farms have the general objective of reducing both cow- and herd-level prevalence of Mycobacterium avium ssp. paratuberculosis (MAP). An important aspect of many programs is herd testing for MAP to determine the infection status of participating farms. However, it is uncertain whether MAP herd-level prevalence on farms voluntarily participating in a JD control program is different from that on nonparticipating farms. Therefore, the aim was to compare MAP infection status of participants and nonparticipants in the Alberta Johne's Disease Initiative (AJDI), a voluntary JD control program initiated in 2010 in Alberta, Canada. Between September 2012 and August 2013, environmental fecal samples were collected from 93 randomly selected farms not enrolled in the AJDI. Additionally, 81 farms that initially enrolled in the AJDI during the same time interval were also sampled. Samples were collected from 6 defined locations on each farm and cultured for MAP. Results were confirmed using conventional IS900 PCR and F 0285 quantitative PCR. Overall, 51% of participating and 51% of nonparticipating farms were identified as being MAP-infected. Furthermore, based on multivariable logistic regression, the number of MAP-positive samples was not associated with AJDI participation (taking herd size into account as a potentially modifying or confounding variable). In conclusion, there was no indication that voluntary participation in the AJDI was associated with herd-level MAP prevalence. PMID:26778309

  19. Anthocyanin Accumulation, Antioxidant Ability and Stability, and a Transcriptional Analysis of Anthocyanin Biosynthesis in Purple Heading Chinese Cabbage (Brassica rapa L. ssp. pekinensis).

    PubMed

    He, Qiong; Zhang, Zhanfeng; Zhang, Lugang

    2016-01-13

    Heading Chinese cabbage (Brassica rapa L. ssp. pekinensis) is a significant dietary vegetable for its edible heading leaves in Asia countries. The new purple anthocyanin-rich pure line (11S91) was successfully bred, and the anthocyanins were mainly distributed in 2-3 cell layers beneath the leaf epidermis, whereas siliques and stems accumulated only a cell layer of anthocyanins. The anthocyanins of 11S91 were more stable at pHs below 3.0 and temperatures below 45 °C. The total antioxidant ability was highly positive correlated with the anthocyanin content in 11S91. Thirty-two anthocyanins were separated and identified, and 70% of them were glycosylated and acylated cyanidins. The four major anthocyanins present were cyanidin-3-sophoroside(p-coumaroyl)-5-glucoside(malonyl), cyanidin-3-sophoroside(ferulyl)-5-glucoside(malonyl), cyanidin-3-sophoroside(sinapyl-p-coumaroyl)-5-glucoside(malonyl), and cyanidin-3-sophoroside-(sinapyl-ferulyl)-5-glucoside(malonyl). According to the expression of biosynthetic genes and the component profile of anthocyanins in 11S91 and its parents, regulatory genes BrMYB2 and BrTT8 probably activate the anthocyanin biosynthesis but other factors may govern the primary anthocyanins and the distribution.

  20. Discrimination of cabbage (Brassica rapa ssp. pekinensis) cultivars grown in different geographical areas using ¹H NMR-based metabolomics.

    PubMed

    Kim, Jahan; Jung, Youngae; Song, Byeongyeol; Bong, Yeon-Sik; Ryu, Do Hyun; Lee, Kwang-Sik; Hwang, Geum-Sook

    2013-04-15

    Cabbage (Brassica rapa ssp. pekinensis) is one of the most popular foods in Asia and is widely cultivated in many countries for the production of lightly fermented vegetables. In this study, metabolomic analysis was performed to distinguish two cultivars of cabbage grown in different geographical areas, Korea and China, using ¹H NMR spectroscopy coupled with multivariate statistical analysis. Principal component analysis (PCA) showed clear discrimination between extracts of cabbage grown in Korea and China for two different cultivars (Chunmyeong and Chunjung). The major biochemicals (metabolites) that contributed to discrimination between cabbages grown in the two regions were 4-aminobutyrate (GABA), acetate, asparagine, leucine, isoleucine, O-phosphocholine, phenylacetate, phenylalanine, succinate, sucrose, tyrosine, and valine. These results suggest that the levels of the major metabolites that differ significantly between cabbages grown in these two areas were influenced by environmental factors such as climate and geology. Our study demonstrates that ¹H NMR based on metabolomics, coupled with multivariate statistics, can be applied to identify the regions of cultivation of various cabbage cultivars.