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Sample records for alternaria stem canker

  1. Phomopsis stem canker: a re-emerging threat to sunflowers (Helianthus annuus) in the United States

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Phomopsis stem canker frequently causes yield reductions on sunflowers (Helianthus annuus L.) on several continents, including Australia, Russia, Europe and North America. Between 2001 and 2012, the incidence of Phomopsis stem canker has increased 16 fold in the Northern Great Plains of the United...

  2. Phomopsis Stem Canker: A Reemerging Threat to Sunflower (Helianthus annuus) in the United States

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Phomopsis stem canker causes yield reductions on sunflower (Helianthus annuus L.) on several continents, including Australia, Europe, and North America. In the United States, Phomopsis stem canker incidence has increased 16-fold in the Northern Great Plains between 2001 and 2012. Although Diaporthe ...

  3. Phomopsis Stem Canker: A Reemerging Threat to Sunflower (Helianthus annuus) in the United States.

    PubMed

    Mathew, Febina M; Alananbeh, Kholoud M; Jordahl, James G; Meyer, Scott M; Castlebury, Lisa A; Gulya, Thomas J; Markell, Samuel G

    2015-07-01

    Phomopsis stem canker causes yield reductions on sunflower (Helianthus annuus L.) on several continents, including Australia, Europe, and North America. In the United States, Phomopsis stem canker incidence has increased 16-fold in the Northern Great Plains between 2001 and 2012. Although Diaporthe helianthi was assumed to be the sole causal agent in the United States, a newly described species, D. gulyae, was found to be the primary cause of Phomopsis stem canker in Australia. To determine the identity of Diaporthe spp. causing Phomopsis stem canker in the Northern Great Plains, 275 infected stems were collected between 2010 and 2012. Phylogenetic analyses of sequences of the ribosomal DNA internal transcribed spacer region, elongation factor subunit 1-α, and actin gene regions of representative isolates, in comparison with those of type specimens, confirmed two species (D. helianthi and D. gulyae) in the United States. Differences in aggressiveness between the two species were determined using the stem-wound method in the greenhouse; overall, D. helianthi and D. gulyae did not vary significantly (P≤0.05) in their aggressiveness at 10 and 14 days after inoculation. These findings indicate that both Diaporthe spp. have emerged as sunflower pathogens in the United States, and have implications on the management of this disease.

  4. Draft genome sequence of diaporthe aspalathi isolate ms-ssc91 a fungus causing stem canker in soybean

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Diaporthe aspalathi (formerly D. phaseolorum var. meridionalis) is the causal agent of the southern stem canker disease in soybean. This disease can kill plants from the middle to the end of the growing season resulting in severe yield loss. The mechanisms of disease development and pathogen invasi...

  5. Assessing quantitative resistance against Leptosphaeria maculans (phoma stem canker) in Brassica napus (oilseed rape) in young plants.

    PubMed

    Huang, Yong-Ju; Qi, Aiming; King, Graham J; Fitt, Bruce D L

    2014-01-01

    Quantitative resistance against Leptosphaeria maculans in Brassica napus is difficult to assess in young plants due to the long period of symptomless growth of the pathogen from the appearance of leaf lesions to the appearance of canker symptoms on the stem. By using doubled haploid (DH) lines A30 (susceptible) and C119 (with quantitative resistance), quantitative resistance against L. maculans was assessed in young plants in controlled environments at two stages: stage 1, growth of the pathogen along leaf veins/petioles towards the stem by leaf lamina inoculation; stage 2, growth in stem tissues to produce stem canker symptoms by leaf petiole inoculation. Two types of inoculum (ascospores; conidia) and three assessment methods (extent of visible necrosis; symptomless pathogen growth visualised using the GFP reporter gene; amount of pathogen DNA quantified by PCR) were used. In stage 1 assessments, significant differences were observed between lines A30 and C119 in area of leaf lesions, distance grown along veins/petioles assessed by visible necrosis or by viewing GFP and amount of L. maculans DNA in leaf petioles. In stage 2 assessments, significant differences were observed between lines A30 and C119 in severity of stem canker and amount of L. maculans DNA in stem tissues. GFP-labelled L. maculans spread more quickly from the stem cortex to the stem pith in A30 than in C119. Stem canker symptoms were produced more rapidly by using ascospore inoculum than by using conidial inoculum. These results suggest that quantitative resistance against L. maculans in B. napus can be assessed in young plants in controlled conditions. Development of methods to phenotype quantitative resistance against plant pathogens in young plants in controlled environments will help identification of stable quantitative resistance for control of crop diseases.

  6. Assessing Quantitative Resistance against Leptosphaeria maculans (Phoma Stem Canker) in Brassica napus (Oilseed Rape) in Young Plants

    PubMed Central

    Huang, Yong-Ju; Qi, Aiming; King, Graham J.; Fitt, Bruce D. L.

    2014-01-01

    Quantitative resistance against Leptosphaeria maculans in Brassica napus is difficult to assess in young plants due to the long period of symptomless growth of the pathogen from the appearance of leaf lesions to the appearance of canker symptoms on the stem. By using doubled haploid (DH) lines A30 (susceptible) and C119 (with quantitative resistance), quantitative resistance against L. maculans was assessed in young plants in controlled environments at two stages: stage 1, growth of the pathogen along leaf veins/petioles towards the stem by leaf lamina inoculation; stage 2, growth in stem tissues to produce stem canker symptoms by leaf petiole inoculation. Two types of inoculum (ascospores; conidia) and three assessment methods (extent of visible necrosis; symptomless pathogen growth visualised using the GFP reporter gene; amount of pathogen DNA quantified by PCR) were used. In stage 1 assessments, significant differences were observed between lines A30 and C119 in area of leaf lesions, distance grown along veins/petioles assessed by visible necrosis or by viewing GFP and amount of L. maculans DNA in leaf petioles. In stage 2 assessments, significant differences were observed between lines A30 and C119 in severity of stem canker and amount of L. maculans DNA in stem tissues. GFP-labelled L. maculans spread more quickly from the stem cortex to the stem pith in A30 than in C119. Stem canker symptoms were produced more rapidly by using ascospore inoculum than by using conidial inoculum. These results suggest that quantitative resistance against L. maculans in B. napus can be assessed in young plants in controlled conditions. Development of methods to phenotype quantitative resistance against plant pathogens in young plants in controlled environments will help identification of stable quantitative resistance for control of crop diseases. PMID:24454767

  7. Cutaneous phaeohyphomycosis in a hematopoietic stem cell transplant patient caused by Alternaria rosae: First case report.

    PubMed

    Liu, Amy W; Bateman, Allen C; Greenbaum, Adam; Garvin, Kanishka; Clarridge, Jill; Grim, Jonathan

    2017-03-13

    Alternaria species have been reported as a rare cause of fungal infection in organ and stem cell transplant recipients, but to date no reports have been published of infection in humans caused by Alternaria rosae. Here, we report cutaneous A. rosae infection in a 66-year-old farmer with a history of primary myelofibrosis who had undergone allogeneic unrelated donor hematopoietic stem cell transplantation. Forty-nine days post transplant, he presented with a nodule on the thumb with no findings suggestive of disseminated infection. Pathology, culture, and molecular speciation showed the nodule was caused by cutaneous A. rosae. He had been on voriconazole as antifungal prophylaxis, but was found to have a subtherapeutic voriconazole level. He was switched to posaconazole based on published in vitro data showing its superior efficacy in Alternaria treatment. Susceptibility testing showed that the A. rosae isolate was indeed susceptible to posaconazole. His cutaneous lesion remained stable, but he died from respiratory failure secondary to lobar pneumonia. At lung autopsy, A. rosae was not identified in the lungs. We believe this to be the first published report, to our knowledge, of A. rosae infection in humans. This article is protected by copyright. All rights reserved.

  8. Comparative genomic analysis of duplicated homoeologous regions involved in the resistance of Brassica napus to stem canker.

    PubMed

    Fopa Fomeju, Berline; Falentin, Cyril; Lassalle, Gilles; Manzanares-Dauleux, Maria J; Delourme, Régine

    2015-01-01

    All crop species are current or ancient polyploids. Following whole genome duplication, structural and functional modifications result in differential gene content or regulation in the duplicated regions, which can play a fundamental role in the diversification of genes underlying complex traits. We have investigated this issue in Brassica napus, a species with a highly duplicated genome, with the aim of studying the structural and functional organization of duplicated regions involved in quantitative resistance to stem canker, a disease caused by the fungal pathogen Leptosphaeria maculans. Genome-wide association analysis on two oilseed rape panels confirmed that duplicated regions of ancestral blocks E, J, R, U, and W were involved in resistance to stem canker. The structural analysis of the duplicated genomic regions showed a higher gene density on the A genome than on the C genome and a better collinearity between homoeologous regions than paralogous regions, as overall in the whole B. napus genome. The three ancestral sub-genomes were involved in the resistance to stem canker and the fractionation profile of the duplicated regions corresponded to what was expected from results on the B. napus progenitors. About 60% of the genes identified in these duplicated regions were single-copy genes while less than 5% were retained in all the duplicated copies of a given ancestral block. Genes retained in several copies were mainly involved in response to stress, signaling, or transcription regulation. Genes with resistance-associated markers were mainly retained in more than two copies. These results suggested that some genes underlying quantitative resistance to stem canker might be duplicated genes. Genes with a hydrolase activity that were retained in one copy or R-like genes might also account for resistance in some regions. Further analyses need to be conducted to indicate to what extent duplicated genes contribute to the expression of the resistance phenotype.

  9. Multi-gene phylogenies and phenotypic characters distinguish two species within the Colletogloeopsis zuluensis complex associated with Eucalyptus stem cankers

    PubMed Central

    Cortinas, Maria-Noel; Crous, Pedro W.; Wingfield, Brenda D.; Wingfield, Michael J.

    2006-01-01

    Colletogloeopsis zuluensis, previously known as Coniothyrium zuluense, causes a serious stem canker disease on Eucalyptus spp. grown as non-natives in many tropical and sub-tropical countries. This stem canker disease was first reported from South Africa and it has subsequently been found on various species and hybrids of Eucalyptus in other African countries as well as in countries of South America and South-East Asia. In previous studies, phylogenetic analyses based on DNA sequence data of the ITS region suggested that all material of C. zuluensis was monophyletic. However, the occurrence of the fungus in a greater number of countries, and analyses of DNA sequences with additional isolates has challenged the notion that a single species is involved with Coniothyrium canker. The aim of this study was to consider the phylogenetic relationships amongst C. zuluensis isolates from all available locations and to support these analyses with phenotypic and morphological comparisons. Individual and combined phylogenies were constructed using DNA sequences from the ITS region, exons 3 through 6 of the β-tubulin gene, the intron of the translation elongation factor 1-α gene, and a partial sequence of the mitochondrial ATPase 6 gene. Both phylogenetic data and morphological characteristics showed clearly that isolates of C. zuluensis represent at least two taxa. One of these is C. zuluensis as it was originally described from South Africa, and we provide an epitype for it. The second species occurs in Argentina and Uruguay, and is newly described as C. gauchensis. Both fungi are serious pathogens resulting in identical symptoms. Recognising them as different species has important quarantine consequences. PMID:18490975

  10. Alternaria redefined

    PubMed Central

    Woudenberg, J.H.C.; Groenewald, J.Z.; Binder, M.; Crous, P.W.

    2013-01-01

    Alternaria is a ubiquitous fungal genus that includes saprobic, endophytic and pathogenic species associated with a wide variety of substrates. In recent years, DNA-based studies revealed multiple non-monophyletic genera within the Alternaria complex, and Alternaria species clades that do not always correlate to species-groups based on morphological characteristics. The Alternaria complex currently comprises nine genera and eight Alternaria sections. The aim of this study was to delineate phylogenetic lineages within Alternaria and allied genera based on nucleotide sequence data of parts of the 18S nrDNA, 28S nrDNA, ITS, GAPDH, RPB2 and TEF1-alpha gene regions. Our data reveal a Pleospora/Stemphylium clade sister to Embellisia annulata, and a well-supported Alternaria clade. The Alternaria clade contains 24 internal clades and six monotypic lineages, the assemblage of which we recognise as Alternaria. This puts the genera Allewia, Brachycladium, Chalastospora, Chmelia, Crivellia, Embellisia, Lewia, Nimbya, Sinomyces, Teretispora, Ulocladium, Undifilum and Ybotromyces in synonymy with Alternaria. In this study, we treat the 24 internal clades in the Alternaria complex as sections, which is a continuation of a recent proposal for the taxonomic treatment of lineages in Alternaria. Embellisia annulata is synonymised with Dendryphiella salina, and together with Dendryphiella arenariae, are placed in the new genus Paradendryphiella. The sexual genera Clathrospora and Comoclathris, which were previously associated with Alternaria, cluster within the Pleosporaceae, outside Alternaria s. str., whereas Alternariaster, a genus formerly seen as part of Alternaria, clusters within the Leptosphaeriaceae. Paradendryphiella is newly described, the generic circumscription of Alternaria is emended, and 32 new combinations and 10 new names are proposed. A further 10 names are resurrected, while descriptions are provided for 16 new Alternaria sections. Taxonomic novelties: New

  11. Effect of climate change on sporulation of the teleomorphs of Leptosphaeria species causing stem canker of brassicas.

    PubMed

    Kaczmarek, Joanna; Kedziora, Andrzej; Brachaczek, Andrzej; Latunde-Dada, Akinwunmi O; Dakowska, Sylwia; Karg, Grzegorz; Jedryczka, Małgorzata

    Leptosphaeria maculans and L. biglobosa are closely related sibling fungal pathogens that cause phoma leaf spotting, stem canker (blackleg) and stem necrosis of oilseed rape (Brassica napus). The disease is distributed worldwide, and it is one of the main causes of considerable decrease in seed yield and quality. Information about the time of ascospore release at a particular location provides important data for decision making in plant protection, thereby enabling fungicides to be used only when necessary and at optimal times and doses. Although the pathogens have been studied very extensively, the effect of climate change on the frequencies and distributions of their aerially dispersed primary inoculum has not been reported to date. We have collected a large dataset of spore counts from Poznan, located in central-west part of Poland, and studied the relationships between climate and the daily concentrations of airborne propagules over a period of 17 years (1998-2014). The average air temperature and precipitation for the time of development of pseudothecia and ascospore release (July-November), increased during the years under study at the rates of 0.1 °C and 6.3 mm per year. The day of the year (DOY) for the first detection of spores, as well as the date with maximum of spores, shifted from 270 to 248 DOY, and from 315 to 265 DOY, respectively. The acceleration of the former parameter by 22 days and the latter by 50 days has great influence on the severity of stem canker of oilseed rape.

  12. Canker Sores

    MedlinePlus

    Canker sores are small, round sores in your mouth. They can be on the inside of your cheek, under your tongue, or in the back of your throat. They usually have a red edge and a gray center. They can be quite painful. They are ...

  13. Ethanol attracts scolytid beetles to Phytophthora ramorum cankers on coast live oak

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Technical abstract: Ethanol in sapwood was analyzed along vertical transects, through small spot cankers and larger basal cankers, of Phytophthora ramorum-infected stems of Quercus agrifolia at three sites in California. Trees with large basal cankers, known to attract scolytid beetles, had a 4.3 ti...

  14. Canker Sores (For Parents)

    MedlinePlus

    ... Lessons? Visit KidsHealth in the Classroom What Other Parents Are Reading Your Child's Development (Birth to 3 Years) Feeding Your 1- to 3-Month-Old Feeding Your 4- to 7-Month-Old Feeding Your 8- to 12-Month-Old Feeding Your 1- to 2-Year-Old Canker ... > For Parents > Canker Sores Print A A A What's in ...

  15. Visualization of wound periderm and hyphal profiles in pine stems inoculated with the pitch canker fungus Fusarium circinatum.

    PubMed

    Kim, Ki Woo; Lee, In Jung; Thoungchaleun, Vilakon; Kim, Chang Soo; Lee, Don Koo; Park, Eun Woo

    2009-12-01

    Postpenetration behavior of Fusarium circinatum in stems of pine species was investigated with light and transmission electron microscopy. Two-year-old stems of Pinus rigida and P. densiflora were wound-inoculated with the fungal conidial suspension and subjected to 25 degrees C for up to 30 days. It was common to observe the formation of wound periderm on each pine species, recovering wounded sites with newly formed tissues. The outermost thick layer of wound periderm was pink to red colored with the phloroglucinol-EtOH staining, indicating heavy deposition of lignin in wound periderm. The cork layers in the wound periderm of the two pine species consisted of cells that were mostly devoid of cellular contents in cytoplasm. The cork cells showed convoluted cell walls with different electron density (lamellations), which was seemingly more prevalent in P. densiflora than P. rigida. Hyphae of F. circinatum appeared normal with typical eucaryotic cytoplasm in P. rigida on ultrathin sections. Meanwhile, hyphae in P. densiflora were found to possess highly vacuolated cytoplasm, implying hyphal weakening and disintegration. Hyphal cytoplasm appeared to be a thin layer between the vacuole and the plasma membrane surrounded by cell wall. In addition, intrahyphal hyphae and concentric bodies were observed in hyphal cytoplasm. These results suggest that the architecture of wound periderm may be responsible for different responses of pine species to the invasion of F. circinatum.

  16. Management of apple anthracnose canker

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Apple anthracnose (caused by Neofabraea malicorticis anamorph Cryptosporiopsis curvispora) is a fungal disease that causes cankers on trees and ‘Bull’s-eye rot’ on fruit. In western Washington, it is the canker phase of apple anthracnose that is considered most serious as it can result in death of ...

  17. Water activity and temperature effects on growth of Alternaria arborescens on tomato medium.

    PubMed

    Vaquera, Sandra; Patriarca, Andrea; Fernández Pinto, Virginia

    2014-08-18

    Alternaria arborescens is the causal agent of tomato stem canker, a disease frequently responsible of substantial economic losses. A. arborescens can produce several mycotoxins, such as alternariol, alternariol monomethyl ether and tenuazonic acid and phytotoxins such as the AAL toxins. The objective of this study was to determine the effect of water activity (aw, 0.950, 0.975, 0.995) and temperature (6, 15, 20, 25 and 30°C) on the germination and radial growth rate of A. arborescens on a synthetic tomato medium. Germination followed by growth was observed at all temperatures and aw levels analyzed. The shortest germination time (0.5 days) was observed at 0.995 aw, both at 25°C and at 30°C. The germination time increased with a reduction of aw and temperature. The highest growth rate was registered at 0.995 aw and 30°C (7.21 mm/day) while the lowest occurred at 0.950 aw and 6°C (0.52 mm/day), conditions at which the longest lag phase was observed (8 days). Growth rates increased with aw and temperature. Knowledge of the ecophysiology of the fungus in this substrate is necessary to formulate future strategies to prevent its development and evaluate the consumer health risk posed by potential exposure to the toxins.

  18. Pitch canker disease of pines.

    PubMed

    Gordon, T R

    2006-06-01

    ABSTRACT Pitch canker, caused by Fusarium circinatum, is a disease affecting pines in many locations throughout the world. The pathosystem was originally described in the southeastern (SE) United States and was identified in California in 1986. Limited vegetative compatibility group (VCG) diversity in the California population of F. circinatum, relative to the SE United States, suggests the former is a recently established and clonally propagating population. Although the much greater VCG diversity found in the SE United States is suggestive of out-crossing, molecular markers indicate that many vegetatively incompatible isolates are clonally related. This implies that VCG diversity may derive, at least in part, from somatic mutations rather than sexual reproduction. Pitch canker is damaging to many pine species and one at particular risk is Monterey pine (Pinus radiata), which is widely grown in plantations and is highly susceptible to pitch canker. However, some Monterey pines are resistant to pitch canker and some severely diseased trees have been observed to recover. The absence of new infections on these trees reflects the operation of systemic induced resistance, apparently in response to repeated infection by the pitch canker pathogen.

  19. Integrated Management of Citrus Canker

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Fruit losses due to citrus canker, caused by Xanthomonas citri subsp. citri (Xcc), vary each crop season depending on citrus variety, tree age, flushing condition, leafminer control, and coincidence of weather events with occurrence of susceptible fruit and foliage. In 2012, crop losses in Hamlin f...

  20. Alternaria section Alternaria: Species, formae speciales or pathotypes?

    PubMed Central

    Woudenberg, J.H.C.; Seidl, M.F.; Groenewald, J.Z.; de Vries, M.; Stielow, J.B.; Thomma, B.P.H.J.; Crous, P.W.

    2015-01-01

    The cosmopolitan fungal genus Alternaria consists of multiple saprophytic and pathogenic species. Based on phylogenetic and morphological studies, the genus is currently divided into 26 sections. Alternaria sect. Alternaria contains most of the small-spored Alternaria species with concatenated conidia, including important plant, human and postharvest pathogens. Species within sect. Alternaria have been mostly described based on morphology and / or host-specificity, yet molecular variation between them is minimal. To investigate whether the described morphospecies within sect. Alternaria are supported by molecular data, whole-genome sequencing of nine Alternaria morphospecies supplemented with transcriptome sequencing of 12 Alternaria morphospecies as well as multi-gene sequencing of 168 Alternaria isolates was performed. The assembled genomes ranged in size from 33.3–35.2 Mb within sect. Alternaria and from 32.0–39.1 Mb for all Alternaria genomes. The number of repetitive sequences differed significantly between the different Alternaria genomes; ranging from 1.4–16.5 %. The repeat content within sect. Alternaria was relatively low with only 1.4–2.7 % of repeats. Whole-genome alignments revealed 96.7–98.2 % genome identity between sect. Alternaria isolates, compared to 85.1–89.3 % genome identity for isolates from other sections to the A. alternata reference genome. Similarly, 1.4–2.8 % and 0.8–1.8 % single nucleotide polymorphisms (SNPs) were observed in genomic and transcriptomic sequences, respectively, between isolates from sect. Alternaria, while the percentage of SNPs found in isolates from different sections compared to the A. alternata reference genome was considerably higher; 8.0–10.3 % and 6.1–8.5 %. The topology of a phylogenetic tree based on the whole-genome and transcriptome reads was congruent with multi-gene phylogenies based on commonly used gene regions. Based on the genome and transcriptome data, a set of core

  1. Dynamics of Cryphonectria hypovirus infection in chestnut blight cankers.

    PubMed

    Bryner, Sarah Franziska; Prospero, Simone; Rigling, Daniel

    2014-09-01

    Virulent strains of the chestnut blight fungus Cryphonectria parasitica cause lethal bark cankers on chestnut trees. Infection of C. parasitica with Cryphonectria hypovirus 1 in Europe biologically controls this disease, leading to nonlethal and inactive cankers. Unexpectedly, virus-free C. parasitica strains have been isolated from inactive cankers. In this study, we compared the virulence of virus-infected and virus-free C. parasitica strains isolated from either inactive or active cankers on chestnut seedlings and sprouts. In the seedling experiment, we assessed canker growth and seedling mortality. In the sprout experiment, we also assessed canker growth and made fungal reisolations to determine virus infection and immigration of foreign vegetative compatibility (vc) types over a period of 13 years in a coppice forest. Overall, the virulence of virus-free C. parasitica strains isolated from inactive versus active cankers did not differ. Significant differences were only attributed to virus infection. Virus infection and fungal strain composition in cankers changed over time. Foreign vc types immigrated into cankers and virus-free cankers became virus-infected within a few years. Most of the cankers were callused over time and became inactive. However, we observed that the virus did not always persist in these cankers. This study demonstrates that virus spread occurs effectively in European chestnut forests and that this biocontrol system is highly dynamic.

  2. Ethanol attracts scolytid beetles to Phytophthora ramorum cankers on coast live oak.

    PubMed

    Kelsey, Rick G; Beh, Maia M; Shaw, David C; Manter, Daniel K

    2013-04-01

    Ethanol in sapwood was analyzed along vertical transects, through small spot cankers and larger basal cankers, of Phytophthora ramorum-infected stems of Quercus agrifolia at three sites in California. Trees with large basal cankers, known to attract scolytid beetles, had a 4.3 times higher ethanol level than trees with spot cankers that attract fewer beetles. Ethanol concentrations inside cankers, where scolytid beetles preferentially attack, varied by about four orders of magnitude among samples, with a median level of 16.0 μg.g(-1) fresh mass. This concentration was 4.3 and 15.5 times greater, respectively, than the concentrations at 1 cm or 15-30 cm outside the canker boundaries. In the laboratory, we demonstrated that ethanol escaped through the bark of a Q. garryana log just 3 days after it was added to the sapwood. At the three study sites, traps baited with ethanol captured more Xyleborinus saxesenii, Pseudopityophthorus pubipennis, and Monarthrum dentiger (all Coleoptera: Curculionidae: Scolytinae) than traps baited with ethanol plus (-)-α-pinene, or ethanol plus 4-allylanisole (4AA). Logs of Q. agrifolia with a 50 % ethanol solution added to the sapwood were placed at the study sites, with or without additional bark treatments above the ethanol. The number of scolytid beetle gallery holes above the ethanol-infused sapwood was 4.4 times greater than that on the opposite side of the log where no ethanol was added. Attachment of ultra-high release (-)-α-pinene pouches to the bark surface above the 50 % ethanol solution reduced scolytid attacks to a density of 19.1 % that of logs without this treatment. We conclude that ethanol in P. ramorum cankers functions as a primary host attractant for scolytid beetles and is an important link in colonization of these cankers and accelerated mortality of Q. agrifolia. The results of this research shed light on the chemical ecology behind the focused scolytid attacks on P. ramorum-infected coast live oaks, and lay the

  3. The HPLC-Fluorescence Detection of Coumarins in ‘Hamlin’ Sweet Orange and ‘Marsh’ Grapefruit Leaf Cankers

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Canker is a devastating disease for the citrus fresh fruit market and is caused by the pathogenic bacterium Xanthomonas citri var. citri (Xcc). Infection occurs by bacterial penetration through physical damage of leaves, peel and stems, and also by bacterial entry through the stomates of these photo...

  4. Reasons for inconsistent citrus canker control

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Crop losses from citrus canker in 2014 for Hamlin due to premature fruit drop, or for grapefruit from unacceptable severity of fruit lesions, were highly variable due to periodic rains that in certain locations were coincident with grapefruit flushes in February-March or with early Hamlin fruit deve...

  5. Research progress for integrated canker management

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Fruit losses due to citrus canker, caused by Xanthomonas citri subsp. citri (Xcc), vary each crop season depending on citrus variety, tree age, flushing condition, leafminer control, and coincidence of weather events with occurrence of susceptible fruit and foliage. In 2013, crop losses in Hamlin f...

  6. Hyperspectral reflectance imaging for detecting citrus canker based on dual-band ratio image classification method

    NASA Astrophysics Data System (ADS)

    Li, Jiangbo; Rao, Xiuqin; Guo, Junxian; Ying, Yibin

    2010-10-01

    Citrus are one of the major fruit produced in China. Most of this production is exported to Europe for fresh consumption, where consumers increasingly demand best quality. Citrus canker is one of the most devastating diseases that threaten peel of most commercial citrus varieties. The aim of this research was to investigate the potential of using hyperspectral imaging technique for detecting canker lesions on citrus fruit. Navel oranges with cankerous, normal and various common diseased skin conditions including wind scar, thrips scarring, scale insect, dehiscent fruit, phytotoxicity, heterochromatic stripe, and insect damage were studied. The imaging system (400-1000 nm) was established to acquire reflectance images from samples. Region of interest (ROI) spectral feature of various diseased peel areas was analyzed and characteristic wavebands (630, 685, and 720 nm) were extracted. The dual-band reflectance ratio (such as Q720/685) algorithm was performed on the hyperspectral images of navel oranges for differentiating canker from normal fruit skin and other surface diseases. The overall classification success rate was 96.84% regardless of the presence of other confounding diseases. The presented processing approach overcame the presence of stem/navel on navel oranges that typically has been a problematic source for false positives in the detection of defects. Because of the limited sample size, delineation of an optimal detection scheme is beyond the scope of the current study. However, the results showed that two-band ratio (Q685/630) along with the use of a simple threshold value segmentation method for discriminating canker on navel oranges from other peel diseases may be feasible.

  7. First Report and Characterization of Pestalotiopsis ellipsospora Causing Canker on Acanthopanax divaricatus

    PubMed Central

    Yun, Yeo Hong; Ahn, Geum Ran

    2015-01-01

    Acanthopanax divaricatus, a member of the Araliaceae family, has been used as an invigorant in traditional Korean medicine. During disease monitoring, a stem with small, irregular, brown lesions was sampled at a farm in Cheonan in 2011. The symptoms seen were sunken cankers and reddish-brown needles on the infected twig. The isolated fungal colonies were whitish, having crenated edges and aerial mycelium on the surface, and with black gregarious fruiting bodies. The reverse plate was creamy white. Conidia were 17~22 × 3.5~4.2 µm, fusiform, 4-septate, and straight to slightly curved. The nucleotide sequence of the partial translation elongation factor 1 alpha gene of the fungal isolate, shares 99% sequence identity with that of known Pestalotiopsis ellipsospora. Based on the results of the morphological and molecular analyses, the fungal isolate was identified as P. ellipsospora. In Korea, this is the first report of canker on A. divaricatus. PMID:26539058

  8. Alternaria Species and Their Associated Mycotoxins.

    PubMed

    Pinto, Virginia Elena Fernández; Patriarca, Andrea

    2017-01-01

    The genus Alternaria includes more than 250 species. The traditional methods for identification of Alternaria species are based on morphological characteristics of the reproductive structures and sporulation patterns under controlled culture conditions. Cladistics analyses of "housekeeping genes" commonly used for other genera, failed to discriminate among the small-spored Alternaria species. The development of molecular methods achieving a better agreement with morphological differences is still needed. The production of secondary metabolites has also been used as a means of classification and identification. Alternaria spp. can produce a wide variety of toxic metabolites. These metabolites belong principally to three different structural groups: (1) the dibenzopyrone derivatives, alternariol (AOH), alternariol monomethyl ether (AME), and altenuene (ALT); (2) the perylene derivative altertoxins (ATX-I, ATX-II, and ATX II); and (3) the tetramic acid derivative, tenuazonic acid (TeA). TeA, AOH, AME, ALT, and ATX-I are the main. Certain species in the genus Alternaria produce host-specific toxins (HSTs) that contribute to their pathogenicity and virulence. Alternaria species are plant pathogens that cause spoilage of agricultural commodities with consequent mycotoxin accumulation and economic losses. Vegetable foods infected by Alternaria rot could introduce high amounts of these toxins to the human diet. More investigations on the toxic potential of these toxins and their hazard for human consumption are needed to make a reliable risk assessment of dietary exposure.

  9. Apple anthracnose canker life cycle and disease cycle

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Apple anthracnose [caused by Neofabraea malicorticis (H.S. Jacks) anamorph Cryptosporiopsis curvispora (Peck)] is a fungal disease that impacts apple production. The pathogen produces cankers on trees as well as a rot on the fruit known as ‘Bull’s-eye rot’. The cankers cause severe damage to trees...

  10. Research promises earlier warning for grapevine canker diseases

    Technology Transfer Automated Retrieval System (TEKTRAN)

    When it comes to detecting and treating vineyards for grapevine canker diseases (also called trunk diseases), like Botryosphaeria dieback (Bot canker), Esca, Eutypa dieback and Phomopsis dieback, the earlier the better, says plant pathologist Kendra Baumgartner, with the USDA’s Agricultural Research...

  11. Toxigenic Alternaria species from Argentinean blueberries.

    PubMed

    Greco, M; Patriarca, A; Terminiello, L; Fernández Pinto, V; Pose, G

    2012-03-15

    Blueberries are traditionally consumed in North America, some European countries and Japan. In Argentina, the blueberry crop is profitable because production starts in November, when the northern hemisphere lacks fresh fruit. Fungal contaminants can grow and produce mycotoxins in fresh fruit. The aims of this work were to identify the main genera of the mycobiota of blueberries grown in Argentina and to determine the toxicogenic potential, pathogenicity and host specificity of the species isolated. The genus Alternaria was the main component of the blueberry mycobiota (95%); minor proportions of Phoma spp. (4%) and Penicillium spp. (1%) were also isolated. According to their sporulation patterns, 127 Alternaria isolates belonged to the Alternaria tenuissima species-group, 5 to the Alternaria alternata species-group and 2 to the Alternaria arborescens species-group. The last mentioned species-group was not isolated at 5°C. Of the 134 isolates, 61% were toxicogenic in autoclaved rice; 97% of these produced alternariol (AOH) in a range from 0.14 to 119.18 mg/kg, 95% produced alternariol methylether (AME) in a range from 1.23 to 901.74 mg/kg and 65% produced tenuazonic acid (TA) in a range from 0.13 to 2778 mg/kg. Fifty two isolates co-produced the three mycotoxins. According to the size of the lesion that they caused on blueberries, the isolates were classified as slightly pathogenic, moderately pathogenic and very pathogenic. No significant differences in pathogenicity were found on different blueberry varieties. In this work, high incidence and toxicogenic potential of the Alternaria isolates from blueberries were demonstrated. Thus, more studies should be done to evaluate the health risk posed by the presence of the Alternaria toxins in blueberries and in the manufactured by-products.

  12. Copper Sprays and Windbreaks for Control of Citrus Canker on Young Orange Trees in Southern Brazil

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The benefit of windbreaks and copper sprays for control of citrus canker caused by Xanthomonas axonopodis pv. citri was investigated in a commercial citrus orchard located in a citrus canker endemic area in southern Brazil. Control of canker was evaluated as incidence and severity of lesions on foli...

  13. Horsfall-Barratt recalibration and replicated severity estimates of citrus canker

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus canker is a serious disease of citrus in tropical and subtropical citrus growing regions. Accurate and precise assessment of citrus canker and other plant pathogens is needed to obtain good quality data. Citrus canker assessment data were used to ascertain some of the mechanics of the Horsfal...

  14. Pruning for prevention and management of canker diseases

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Trunk diseases (wood-canker diseases) threaten all California vineyards due to widespread distribution of the fungal pathogens. The infections are chronic and occur each year. Trunk diseases in mature vineyards reduce yields and increase management costs to the point where the vineyard is no longer ...

  15. 76 FR 81359 - European Larch Canker; Expansion of Regulated Areas

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-12-28

    ... From the Federal Register Online via the Government Publishing Office DEPARTMENT OF AGRICULTURE Animal and Plant Health Inspection Service 7 CFR Part 301 European Larch Canker; Expansion of Regulated Areas AGENCY: Animal and Plant Health Inspection Service, USDA. ACTION: Affirmation of interim rule...

  16. 76 FR 52543 - European Larch Canker; Expansion of Regulated Areas

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-08-23

    ... From the Federal Register Online via the Government Publishing Office DEPARTMENT OF AGRICULTURE Animal and Plant Health Inspection Service 7 CFR Part 301 European Larch Canker; Expansion of Regulated Areas AGENCY: Animal and Plant Health Inspection Service, USDA. ACTION: Interim rule and request...

  17. Citrus diseases with global ramifications including citrus canker and huanglongbing

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Although there are a number of diseases that plague citrus production worldwide, two bacterial diseases are particularly problematic. Both are of Asian origin and currently cause severe economic damage: Asiatic citrus canker (ACC) and citrus huanglongbing (HLB). Although ACC has been found in the ...

  18. Developing Transgenic Citrus for Resistance to Huanglongbing and Citrus Canker

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Huanglongbing (HLB) and Citrus Bacterial Canker (CBC) are serious threats to citrus production, and resistant transgenic citrus is desirable. Genes for antimicrobial peptides (AMPs) with diverse promoters have been used to generate thousands of rootstock and scion transformants. D35S::D4E1 transfor...

  19. Comparative Genomics of Pathogens Causing Brown Spot Disease of Tobacco: Alternaria longipes and Alternaria alternata

    PubMed Central

    Wan, Wenting; Long, Ni; Zhang, Jing; Tan, Yuntao; Duan, Shengchang; Zeng, Yan; Dong, Yang

    2016-01-01

    The genus Alternaria is a group of infectious/contagious pathogenic fungi that not only invade a wide range of crops but also induce severe allergic reactions in a part of the human population. In this study, two strains Alternaria longipes cx1 and Alternaria alternata cx2 were isolated from different brown spot lesions on infected tobacco leaves. Their complete genomes were sequenced, de novo assembled, and comparatively analyzed. Phylogenetic analysis revealed that A. longipes cx1 and A. alternata cx2 diverged 3.3 million years ago, indicating a recent event of speciation. Seventeen non-ribosomal peptide synthetase (NRPS) genes and 13 polyketide synthase (PKS) genes in A. longipes cx1 and 13 NRPS genes and 12 PKS genes in A. alternata cx2 were identified in these two strains. Some of these genes were predicted to participate in the synthesis of non-host specific toxins (non-HSTs), such as tenuazonic acid (TeA), alternariol (AOH) and alternariol monomethyl ether (AME). By comparative genome analysis, we uncovered that A. longipes cx1 had more genes putatively involved in pathogen-plant interaction, more carbohydrate-degrading enzymes and more secreted proteins than A. alternata cx2. In summary, our results demonstrate the genomic distinction between A. longipes cx1 and A. altenata cx2. They will not only improve the understanding of the phylogenetic relationship among genus Alternaria, but more importantly provide valuable genomic resources for the investigation of plant-pathogen interaction. PMID:27159564

  20. Presence of adhesive vesicles in the mycoherbicide Alternaria helianthi

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Alternaria helianthi conidia have been shown to cause disease on common cocklebur. Conidia were applied to slides made hydrophobic by coating with dimethyldicholorosilane (mimics leaf surface), then rinsed and treated with FITC-Con A to stain the adhesive material. Alternaria helianthi coni...

  1. Alternaria in Food: Ecophysiology, Mycotoxin Production and Toxicology

    PubMed Central

    Patriarca, Andrea; Magan, Naresh

    2015-01-01

    Alternaria species are common saprophytes or pathogens of a wide range of plants pre- and post-harvest. This review considers the relative importance of Alternaria species, their ecology, competitiveness, production of mycotoxins and the prevalence of the predominant mycotoxins in different food products. The available toxicity data on these toxins and the potential future impacts of Alternaria species and their toxicity in food products pre- and post-harvest are discussed. The growth of Alternaria species is influenced by interacting abiotic factors, especially water activity (aw), temperature and pH. The boundary conditions which allow growth and toxin production have been identified in relation to different matrices including cereal grain, sorghum, cottonseed, tomato, and soya beans. The competitiveness of Alternaria species is related to their water stress tolerance, hydrolytic enzyme production and ability to produce mycotoxins. The relationship between A. tenuissima and other phyllosphere fungi has been examined and the relative competitiveness determined using both an Index of Dominance (ID) and the Niche Overlap Index (NOI) based on carbon-utilisation patterns. The toxicology of some of the Alternaria mycotoxins have been studied; however, some data are still lacking. The isolation of Alternaria toxins in different food products including processed products is reviewed. The future implications of Alternaria colonization/infection and the role of their mycotoxins in food production chains pre- and post-harvest are discussed. PMID:26190916

  2. Fungicidal activity of silver nanoparticles against Alternaria brassicicola

    NASA Astrophysics Data System (ADS)

    Gupta, Deepika; Chauhan, Pratima

    2016-04-01

    This work highlighted the fungicidal properties of silver nanoparticles against Alternaria brassicicola. Alternaria brassicicola causes Black spot of Cauliflower, radish, cabbage, kale which results in sever agricultural loss. We treat the synthesised silver nanoparticles (AgNPs) of 10, 25, 50, 100 and 110 ppm concentrations against Alternaria brassicicola on PDA containing Petri dish. We calculated inhibitory rate (%) in order to evaluate the antifungal efficacy of silver nanoparticles against pathogens. Treatment with 100ppm AgNPs resulted in maximum inhibition of Alternaria brassicicola i.e.92.2%. 110ppm of AgNPS also shows the same result, therefore 100ppm AgNPs was treated as optimize concentration. AgNPs effectively inhibited the growth of a Alternaria brassicicola, which suggests that AgNPs could be used as fungicide in plant disease management. Further research and development are necessary to translate this technology into plant disease management strategies.

  3. First Record of Alternaria simsimi Causing Leaf Spot on Sesame (Sesamum indicum L.) in Korea.

    PubMed

    Choi, Young Phil; Paul, Narayan Chandra; Lee, Hyang Burm; Yu, Seung Hun

    2014-12-01

    Leaf spot disease was observed in sesame (Sesamum indicum L.) during 2009 and 2010 in Korea. The pathogen was identified as Alternaria simsimi based on morphological and cultural characteristics. The morphological identification was well supported by phylogenetic analysis of the ribosomal DNA-internal transcribed spacer region. A. simsimi isolates caused spot symptoms on leaves and stems of sesame plants 2 wk after artificial inoculation, which were similar to those observed in the field. This is the first record of leaf spot disease in Korea caused by A. simsimi.

  4. Transcriptomic profile of tobacco in response to Alternaria longipes and Alternaria alternata infections

    PubMed Central

    Duan, Shengchang; Ma, Xiao; Chen, Wei; Wan, Wenting; He, Yuqi; Ma, Xiaoqin; Ma, Yujin; Long, Ni; Tan, Yuntao; Wang, Yangzi; Hou, Yujie; Dong, Yang

    2016-01-01

    Tobacco brown spot caused by Alternaria fungal species is one of the most damaging diseases, and results in significant yield losses. However, little is known about the systematic response of tobacco to this fungal infection. To fill this knowledge gap, de novo assemblies of tobacco leaf transcriptomes were obtained in cultivars V2 and NC89 after the inoculation of either Alternaria longipes (AL) or Alternaria alternata (AA) at three different time points. We studied the gene expression profile of each cultivar-pathogen combination, and identified eight differentially expressed genes shared among all combinations. Gene ontology enrichment analysis of the differentially expressed genes revealed key components during the fungal infection, which included regulation of gene expression (GO:0010468), regulation of RNA metabolic process (GO:0051252), tetrapyrrole binding (GO:0046906), and external encapsulating structure (GO:0030312). Further analyses of the continuously upregulated/downregulated genes and the resistance genes demonstrated that the gene expression profile upon fungal infection was contingent on the specific cultivar and pathogen. In conclusion, this study provides a solid foundation for the investigation of plant-pathogen interaction, and is of great importance for disease prevention and molecular breeding. PMID:27157477

  5. Large-spored Alternaria pathogens in section Porri disentangled

    PubMed Central

    Woudenberg, J.H.C.; Truter, M.; Groenewald, J.Z.; Crous, P.W.

    2014-01-01

    The omnipresent fungal genus Alternaria was recently divided into 24 sections based on molecular and morphological data. Alternaria sect. Porri is the largest section, containing almost all Alternaria species with medium to large conidia and long beaks, some of which are important plant pathogens (e.g. Alternaria porri, A. solani and A. tomatophila). We constructed a multi-gene phylogeny on parts of the ITS, GAPDH, RPB2, TEF1 and Alt a 1 gene regions, which, supplemented with morphological and cultural studies, forms the basis for species recognition in sect. Porri. Our data reveal 63 species, of which 10 are newly described in sect. Porri, and 27 species names are synonymised. The three known Alternaria pathogens causing early blight on tomato all cluster in one clade, and are synonymised under the older name, A. linariae. Alternaria protenta, a species formerly only known as pathogen on Helianthus annuus, is also reported to cause early blight of potato, together with A. solani and A. grandis. Two clades with isolates causing purple blotch of onion are confirmed as A. allii and A. porri, but the two species cannot adequately be distinguished based on the number of beaks and branches as suggested previously. This is also found among the pathogens of Passifloraceae, which are reduced from four to three species. In addition to the known pathogen of sweet potato, A. bataticola, three more species are delineated of which two are newly described. A new Alternaria section is also described, comprising two large-spored Alternaria species with concatenate conidia. PMID:25492985

  6. 78 FR 58992 - Notice of Request for Extension of Approval of an Information Collection; Citrus Canker...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-09-25

    ... Collection; Citrus Canker; Interstate Movement of Regulated Nursery Stock and Fruit From Quarantined Areas... nursery stock and fruit from quarantined areas to prevent the spread of citrus canker. DATES: We will...: For information on the regulations for the interstate movement of regulated nursery stock and...

  7. Effect of windbreaks on wind speed and canker incidence and severity on grapefruit

    Technology Transfer Automated Retrieval System (TEKTRAN)

    For fresh grapefruit the goal is to maximize pack-out by minimizing canker lesions on fruit. The objective of these trials was to determine the relationship between wind speed and incidence and severity of canker on 5 to 7 yr-old Ruby Red grapefruit trees located in two trial blocks (~4.5 ha) surrou...

  8. Effect of windbreaks on wind speed and citrus canker incidence on grapefruit

    Technology Transfer Automated Retrieval System (TEKTRAN)

    For fresh grapefruit the goal is to maximize pack-out by minimizing canker lesions on fruit. The objective of these trials was to determine the relationship between wind speed and incidence and severity of citrus canker on 5 to 7 yr-old ‘Ruby Red’ grapefruit trees located in two trial blocks (~4.5 h...

  9. A novel Fusarium species causes a canker disease of the critically endangered conifer, Torreya taxifolia

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A canker disease of Florida torreya (Torreya taxifolia), here designated CDFT, has been implicated in the decline of this critically endangered species in its native range of northern Florida and southeastern Georgia. In our current surveys of eight Florida torreya sites, cankers were present on all...

  10. First report of citrus canker caused by Xanthomonas citri in Somalia

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Xanthomonas citri, causal agent of citrus canker, has been reported in several countries in Africa, but not Somalia. During 2006 and 2007, hyperplasia-type lesions, often surrounded by a water-soaked margin and yellow halo, typical of citrus canker caused by X. citri, were found on 8-10 year-old gr...

  11. Volatilization of Selenium by Alternaria alternata

    PubMed Central

    Thompson-Eagle, E. T.; Frankenberger, W. T.; Karlson, U.

    1989-01-01

    Seleniferous water continues to be a serious problem to wildlife in the central valley of California. Water samples collected from Kesterson Reservoir, Peck Ranch, and Lost Hills evaporation pond facilities contained between 0.005 and 5 mg of Se per liter. The objective of this study was to isolate Se-methylating organisms in evaporation pond water and to assess, through enrichment and manipulation of their optimal growth parameters, the environmental factors which govern microbial Se methylation. Alternaria alternata was isolated as an active Se-methylating organism. The volatile product was identified as dimethylselenide. The effects of pH, temperature, Se substrates, and methyl donors on the ability of A. alternata to methylate Se were investigated in liquid medium containing 100 mg of Se per liter. The optimum pH and temperature for methylation were 6.5 and 30°C, respectively. Selenate and selenite were methylated more rapidly than selenium sulfide and various organic Se compounds (6-selenoguanosine, 6-selenoinosine, seleno-dl-methionine, and 6-selenopurine). l-Methionine and methyl cobalamine (0.1 μM) stimulated dimethylselenide production. This study demonstrates that Se-methylating organisms are present in evaporation pond water and are capable of liberating substantial quantities of Se in the volatile dimethylselenide form. By determining the optimum environmental conditions which stimulate volatilization, it may be possible to design a way to remove Se from seleniferous water in situ. PMID:16347933

  12. Characterization of Alternaria infectoria extracellular vesicles

    PubMed Central

    Silva, Branca M.A.; Prados-Rosales, Rafael; Espadas-Moreno, Javier; Wolf, Julie M.; Luque-Garcia, Jose L.; Gonçalves, Teresa; Casadevall, Arturo

    2015-01-01

    Many fungi use membrane vesicles to transport complex molecules across their cell walls. Like mammalian exosomes, fungal vesicles contain lipids, proteins, and polysaccharides, many of which are associated with virulence. Here we identify and characterize extracellular vesicles (EVs) in Alternaria infectoria, a ubiquitous, environmental filamentous fungus that is also an opportunistic human pathogen. Examination of the A. infectoria EVs revealed a morphology similar to that of vesicles described in other fungal species. Of note, proteomic analysis detected a reduced number of vesicle-associated proteins. There were two prevalent categories among the 20 identified proteins, including the polysaccharide metabolism group, probably related to plant host invasion or biosynthesis/degradation of cell wall components, and the nuclear proteins, especially DNA repair enzymes. We also found enzymes related to pigment synthesis, adhesion to the host cell, and trafficking of vesicles/organelles/molecules. This is the first time EV secretions have been identified in a filamentous fungus. We believe that these vesicles might have a role in virulence. PMID:24576997

  13. Characterization of Citrus-Associated Alternaria Species in Mediterranean Areas

    PubMed Central

    Garganese, Francesca; Schena, Leonardo; Siciliano, Ilenia; Prigigallo, Maria Isabella; Spadaro, Davide; De Grassi, Anna; Ippolito, Antonio

    2016-01-01

    Alternaria brown spot is one of the most important diseases of tangerines and their hybrids worldwide. Recently, outbreaks in Mediterranean areas related to susceptible cultivars, refocused attention on the disease. Twenty representatives were selected from a collection of 180 isolates of Alternaria spp. from citrus leaves and fruit. They were characterized along with reference strains of Alternaria spp. Micro- and macroscopic characteristics separated most Alternaria isolates into six morphotypes referable to A. alternata (5) and A. arborescens (1). Phylogenetic analyses, based on endopolygalacturonase (endopg) and internal transcribed spacer (ITS), confirmed this finding. Moreover, a five-gene phylogeny including two anonymous genomics regions (OPA 1–3 and OPA 2–1), and the beta-tubulin gene (ß-tub), produced a further clustering of A. alternata into three clades. This analysis suggested the existence of intra-species molecular variability. Investigated isolates showed different levels of virulence on leaves and fruit. In particular, the pathogenicity on fruit seemed to be correlated with the tissue of isolation and the clade. The toxigenic behavior of Alternaria isolates was also investigated, with tenuazonic acid (TeA) being the most abundant mycotoxin (0.2–20 mg/L). Isolates also synthesized the mycotoxins alternariol (AOH), its derivate alternariol monomethyl ether (AME), and altenuene (ALT), although to a lesser extent. AME production significantly varied among the six morphotypes. The expression of pksJ/pksH, biosynthetic genes of AOH/AME, was not correlated with actual toxin production, but it was significantly different between the two genotypes and among the four clades. Finally, ten isolates proved to express the biosynthetic genes of ACTT1 phytotoxin, and thus to be included in the Alternaria pathotype tangerine. A significant correlation between pathogenicity on leaves and ACTT1 gene expression was recorded. The latter was significantly

  14. [Equine exudative canker: an (auto-)immune disease?].

    PubMed

    Jongbloets, A M C; Sloet van Oldruitenborgh-Oosterbaan, M M; Meeus, P J H M; Back, W

    2005-02-15

    Equine canker is a chronic, hyperplastic, exudative pododermatitis affecting one or more feet. Although many causes and treatments have been suggested, the cause of the disease is still unknown and most probably multifactorial. Local treatments include radical surgical debridement of the diseased hoof tissue and application of caustic substances, antibiotics, and pressure bandaging. Nevertheless, the number of recurrences is high (45%). This article presents a 3-year-old New Forest pony-cross mare in which all horny structures (frogs, coronets, spurs, chestnuts) of all feet were affected. Bacteriological and fungal cultures of the frogs were found negative for the pathogens tested. Papilloma virus was not found. Clinical findings raised the hypothesis that the non-specific hyperplastic inflammation of these horn-like structures might have been caused by an (auto-)immune reaction. On the basis of the clinical findings, the pony was treated with surgical debridement of the frogs of a diagonal pair of feet and oral administration of prednisolone (1 mg/kg sid). The frogs, coronets, spurs, and chestnuts of all four feet healed completely within 8 weeks, thus making an (auto-)immune reaction more likely. In conclusion, this case report raised the hypothesis that an aspecific, hyperplastic inflammation of all four feet ('equine canker') and other horny structures may be caused by an (auto-)immune reaction, and that corticosteroids (prednisolone 1 mg/kg sid per os) are effective as treatment.

  15. 40 CFR 180.1256 - Alternaria destruens strain 059; exemption from the requirement of a tolerance.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... RESIDUES IN FOOD Exemptions From Tolerances § 180.1256 Alternaria destruens strain 059; exemption from the... the microbial pesticide Alternaria destruens Strain 059 when used in or on all raw...

  16. 40 CFR 180.1256 - Alternaria destruens strain 059; exemption from the requirement of a tolerance.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... RESIDUES IN FOOD Exemptions From Tolerances § 180.1256 Alternaria destruens strain 059; exemption from the... the microbial pesticide Alternaria destruens Strain 059 when used in or on all raw...

  17. Allergic bronchopulmonary mycosis due to Alternaria: Case report and review.

    PubMed

    Singh, Bhagteshwar; Denning, David W

    2012-01-01

    While allergic bronchopulmonary aspergillosis and mycosis are well recognised, no cases have been described related to Alternaria spp. Alternaria is a common sensitising fungus in asthmatics and related to thunderstorm asthma. We report a case of an asthmatic who presented with worsening asthma control, mild eosinophilia on high dose inhaled corticosteroids (800 μg/day), a total IgE of 3800 KIU/L, an Alternaria-specific IgE of 21.3 KUa/L and positive skin prick test, negative specific IgE and skin prick test to Aspergillus fumigatus, Penicillium spp., Cladosporium spp., Trichophyton spp. and a normal CT scan of the thorax. He responded well to a short course of oral prednisolone and then oral itraconazole, given over 17 months but relapsed 1 month after stopping it.

  18. 40 CFR 180.1256 - Alternaria destruens strain 059; exemption from the requirement of a tolerance.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 24 2011-07-01 2011-07-01 false Alternaria destruens strain 059... RESIDUES IN FOOD Exemptions From Tolerances § 180.1256 Alternaria destruens strain 059; exemption from the... the microbial pesticide Alternaria destruens Strain 059 when used in or on all raw...

  19. A Novel Alternaria Species Isolated from Peucedanum japonicum in Korea

    PubMed Central

    Deng, Jian Xin; Cho, Hye Sun; Paul, Narayan Chandra; Lee, Hyang Burm

    2014-01-01

    We isolated and examined a new Alternaria sp., which causes leaf spots on Peucedanum japonicum in Korea, by using molecular and morphological methods. Phylogenetic analysis based on a combined internal transcribed spacer region analysis and two protein-coding genes (gpd and Alt a1) demonstrated that the causal fungus was most closely related to A. cinerariae and A. sonchi, and relevant to A. brassicae. However, conidial morphology indicated that it is a novel species within the genus Alternaria, and therefore we have assigned the fungus a new name in this study. PMID:24808728

  20. A Novel Alternaria Species Isolated from Peucedanum japonicum in Korea.

    PubMed

    Deng, Jian Xin; Cho, Hye Sun; Paul, Narayan Chandra; Lee, Hyang Burm; Yu, Seung Hun

    2014-03-01

    We isolated and examined a new Alternaria sp., which causes leaf spots on Peucedanum japonicum in Korea, by using molecular and morphological methods. Phylogenetic analysis based on a combined internal transcribed spacer region analysis and two protein-coding genes (gpd and Alt a1) demonstrated that the causal fungus was most closely related to A. cinerariae and A. sonchi, and relevant to A. brassicae. However, conidial morphology indicated that it is a novel species within the genus Alternaria, and therefore we have assigned the fungus a new name in this study.

  1. Development of a qPCR technique to screen for resistance to Asiatic citrus canker

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Asiatic citrus canker (Acc) (causal organism Xanthomonas citri subspc. citri (Xcc) is threatening sustainability of the Florida citrus industry. Resistant cultivars, whether developed through conventional breeding or genetic transformation, will be he best solution for dealint with Acc. In Florida...

  2. Widespread Distribution of Fungivorus Aphelenchoides spp. in Blight Cankers on American Chestnut Trees.

    PubMed

    Griffin, G J; Eisenback, J D; Oldham, K

    2012-12-01

    Previously we showed in laboratory studies that the fungivorus nematode, Aphelenchoides hylurgi, was attracted to and fed upon the chestnut blight fungus, Cryphonectria parasitica, from American chestnut bark cankers and was a carrier of biocontrol, white hypovirulent C. parasitica strains. In the present field study, we recovered Aphelenchoides spp. in almost all (97.0 %) of 133 blight canker tissue assays (three 5-g samples each) from four eastern states. High mean population densities (227 to 474 nematodes per 5 g tissue) of Aphelenchoides spp. were recovered from cankers in Virginia, West Virginia, and Tennessee but not from New Hampshire (mean = 75 nematodes per 5 g tissue). Overall, most canker assays yielded population densities less than 200 nematodes per 5 g tissue. All of 12 very small or young cankers yielded a few to many Aphelenchoides spp. Regression analysis indicated greatest recovery of Aphelenchoides spp. occurred in the month of May (r = 0.94). The results indicate that Aphelenchoides spp. appear to be widespread in blight cankers on American chestnut trees and could play a role in biocontrol of chestnut blight.

  3. Genetic variability in the pistachio late blight fungus, Alternaria alternata

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Genetic variation in the pistachio late blight fungus, Alternaria alternata, was investigated by restriction fragment length polymorphism (RFLP) in the rDNA region. Southern hybridization of EcoRI, HindIII, and Xbal digested fungal DNA with a RNA probe derived from Alt1, an rDNA clone isolated from ...

  4. Airborne fungal spores of Alternaria, meteorological parameters and predicting variables

    NASA Astrophysics Data System (ADS)

    Filali Ben Sidel, Farah; Bouziane, Hassan; del Mar Trigo, Maria; El Haskouri, Fatima; Bardei, Fadoua; Redouane, Abdelbari; Kadiri, Mohamed; Riadi, Hassane; Kazzaz, Mohamed

    2015-03-01

    Alternaria is frequently found as airborne fungal spores and is recognized as an important cause of respiratory allergies. The aerobiological monitoring of fungal spores was performed using a Burkard volumetric spore traps. To establish predicting variables for daily and weakly spore counts, a stepwise multiple regression between spore concentrations and independent variables (meteorological parameters and lagged values from the series of spore concentrations: previous day or week concentration (Alt t - 1) and mean concentration of the same day or week in other years ( C mean)) was made with data obtained during 2009-2011. Alternaria conidia are present throughout the year in the atmosphere of Tetouan, although they show important seasonal fluctuations. The highest levels of Alternaria spores were recorded during the spring and summer or autumn. Alternaria showed maximum daily values in April, May or October depending on year. When the spore variables of Alternaria, namely C mean and Alt t - 1, and meteorological parameters were included in the equation, the resulting R 2 satisfactorily predict future concentrations for 55.5 to 81.6 % during the main spore season and the pre-peak 2. In the predictive model using weekly values, the adjusted R 2 varied from 0.655 to 0.676. The Wilcoxon test was used to compare the results from the expected values and the pre-peak spore data or weekly values for 2012, indicating that there were no significant differences between series compared. This test showed the C mean, Alt t - 1, frequency of the wind third quadrant, maximum wind speed and minimum relative humidity as the most efficient independent variables to forecast the overall trend of this spore in the air.

  5. [Colonization characteristics of endophytic bacteria NJ13 in Panax ginseng and its biocontrol efficiency against Alternaria leaf spot of ginseng].

    PubMed

    Chen, Chang-Qing; Li, Tong; Li, Xin-Lian; Jiang, Yun; Tian, Lei; Xu, Peng

    2014-05-01

    To reveal the colonization characteristics in host of endophytic biocontrol bacteria NJ13 isolated from Panax ginseng, this study obtained the marked strain NJ13-R which was double antibiotic resistant to rifampicin and streptomycin through enhancing the method of inducing antibiotic. The colonization characteristics in ginseng and its biocontrol efficiency against Alternaria spot of ginseng in the field were studied. The results showed that the strain could colonize in root, stem and leaf of ginseng and the colonization amount was positive correlated with inoculation concentration. Meanwhile, the strain could infect and then transfer in different tissues of ginseng The colonization amount of strain in roots and leaves of ginseng increased first and then decreased. However, the tendency of colonization amount of strain in stems was ascend at first and then descend slowly, and was more than that in roots and leaves along with time, which had a preference to specific tissue of its host. In field experiment, the endophytic bacteria NJ13 was proved to be effective in controlling Alternaria leaf spot of ginseng. The biocontrol efficiency of fermentation broth at the concentration of 0.76 x 10(8) cfu x mL(-1) reached 75.62%, which was close to the controlling level (73.06%) of 0.67 mg x L(-1) 50% cyprodinil WG.

  6. Oak Tree Canker Disease Supports Arthropod Diversity in a Natural Ecosystem

    PubMed Central

    Lee, Yong-Bok; An, Su Jung; Park, Chung Gyoo; Kim, Jinwoo; Han, Sangjo; Kwak, Youn-Sig

    2014-01-01

    Microorganisms have many roles in nature. They may act as decomposers that obtain nutrients from dead materials, while some are pathogens that cause diseases in animals, insects, and plants. Some are symbionts that enhance plant growth, such as arbuscular mycorrhizae and nitrogen fixation bacteria. However, roles of plant pathogens and diseases in natural ecosystems are still poorly understood. Thus, the current study addressed this deficiency by investigating possible roles of plant diseases in natural ecosystems, particularly, their positive effects on arthropod diversity. In this study, the model system was the oak tree (Quercus spp.) and the canker disease caused by Annulohypoxylon truncatum, and its effects on arthropod diversity. The oak tree site contained 44 oak trees; 31 had canker disease symptoms while 13 were disease-free. A total of 370 individual arthropods were detected at the site during the survey period. The arthropods belonged to 25 species, 17 families, and seven orders. Interestingly, the cankered trees had significantly higher biodiversity and richness compared with the canker-free trees. This study clearly demonstrated that arthropod diversity was supported by the oak tree canker disease. PMID:25288984

  7. Oak tree canker disease supports arthropod diversity in a natural ecosystem.

    PubMed

    Lee, Yong-Bok; An, Su Jung; Park, Chung Gyoo; Kim, Jinwoo; Han, Sangjo; Kwak, Youn-Sig

    2014-03-01

    Microorganisms have many roles in nature. They may act as decomposers that obtain nutrients from dead materials, while some are pathogens that cause diseases in animals, insects, and plants. Some are symbionts that enhance plant growth, such as arbuscular mycorrhizae and nitrogen fixation bacteria. However, roles of plant pathogens and diseases in natural ecosystems are still poorly understood. Thus, the current study addressed this deficiency by investigating possible roles of plant diseases in natural ecosystems, particularly, their positive effects on arthropod diversity. In this study, the model system was the oak tree (Quercus spp.) and the canker disease caused by Annulohypoxylon truncatum, and its effects on arthropod diversity. The oak tree site contained 44 oak trees; 31 had canker disease symptoms while 13 were disease-free. A total of 370 individual arthropods were detected at the site during the survey period. The arthropods belonged to 25 species, 17 families, and seven orders. Interestingly, the cankered trees had significantly higher biodiversity and richness compared with the canker-free trees. This study clearly demonstrated that arthropod diversity was supported by the oak tree canker disease.

  8. Development and validation of standard area diagrams as assessment aids for estimating the severity of citrus canker on unripe oranges

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Canker (caused by Xanthomonas citri subsp. citri) is an important disease of citrus in Brazil and elsewhere in the world, and can cause severe disease on the fruit. The severity of citrus canker of fruit must often be estimated visually. The objective of this research was to construct and validate s...

  9. Comparison of enzyme immunoassay–based assays for environmental Alternaria alternata

    PubMed Central

    Barnes, Charles; Portnoy, Jay; Sever, Michelle; Arbes, Samuel; Vaughn, Ben; Zeldin, Darryl C.

    2007-01-01

    Background Alternaria alternata–derived allergenic materials are causes of human disease. Several immunoassays exist to quantify these materials. Objective To compare methods for evaluating Alternaria content. Methods Four methods, including 1 monoclonal antibody (MAb)–based assay specific for recombinant Alt a 1, 1 MAb-based assay for chromatographically purified Alt a 1, 1 polyclonal antibody (PAb)–based assay for chromatographically purified Alt a 1, and 1 PAb-based assay for whole Alternaria extract, were evaluated. Environmental samples collected as part of the National Survey of Lead and Allergens in Housing were examined. Alternaria spore counts were determined in dust by observation. Results The MAb-based assay for recombinant Alt a 1 detected Alternaria in few samples (25%); the PAb-based assay for whole Alternaria proteins detected antigen in 97% of the samples. The PAb- and MAb-based assays for purified Alt a 1 detected antigen in 100% of the samples. There was a significant positive correlation between the 2 assays directed against purified Alt a 1. There was a positive correlation between the PAb-based assay for whole Alternaria and the PAb-based assay for Alt a 1. Nearly all the dust samples contained Alternaria spores, and there was a strong positive correlation between counts and all assays. Conclusion Because of the multifaceted nature of Alternaria, the disparities between methods for quantifying Alternaria, the cross-reactivity between fungal allergens, and the documented genetic promiscuity of this fungus, enzyme immunoassays using PAbs against a range of Alternaria proteins will probably produce the most reliable estimation of overall Alternaria exposure in house dust. PMID:17042141

  10. STEM?!?!

    ERIC Educational Resources Information Center

    Merrill, Jen

    2012-01-01

    The author's son has been an engineer since birth. He never asked "why" as a toddler, it was always "how's it work?" So that he wanted a STEM-based home education was no big surprise. In this article, the author considers what kind of curricula would work best for her complex kid.

  11. Daily variations of Alternaria spores in the city of Murcia (semi-arid southeastern Spain)

    NASA Astrophysics Data System (ADS)

    Munuera Giner, M.; Carrión García, J. S.

    1995-12-01

    Annual variations in the abundance of Alternaria spores were related to the length of the spore period for data from Murcia (southeastern Spain). To understand the relationship between the number of spores and climatic factors, Alternaria spore counts for March 1993 to February 1994 were examined by means of correlation and regression analyses with fourteen different weather parameters. The results indicated that there was a tendency for Alternaria spore concentrations to increase with increases in temperature, wind speed and hours of sunshine. Negative correlations were observed with air pressure, wind direction and humidity. Theoretical curves for Alternaria spore counts are given in relation to temperatures during the period studied.

  12. Air-borne fungi in the air of Barcelona (Spain). II. The genus alternaria.

    PubMed

    Calvo, M A; Guarro, J; Suarez, G; Ramírez, C

    1979-12-28

    The genus Alternaria Nees formed one of the more important components of the fungus population of the air at Barcelone (Spain), during a two-year study, from February 1976 through January 1978. Results were based only on studied of colonies obtained by gravity-exposed plates. The occurrence of this genus was greatly affected by climatic conditions. In general. however, Alternaria appeared to prefer warmer weather. A total of six species of Alternaria was identified, of which, Alternaria alternata (Fries) Keissler was by far the commonest, representing 71.3% of all colonies.

  13. Detection of citrus canker and Huanglongbing using fluorescence imaging spectroscopy and support vector machine technique.

    PubMed

    Wetterich, Caio Bruno; Felipe de Oliveira Neves, Ruan; Belasque, José; Marcassa, Luis Gustavo

    2016-01-10

    Citrus canker and Huanglongbing (HLB) are citrus diseases that represent a serious threat to the citrus production worldwide and may cause large economic losses. In this work, we combined fluorescence imaging spectroscopy (FIS) and a machine learning technique to discriminate between these diseases and other ordinary citrus conditions that may be present at citrus orchards, such as citrus scab and zinc deficiency. Our classification results are highly accurate when discriminating citrus canker from citrus scab (97.8%), and HLB from zinc deficiency (95%). These results show that it is possible to accurately identify citrus diseases that present similar symptoms.

  14. Different Transcriptional Response to Xanthomonas citri subsp. citri between Kumquat and Sweet Orange with Contrasting Canker Tolerance

    PubMed Central

    Fu, Xing-Zheng; Gong, Xiao-Qing; Zhang, Yue-Xin; Wang, Yin; Liu, Ji-Hong

    2012-01-01

    Citrus canker disease caused by Xanthomonas citri subsp. citri (Xcc) is one of the most devastating biotic stresses affecting the citrus industry. Meiwa kumquat (Fortunella crassifolia) is canker-resistant, while Newhall navel orange (Citrus sinensis Osbeck) is canker-sensitive. To understand the molecular mechanisms underlying the differences in responses to Xcc, transcriptomic profiles of these two genotypes following Xcc attack were compared by using the Affymetrix citrus genome GeneChip. A total of 794 and 1324 differentially expressed genes (DEGs) were identified as canker-responsive genes in Meiwa and Newhall, respectively. Of these, 230 genes were expressed in common between both genotypes, while 564 and 1094 genes were only significantly expressed in either Meiwa or Newhall. Gene ontology (GO) annotation and Singular Enrichment Analysis (SEA) of the DEGs showed that genes related to the cell wall and polysaccharide metabolism were induced for basic defense in both Meiwa and Newhall, such as chitinase, glucanase and thaumatin-like protein. Moreover, apart from inducing basic defense, Meiwa showed specially upregulated expression of several genes involved in the response to biotic stimulus, defense response, and cation binding as comparing with Newhall. And in Newhall, abundant photosynthesis-related genes were significantly down-regulated, which may be in order to ensure the basic defense. This study revealed different molecular responses to canker disease in Meiwa and Newhall, affording insight into the response to canker and providing valuable information for the identification of potential genes for engineering canker tolerance in the future. PMID:22848606

  15. Influence of environmental parameters on mycotoxin production by Alternaria arborescens.

    PubMed

    Vaquera, Sandra; Patriarca, Andrea; Fernández Pinto, V

    2016-02-16

    Alternaria arborescens has been reported as a common fungal species invading tomatoes and is capable of producing several mycotoxins in infected plants, fruits and in agricultural commodities. Alternariol (AOH), alternariol monomethyl ether (AME), and tenuazonic acid (TeA) are some of the main Alternaria mycotoxins that can be found as contaminants of food. This species can produce these toxic metabolites together with AAL toxins (Alternaria alternata f. sp. lycopersicum toxins), which can act as inhibitors of sphingolipid biosynthesis. The objective of this study was to determine the effect of water activity (aw, 0.995, 0.975, 0.950) and temperature (6, 15, 20, 25 and 30 °C) on mycotoxin production by A. arborescens on a synthetic tomato medium. The optimum production of AOH and AME occurred at 0.975 aw after 40 days of incubation at 30 °C. The maximum TeA accumulation was observed at 0.975 aw and 25 °C and at 0.950 aw and 30 °C. AAL TA was produced in higher quantities at 0.995 aw and 30 °C. At 6 °C no quantifiable levels of AOH or AME were detected, but significant amounts of TeA were produced at 0.975 aw. In general, high aw levels and high temperatures were favorable for mycotoxin production. The greatest accumulation of all four toxins occurred at 0.975 aw and 30 °C. The results obtained here could be extrapolated to evaluate the risk of tomato fruits and tomato products contamination caused by these toxins.

  16. Alternaria roseogrisea, a new species from achenes of Helianthus annuus (sunflower)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Alternaria roseogrisea was isolated from the achenes of Helianthus annuus L. during studies conducted in 1983-85 to document the diversity of fungi occurring in sunflower seeds and the possible role these fungi play in degradation of oilseed quality. A. roseogrisea was reported as "Alternaria sp. 2...

  17. Complete DNA Sequence of Pseudomonas syringae pv. actinidiae, the Causal Agent of Kiwifruit Canker Disease.

    PubMed

    Templeton, Matthew D; Warren, Benjamin A; Andersen, Mark T; Rikkerink, Erik H A; Fineran, Peter C

    2015-09-17

    Pseudomonas syringae pv. actinidiae is the causal agent of bacterial canker of kiwifruit, a disease that has rapidly spread worldwide. We have fully sequenced and assembled the chromosomal and plasmid DNA from P. syringae pv. actinidiae ICMP 18884 using the PacBio RS II platform.

  18. Rapid screening for citrus canker resistance employing pattern-triggered immunity (PTI) responses

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus canker, caused by the bacterial pathogen Xanthomonas citri ssp. citri (Xcc), has been attributed to millions of dollars in loss or damage to commercial citrus crops in subtropical production areas of the world. Since identification of resistant plants is one of the most effective methods of d...

  19. Optimal strategies for the eradication of Asiatic citrus canker in heterogeneous host landscapes

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The eradication of non-native plant pathogens is a key challenge in plant disease epidemiology. Asiatic citrus canker is an economically significant disease of citrus caused by the bacterial plant pathogen Xanthomonas citri subsp. citri. The pathogen is a major exotic disease problem in many citru...

  20. Wind speed and wind-associated leaf injury affect severity of citrus canker on Swingle citrumelo

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus canker (caused by the bacterial pathogen Xanthomonas citri subsp. citri, Xcc) can cause severe damage to citrus. It is endemic in Florida, and occurs in other citrus growing regions. The bacterium is dispersed predominantly in rain splash. To simulate dispersal in splash, and to investigate t...

  1. Commercial postharvest practices used to handle fresh citrus fruit with canker symptoms

    Technology Transfer Automated Retrieval System (TEKTRAN)

    To assist in developing best postharvest practices for handling fruit with canker lesions, a survey was distributed in summers of 2008 and 2009 to better understand current practices. Approximately 60% of the surveys were returned each year representing about 55% of total fresh fruit shipments. As e...

  2. Evaluation of resistance to asiatic citrus canker among selections of pera sweet orange (Citrus sinensis)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Asiatic citrus canker (ACC, caused by the bacterium Xanthomonas citri subsp. citri) is a destructive disease of citrus in Brazil and in several other citrus-producing countries. ACC management is problematic, and bactericides such as copper can be reasonably efficacious but do not completely control...

  3. Thousand cankers disease complex: a forest health issue across the U.S.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Thousand Cankers Disease (TCD) is a disease complex wherein the fungus (Geosmithia morbida), is vectored by the walnut twig beetle (WTB, Pityophthorus juglandis). Disease causes mortality primarily of eastern black walnut (Juglans nigra), though other walnut species are also susceptible. Eastern bla...

  4. New canker disease of Incense-cedar in Oregon caused by Phaeobotryon cupressi.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Incense-cedar (Calocedrus decurrens) is a native tree occurring in Oregon and California. Since the early 2000’s, a new canker disease has been observed with increasing frequency on ornamental and windbreak trees planted in the Willamette Valley of Oregon. Symptoms appear as dead, flagging, small-di...

  5. Resistance evaluation of Pera (Citrus sinensis) genotypes to citrus canker in greenhouse conditions

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus canker, caused by the bacterium Xanthomonas citri subsp. citri results in serious yield losses and phytoregulation penalties. The use of resistant genotypes is recognized as an important tool to facilitate control of the pathogen. Studies have show that artificial inoculation results in typic...

  6. Exacerbation of citrus canker by citrus leafminer, Phyllocnistis citrella in Florida

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus canker (caused by Xanthomonas citri subsp. citri, Xcc) is an important bacterial disease of citrus that is spread naturally by rain and wind. Damage to citrus leaves by the citrus leafminer (CLM) , Phyllocnistis citrella Stainton (Lepidoptera: Gracillariidae), has been shown to promote infect...

  7. Canker and twig dieback of blueberry caused by Pestalotiopsis spp. and a Truncatella sp. in Chile

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Blueberry (Vaccinium spp.) has great economic importance in Chile, currently with about 8,500 ha being cultivated. Recently, the presence of canker and dieback symptoms has been observed along the productive blueberry zone of Chile extending from the V Region (32º49´ South lat.) in the north to the ...

  8. First report of Phaeobotryon cupressi causing canker of Calocedrus decurrens in Oregon.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Since the early 2000’s a canker disease has been noticed with increasing frequency on landscape specimens of native incense cedar (Calocedrus decurrens) planted throughout the Willamette Valley (from Portland south to Eugene) in western Oregon. Symptoms initially appear as dead and flagging small-di...

  9. Overexpression of a Modified Plant Thionin Enhances Disease Resistance to Citrus Canker and Huanglongbing (HLB).

    PubMed

    Hao, Guixia; Stover, Ed; Gupta, Goutam

    2016-01-01

    Huanglongbing (HLB or citrus greening disease) caused by Candidatus Liberibacter asiaticus (Las) is a great threat to the US citrus industry. There are no proven strategies to eliminate HLB disease and no cultivar has been identified with strong HLB resistance. Citrus canker is also an economically important disease associated with a bacterial pathogen (Xanthomonas citri). In this study, we characterized endogenous citrus thionins and investigated their expression in different citrus tissues. Since no HLB-resistant citrus cultivars have been identified, we attempted to develop citrus resistant to both HLB and citrus canker through overexpression of a modified plant thionin. To improve effectiveness for disease resistance, we modified and synthesized the sequence encoding a plant thionin and cloned into the binary vector pBinPlus/ARS. The construct was then introduced into Agrobacterium strain EHA105 for citrus transformation. Transgenic Carrizo plants expressing the modified plant thionin were generated by Agrobacterium-mediated transformation. Successful transformation and transgene gene expression was confirmed by molecular analysis. Transgenic Carrizo plants expressing the modified thionin gene were challenged with X. citri 3213 at a range of concentrations, and a significant reduction in canker symptoms and a decrease in bacterial growth were demonstrated compared to nontransgenic plants. Furthermore, the transgenic citrus plants were challenged with HLB via graft inoculation. Our results showed significant Las titer reduction in roots of transgenic Carrizo compared with control plants and reduced scion Las titer 12 months after graft inoculation. These data provide promise for engineering citrus disease resistance against HLB and canker.

  10. Production of transgenic citrus resistant to citrus canker and Huanglongbing diseases

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Huanglongbing (HLB or citrus greening disease) caused by Candidatus Liberibacter asiaticus (Las) is a great threat to the U.S. citrus industry. There are no proven strategies to eliminate HLB disease and no cultivars identified with strong HLB resistance. Citrus canker is also an economically import...

  11. Overexpression of a modified plant thionin enhances disease resistance to citrus canker and Huanglongbing (HLB)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Huanglongbing (HLB or citrus greening disease) caused by Candidatus Liberibacter asiaticus (Las) is a great threat to the United States citrus industry. There are no proven strategies to eliminate HLB disease and no cultivar has been identified with strong HLB resistance. Citrus canker is also an ec...

  12. Carrier-bound Alt a 1 peptides without allergenic activity for vaccination against Alternaria alternata allergy

    PubMed Central

    Twaroch, T. E.; Focke, M.; Fleischmann, K.; Balic, N.; Lupinek, C.; Blatt, K.; Ferrara, R.; Mari, A.; Ebner, C.; Valent, P.; Spitzauer, S.; Swoboda, I.; Valenta, R.

    2017-01-01

    Background The mould Alternaria alternata is a major elicitor of allergic asthma. Diagnosis and specific immunotherapy (SIT) of Alternaria allergy are often limited by the insufficient quality of natural mould extracts. Objective To investigate whether recombinant Alt a 1 can be used for reliable diagnosis of Alternaria alternata allergy and to develop a safe, non-allergenic vaccine for SIT of Alternaria allergy. Methods The qualitative sensitization profile of 80 Alternaria-allergic patients from Austria and Italy was investigated using an allergen micro-array and the amount of Alternaria-specific IgE directed to rAlt a 1 was quantified by ImmunoCAP measurements. Peptides spanning regions of predicted high surface accessibility of Alt a 1 were synthesized and tested for IgE reactivity and allergenic activity, using sera and basophils from allergic patients. Carrier-bound peptides were studied for their ability to induce IgG antibodies in rabbits which recognize Alt a 1 and inhibit allergic patients’ IgE reactivity to Alt a 1. Results rAlt a 1 allowed diagnosis of Alternaria allergy in all tested patients, bound the vast majority (i.e. >95%) of Alternaria-specific IgE and elicited basophil activation already at a concentration of 0.1 ng/mL. Four non-allergenic peptides were synthesized which, after coupling to the carrier protein keyhole limpet hemocyanin, induced Alt a 1-specific IgG and inhibited allergic patients’ IgE binding to Alt a 1. Conclusions and clinical relevance rAlt a 1 is a highly allergenic molecule allowing sensitive diagnosis of Alternaria allergy. Carrier-bound non-allergenic Alt a 1 peptides are candidates for safe SIT of Alternaria allergy. PMID:22909168

  13. Diversity and movement of indoor Alternaria alternata across the mainland USA.

    PubMed

    Woudenberg, J H C; van der Merwe, N A; Jurjević, Ž; Groenewald, J Z; Crous, P W

    2015-08-01

    Alternaria spp. from sect. Alternaria are frequently associated with hypersensitivity pneumonitis, asthma and allergic fungal rhinitis and sinusitis. Since Alternaria is omnipresent in the outdoor environment, it is thought that the indoor spore concentration is mainly influenced by the outdoor spore concentration. However, few studies have investigated indoor Alternaria isolates, or attempted a phylogeographic or population genetic approach to investigate their movement. Therefore, the aim of the current study was to investigate the molecular diversity of indoor Alternaria isolates in the USA, and to test for recombination, using these approaches. Alternaria isolates collected throughout the USA were identified using ITS, gapdh and endoPG gene sequencing. This was followed by genotyping and population genetic inference of isolates belonging to Alternaria sect. Alternaria together with 37 reference isolates, using five microsatellite markers. Phylogenetic analyses revealed that species of Alternaria sect. Alternaria represented 98% (153 isolates) of the indoor isolates collected throughout the USA, of which 137 isolates could be assigned to A. alternata, 15 to the A. arborescens species complex and a single isolate to A. burnsii. The remaining 2% (3 isolates) represented sect. Infectoriae (single isolate) and sect. Pseudoulocladium (2 isolates). Population assignment analyses of the 137 A. alternata isolates suggested that subpopulations did not exist within the sample. The A. alternata isolates were thus divided into four artificial subpopulations to represent four quadrants of the USA. Forty-four isolates representing the south-western quadrant displayed the highest level of uniqueness based on private alleles, while the highest level of gene flow was detected between the south-eastern (32 isolates) and south-western quadrants. Genotypic diversity was high for all quadrants, and a test for linkage disequilibrium suggested that A. alternata has a cryptic sexual

  14. Activation of Melanin Synthesis in Alternaria infectoria by Antifungal Drugs

    PubMed Central

    Fernandes, Chantal; Prados-Rosales, Rafael; Silva, Branca M. A.; Nakouzi-Naranjo, Antonio; Zuzarte, Mónica; Chatterjee, Subhasish; Stark, Ruth E.; Casadevall, Arturo

    2015-01-01

    The importance of Alternaria species fungi to human health ranges from their role as etiological agents of serious infections with poor prognoses in immunosuppressed individuals to their association with respiratory allergic diseases. The present work focuses on Alternaria infectoria, which was used as a model organism of the genus, and was designed to unravel melanin production in response to antifungals. After we characterized the pigment produced by A. infectoria, we studied the dynamics of 1,8-dihydroxynaphthalene (DHN)-melanin production during growth, the degree of melanization in response to antifungals, and how melanization affected susceptibility to several classes of therapeutic drugs. We demonstrate that A. infectoria increased melanin deposition in cell walls in response to nikkomycin Z, caspofungin, and itraconazole but not in response to fluconazole or amphotericin B. These results indicate that A. infectoria activates DHN-melanin synthesis in response to certain antifungal drugs, possibly as a protective mechanism against these drugs. Inhibition of DHN-melanin synthesis by pyroquilon resulted in a lower minimum effective concentration (MEC) of caspofungin and enhanced morphological changes (increased hyphal balloon size), characterized by thinner and less organized A. infectoria cell walls. In summary, A. infectoria synthesizes melanin in response to certain antifungal drugs, and its susceptibility is influenced by melanization, suggesting the therapeutic potential of drug combinations that affect melanin synthesis. PMID:26711773

  15. Mapping Resistance to Alternaria cucumerina in Cucumis melo.

    PubMed

    Daley, James; Branham, Sandra; Levi, Amnon; Hassell, Richard; Wechter, Patrick

    2017-04-01

    Infection with Alternaria cucumerina causes Alternaria leaf blight (ALB), a disease characterized by lesion formation on leaves, leading to substantial yield and quality losses in Cucumis melo (melon). Although fungicides are effective against ALB, reduction in the frequency of application would be economically and environmentally beneficial. Resistant melon lines have been identified but the genetic basis of this resistance has not been determined. A saturated melon genetic map was constructed with markers developed through genotyping by sequencing of a recombinant inbred line population (F6 to F10; n = 82) derived from single-seed descent of a F2 population from a cross between the ALB-resistant parent MR-1 and the ALB-susceptible parent Ananas Yokneum. The population was evaluated for A. cucumerina resistance with an augmented block greenhouse study using inoculation with the wounded-leaf method. Multiple quantitative trait loci (QTL) mapping identified two QTL that explained 33.9% of variation in lesion area. Several candidate genes within range of these QTL were identified using the C. melo v3.5 genome. Markers linked to these QTL will be used to accelerate efforts to breed melon cultivars resistant to ALB.

  16. Activation of Melanin Synthesis in Alternaria infectoria by Antifungal Drugs.

    PubMed

    Fernandes, Chantal; Prados-Rosales, Rafael; Silva, Branca M A; Nakouzi-Naranjo, Antonio; Zuzarte, Mónica; Chatterjee, Subhasish; Stark, Ruth E; Casadevall, Arturo; Gonçalves, Teresa

    2015-12-28

    The importance of Alternaria species fungi to human health ranges from their role as etiological agents of serious infections with poor prognoses in immunosuppressed individuals to their association with respiratory allergic diseases. The present work focuses on Alternaria infectoria, which was used as a model organism of the genus, and was designed to unravel melanin production in response to antifungals. After we characterized the pigment produced by A. infectoria, we studied the dynamics of 1,8-dihydroxynaphthalene (DHN)-melanin production during growth, the degree of melanization in response to antifungals, and how melanization affected susceptibility to several classes of therapeutic drugs. We demonstrate that A. infectoria increased melanin deposition in cell walls in response to nikkomycin Z, caspofungin, and itraconazole but not in response to fluconazole or amphotericin B. These results indicate that A. infectoria activates DHN-melanin synthesis in response to certain antifungal drugs, possibly as a protective mechanism against these drugs. Inhibition of DHN-melanin synthesis by pyroquilon resulted in a lower minimum effective concentration (MEC) of caspofungin and enhanced morphological changes (increased hyphal balloon size), characterized by thinner and less organized A. infectoria cell walls. In summary, A. infectoria synthesizes melanin in response to certain antifungal drugs, and its susceptibility is influenced by melanization, suggesting the therapeutic potential of drug combinations that affect melanin synthesis.

  17. Chemical constituents of marine mangrove-derived endophytic fungus Alternaria tenuissima EN-192

    NASA Astrophysics Data System (ADS)

    Sun, Hong; Gao, Shushan; Li, Xiaoming; Li, Chunshun; Wang, Bingui

    2013-03-01

    A chemical investigation of the ethyl acetate extract of the fermentation broth of Alternaria tenuissima EN-192, an endophytic fungus obtained from the stems of the marine mangrove plant Rhizophora stylosa, resulted in the isolation of nine known secondary metabolites, including four indole-diterpenoids: penijanthine A ( 1), paspaline ( 2), paspalinine ( 3), and penitrem A ( 4); three tricycloalternarene derivatives: tricycloalternarene 3a ( 5), tricycloalternarene 1b ( 6), and tricycloalternarene 2b ( 7); and two alternariol congeners: djalonensone ( 8) and alternariol ( 9). The chemical structures of these metabolites were characterized through a combination of detailed spectroscopic analyses and their comparison with reports from the literature. The inhibitory activities of each isolated compound against four bacteria were evaluated and compounds 5 and 8 displayed moderate activity against the aquaculture pathogenic bacterium Vibrio anguillarum, with inhibition zone diameters of 8 and 9 mm, respectively, at 100 μg/disk. To the best of our knowledge, this is the first report on the secondary metabolites of mangrove-derived A lternaria tenuissima and also the first report of the isolation of indole-diterpenoids from fungal genus A lternaria.

  18. Cytospora species associated with walnut canker disease in China, with description of a new species C. gigalocus.

    PubMed

    Fan, Xinlei; Hyde, Kevin D; Liu, Min; Liang, Yingmei; Tian, Chengming

    2015-05-01

    Cytospora species associated with canker disease are presently difficult to identify because of lack of ex-types cultures with molecular data, few distinguishable characters, and only Internal transcribed spacer (ITS) sequence data is available for most Cytospora strains in GenBank. We report on Cytospora species from the walnut tree in China. Collections were subjected to morphological and phylogenetic study. The relatedness of species associated with walnut canker were established using combined ITS, nrLSU, β-tubulin, and actin gene sequence data. Cytospora atrocirrhata, Cytospora chrysosperma, Cytospora sacculus, and a new species, Cytospora gigalocus, were identified causing canker disease of walnut. Cytospora gigalocus is formally described and compared with most similar species. Cytospora chrysosperma and C. sacculus have previously been recorded from walnut, whereas C. atrocirrhata is reported as associated with walnut canker for the first time. This is the first study that has established the Cytospora species causing walnut canker in China using a multi-phasic approach. All species are recorded as being associated with walnut canker disease in China for the first time.

  19. Phylogenetic, Morphological, and Pathogenic Characterization of Alternaria Species Associated with Fruit Rot of Blueberry in California.

    PubMed

    Zhu, X Q; Xiao, C L

    2015-12-01

    Fruit rot caused by Alternaria spp. is one of the most important factors affecting the postharvest quality and shelf life of blueberry fruit. The aims of this study were to characterize Alternaria isolates using morphological and molecular approaches and test their pathogenicity to blueberry fruit. Alternaria spp. isolates were collected from decayed blueberry fruit in the Central Valley of California during 2012 and 2013. In total, 283 isolates were obtained and five species of Alternaria, including Alternaria alternata, A. tenuissima, A. arborescens, A. infectoria, and A. rosae, were identified based on DNA sequences of the plasma membrane ATPase, Alt a1 and Calmodulin gene regions in combination with morphological characters of the culture and sporulation. Of the 283 isolates, 61.5% were identified as A. alternata, 32.9% were A. arborescens, 5.0% were A. tenuissima, and only one isolate of A. infectoria and one isolate of A. rosae were found. These fungi were able to grow at temperatures from 0 to 35°C, and mycelial growth was arrested at 40°C. Optimal radial growth occurred between 20 to 30°C. Pathogenicity tests showed that all five Alternaria spp. were pathogenic on blueberry fruit at 0, 4, and 20°C, with A. alternata, A. arborescens, and A. tenuissima being the most virulent species, followed by A. infectoria and A. rosae. Previously A. tenuissima has been reported to be the primary cause of Alternaria fruit rot of blueberry worldwide. Our results indicated that the species composition of Alternaria responsible for Alternaria fruit rot in blueberry can be dependent on geographical region. A. alternata, A. arborescens, A. infectoria, and A. rosae are reported for the first time on blueberry in California. This is also the first report of A. infectoria and A. rosae infecting blueberry fruit.

  20. Mold and Alternaria skin test reactivity and asthma in children in Connecticut

    PubMed Central

    Lyons, Todd W.; Wakefield, Dorothy B.; Cloutier, Michelle M.

    2013-01-01

    Background Sensitivity to mold has been associated with asthma incidence, persistence, and severity. Objective To examine the relationship between skin test reactivity (STR) to molds and specifically to Alternaria and asthma severity in a group of ethnically diverse children in Connecticut. Methods Demographics and STR to 14 local allergens, including Alternaria, Penicillium, and mold mix, were obtained for 914 Puerto Rican, African American, and non-Hispanic white children. Results A total of 126 children (14%) had a positive skin test result to mold, and 58 (6%) demonstrated STR to Alternaria. Compared with non-Hispanic white children, there was no difference in the likelihood of being sensitized to Alternaria for Puerto Rican and African American children (odds ratio [OR], 0.7; 95% confidence interval [CI], 0.3–1.5; and OR, 0.9; 95% CI, 0.4–2.2; respectively). In an adjusted analysis, Alternaria STR was associated with severe, persistent asthma (OR, 3.4; 95% CI, 1.2–8.6) but did not predict increasing asthma severity. STR to cat (OR, 2.5; 95% CI, 1.3–4.9) and dog (OR, 2.9; 95% CI, 1.3–6.0) was also associated with severe persistent asthma. Alternaria STR was associated with severe persistent asthma independent of the total number of positive skin test results. Conclusions Mold and Alternaria STR were uncommon among children in Connecticut. Alternaria STR was not associated with increasing asthma severity but was associated with severe, persistent asthma independent of the total number of positive skin test results. There was no association between ethnicity and Alternaria STR. PMID:21457878

  1. Draft Genome Sequences of Pectobacterium carotovorum subsp. actinidiae ICMP 19971 and ICMP 19972, Two Strains Isolated from Actinidia chinensis with Symptoms of Summer Canker in South Korea.

    PubMed

    Visnovsky, Sandra B; Panda, Preetinanda; Taylor, Robert; Pitman, Andrew R

    2017-04-06

    Pectobacterium carotovorum subsp. actinidiae is the causal agent of summer canker in kiwifruit plants in South Korea. We report here the draft genome sequences of two P. carotovorum subsp. actinidiae strains, ICMP 19971 and ICMP 19972, which were originally isolated from Actinidia chinensis with symptoms of summer canker. These genome sequences will aid in the identification of genetic traits associated with their unusual capacity to cause canker and help understanding of the threat these exotic enterobacteria pose to the New Zealand kiwifruit industry.

  2. Morphogenetic Alterations of Alternaria alternata Exposed to Dicarboximide Fungicide, Iprodione

    PubMed Central

    Kim, Eunji; Lee, Hye Min; Kim, Young Ho

    2017-01-01

    Fungicide-resistant Alternaria alternata impede the practical control of the Alternaria diseases in crop fields. This study aimed to investigate cytological fungicide resistance mechanisms of A. alternata against dicarboximide fungicide iprodione. A. alternata isolated from cactus brown spot was cultured on potato-dextrose agar (PDA) with or without iprodione, and the fungal cultures with different growth characteristics from no, initial and full growth were observed by light and electron microscopy. Mycelia began to grow from one day after incubation (DAI) and continued to be in full growth (control-growth, Con-G) on PDA without fungicide, while on PDA with iprodione, no fungal growth (iprodione-no growth, Ipr-N) occurred for the first 3 DAI, but once the initial growth (iprodione-initial growth, Ipr-I) began at 4–5 DAI, the colonies grew and expanded continuously to be in full growth (iprodione-growth, Ipr-G), suggesting Ipr-I may be a turning moment of the morphogenetic changes resisting fungicidal toxicity. Con-G formed multicellular conidia with cell walls and septa and intact dense cytoplasm. In Ipr-N, fungal sporulation was inhibited by forming mostly undeveloped unicellular conidia with degraded and necrotic cytoplasm. However, in Ipr-I, conspicuous cellular changes occurred during sporulation by forming multicellular conidia with double layered (thickened) cell walls and accumulation of proliferated lipid bodies in the conidial cytoplasm, which may inhibit the penetration of the fungicide into conidial cells, reducing fungicide-associated toxicity, and may be utilized as energy and nutritional sources, respectively, for the further fungal growth to form mature colonies as in Ipr-G that formed multicellular conidia with cell walls and intact cytoplasm with lipid bodies as in Con-G. PMID:28167893

  3. Establishment of CRISPR/Cas9 in Alternaria alternata.

    PubMed

    Wenderoth, Maximilian; Pinecker, Christoph; Voß, Benjamin; Fischer, Reinhard

    2017-03-10

    The filamentous fungus Alternaria alternata is a potent producer of many secondary metabolites, some of which like alternariol or alternariol-methyl ether are toxic and/or cancerogenic. Many Alternaria species do not only cause post-harvest losses of food and feed, but are aggressive plant pathogens. Despite the great economic importance and the large number of research groups working with the fungus, the molecular toolbox is rather underdeveloped. Gene deletions often result in heterokaryotic strains and therefore, gene-function analyses are rather tedious. In addition, A. alternata lacks a sexual cycle and classical genetic approaches cannot be combined with molecular biological methods. Here, we show that CRISPR/Cas9 can be efficiently used for gene inactivation. Two genes of the melanin biosynthesis pathway, pksA and brm2, were chosen as targets. Several white mutants were obtained after several rounds of strain purification through protoplast regeneration or spore inoculation. Mutation of the genes was due to deletions from 1bp to 1.5kbp. The CRISPR/Cas9 system was also used to inactivate the orotidine-5-phosphate decarboxylase gene pyrG to create a uracil-auxotrophic strain. The strain was counter-selected with fluor-orotic acid and could be re-transformed with pyrG from Aspergillus fumigatus and pyr-4 from Neurospora crassa. In order to test the functioning of GFP, the fluorescent protein was fused to a nuclear localization signal derived from the StuA transcription factor of Aspergillus nidulans. After transformation bright nuclei were visible.

  4. Morphogenetic Alterations of Alternaria alternata Exposed to Dicarboximide Fungicide, Iprodione.

    PubMed

    Kim, Eunji; Lee, Hye Min; Kim, Young Ho

    2017-02-01

    Fungicide-resistant Alternaria alternata impede the practical control of the Alternaria diseases in crop fields. This study aimed to investigate cytological fungicide resistance mechanisms of A. alternata against dicarboximide fungicide iprodione. A. alternata isolated from cactus brown spot was cultured on potato-dextrose agar (PDA) with or without iprodione, and the fungal cultures with different growth characteristics from no, initial and full growth were observed by light and electron microscopy. Mycelia began to grow from one day after incubation (DAI) and continued to be in full growth (control-growth, Con-G) on PDA without fungicide, while on PDA with iprodione, no fungal growth (iprodione-no growth, Ipr-N) occurred for the first 3 DAI, but once the initial growth (iprodione-initial growth, Ipr-I) began at 4-5 DAI, the colonies grew and expanded continuously to be in full growth (iprodione-growth, Ipr-G), suggesting Ipr-I may be a turning moment of the morphogenetic changes resisting fungicidal toxicity. Con-G formed multicellular conidia with cell walls and septa and intact dense cytoplasm. In Ipr-N, fungal sporulation was inhibited by forming mostly undeveloped unicellular conidia with degraded and necrotic cytoplasm. However, in Ipr-I, conspicuous cellular changes occurred during sporulation by forming multicellular conidia with double layered (thickened) cell walls and accumulation of proliferated lipid bodies in the conidial cytoplasm, which may inhibit the penetration of the fungicide into conidial cells, reducing fungicide-associated toxicity, and may be utilized as energy and nutritional sources, respectively, for the further fungal growth to form mature colonies as in Ipr-G that formed multicellular conidia with cell walls and intact cytoplasm with lipid bodies as in Con-G.

  5. Overexpression of a Modified Plant Thionin Enhances Disease Resistance to Citrus Canker and Huanglongbing (HLB)

    PubMed Central

    Hao, Guixia; Stover, Ed; Gupta, Goutam

    2016-01-01

    Huanglongbing (HLB or citrus greening disease) caused by Candidatus Liberibacter asiaticus (Las) is a great threat to the US citrus industry. There are no proven strategies to eliminate HLB disease and no cultivar has been identified with strong HLB resistance. Citrus canker is also an economically important disease associated with a bacterial pathogen (Xanthomonas citri). In this study, we characterized endogenous citrus thionins and investigated their expression in different citrus tissues. Since no HLB-resistant citrus cultivars have been identified, we attempted to develop citrus resistant to both HLB and citrus canker through overexpression of a modified plant thionin. To improve effectiveness for disease resistance, we modified and synthesized the sequence encoding a plant thionin and cloned into the binary vector pBinPlus/ARS. The construct was then introduced into Agrobacterium strain EHA105 for citrus transformation. Transgenic Carrizo plants expressing the modified plant thionin were generated by Agrobacterium-mediated transformation. Successful transformation and transgene gene expression was confirmed by molecular analysis. Transgenic Carrizo plants expressing the modified thionin gene were challenged with X. citri 3213 at a range of concentrations, and a significant reduction in canker symptoms and a decrease in bacterial growth were demonstrated compared to nontransgenic plants. Furthermore, the transgenic citrus plants were challenged with HLB via graft inoculation. Our results showed significant Las titer reduction in roots of transgenic Carrizo compared with control plants and reduced scion Las titer 12 months after graft inoculation. These data provide promise for engineering citrus disease resistance against HLB and canker. PMID:27499757

  6. Biofilm formation, epiphytic fitness, and canker development in Xanthomonas axonopodis pv. citri.

    PubMed

    Rigano, Luciano A; Siciliano, Florencia; Enrique, Ramón; Sendín, Lorena; Filippone, Paula; Torres, Pablo S; Qüesta, Julia; Dow, J Maxwell; Castagnaro, Atilio P; Vojnov, Adrián A; Marano, María Rosa

    2007-10-01

    The phytopathogenic bacterium Xanthomonas axonopodis pv. citri is responsible for the canker disease affecting citrus plants throughout the world. Here, we have evaluated the role of bacterial attachment and biofilm formation in leaf colonization during canker development on lemon leaves. Crystal violet staining and confocal laser scanning microscopy analysis of X. axonopodis pv. citri strains expressing the green fluorescent protein were used to evaluate attachment and biofilm formation on abiotic and biotic (leaf) surfaces. Wild-type X. axonopodis pv. citri attached to and formed a complex, structured biofilm on glass in minimal medium containing glucose. Similar attachment and structured biofilm formation also were seen on lemon leaves. An X. axonopodis pv. citri gumB mutant strain, defective in production of the extracellular polysaccharide xanthan, did not form a structured biofilm on either abiotic or biotic surfaces. In addition, the X. axonopodis pv. citri gumB showed reduced growth and survival on leaf surfaces and reduced disease symptoms. These findings suggest an important role for formation of biofilms in the epiphytic survival of X. axonopodis pv. citri prior to development of canker disease.

  7. Rapid screening for citrus canker resistance employing pathogen-associated molecular pattern-triggered immunity responses

    PubMed Central

    Pitino, Marco; Armstrong, Cheryl M; Duan, Yongping

    2015-01-01

    Citrus canker, caused by the bacterial pathogen Xanthomonas citri ssp. citri (Xcc), has been attributed to millions of dollars in loss or damage to commercial citrus crops in subtropical production areas of the world. Since identification of resistant plants is one of the most effective methods of disease management, the ability to screen for resistant seedlings plays a key role in the production of a long-term solution to canker. Here, an inverse correlation between reactive oxygen species (ROS) production by the plant and the ability of Xcc to grow and form lesions on infected plants is reported. Based on this information, a novel screening method that can rapidly identify citrus seedlings that are less susceptible to early infection by Xcc was devised by measuring ROS accumulation triggered by a 22-amino acid sequence of the conserved N-terminal part of flagellin (flg22) from X. citri ssp. citri (Xcc-flg22). In addition to limiting disease symptoms, ROS production was also correlated with the expression of basal defense-related genes such as the pattern recognition receptors LRR8 and FLS2, the leucine-rich repeat receptor-like protein RLP12, and the defense-related gene PR1, indicating an important role for pathogen-associated molecular pattern-triggered immunity (PTI) in determining resistance to citrus canker. Moreover, the differential expression patterns observed amongst the citrus seedlings demonstrated the existence of genetic variations in the PTI response among citrus species/varieties. PMID:26504581

  8. Occurrence of Leaf Blight on Cosmos Caused by Alternaria cosmosa in Korea.

    PubMed

    Deng, Jian Xin; Lee, Ji Hye; Paul, Narayan Chandra; Cho, Hye Sun; Lee, Hyang Burm; Yu, Seung Hun

    2015-03-01

    In 2011, a leaf blight disease was observed on cosmos (Cosmos bipinnatus) leaves in Nonsan, Korea. The causal pathogen was isolated and identified based on morphological and molecular approaches. Morphological characteristics of the pathogen matched well with the Alternaria cosmosa and also easily distinguishable from Alternaria zinniae reported from cosmos seeds by producing branched beak. Phylogenetically, the pathogen could not be distinguished from A. passiflorae based on the sequence analysis of a combined data set of Alt a1 and gpd genes. However, A. passiflorae was distinguished from the present species by having conidiophores with 4 to 5 conidiogenous loci. The results indicate that the present Alternaria species is A. cosmosa. Pathogenicity tests revealed that the isolate was pathogenic to the leaves of Cosmos bipinnatus. This is the first report of Alternaria blight disease caused by A. cosmosa on cosmos in Korea.

  9. The dynamics of the fungal aerospores Alternaria sp. and Cladosporium sp. in Parisian atmospheric air, in France

    NASA Astrophysics Data System (ADS)

    Brezoczki, V. M.

    2016-08-01

    The bioallergens occurring naturally in the atmospheric air are microorganisms, pollen grains, plant seeds, leaf and stem scrap, or their protein molecules. The presence of various airborne fungal spores determines a high allergenic potential for public health. This effect is due to the high number of produced spores, which under favourable meteorological conditions (dry weather and wind) reach the surrounding air. This paper traces the dynamics of two types of fungi, Alternaria sp and Cladosporium sp, fungi which can be found outdoors, in the surrounding air, as well as indoors, inside houses (especially the conidia of Cladosporium sp). The effects of these fungal spores on human health are varies, ranging from seasonal allergies (hay fever, rhinitis, sinusitis etc.) to sever afflictions of the respiratory system, onset of asthma, disfunctionalities of the nervous systems, of the immune system, zymoses etc. The monitoring of the dynamics of the aerospores Alternaria sp and Cladosporium sp was carried out between 2010 and 2013, over a period of 42 weeks during one calendar year, from February to the end of September, in the surrounding air in the French capital, Paris. The regional and global climate and meteorological conditions are directly involved in the occurrence and development of fungi colonies, the transportation and dispersion of fungal spores in the atmospheric air, as well as in the creation of the environment required for the interaction of chemical and biological components in the air. Knowledge of the dynamics of the studied fungal aerospores, coupled with climate and meteorological changes, offers a series of information on the magnitude of the allergenic potential these airborne spores can determine. Legal regulations in this domain set the allergen risk threshold for the Alternaria sp aerospores at 3500 ÷ 7000 spores/m3 air/week, and for the Cladosporium sp aerospores at 56,000 spores/m3 air/week. Besides these regulations there exist a series of

  10. First report of stem canker of Salsola tragus caused by Diaporthe eres in Russia

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Salsola tragus L. (Russian thistle, tumbleweed), family Chenopodiaceae, is a problematic invasive weed in the western United States and a target of biological control efforts. In September of 2007, dying Salsola tragus plants were found along the Azov Sea at Chushka, Russia. About 30 plants in the...

  11. Characterization of Alternaria strains from Argentinean blueberry, tomato, walnut and wheat.

    PubMed

    Andersen, Birgitte; Nielsen, Kristian F; Fernández Pinto, Virginia; Patriarca, Andrea

    2015-03-02

    Alternaria species have the ability to produce a variety of secondary metabolite, which plays important roles in food safety. Argentina is the second largest exporter of fresh and processed food products to Europe, however, few studies on Alternaria mycotoxins and other bioactive secondary metabolites have been carried out on Argentinean cereals, fruit and vegetables. Knowing the full chemical potential and the distribution of Alternaria spp. on crops, it is necessary to establish a toxicological risk assessment for food products for human consumption. In the present study, 87 Alternaria strains from different substrates (tomato, wheat, blueberries and walnuts) were characterized according to morphology and metabolite production. Aggressive dereplication (accurate mass, isotopic patterns and lists of all described compounds from Alternaria) was used for high-throughput evaluation of the chemical potential. Four strains belonged to the Alternaria infectoria sp.-grp., 6 to the Alternaria arborescens sp.-grp., 6 showed a sporulation pattern similar to that of "M" according to Simmons, 1 to that of Alternaria vaccinii, and the remaining 70 constituted a diverse group belonging to morphological groups "G" and "H". The cluster analysis yielded 16 almost identical dendrograms and grouped the Alternaria strains into four clusters and 11 singletons and outlier groups. The chemical analysis showed that AOH and AME were the most common metabolites produced, followed by TEN, ALXs and TeA. The A. infectoria sp.-grp. had no metabolites in common with the rest of the strains. Several secondary metabolites isolated from large-spored Alternaria species or other fungal genera were detected, such as dehydrocurvularin, pyrenochaetic acid and alternarienonic acid. The strains isolated from tomato produced lower amounts of metabolites than strains from blueberries, walnut and wheat, although individual strains from tomato produced the highest amount of some metabolites. The A

  12. Dustborne Alternaria alternata antigens in U.S. homes: Results from the National Survey of Lead and Allergens in Housing

    PubMed Central

    Salo, Päivi M.; Yin, Ming; Arbes, Samuel J.; Cohn, Richard D.; Sever, Michelle; Muilenberg, Michael; Burge, Harriet A.; London, Stephanie J.; Zeldin, Darryl C.

    2005-01-01

    Background: Alternaria alternata is one of the most common fungi associated with allergic disease. However, Alternaria exposure in indoor environments is not well characterized. Objective: The primary goals of this study were to examine the prevalence of Alternaria exposure and identify independent predictors of Alternaria antigen concentrations in U.S. homes. Methods: Data for this cross-sectional study were obtained from the National Survey of Lead and Allergens in Housing. A nationally representative sample of 831 housing units in 75 different locations throughout the U.S. completed the survey. Information on housing and household characteristics was obtained by questionnaire and environmental assessments. Concentrations of Alternaria antigens in dust collected from various indoor sites were assessed with a polyclonal anti-Alternaria antibody assay. Results: Alternaria antigens were detected in most (95-99%) of the dust samples. The geometric mean concentration, reflecting the average Alternaria concentration in homes, was 4.88 μg/g (SE=0.13 μg/g). In the multivariable linear regression analysis, the age of the housing unit, geographic region, urbanization, poverty, family race, observed mold and moisture problems, use of dehumidifier, and presence of cats and dogs were independent predictors of Alternaria antigen concentrations. Less frequent cleaning and smoking indoors also contributed to higher Alternaria antigen levels in homes. Conclusion: Exposure to Alternaria alternata antigens in U.S. homes is common. Antigen levels in homes are not only influenced by regional factors but also by residential characteristics. Preventing mold and moisture problems, avoiding smoking indoors, and regular household cleaning may help reduce exposure to Alternaria antigens indoors. PMID:16159634

  13. The effect of wind on dispersal of splash-borne Xanthomonas citri subsp citri at different heights and distances downwind of canker-infected grapefruit trees

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Xanthomonas citri subsp citri (Xcc), which causes citrus canker, is a major pathogen of grapefruit and other canker-susceptible citrus species and cultivars grown in Florida and elsewhere. It is dispersed by rain splash, and wind promotes the dispersal of the pathogen. The aim of this study was to e...

  14. Hyperspectral and Thermal Imaging of Oilseed Rape (Brassica napus) Response to Fungal Species of the Genus Alternaria

    PubMed Central

    Baranowski, Piotr; Jedryczka, Malgorzata; Mazurek, Wojciech; Babula-Skowronska, Danuta; Siedliska, Anna; Kaczmarek, Joanna

    2015-01-01

    In this paper, thermal (8-13 µm) and hyperspectral imaging in visible and near infrared (VNIR) and short wavelength infrared (SWIR) ranges were used to elaborate a method of early detection of biotic stresses caused by fungal species belonging to the genus Alternaria that were host (Alternaria alternata, Alternaria brassicae, and Alternaria brassicicola) and non-host (Alternaria dauci) pathogens to oilseed rape (Brassica napus L.). The measurements of disease severity for chosen dates after inoculation were compared to temperature distributions on infected leaves and to averaged reflectance characteristics. Statistical analysis revealed that leaf temperature distributions on particular days after inoculation and respective spectral characteristics, especially in the SWIR range (1000-2500 nm), significantly differed for the leaves inoculated with A. dauci from the other species of Alternaria as well as from leaves of non-treated plants. The significant differences in leaf temperature of the studied Alternaria species were observed in various stages of infection development. The classification experiments were performed on the hyperspectral data of the leaf surfaces to distinguish days after inoculation and Alternaria species. The second-derivative transformation of the spectral data together with back-propagation neural networks (BNNs) appeared to be the best combination for classification of days after inoculation (prediction accuracy 90.5%) and Alternaria species (prediction accuracy 80.5%). PMID:25826369

  15. Abscisic acid enhances resistance to Alternaria solani in tomato seedlings.

    PubMed

    Song, Weiwei; Ma, Xinrong; Tan, Hong; Zhou, Jinyan

    2011-07-01

    The plant hormone abscisic acid (ABA) is an important regulator in many aspects of plant growth and development, as well as stress resistance. Here, we investigated the effects of exogenous ABA application on the interaction between tomato (Solanum lycopersicon L.) and Alternaria solani (early blight). Foliar spraying of 7.58 μM ABA was effective in reducing disease severity in tomato plants. Previously, increased activities of phenylalanine ammonia-lyase (PAL), polyphenol oxidase (PPO) and peroxidase (POD) were observed in exogenous ABA-treated tomato leaves. Moreover, these enzyme activities were maintained at higher levels in ABA-pretreated and A. solani challenged tomato plants. Tomato defense genes, such as PR1, β-1, 3-glucanase (GLU), PPO, POD, and superoxide dismutase (SOD), were rapidly and significantly up-regulated by exogenous ABA treatment. Furthermore, a subsequent challenge of ABA-pretreated plants with the pathogen A. solani resulted in higher expression of defense genes, compared to water-treated or A. solani inoculated plants. Therefore, our results suggest that exogenous ABA could enhance disease resistance against A. solani infection in tomato through the activation of defense genes and via the enhancement of defense-related enzymatic activities.

  16. Evaluation of Alternaria mycotoxins in strawberries: quantification and storage condition.

    PubMed

    Juan, Cristina; Oueslati, Souheib; Mañes, Jordi

    2016-05-01

    Alternariol (AOH), alternariol methyl ether (AME) and tentoxin (TEN) are Alternaria mycotoxins produced by the most common post-harvest pathogens of fruits. The production of these metabolites depends on several environmental factors, mainly temperature, water activity, pH and the technological treatments that have been applied to the product. In this study, the occurrence of AOH, AME and TEN was evaluated in strawberries samples stored at different temperatures ranges (at 22 ± 2 or 6 ± 2°C) and different periods (up to 1 month) simulating the current practice of consumer's storage conditions. Sample extraction was performed using a liquid-liquid extraction method prior to LC-MS/MS analysis. AOH was the most prevalent mycotoxins with a 42% at strawberries stored at (22 ± 2)°C and 37% stored at (6 ± 2)°C. The highest AOH levels were found in samples conserved at (22 ± 2)°C ranging between 26 and 752 ng g(-1). AME levels ranged between 11 and 137 ng g(-)(1), which were found mainly in stored samples at (6 ± 2)°C for more than 28 days. None sample presented levels of TEN in either of the studied conditions.

  17. Bacterial canker of plum trees, caused by Pseudomonas syringae pathovars, as a serious threat for plum production in the Netherlands.

    PubMed

    Wenneker, M; Janse, J D; De Bruine, J A

    2011-01-01

    In the Netherlands, bacterial canker in plum trees (Prunus domestica) is a serious and recent problem in plum production. It is caused by Pseudomonas syringae pathovars syringae and morsprunorum. The trunks of the affected plum trees are girdled by bacterial cankers resulting in sudden death of infected trees in 3-4 years after planting. Disease incidences can be very high, and sometimes complete orchards have to be removed. Recently, plum cultivation in the Netherlands has changed from a relatively extensive into an intensive cultivation. However, due to the risks of losses of trees due to bacterial canker, growers are reluctant to plant new plum orchards. In general nurseries and fruit growers are not familiar with bacterial diseases and lack knowledge in order to prevent infections. Therefore, control strategies to manage plum decline have to be developed.

  18. Multiple Ceratocystis smalleyi infections associated with reduced stem water transport in bitternut hickory.

    PubMed

    Park, J-H; Juzwik, J; Cavender-Bares, J

    2013-06-01

    Hundreds of cankers caused by Ceratocystis smalleyi are associated with hickory bark beetle-attacked bitternut hickory exhibiting rapid crown decline in the north-central and northeastern United States. Discolored sapwood colonized by the fungus commonly underlies the cankers. Field studies were conducted to test the hypothesis that C. smalleyi infections cause vascular system dysfunction in infected trees. Fifty C. smalleyi inoculations made at 1.8 to 3.8 m in height on stems of healthy bitternut hickory trees (13 to 28 cm in diameter at 1.4 m in height) resulted in extensive canker formation and sapwood discoloration 12 to 14 months after treatment compared with water-inoculated and noninoculated controls. Sap flow velocity (midday) was significantly lower in the infected trees compared with that in the controls. Sap flow velocity also was inversely correlated with the proportion of bark area with cankered tissues and with tylose abundance in the youngest two growth rings. Tylose formation in current-year vessels associated with C. smalleyi infections is likely responsible for much of the water transport disruption. It is hypothesized that multiple stem infections of C. smalleyi and the resulting xylem dysfunction contribute to crown wilt development in bitternut hickory exhibiting rapid crown decline.

  19. Amplification of DNA of Xanthomonas axonopodis pv. citri from historic citrus canker herbarium specimens.

    PubMed

    Li, Wenbin; Brlansky, Ronald H; Hartung, John S

    2006-05-01

    Herbaria are important resources for the study of the origins and dispersal of plant pathogens, particularly bacterial plant pathogens that incite local lesions in which large numbers of pathogen genomes are concentrated. Xanthomonas axonopodis pv. citri (Xac), the causal agent of citrus bacterial canker disease, is a notable example of such a pathogen. The appearance of novel strains of the pathogen in Florida and elsewhere make it increasingly important to understand the relationships among strains of this pathogen. USDA-ARS at Beltsville, Maryland maintains approximately 700 herbarium specimens with citrus canker disease lesions up to 90 years old, originally collected from all over the world, and so is an important resource for phytogeographic studies of this bacterium. Unfortunately, DNA in herbarium specimens is degraded and may contain high levels of inhibitors of PCR. In this study, we compared a total of 23 DNA isolation techniques in combination with 31 novel primer pairs in order to develop an efficient protocol for the analysis of Xac DNA in herbarium specimens. We identified the most reliable extraction method, identified in terms of successful amplification by our panel of 31 primer pairs. We also identified the most robust primer pairs, identified as successful in the largest number of extracts prepared by different methods. We amplified Xac genomic sequences up to 542 bp long from herbarium samples up to 89 years old. Primers varied in effectiveness, with some primer pairs amplifying Xac DNA from a 1/10,000 dilution of extract from a single lesion from a citrus canker herbarium specimen. Our methodology will be useful to identify pathogens and perform molecular analyses of bacterial and possibly fungal genomes from herbarium specimens.

  20. Positive selection is the main driving force for evolution of citrus canker-causing Xanthomonas.

    PubMed

    Zhang, Yunzeng; Jalan, Neha; Zhou, Xiaofeng; Goss, Erica; Jones, Jeffrey B; Setubal, João C; Deng, Xiaoling; Wang, Nian

    2015-10-01

    Understanding the evolutionary history and potential of bacterial pathogens is critical to prevent the emergence of new infectious bacterial diseases. Xanthomonas axonopodis subsp. citri (Xac) (synonym X. citri subsp. citri), which causes citrus canker, is one of the hardest-fought plant bacterial pathogens in US history. Here, we sequenced 21 Xac strains (14 XacA, 3 XacA* and 4 XacA(w)) with different host ranges from North America and Asia and conducted comparative genomic and evolutionary analyses. Our analyses suggest that acquisition of beneficial genes and loss of detrimental genes most likely allowed XacA to infect a broader range of hosts as compared with XacA(w) and XacA*. Recombination was found to have occurred frequently on the relative ancient branches, but rarely on the young branches of the clonal genealogy. The ratio of recombination/mutation ρ/θ was 0.0790±0.0005, implying that the Xac population was clonal in structure. Positive selection has affected 14% (395 out of 2822) of core genes of the citrus canker-causing Xanthomonas. The genes affected are enriched in 'carbohydrate transport and metabolism' and 'DNA replication, recombination and repair' genes (P<0.05). Many genes related to virulence, especially genes involved in the type III secretion system and effectors, are affected by positive selection, further highlighting the contribution of positive selection to the evolution of citrus canker-causing Xanthomonas. Our results suggest that both metabolism and virulence genes provide advantages to endow XacA with higher virulence and a wider host range. Our analysis advances our understanding of the genomic basis of specialization by positive selection in bacterial evolution.

  1. Pseudomonas protegens CS1 from the lemon phyllosphere as a candidate for citrus canker biocontrol agent.

    PubMed

    Michavila, G; Adler, C; De Gregorio, P R; Lami, M J; Caram Di Santo, M C; Zenoff, A M; de Cristobal, R E; Vincent, P A

    2017-02-14

    Citrus canker is a worldwide-distributed disease caused by Xanthomonas citri subsp. citri. One of the most used strategies to control the disease is centred on copper-based compounds that cause environmental problems. Therefore, it is of interest to develop new strategies to manage the disease. Previously, we reported the ability of the siderophore pyochelin, produced by the opportunistic human pathogen Pseudomonas aeruginosa, to inhibit in vitro several bacterial species, including X. citri subsp. citri. The action mechanism, addressed with the model bacterium Escherichia coli, was connected to the generation of reactive oxygen species (ROS). This work aimed to find a non-pathogenic strain from the lemon phyllosphere that would produce pyochelin and therefore serve in canker biocontrol. An isolate that retained its capacity to colonise the lemon phyllosphere and inhibit X. citri subsp. citri was selected and characterised as Pseudomonas protegens CS1. From a liquid culture of this strain, the active compound was purified and identified as the pyochelin enantiomer, enantio-pyochelin. Using the producing strain and the pure compound, both in vitro and in vivo, we determined that the action mechanism of X. citri subsp. citri inhibition also involved the generation of ROS. Finally, the potential application of P. protegens CS1 was evaluated by spraying the bacterium in a model that mimics the natural X. citri subsp. citri infection. The ability of P. protegens CS1 to reduce canker formation makes this strain an interesting candidate as a biocontrol agent.

  2. Genetic variability of Brazilian isolates of Alternaria alternata detected by AFLP and RAPD techniques

    PubMed Central

    Dini-Andreote, Francisco; Pietrobon, Vivian Cristina; Andreote, Fernando Dini; Romão, Aline Silva; Spósito, Marcel Bellato; Araújo, Welington Luiz

    2009-01-01

    The Alternaria brown spot (ABS) is a disease caused in tangerine plants and its hybrids by the fungus Alternaria alternata f. sp. citri which has been found in Brazil since 2001. Due to the recent occurrence in Brazilian orchards, the epidemiology and genetic variability of this pathogen is still an issue to be addressed. Here it is presented a survey about the genetic variability of this fungus by the characterization of twenty four pathogenic isolates of A. alternata f. sp. citri from citrus plants and four endophytic isolates from mango (one Alternaria tenuissima and three Alternaria arborescens). The application of two molecular markers Random Amplified Polymorphic DNA (RAPD) and Amplified Fragment Length Polymorphism (AFLP) had revealed the isolates clustering in distinct groups when fingerprintings were analyzed by Principal Components Analysis (PCA). Despite the better assessment of the genetic variability through the AFLP, significant modifications in clusters components were not observed, and only slight shifts in the positioning of isolates LRS 39/3 and 25M were observed in PCA plots. Furthermore, in both analyses, only the isolates from lemon plants revealed to be clustered, differently from the absence of clustering for other hosts or plant tissues. Summarizing, both RAPD and AFLP analyses were both efficient to detect the genetic variability within the population of the pathogenic fungus Alternaria spp., supplying information on the genetic variability of this species as a basis for further studies aiming the disease control. PMID:24031413

  3. Natural Occurrence of Alternaria Toxins in the 2015 Wheat from Anhui Province, China

    PubMed Central

    Xu, Wenjing; Han, Xiaomin; Li, Fengqin; Zhang, Lishi

    2016-01-01

    The exposure to Alternaria toxins from grain and grain-based products has been reported to be related to human esophageal cancer in China. In this study, a total of 370 freshly harvested wheat kernel samples collected from Anhui province of China in 2015 were analyzed for the four Alternaria toxins tenuazonic acid (TeA), tentoxin (TEN), alternariol (AOH) and alternariol monomethyl ether (AME) by high performance liquid chromatography-tandem mass spectrometry method (HPLC-MS/MS). TeA was the predominant toxin detected followed by TEN, AOH and AME. The concentrations of the four Alternaria toxins varied geographically. The samples from Fuyang district showed higher TEN concentration levels than the other regions studied (p < 0.05). Furthermore, 95% (352/370) of the wheat samples were positive for more than one type of Alternaria toxins. Positive correlation was observed between concentration levels of TeA and TEN, AOH and AME, TeA and AOH, and the total dibenzopyrone derivatives (AOH + AME) and TeA. Results indicate that there is a need to set the tolerance limit for Alternaria toxins in China, and more data on the contamination of these toxins in agro-products is required. PMID:27792182

  4. Alternaria toxins in wheat during the 2004 to 2005 Argentinean harvest.

    PubMed

    Azcarate, M P; Patriarca, A; Terminiello, L; Fernández Pinto, V

    2008-06-01

    The natural occurrence of Alternaria mycotoxins in Argentinean wheat from the zone 5 South during the 2004 to 2005 harvest was investigated in 64 wheat samples. All samples were highly contaminated with a wide range of fungal species. Alternaria was found as the main component of the mycota, with an infection percentage of 100%. Three mycotoxins produced by species of Alternaria were determined in wheat: alternariol, alternariol monomethyl ether, and tenuazonic acid. Alternariol was detected in 4 (6%) of 64 samples, with a range of 645 to 1,388 microg/kg (mean of 1,054 microg/kg); alternariol monomethyl ether, with a range of 566 to 7,451 microg/kg (mean of 2,118 microg/kg) in 15 (23%) of 64 samples; and tenuazonic acid in 12 (19%) of 64 samples, with a range of 1,001 to 8,814 microg/kg (mean, 2,313 microg/kg). Alternariol monomethyl ether was the predominant toxin, but tenuazonic acid was detected in higher concentrations. Alternariol was present in fewer samples and in lower levels than were the other toxins. Tenuazonic acid and alternariol monomethyl ether occurred together in four samples, while tenuazonic acid and alternariol co-occurred in one sample. This the first report of the natural occurrence of Alternaria mycotoxins in Argentinean wheat. Toxin levels were high, probably due to the heavy infection with Alternaria species found in the samples.

  5. Isolation and characterization of the grain mold fungi, Cochliobolus and Alternaria spp., from sorghum using semi-selective media and DNA sequence analyses

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Mold diseases, caused by fungal complexes including Alternaria, Cochliobolus and Fusarium species, limit sorghum grain production. Media were tested by plating Fusarium thapsinum, Alternaria sp. and Curvularia lunata, individually and competitively. Dichloran chloramphenicol rose bengal (DRBC) and m...

  6. Synergistic estrogenic effects of Fusarium and Alternaria mycotoxins in vitro.

    PubMed

    Vejdovszky, Katharina; Hahn, Kathrin; Braun, Dominik; Warth, Benedikt; Marko, Doris

    2017-03-01

    Mycotoxins are toxic secondary metabolites formed by various fungal species that are found as natural contaminants in food. This very heterogeneous group of compounds triggers multiple toxic mechanisms, including endocrine disruptive potential. Current risk assessment of mycotoxins, as for most chemical substances, is based on the effects of single compounds. However, concern on a potential enhancement of risks by interactions of single substances in naturally occurring mixtures has greatly increased recently. In this study, the combinatory effects of three mycoestrogens were investigated in detail. This includes the endocrine disruptors zearalenone (ZEN) and α-zearalenol (α-ZEL) produced by Fusarium fungi and alternariol (AOH), a cytotoxic and estrogenic mycotoxin formed by Alternaria species. For evaluation of effects, estrogen-dependent activation of alkaline phosphatase (AlP) and cell proliferation were tested in the adenocarcinoma cell line Ishikawa. The estrogenic potential varied among the single substances. Half maximum effect concentrations (EC50) for AlP activation were evaluated for α-ZEL, ZEN and AOH as 37 pM, 562 pM and 995 nM, respectively. All three mycotoxins were found to act as partial agonists. The majority of binary combinations, even at very low concentrations in the case of α-ZEL, showed strong synergism in the AlP assay. These potentiating phenomena of mycotoxin mixtures highlight the urgent need to incorporate combinatory effects into future risk assessment, especially when endocrine disruptors are involved. To the best of our knowledge, this study presents the first investigation on synergistic effects of mycoestrogens.

  7. Cutaneous infection with Alternaria triticina in a Bilateral lung transplant recipient.

    PubMed

    González-Vela, M C; Armesto, S; Unda-Villafuerte, F; Val-Bernal, J F

    2014-10-01

    We report the case of a 60-year-old man who was receiving immunosuppressive therapy for a bilateral lung transplant and presented with a crusted, violaceous plaque on the left hand. Based on histopathology and microbiological culture the patient was diagnosed with infection by Alternaria species. Treatment with itraconazole led to complete resolution of the skin lesion. Forty months later he developed four reddish, nodular, skin lesions on the left leg. Analysis of a biopsy from one of these lesions using histopathologic and molecular techniques identified a mold that shared 98% homology with a strain of Alternaria triticina. Alternaria species belong to a group of dematiaceous fungi that cause opportunistic infections in humans. The incidence of these infections is increasing, mainly in transplant centers. To the best of our knowledge, this is the first reported case of a human infection caused by A. triticina.

  8. Gnomoniopsis smithogilvyi causes chestnut canker symptoms in Castanea sativa shoots in Switzerland.

    PubMed

    Pasche, Sabrina; Calmin, Gautier; Auderset, Guy; Crovadore, Julien; Pelleteret, Pegah; Mauch-Mani, Brigitte; Barja, François; Paul, Bernard; Jermini, Mauro; Lefort, François

    2016-02-01

    A screening of Castanea sativa scions for grafting for the presence of endophytes showed that the opportunistic fungal pathogen Gnomoniopsis smithogilvyi was the most abundant member of the endophytic flora. This fungus is known as a pathogen affecting chestnut fruits in Italy and Australia. Here, we present evidence that it causes cankers very similar to the ones due to Cryphonectria parasitica infection on twigs and scions of chestnut trees. We found natural infections of G. smithogilvyi in healthy grafted plants as well as in scions from chestnut trees. The identity of the fungus isolated from asymptomatic tissues was verified by applying Koch's postulates and corroborated by DNA sequencing of four different gene regions. In contrast to C. parasitica that appears on the bark as yellow to orange pycnidia, stromata and slimy twisted tendrils, G. smithogilvyi forms orange to red and black pycnidia, gray stromata and cream-colored to beige slimy twisted tendrils on the bark. These Swiss strains are closely related to G. smithogilvyi strains from Australia and from New Zealand, Gnomoniopsis sp. and Gnomoniopsis castanea from New Zealand, Italy, France and Switzerland. While the strains from Ticino are genetically very close to G. smithogilvyi and G. castanea from Italy, the differences between the strains from Ticino and Geneva suggest two different origins. The present study supports the hypothesis that a single species named G. smithogilvyi, which is known to be the agent of chestnut rot, also causes wood cankers on chestnut.

  9. Unconventional Recombination in the Mating Type Locus of Heterothallic Apple Canker Pathogen Valsa mali

    PubMed Central

    Yin, Zhiyuan; Ke, Xiwang; Li, Zhengpeng; Chen, Jiliang; Gao, Xiaoning; Huang, Lili

    2017-01-01

    Sexual reproduction in filamentous ascomycetes is controlled by the mating type (MAT) locus, including two idiomorphs MAT1-1 and MAT1-2. Understanding the MAT locus can provide clues for unveiling the sexual development and virulence factors for fungal pathogens. The genus Valsa (Sordariomycetes, Diaporthales) contains many tree pathogens responsible for destructive canker diseases. The sexual stage of these ascomycetes is occasionally observed in nature, and no MAT locus has been reported to date. Here, we identified the MAT locus of the apple canker pathogen Valsa mali, which causes extensive damage, and even death, to trees. V. mali is heterothallic in that each isolate carries either the MAT1-1 or MAT1-2 idiomorph. However, the MAT structure is distinct from that of many other heterothallic fungi in the Sordariomycetes. Two flanking genes, COX13 and APN2, were coopted into the MAT locus, possibly by intrachromosomal rearrangement. After the acquisition of foreign genes, unequal recombination occurred between MAT1-1/2 idiomorphs, resulting in a reverse insertion in the MAT1-2 idiomorph. Evolutionary analysis showed that the three complete MAT1-1-2, COX13, and APN2 genes in this region diverged independently due to different selection pressure. Null hypothesis tests of a 1:1 MAT ratio of 86 V. mali isolates from four different provinces showed a relatively balanced distribution of the two idiomorphs in the fields. These results provide insights into the evolution of the mating systems in Sordariomycetes. PMID:28228472

  10. Occurrence of shallow bark canker of walnut (Juglans regia) in southern provinces of Iran.

    PubMed

    Yousefikopaei, F; Taghavi, S M; Banihashemi, Z

    2007-05-01

    From April 2001 to November 2002, samples of walnut branches and trunks with symptoms of shallow bark canker were collected from Fars and Kohgiluyeh-va-Boyerahmad provinces. Symptoms of the disease were small cracks in the bark of the trunk and scaffold branches of mature trees with dark watery exudates which stained the affected trunk or limb. By removal of phelloderm, extensive necrosis of the underlying tissues was observed. In some cases, necrosis extended to cambium and outer xylem. Sixty-one strains of a bacterium were isolated from infected tissues using EMB and YDC media. On the basis of standard biochemical and physiological tests the bacterium was identified as Brenneria nigrifluens. The pathogen was found to be wide-spread in the provinces. Isolates were compared by physiological and biochemical characters, antibiotic sensitivity and protein electrophoretic pattern. Most of the strains were fairly similar in phenotypic features and electrophoretic profiles ofwhole-cell proteins were similar to each other and to reference strain (B. nigrifluens 5D313). Inoculation of 1-2 years-old walnut seedlings in May and June produced blackening symptoms and the bacterium survived for long period in infected tissues. This is the first report of the shallow bark canker of walnut in southern Iran.

  11. Isolation and Characterization of Bacteriophages Against Pseudomonas syringae pv. actinidiae Causing Bacterial Canker Disease in Kiwifruit.

    PubMed

    Yu, Ji-Gang; Lim, Jeong-A; Song, Yu-Rim; Heu, Sunggi; Kim, Gyoung Hee; Koh, Young Jin; Oh, Chang-Sik

    2016-02-01

    Pseudomonas syringae pv. actinidiae causes bacterial canker disease in kiwifruit. Owing to the prohibition of agricultural antibiotic use in major kiwifruit-cultivating countries, alternative methods need to be developed to manage this disease. Bacteriophages are viruses that specifically infect target bacteria and have recently been reconsidered as potential biological control agents for bacterial pathogens owing to their specificity in terms of host range. In this study, we isolated bacteriophages against P. syringae pv. actinidiae from soils collected from kiwifruit orchards in Korea and selected seven bacteriophages for further characterization based on restriction enzyme digestion patterns of genomic DNA. Among the studied bacteriophages, two belong to the Myoviridae family and three belong to the Podoviridae family, based on morphology observed by transmission electron microscopy. The host range of the selected bacteriophages was confirmed using 18 strains of P. syringae pv. actinidiae, including the Psa2 and Psa3 groups, and some were also effective against other P. syringae pathovars. Lytic activity of the selected bacteriophages was sustained in vitro until 80 h, and their activity remained stable up to 50°C, at pH 11, and under UV-B light. These results indicate that the isolated bacteriophages are specific to P. syringae species and are resistant to various environmental factors, implying their potential use in control of bacterial canker disease in kiwifruits.

  12. Proteome Analysis of Pathogen-Responsive Proteins from Apple Leaves Induced by the Alternaria Blotch Alternaria alternata.

    PubMed

    Zhang, Cai-xia; Tian, Yi; Cong, Pei-hua

    2015-01-01

    Understanding the defence mechanisms used by apple leaves against Alternaria alternate pathogen infection is important for breeding purposes. To investigate the ultrastructural differences between leaf tissues of susceptible and resistant seedlings, in vitro inoculation assays and transmission electron microscopy (TEM) analysis were conducted with two different inoculation assays. The results indicated that the resistant leaves may have certain antifungal activity against A. alternate that is lacking in susceptible leaves. To elucidate the two different host responses to A. alternate infection in apples, the proteomes of susceptible and resistant apple leaves that had or had not been infected with pathogen were characterised using two-dimensional electrophoresis (2-DE) and matrix-assisted laser desorption/ionisation time-of-flight tandem mass spectrometry (MALDI-TOF-TOF MS). MS identified 43 differentially expressed proteins in two different inoculation assays. The known proteins were categorised into 5 classes, among these proteins, some pathogenesis-related (PR) proteins, such as beta-1,3-glucanase, ascorbate peroxidase (APX), glutathione peroxidase (GPX) and mal d1, were identified in susceptible and resistant hosts and were associated with disease resistance of the apple host. In addition, the different levels of mal d1 in susceptible and resistant hosts may contribute to the outstanding anti-disease properties of resistant leaves against A. alternate. Taken together, the resistance mechanisms of the apple host against A. alternate may be a result of the PR proteins and other defence-related proteins. Given the complexity of the biology involved in the interaction between apple leaves and the A. alternate pathogen, further investigation will yield more valuable insights into the molecular mechanisms of suppression of the A. alternate pathogen. Overall, we outline several novel insights into the response of apple leaves to pathogen attacks. These findings

  13. Proteome Analysis of Pathogen-Responsive Proteins from Apple Leaves Induced by the Alternaria Blotch Alternaria alternata

    PubMed Central

    Zhang, Cai-xia; Tian, Yi; Cong, Pei-hua

    2015-01-01

    Understanding the defence mechanisms used by apple leaves against Alternaria alternate pathogen infection is important for breeding purposes. To investigate the ultrastructural differences between leaf tissues of susceptible and resistant seedlings, in vitro inoculation assays and transmission electron microscopy (TEM) analysis were conducted with two different inoculation assays. The results indicated that the resistant leaves may have certain antifungal activity against A. alternate that is lacking in susceptible leaves. To elucidate the two different host responses to A. alternate infection in apples, the proteomes of susceptible and resistant apple leaves that had or had not been infected with pathogen were characterised using two-dimensional electrophoresis (2-DE) and matrix-assisted laser desorption/ionisation time-of-flight tandem mass spectrometry (MALDI-TOF-TOF MS). MS identified 43 differentially expressed proteins in two different inoculation assays. The known proteins were categorised into 5 classes, among these proteins, some pathogenesis-related (PR) proteins, such as beta-1,3-glucanase, ascorbate peroxidase (APX), glutathione peroxidase (GPX) and mal d1, were identified in susceptible and resistant hosts and were associated with disease resistance of the apple host. In addition, the different levels of mal d1 in susceptible and resistant hosts may contribute to the outstanding anti-disease properties of resistant leaves against A. alternate. Taken together, the resistance mechanisms of the apple host against A. alternate may be a result of the PR proteins and other defence-related proteins. Given the complexity of the biology involved in the interaction between apple leaves and the A. alternate pathogen, further investigation will yield more valuable insights into the molecular mechanisms of suppression of the A. alternate pathogen. Overall, we outline several novel insights into the response of apple leaves to pathogen attacks. These findings

  14. Alternaria toxins in wheat from the Autonomous Province of Vojvodina, Serbia: a preliminary survey.

    PubMed

    Janić Hajnal, Elizabet; Orčić, Dejan; Torbica, Aleksandra; Kos, Jovana; Mastilović, Jasna; Škrinjar, Marija

    2015-01-01

    Although Fusarium species remain a main source of mycotoxin contamination of wheat, in recent years, due to the evident climatic changes, other mycotoxigenic fungi have been recognised as important wheat contaminants. Alternaria species, especially A. alternata, have been found as contaminants of wheat as well as wheat-based products. Under favourable conditions A. alternata very often produce alternariol (AOH), alternariol monomethyl ether (AME), tenuazonic acid (TeA) and others Alternaria toxins. The aim of the present study was to examine the presence of three Alternaria toxins (AOH, AME and TeA) in wheat samples harvested during three years (2011-13). To this end, 92 samples were collected during wheat harvesting from different growing regions of the Autonomous Province of Vojvodina, which represents the most important wheat-growing area in Serbia. The presence of Alternaria toxins was analysed by HPLC with electrospray ionisation triple quadrupole mass spectrometry (LC-ESI-MS/MS). Among all the analysed wheat samples, 63 (68.5%) were contaminated with TeA, 11 (12.0%) with AOH and 6 (6.5%) with AME. Furthermore, the maximum and mean toxin concentrations were 2676 and 92.4 µg kg(-1), 48.9 and 18.6 µg kg(-1), and 70.2 and 39.0 µg kg(-1) for TeA, AOH and AME, respectively. Co-occurrence of three Alternaria toxins in wheat samples was detected in six samples; a combination of two toxins was found in two samples; and 64 samples contained one toxin. The results showed that among 92 analysed wheat samples, only 20 (21.7%) samples were without Alternaria toxins. The presence of Alternaria toxins was also investigated in terms of weather conditions recorded during the period of investigation, as well as with the sampling region. This study represents the first preliminary report of the natural occurrence of Alternaria toxins in wheat (Triticum aestivum) from Serbia.

  15. Primary subcutaneous Alternaria alternata infection of the hand in an immunocompromised host.

    PubMed

    Kpodzo, Dzifa S; Calderwood, Michael S; Ruchelsman, David E; Abramson, Jeremy S; Piris, Adriano; Winograd, Jonathan M; Kotton, Camille N

    2011-07-01

    We describe a case of a progressive subcutaneous Alternaria alternata infection in the hand of a patient with chronic lymphocytic leukemia (CLL). The diagnosis was based upon the examination of tissue biopsy and isolation of the etiologic agent in culture. The identity of the isolate was determined by phenotypic characteristics and by sequencing the ITS and D1/D2 regions of the rDNA. Despite combination therapy with voriconazole and micafungin, the lesion continued to progress. Posaconazole therapy, along with surgical excision of the infected tissue, resulted in the eradication of infection. The limitations of the clinical management of invasive Alternaria infections are discussed.

  16. Additive roles of PthAs in bacterial growth and pathogenicity associated with nucleotide polymorphisms in effector-binding elements of citrus canker susceptibility genes.

    PubMed

    Abe, Valeria Yukari; Benedetti, Celso Eduardo

    2016-10-01

    Citrus canker, caused by Xanthomonas citri, affects most commercial citrus varieties. All X. citri strains possess at least one transcription activator-like effector of the PthA family that activates host disease susceptibility (S) genes. The X. citri strain 306 encodes four PthA effectors; nevertheless, only PthA4 is known to elicit cankers on citrus. As none of the PthAs act as avirulence factors on citrus, we hypothesized that PthAs 1-3 might also contribute to pathogenicity on certain hosts. Here, we show that, although PthA4 is indispensable for canker formation in six Brazilian citrus varieties, PthAs 1 and 3 contribute to canker development in 'Pera' sweet orange, but not in 'Tahiti' lemon. Deletions in two or more pthA genes reduce bacterial growth in planta more pronouncedly than single deletions, suggesting an additive role of PthAs in pathogenicity and bacterial fitness. The contribution of PthAs 1 and 3 in canker formation in 'Pera' plants does not correlate with the activation of the canker S gene, LOB1 (LATERAL ORGAN BOUNDARIES 1), but with the induction of other PthA targets, including LOB2 and citrus dioxygenase (DIOX). LOB1, LOB2 and DIOX show differential PthA-dependent expression between 'Pera' and 'Tahiti' plants that appears to be associated with nucleotide polymorphisms found at or near PthA-binding sites. We also present evidence that LOB1 activation alone is not sufficient to elicit cankers on citrus, and that DIOX acts as a canker S gene in 'Pera', but not 'Tahiti', plants. Our results suggest that the activation of multiple S genes, such as LOB1 and DIOX, is necessary for full canker development.

  17. Transcriptional Profiling of Canker-Resistant Transgenic Sweet Orange (Citrus sinensis Osbeck) Constitutively Overexpressing a Spermidine Synthase Gene

    PubMed Central

    Fu, Xing-Zheng; Liu, Ji-Hong

    2013-01-01

    Citrus canker disease caused by Xanthomonas citri subsp. citri (Xcc) is one of the most devastating diseases affecting the citrus industry worldwide. In our previous study, the canker-resistant transgenic sweet orange (Citrus sinensis Osbeck) plants were produced via constitutively overexpressing a spermidine synthase. To unravel the molecular mechanisms underlying Xcc resistance of the transgenic plants, in the present study global transcriptional profiling was compared between untransformed line (WT) and the transgenic line (TG9) by hybridizing with Affymetrix Citrus GeneChip. In total, 666 differentially expressed genes (DEGs) were identified, 448 upregulated, and 218 downregulated. The DEGs were classified into 33 categories after Gene ontology (GO) annotation, in which 68 genes are in response to stimulus and involved in immune system process, 12 genes are related to cell wall, and 13 genes belong to transcription factors. These genes and those related to starch and sucrose metabolism, glutathione metabolism, biosynthesis of phenylpropanoids, and plant hormones were hypothesized to play major roles in the canker resistance of TG9. Semiquantitative RT-PCR analysis showed that the transcript levels of several candidate genes in TG9 were significantly higher than in WT both before and after Xcc inoculation, indicating their potential association with canker disease. PMID:23509803

  18. Characteristics of the perception of different severity measures of citrus canker and the relations between the various symptom types

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus canker is a disease of citrus and is caused by the bacterial pathogen Xanthomonas axonopodis pv citri (Xac). Ways of managing the disease are being sought, and accurate, precise, reproducible disease assessment is needed for monitoring epidemics. The objective of this study was to investigate...

  19. Visual rating and the use of image analysis for assessing different symptoms of citrus canker on grapefruit leaves

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus canker is caused by the bacterial pathogen Xanthomonas axonopodis pv citri (Xac) and infects several citrus species in wet tropical and subtropical citrus growing regions. Accurate, precise and reproducible disease assessment is needed for monitoring epidemics and disease response in breeding...

  20. Overexpression of a modified plant thionin enhances disease resistance to citrus canker and huanglongbing (HLB, citrus greening)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Huanglongbing (HLB or citrus greening disease) caused by Candidatus Liberibacter asiaticus (Las) is a great threat to the United States citrus industry. Citrus canker is also an economically important disease associated with a bacterial pathogen (Xanthomonas citri). In this study, we characterized e...

  1. Under severe HLB and citrus canker pressure, 'Triumph' and 'Jackson' perform better than 'Flame' and 'Marsh' grapefruit

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Huanglongbing (HLB) and Citrus Canker (CC) threaten the viability of Florida grapefruit production. ‘Triumph’ (T), reportedly a grapefruit/sweet orange hybrid, is similar to seedy white grapefruit with earlier maturity and lower bitterness. ‘Jackson’ (J) is a low-seeded budsport of ‘Triumph’. Tree h...

  2. Under severe citrus canker and HLB (Huanglongbing) pressure, Triumph and Jackson perform better than Flame and Marsh grapefruit

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Huanglongbing (HLB) and Citrus Canker (CC) threaten the viability of Florida grapefruit production. Triumph (T), reportedly a grapefruit/sweet orange hybrid, is similar to seedy white grapefruit with earlier maturity and lower bitterness. Jackson (J) is a low-seeded budsport of Triumph. Tree health ...

  3. Transcriptional profiling of canker-resistant transgenic sweet orange (Citrus sinensis Osbeck) constitutively overexpressing a spermidine synthase gene.

    PubMed

    Fu, Xing-Zheng; Liu, Ji-Hong

    2013-01-01

    Citrus canker disease caused by Xanthomonas citri subsp. citri (Xcc) is one of the most devastating diseases affecting the citrus industry worldwide. In our previous study, the canker-resistant transgenic sweet orange (Citrus sinensis Osbeck) plants were produced via constitutively overexpressing a spermidine synthase. To unravel the molecular mechanisms underlying Xcc resistance of the transgenic plants, in the present study global transcriptional profiling was compared between untransformed line (WT) and the transgenic line (TG9) by hybridizing with Affymetrix Citrus GeneChip. In total, 666 differentially expressed genes (DEGs) were identified, 448 upregulated, and 218 downregulated. The DEGs were classified into 33 categories after Gene ontology (GO) annotation, in which 68 genes are in response to stimulus and involved in immune system process, 12 genes are related to cell wall, and 13 genes belong to transcription factors. These genes and those related to starch and sucrose metabolism, glutathione metabolism, biosynthesis of phenylpropanoids, and plant hormones were hypothesized to play major roles in the canker resistance of TG9. Semiquantitative RT-PCR analysis showed that the transcript levels of several candidate genes in TG9 were significantly higher than in WT both before and after Xcc inoculation, indicating their potential association with canker disease.

  4. Draft Genome Sequences of Pectobacterium carotovorum subsp. actinidiae ICMP 19971 and ICMP 19972, Two Strains Isolated from Actinidia chinensis with Symptoms of Summer Canker in South Korea

    PubMed Central

    Panda, Preetinanda; Taylor, Robert; Pitman, Andrew R.

    2017-01-01

    ABSTRACT Pectobacterium carotovorum subsp. actinidiae is the causal agent of summer canker in kiwifruit plants in South Korea. We report here the draft genome sequences of two P. carotovorum subsp. actinidiae strains, ICMP 19971 and ICMP 19972, which were originally isolated from Actinidia chinensis with symptoms of summer canker. These genome sequences will aid in the identification of genetic traits associated with their unusual capacity to cause canker and help understanding of the threat these exotic enterobacteria pose to the New Zealand kiwifruit industry. PMID:28385839

  5. Genome editing of the disease susceptibility gene CsLOB1 in citrus confers resistance to citrus canker.

    PubMed

    Jia, Hongge; Zhang, Yunzeng; Orbović, Vladimir; Xu, Jin; White, Frank F; Jones, Jeffrey B; Wang, Nian

    2016-12-09

    Citrus is a highly valued tree crop worldwide, while, at the same time, citrus production faces many biotic challenges, including bacterial canker and Huanglongbing (HLB). Breeding for disease-resistant varieties is the most efficient and sustainable approach to control plant diseases. Traditional breeding of citrus varieties is challenging due to multiple limitations, including polyploidy, polyembryony, extended juvenility and long crossing cycles. Targeted genome editing technology has the potential to shorten varietal development for some traits, including disease resistance. Here, we used CRISPR/Cas9/sgRNA technology to modify the canker susceptibility gene CsLOB1 in Duncan grapefruit. Six independent lines, DLOB 2, DLOB 3, DLOB 9, DLOB 10, DLOB 11 and DLOB 12, were generated. Targeted next-generation sequencing of the six lines showed the mutation rate was 31.58%, 23.80%, 89.36%, 88.79%, 46.91% and 51.12% for DLOB 2, DLOB 3, DLOB 9, DLOB 10, DLOB 11 and DLOB 12, respectively, of the cells in each line. DLOB 2 and DLOB 3 showed canker symptoms similar to wild-type grapefruit, when inoculated with the pathogen Xanthomonas citri subsp. citri (Xcc). No canker symptoms were observed on DLOB 9, DLOB 10, DLOB 11 and DLOB 12 at 4 days postinoculation (DPI) with Xcc. Pustules caused by Xcc were observed on DLOB 9, DLOB 10, DLOB 11 and DLOB 12 in later stages, which were much reduced compared to that on wild-type grapefruit. The pustules on DLOB 9 and DLOB 10 did not develop into typical canker symptoms. No side effects and off-target mutations were detected in the mutated plants. This study indicates that genome editing using CRISPR technology will provide a promising pathway to generate disease-resistant citrus varieties.

  6. Natural occurrence of mycotoxins and toxigenic capacity of Alternaria strains from mouldy peppers.

    PubMed

    da Cruz Cabral, Lucía; Terminiello, Laura; Fernández Pinto, Virginia; Fog Nielsen, Kristian; Patriarca, Andrea

    2016-11-07

    Sweet pepper (Capsicum annuum) is an important crop cultivated worldwide, with Argentina being one of the major producers in South America. The fruit is susceptible to several fungal diseases, leading to severe economic losses for producers. In this study, Alternaria was found as the prevalent genus in mouldy peppers (50% fruits infected). Morphological identification revealed that all 64 Alternaria isolates belonged to small-spored species, most of them corresponding to A. tenuissima, A. arborescens and A. alternata species-groups. Their secondary metabolite profile was evaluated in vitro; alternariols were synthesized by most of the isolates (91% for alternariol and 92% for alternariol monomethyl ether). A high number of Alternaria spp. also produced tenuazonic acid (64%), altenuene (84%) and tentoxin (72%). In addition, damaged pepper fruits were analysed for the presence of tenuazonic acid and alternariols. A total 32 out of 48 spoiled pepper fruits were contaminated with at least one of these metabolites. Half of the samples were positive for tenuazonic acid (range 8-11,422μg/kg), while alternariol and its monomethyl ether were less frequently detected (21 and 29%, respectively) and at lower concentrations. This is the first report on the natural occurrence of Alternaria mycotoxins in Argentinean sweet pepper, and highlights a consumer risk when mouldy fruits are used in industrialized products because these compounds are not destroyed by conventional heat treatments.

  7. Effect of Alternaria solani exudates on resistant and susceptible potato cultivars from two different pathogen isolates

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The resistance phenotypes of two potato cultivars to two isolates of Alternaria solani, causal agent of early blight, were studied under greenhouse conditions. The two isolates contain varying degrees of aggressiveness on both susceptible and resistant phenotypes of potatoes. A bioassay was used to ...

  8. Biochemical Evaluation of Resistance Responses of Potato to Different Isolates of Alternaria Solani

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The resistance phenotypes of nine potato cultivars to five isolates of Alternaria solani, causal agent of early blight, were studied after inoculation and growth under greenhouse conditions. We identified potato cultivars with both susceptible and resistant phenotypes as well as A. solani isolates ...

  9. Antileukemic alpha-pyrone derivatives from the endophytic fungus Alternaria phragmospora

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Four new (1–4) and two known (5 and 6)a-pyrone derivatives have been isolated from Alternaria phragmospora, an endophytic fungus from Vinca rosea, leaves. The isolated compounds were chemically identi'ed to be 5-butyl-4-methoxy-6-methyl-2H-pyran-2-one (2) 5-butyl-6-(hydroxymethyl)-4-methoxy-2H-py...

  10. Chronic Leptomeningitis and Spinal Intradural Mass Secondary to Alternaria Infection in a Patient with Ventriculoperitoneal Shunt

    PubMed Central

    Thompson, Russell; Jandial, Rahul; Tegtmeier, Bernard; Chen, Mike Yue

    2016-01-01

    Fungal infection following placement of ventriculostomy or ventriculoperitoneal (VP) shunt is uncommon. We report the first case of Alternaria related central nervous system (CNS) shunt infection in a patient with CNS ependymoma manifesting as leptomeningitis and a spinal intradural mass. This case illustrates the diagnostic and management challenges. PMID:27840750

  11. Meteorological and agricultural effects on airborne Alternaria and Cladosporium spores and clinical aspects in Valladolid (Spain).

    PubMed

    Reyes, Estefanía Sánchez; de la Cruz, David Rodríguez; Merino, Ma Eugenia Sanchís; Sánchez, José Sánchez

    2009-01-01

    The aeropalynological monitoring was carried out from 1 February 2005-31 January 2007. The total number of spores collected during the main spore season (MSS) in 2005 was 4,500 for Alternaria and 93,744 in the case of Cladosporium, whereas in 2006 values were increased (8,385 for Alternaria and 150,144 for Cladosporium), reaching the maximum concentrations on 18 July and 17 June 2006 with 344 and 5,503 spores, respectively. The influence of the main meteorological parameters on spore concentrations was studied, resulting in a positive correlation with temperature. Rainfall, relative humidity and frequency of calms obtained negative correlations in the case of Alternaria, and positive for Cladosporium, the total daily hours of sunshine having an inverse influence on them. The intra-diurnal pattern was very similar for both genera, with a greater representation towards the central hours of the day and at night. Finally, some clinical aspects for the Alternaria spore type were analyzed, with a low percentage of sensitized patients though (9.5%). Only one patient showed positive skin test reaction to Cladosporium.

  12. Draft Genome Sequence of Alternaria alternata Isolated from Onion Leaves in South Africa

    PubMed Central

    Cloete, Michele; Gerrano, Abe Shegro; Oelofse, Dean; Adebola, Patrick

    2016-01-01

    Alternaria alternata (Fr.) Keissler strain PPRI 21032 was isolated from onion leaves collected in Roodeplaat, Pretoria, South Africa. The whole genome of this strain was sequenced and produced a total of 33.12 Mb with a GC content of 50.9%. The whole genome comprises 11,701 predicted coding sequences. PMID:27660793

  13. Sensitization to Alternaria and Cladosporium in asthmatic patients and its in vitro diagnostic confirmation.

    PubMed

    Resano, A; Sanz, M L; Oehling, A

    1998-01-01

    In order to determine the prevalence of airborne mould sensitization and the reliability of the in vitro diagnostic techniques in daily practice (antigen-specific IgE and histamine release test), we performed a 3-year study in 2,200 patients diagnosed with rhinosinusitis and/or bronchial asthma. We found mould sensitization in 101 patients, 20% of whom presented monosensitization against airborne moulds, and the rest associated other sensitizations as follows: 53.7% against Dermatophagoides pteronyssinus, 45% against grass pollen and 30% against Olea europea. The most frequently involved moulds in our patients were Alternaria and Cladosporium. Seventy-six percent of the patients presented sensitization against Alternaria, 56% of whom were monosensitized, 26% presented cosensitization to Cladosporium and the remainder were sensitive to more than two moulds. Regarding Cladosporium, the percentage of patients was similar (66%), although only 23% were monosensitized and 46% presented an associated sensitization against Alternaria. We also observed a correlation between skin tests and both in vitro diagnostic techniques, with a relative sensitivity of the specific IgE determination compared to the skin test of 98% against Alternaria and 90.4% against Cladosporium, whereas the relative sensitivity of the histamine release test was 97.4% for Alternaria and 85% for Cladosporium. In conclusion, we think that in order to confirm the etiopathogenesis of the airborne moulds and before an immunotherapy treatment is indicated, the positive skin reactions should be confirmed by means of reliable laboratory diagnostic techniques, such as antigen-specific IgE determination and histamine release test.

  14. Novel insights into the genomic basis of citrus canker based on the genome sequences of two strains of Xanthomonas fuscans subsp. aurantifolii

    PubMed Central

    2010-01-01

    Background Citrus canker is a disease that has severe economic impact on the citrus industry worldwide. There are three types of canker, called A, B, and C. The three types have different phenotypes and affect different citrus species. The causative agent for type A is Xanthomonas citri subsp. citri, whose genome sequence was made available in 2002. Xanthomonas fuscans subsp. aurantifolii strain B causes canker B and Xanthomonas fuscans subsp. aurantifolii strain C causes canker C. Results We have sequenced the genomes of strains B and C to draft status. We have compared their genomic content to X. citri subsp. citri and to other Xanthomonas genomes, with special emphasis on type III secreted effector repertoires. In addition to pthA, already known to be present in all three citrus canker strains, two additional effector genes, xopE3 and xopAI, are also present in all three strains and are both located on the same putative genomic island. These two effector genes, along with one other effector-like gene in the same region, are thus good candidates for being pathogenicity factors on citrus. Numerous gene content differences also exist between the three cankers strains, which can be correlated with their different virulence and host range. Particular attention was placed on the analysis of genes involved in biofilm formation and quorum sensing, type IV secretion, flagellum synthesis and motility, lipopolysacharide synthesis, and on the gene xacPNP, which codes for a natriuretic protein. Conclusion We have uncovered numerous commonalities and differences in gene content between the genomes of the pathogenic agents causing citrus canker A, B, and C and other Xanthomonas genomes. Molecular genetics can now be employed to determine the role of these genes in plant-microbe interactions. The gained knowledge will be instrumental for improving citrus canker control. PMID:20388224

  15. Selection of Small Synthetic Antimicrobial Peptides Inhibiting Xanthomonas citri subsp. citri Causing Citrus Canker

    PubMed Central

    Choi, Jeahyuk; Park, Euiho; Lee, Se-Weon; Hyun, Jae-Wook; Baek, Kwang-Hyun

    2017-01-01

    Citrus canker disease decreases the fruit quality and yield significantly, furthermore, emerging of streptomycin-resistant pathogens threatens the citrus industry seriously because of a lack of proper control agents. Small synthetic antimicrobial peptides (AMPs) could be a promising alternative. Fourteen hexapeptides were selected by using positional scanning of synthetic peptide combinatorial libraries. Each hexapeptide showed different antimicrobial spectrum against Bacillus, Pseudomonas, Xanthomonas, and Candida species. Intriguingly, BHC10 showed bactericidal activity exclusively on Xanthomonas citri subsp. citri (Xcc), while BHC7 was none-active exclusively against two Pseudomonas spp. at concentration of 100 μg/ml suggesting potential selectivity constrained in hexapeptide frame. Three hexapeptides, BHC02, 06 and 11, showed bactericidal activities against various Xcc strains at concentration of 10 μg/ml. When they were co-infiltrated with pathogens into citrus leaves the disease progress was suppressed significantly. Further study would be needed to confirm the actual disease control capacity of the selected hexapeptides. PMID:28167892

  16. Lonsdalea quercina subsp. populi subsp. nov., isolated from bark canker of poplar trees.

    PubMed

    Tóth, Tímea; Lakatos, Tamás; Koltay, András

    2013-06-01

    Seven Gram-negative bacterial strains were isolated from oozing bark canker of poplar (Populus × euramericana) trees in Hungary. They showed high (>98.3%) 16S rRNA gene sequence similarity to Lonsdalea quercina; however, they differed from this species in several phenotypic characteristics. Multilocus sequence analysis based on three housekeeping genes (gyrB, atpD and infB) revealed, and DNA-DNA hybridization analysis confirmed, that this group of bacterial strains forms a distinct lineage within the species Lonsdalea quercina. A detailed study of phenotypic and physiological characteristics confirmed the separation of isolates from poplars from other subspecies of L. quercina; therefore, a novel subspecies, Lonsdalea quercina subsp. populi, type strain NY060(T) (=DSM 25466(T)=NCAIM B 02483(T)), is proposed.

  17. Extracellular polysaccharides of a bacterium associated with a fungal canker disease of Eucalyptus sp.

    PubMed

    Yang, Byung Yun; Ding, Qiong; Montgomery, Rex

    2002-04-17

    Extracellular polysaccharides (EPSs) produced by an Erwinia sp associated with a fungal canker disease of Eucalyptus were fractionated into one polysaccharide that was identified with that produced by Erwinia chrysanthemi strains SR260, Ech1, and Ech9, and the other distinctively different from any other EPS produced by E. chrysanthemi strains so far studied. Their structures were determined using a combination of chemical and physical techniques including methylation analysis, low pressure gel-filtration, and anion-exchange chromatographies, high-pH anion-exchange chromatography, mass spectrometry and 1D and 2D 1H NMR spectroscopy. The new polysaccharide, identified as EPS Teranera, has the following structure: [structure: see text] The molecular weights of the polysaccharides range from 3.2-6.2 x 10(5) and their hydrodynamic properties are those of polydisperse, polyanionic biopolymers with pseudoplastic, non-thixotropic flow characteristics in aqueous solutions.

  18. Adaptive Potential of Maritime Pine (Pinus pinaster) Populations to the Emerging Pitch Canker Pathogen, Fusarium circinatum

    PubMed Central

    Elvira-Recuenco, Margarita; Iturritxa, Eugenia; Majada, Juan; Alia, Ricardo; Raposo, Rosa

    2014-01-01

    There is a concern on how emerging pests and diseases will affect the distribution range and adaptability of their host species, especially due to different conditions derived from climate change and growing globalization. Fusarium circinatum, which causes pitch canker disease in Pinus species, is an exotic pathogen of recent introduction in Spain that threatens its maritime pine (P. pinaster) stands. To predict the impact this disease will have on the species, we examine host resistance traits and their genetic architecture. Resistance phenotyping was done in a clonal provenance/progeny trial, using three-year-old cuttings artificially inoculated with the pathogen and maintained under controlled environmental conditions. A total number of 670 ramets were assessed, distributed in 10 populations, with a total of 47 families, 2 to 5 half-sibs per family, and 3–7 ramets per clone. High genetic variation was found at the three hierarchical levels studied: population, family and clone, being both additive and non-additive effects important. Narrow-sense and broad-sense heritability estimates were relatively high, with respective values of 0.43–0.58 and 0.51–0.8, depending on the resistance traits measured (lesion length, lesion length rate, time to wilting, and survival). These values suggest the species' high capacity of evolutionary response to the F. circinatum pathogen. A population originated in Northern Spain was the most resistant, while another from Morocco was the most susceptible. The total number of plants that did not show lesion development or presented a small lesion (length<30 mm) was 224 out of 670, indicating a high proportion of resistant trees in the offspring within the analyzed populations. We found large differences among populations and considerable genetic variation within populations, which should allow, through natural or artificial selection, the successful adaptation of maritime pine to pitch canker disease. PMID:25500822

  19. Adaptive potential of maritime pine (Pinus pinaster) populations to the emerging pitch canker pathogen, Fusarium circinatum.

    PubMed

    Elvira-Recuenco, Margarita; Iturritxa, Eugenia; Majada, Juan; Alia, Ricardo; Raposo, Rosa

    2014-01-01

    There is a concern on how emerging pests and diseases will affect the distribution range and adaptability of their host species, especially due to different conditions derived from climate change and growing globalization. Fusarium circinatum, which causes pitch canker disease in Pinus species, is an exotic pathogen of recent introduction in Spain that threatens its maritime pine (P. pinaster) stands. To predict the impact this disease will have on the species, we examine host resistance traits and their genetic architecture. Resistance phenotyping was done in a clonal provenance/progeny trial, using three-year-old cuttings artificially inoculated with the pathogen and maintained under controlled environmental conditions. A total number of 670 ramets were assessed, distributed in 10 populations, with a total of 47 families, 2 to 5 half-sibs per family, and 3-7 ramets per clone. High genetic variation was found at the three hierarchical levels studied: population, family and clone, being both additive and non-additive effects important. Narrow-sense and broad-sense heritability estimates were relatively high, with respective values of 0.43-0.58 and 0.51-0.8, depending on the resistance traits measured (lesion length, lesion length rate, time to wilting, and survival). These values suggest the species' high capacity of evolutionary response to the F. circinatum pathogen. A population originated in Northern Spain was the most resistant, while another from Morocco was the most susceptible. The total number of plants that did not show lesion development or presented a small lesion (length<30 mm) was 224 out of 670, indicating a high proportion of resistant trees in the offspring within the analyzed populations. We found large differences among populations and considerable genetic variation within populations, which should allow, through natural or artificial selection, the successful adaptation of maritime pine to pitch canker disease.

  20. Canker Sores

    MedlinePlus

    ... Birth Control Family HealthInfants and Toddlers Kids and Teens Pregnancy and Childbirth Women Men SeniorsIn The NewsYour Health ... Birth Control Family HealthInfants and Toddlers Kids and Teens Pregnancy and Childbirth Women Men SeniorsIn The NewsYour Health ...

  1. Canker Sores

    MedlinePlus

    ... and Vision Leadership Executive Committee Board of Trustees Governance Past Presidents Staff/Contact History Awards Our Partners ... tell the difference? A: The location is a good clue. Your doctor may recommend performing culture testing ...

  2. Canker sore

    MedlinePlus

    ... which can irritate the area more.) Apply a mixture of half hydrogen peroxide and half water directly ... times a day. Rinse your mouth with a mixture of half Milk of Magnesia and half Benadryl ...

  3. Canker Sore

    MedlinePlus

    ... Policy Notice of Privacy Practices Notice of Nondiscrimination Advertising Mayo Clinic is a not-for-profit organization and proceeds from Web advertising help support our mission. Mayo Clinic does not ...

  4. Canker Sores

    MedlinePlus

    ... SLS is a foaming agent found in most toothpastes and mouthwashes. Finally, not getting the right nutrition, ... you brush your teeth . Brush and rinse with toothpastes and mouthwashes that don't contain sodium lauryl ...

  5. Association of IL-4RA single nucleotide polymorphisms, HLA-DR and HLA-DQ in children with Alternaria-sensitive moderate-severe asthma

    PubMed Central

    2010-01-01

    Background Asthma afflicts 6% to 8% of the United States population, and severe asthma represents approximately 10% of asthmatic patients. Several epidemiologic studies in the United States and Europe have linked Alternaria sensitivity to both persistence and severity of asthma. In order to begin to understand genetic risk factors underlying Alternaria sensitivity and asthma, in these studies we examined T cell responses to Alternaria antigens, HLA Class II restriction and HLA-DQ protection in children with severe asthma. Methods Sixty children with Alternaria-sensitive moderate-severe asthma were compared to 49 children with Alternaria-sensitive mild asthma. We examined HLA-DR and HLA-DQ frequencies in Alternaria-sensitive asthmatic by HLA typing. To determine ratios of Th1/Th2 Alternaria-specific T-cells, cultures were stimulated in media alone, Alternaria alternata extract and Alt a1. Sensitivity to IL-4 stimulation was measured by up-regulation of CD23 on B cells. Results Children with Alternaria-sensitive moderate-severe asthma trended to have increased sensitivities to Cladosporium (46% versus 35%), to Aspergillus (43% versus 28%), and significantly increased sensitivities to trees (78% versus 57%) and to weeds (68% versus 48%). The IL-4RA ile75val polymorphism was significantly increased in Alternaria-sensitive moderate-severe asthmatics, 83% (0.627 allele frequency) compared to Alternaria-sensitive mild asthmatics, 57% (0.388 allele frequency). This was associated with increased sensitivity to IL-4 stimulation measured by significantly increased IL-4 stimulated CD23 expression on CD19+ and CD86+CD19+ B cells of Alternaria-sensitive moderate-severe asthmatics. IL-5 and IL-13 synthesis was significantly increased in Alternaria-sensitive moderate-severe asthmatics compared to mild asthmatics to Alternaria extract and Alt a1 stimulation. The frequency of HLA-DQB1*03 allele was significantly decreased in Alternaria-sensitive moderate-severe asthmatics compared

  6. Effect of X-irradiation on Citrus Canker Pathogen Xanthomonas citri subsp. citri of Satsuma Mandarin Fruits

    PubMed Central

    Song, Min-A; Park, Jae Sin; Kim, Ki Deok; Jeun, Yong Chull

    2015-01-01

    Citrus canker caused by Xanthomonas citri subsp. citri (Xcc) is one of the most important bacterial diseases of citrus. Because citrus canker is not found in many countries including European Union and Australia, Xcc is strictly regulated in order to prevent its spread. In this study, the effects of X-irradiation on Xcc growth either in the suspension or on the surface of citrus fruits were investigated. The suspension containing 1×107 cfu/ml of Xcc was irradiated with different absorbed doses of X-irradiation ranging from 50 to 400 Gy. The results showed that Xcc was fully dead at 400 Gy of X-irradiation. To determine the effect of X-irradiation on quarantine, the Xcc-inoculated citrus fruits were irradiated with different X-ray doses at which Xcc was completely inhibited by an irradiation dose of 250 Gy. The D10 value for Xcc on citrus fruits was found to be 97 Gy, indicating the possibility of direct application on citrus quarantine without any side sterilizer. Beside, presence of Xcc on the surface of asymptomatic citrus fruits obtained from citrus canker-infected orchards was noted. It indicated that the exporting citrus fruits need any treatment so that Xcc on the citrus fruits should be completely eliminated. Based on these results, ionizing radiation can be considered as an alternative method of eradicating Xcc for export of citrus fruits. PMID:26672670

  7. Degradation of the Alternaria mycotoxins alternariol, alternariol monomethyl ether, and altenuene upon bread baking.

    PubMed

    Siegel, David; Feist, Michael; Proske, Matthias; Koch, Matthias; Nehls, Irene

    2010-09-08

    The stability of the Alternaria mycotoxins alternariol, alternariol monomethyl ether, and altenuene upon bread baking was investigated by model experiments using a spiked wholemeal wheat flour matrix. For alternariol and alternariol monomethyl ether, but not for altenuene, degradation products, formed through a sequence of hydrolysis and decarboxylation, could be identified in pilot studies. The simultaneous quantification of alternariol, alternariol monomethyl ether, altenuene, and the degradation products was achieved by a newly developed high performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS) multimethod. The obtained quantitative data indicate that the Alternaria mycotoxins are barely degraded during wet baking, while significant degradation occurs upon dry baking, with the stability decreasing in the order alternariol monomethyl ether>alternariol>altenuene. The novel degradation products could be detected after the wet baking of flour spiked with alternariol and in a sample survey of 24 commercial cereal based baking products.

  8. [Aerobiological study of Alternaria and Cladosporium conidia in the atmosphere of Almeria (SE Spain)].

    PubMed

    Sabariego Ruiz, Silvia; Díaz de la Guardia Guerrero, Consuelo; Alba Sánchez, Francisca

    2004-09-01

    This study analysed the seasonal and intradiurnal behaviour of fungal spores from Alternaria and Cladosporium on air samples collected in the city of Almeria (SE Spain), as well as the influence that meteorological parameters have on the concentration of these type of spores. Aerobiological sampling was made during four years (1998-2001) using a Hirst-type volumetric spore trap. Spores of Alternaria and Cladosporium were detected throughout the year, reaching the highest concentrations from May to October. The diurnal patterns of these taxons reflected a presence similar of spores during a 24 h period, with values horary close to 4% of total sampling daily. The correlations show a positive association with temperature, hours of sunshine and accumulate rainfall, but negative with daily rainfall.

  9. Disruption of Glycosylphosphatidylinositol-Anchored Lipid Transfer Protein Gene Altered Cuticular Lipid Composition, Increased Plastoglobules, and Enhanced Susceptibility to Infection by the Fungal Pathogen Alternaria brassicicola1[W

    PubMed Central

    Lee, Saet Buyl; Go, Young Sam; Bae, Hyun-Jong; Park, Jong Ho; Cho, Sung Ho; Cho, Hong Joo; Lee, Dong Sook; Park, Ohkmae K.; Hwang, Inhwan; Suh, Mi Chung

    2009-01-01

    All aerial parts of vascular plants are covered with cuticular waxes, which are synthesized by extensive export of intracellular lipids from epidermal cells to the surface. Although it has been suggested that plant lipid transfer proteins (LTPs) are involved in cuticular lipid transport, the in planta evidence is still not clear. In this study, a glycosylphosphatidylinositol-anchored LTP (LTPG1) showing higher expression in epidermal peels of stems than in stems was identified from an Arabidopsis (Arabidopsis thaliana) genome-wide microarray analysis. The expression of LTPG1 was observed in various tissues, including the epidermis, stem cortex, vascular bundles, mesophyll cells, root tips, pollen, and early-developing seeds. LTPG1 was found to be localized in the plasma membrane. Disruption of the LTPG1 gene caused alterations of cuticular lipid composition, but no significant changes on total wax and cutin monomer loads were seen. The largest reduction (10 mass %) in the ltpg1 mutant was observed in the C29 alkane, which is the major component of cuticular waxes in the stems and siliques. The reduced content was overcome by increases of the C29 secondary alcohols and C29 ketone wax loads. The ultrastructure analysis of ltpg1 showed a more diffuse cuticular layer structure, protrusions of the cytoplasm into the vacuole in the epidermis, and an increase of plastoglobules in the stem cortex and leaf mesophyll cells. Furthermore, the ltpg1 mutant was more susceptible to infection by the fungus Alternaria brassicicola than the wild type. Taken together, these results indicated that LTPG1 contributed either directly or indirectly to cuticular lipid accumulation. PMID:19321705

  10. Cost-effectiveness of various methods of diagnosing hypersensitivity to Alternaria.

    PubMed

    Escudero, A I; Sánchez-Guerrero, I M; Mora, A M; Soriano, V; López, J D; García, F J; Negro, J M; Hernández, J; Pagán, J A

    1993-01-01

    This study was undertaken for two reasons: 1) It is more difficult to diagnose hypersensitivity to molds than to other allergens, so an evaluation of diagnostic tests was needed. 2) Alternaria is the principal cause of mold sensitization in our area. Sixty-six patients (20 +/- 4 years) were selected and divided into two groups. Group A was made up of patients with rhinitis and/or asthma due to Alternaria sensitization. Group B consisted of patients sensitized to other allergens and patients with nonrespiratory allergic disorders. Skin tests (prick and intradermal), challenge tests (conjunctival, nasal, and bronchial), and specific IgE determination were performed for all patients. A biologically standardized extract of Alternaria tenuis (Alergia e Inmunología Abelló, S. A., Madrid, Spain) obtained from a single batch was used for all tests. Our diagnostic criterion was a clinical history of rhinitis or asthma that coincided with the results of nasal/bronchial challenge. The diagnostic value of the other tests was compared to this criterion. In the group of rhinitic patients, skin tests and conjunctival challenge were more sensitive than specific IgE determination. In asthmatic patients, the most sensitive techniques were nasal and conjunctival challenges, followed by prick and intradermal skin tests, and, lastly, serum specific IgE determination. When rhinitis and asthma were considered jointly, the most sensitive test was conjunctival challenge, followed by skin-prick and intradermal tests. All tests had the same specificity, regardless of disorder. Nasal challenge was positive in all patients. Skin tests are easy to perform, cheap, non-traumatic for the patient, and sufficiently specific and sensitive for the diagnosis of Alternaria hypersensitivity.(ABSTRACT TRUNCATED AT 250 WORDS)

  11. Role of micronutrients in defense to white rust and Alternaria blight infecting Indian mustard.

    PubMed

    Rathi, A S; Singh, Dhiraj; Avtar, Ram; Kumar, Pawan

    2015-03-01

    Field experiments were carried out at Oilseeds Research Area of CCS Haryana Agricultural University, Hisar during rabi, 2008-09 to 2011-12 to find out the possible role of soil application of different micronutrients alone and in combinations in defense to white rust and Alternaria blight diseases in Indian mustard [Brassica juncea (L.) Czern & Coss.]. Among the sole application of micronutrients, minimum disease severity of both white rust (35.0%) and Alternaria blight (31.8%) was observed when S @ 40 kg ha in the form of Gypsum was applied as basal dose in the soil. When Gypsum was supplemented with Borax @10 kg ha(-1) or with ZnSO4 @15 kg ha(-1) the level of tolerance seems to be improved for both the diseases as compared to the sole treatment of each nutrient, i.e., ZnSO4 @ 15 kg/ha, Borax @ 10 kg ha' and Gypsum @ 250 kg ha(-1). Furthermore, minimum disease severity of both white rust (31.3 %) and Alternaria blight (26.3 %) was observed with soil application of ZnSO4 @ 15 kg ha(-1) + Borax @ 10 kg ha(-1) + Gypsum @250 kg ha(-1) as basal dose as compared to the severity of white rust (43.6%) and Alternaria blight (38.6%) in untreated check. Significant increase in seed yield (1612 kg ha(-1)) was also recorded in above mentioned treatment as compared to the yield (1337 kg ha(-1)) in untreated check. These findings will also be helpful in maintaining soil health and minimizing the losses due to both the fungal diseases for eco-friendly sustainability of Indian mustard.

  12. Alternariol 9-methyl ether from the endophytic fungus Alternaria sp. Samif01 and its bioactivities

    PubMed Central

    Lou, Jingfeng; Yu, Ruiting; Wang, Xiaohan; Mao, Ziling; Fu, Linyun; Liu, Yang; Zhou, Ligang

    2016-01-01

    One bioactive compound, identified as alternariol 9-methyl ether, was isolated from the crude extract of the endophytic fungus Alternaria sp. Samif01 residing in the roots of Salvia miltiorrhiza Bunge. Alternariol 9-methyl ether was active against bacteria with minimum inhibitory concentration values ranging from 25 to 75 μg/mL and median inhibitory concentration (IC50) values ranging from 16.00 to 38.27 μg/mL. The IC50 value of alternariol 9-methyl ether against spore germination of Magnaporthe oryzae was 87.18 μg/mL. Alternariol 9-methyl ether also showed antinematodal activity against Bursaphelenchus xylophilus and Caenorhabditis elegans with IC50 values of 98.17 μg/mL and 74.62 μg/mL, respectively. This work is the first report on alternariol 9-methyl ether and its biological activities from the endophytic fungus Alternaria sp. Samif01 derived from S. miltiorrhiza Bunge. The results indicate the potential of Alternaria sp. Samif01 as a source of alternariol 9-methyl ether and also support that alternariol 9-methyl ether is a natural compound with high potential bioactivity against microorganisms. PMID:26887231

  13. Characterization and Pathogenicity of Alternaria burnsii from Seeds of Cucurbita maxima (Cucurbitaceae) in Bangladesh.

    PubMed

    Paul, Narayan Chandra; Deng, Jian Xin; Lee, Hyang Burm; Yu, Seung-Hun

    2015-12-01

    In the course of survey of endophytic fungi from Bangladesh pumpkin seeds in 2011~2012, two strains (CNU111042 and CNU111043) with similar colony characteristics were isolated and characterized by their morphology and by molecular phylogenetic analysis of the internal transcribed spacer, glyceraldehydes-3-phosphate dehydrogenase (gpd), and Alternaria allergen a1 (Alt a1) sequences. Phylogenetic analysis of all three sequences and their combined dataset revealed that the fungus formed a subclade within the A. alternata clade, matching A. burnsi and showing differences with its other closely related Alternaria species, such as A. longipes, A. tomato, and A. tomaticola. Long ellipsoid, obclavate or ovoid beakless conidia, shorter and thinner conidial size (16~60 [90] × 6.5~14 [~16] µm) distinguish this fungus from other related species. These isolates showed more transverse septation (2~11) and less longitudinal septation (0~3) than did other related species. Moreover, the isolate did not produce any diffusible pigment on media. Therefore, our results reveal that the newly recorded fungus from a new host, Cucurbita maxima, is Alternaria burnsii Uppal, Patel & Kamat.

  14. Production of the Allergenic Protein Alt a 1 by Alternaria Isolates from Working Environments

    PubMed Central

    Skóra, Justyna; Otlewska, Anna; Gutarowska, Beata; Leszczyńska, Joanna; Majak, Iwona; Stępień, Łukasz

    2015-01-01

    The aim of the study was to evaluate the ability of Alternaria isolates from workplaces to produce Alt a 1 allergenic protein, and to analyze whether technical materials (cellulose, compost, leather) present within the working environment stimulate or inhibit Alt a 1 production (ELISA test). Studies included identification of the isolated molds by nucleotide sequences analyzing of the ITS1/ITS2 regions, actin, calmodulin and Alt a 1 genes. It has been shown that Alternaria molds are significant part of microbiocenosis in the archive, museum, library, composting plant and tannery (14%–16% frequency in the air). The presence of the gene encoding the Alt a 1 protein has been detected for the strains: Alternaria alternata, A. lini, A. limoniasperae A. nobilis and A. tenuissima. Environmental strains produced Alt a 1 at higher concentrations (1.103–6.528 ng/mL) than a ATCC strain (0.551–0.975 ng/mL). It has been shown that the homogenization of the mycelium and the use of ultrafiltration allow a considerable increase of Alt a 1 concentration. Variations in the production of Alt a 1 protein, depend on the strain and extraction methods. These studies revealed no impact of the technical material from the workplaces on the production of Alt a 1 protein. PMID:25689994

  15. Alternariol 9-methyl ether from the endophytic fungus Alternaria sp. Samif01 and its bioactivities.

    PubMed

    Lou, Jingfeng; Yu, Ruiting; Wang, Xiaohan; Mao, Ziling; Fu, Linyun; Liu, Yang; Zhou, Ligang

    2016-01-01

    One bioactive compound, identified as alternariol 9-methyl ether, was isolated from the crude extract of the endophytic fungus Alternaria sp. Samif01 residing in the roots of Salvia miltiorrhiza Bunge. Alternariol 9-methyl ether was active against bacteria with minimum inhibitory concentration values ranging from 25 to 75μg/mL and median inhibitory concentration (IC50) values ranging from 16.00 to 38.27μg/mL. The IC50 value of alternariol 9-methyl ether against spore germination of Magnaporthe oryzae was 87.18μg/mL. Alternariol 9-methyl ether also showed antinematodal activity against Bursaphelenchus xylophilus and Caenorhabditis elegans with IC50 values of 98.17μg/mL and 74.62μg/mL, respectively. This work is the first report on alternariol 9-methyl ether and its biological activities from the endophytic fungus Alternaria sp. Samif01 derived from S. miltiorrhiza Bunge. The results indicate the potential of Alternaria sp. Samif01 as a source of alternariol 9-methyl ether and also support that alternariol 9-methyl ether is a natural compound with high potential bioactivity against microorganisms.

  16. Production of the allergenic protein Alt a 1 by Alternaria isolates from working environments.

    PubMed

    Skóra, Justyna; Otlewska, Anna; Gutarowska, Beata; Leszczyńska, Joanna; Majak, Iwona; Stępień, Łukasz

    2015-02-16

    The aim of the study was to evaluate the ability of Alternaria isolates from workplaces to produce Alt a 1 allergenic protein, and to analyze whether technical materials (cellulose, compost, leather) present within the working environment stimulate or inhibit Alt a 1 production (ELISA test). Studies included identification of the isolated molds by nucleotide sequences analyzing of the ITS1/ITS2 regions, actin, calmodulin and Alt a 1 genes. It has been shown that Alternaria molds are significant part of microbiocenosis in the archive, museum, library, composting plant and tannery (14%-16% frequency in the air). The presence of the gene encoding the Alt a 1 protein has been detected for the strains: Alternaria alternata, A. lini, A. limoniasperae A. nobilis and A. tenuissima. Environmental strains produced Alt a 1 at higher concentrations (1.103-6.528 ng/mL) than a ATCC strain (0.551-0.975 ng/mL). It has been shown that the homogenization of the mycelium and the use of ultrafiltration allow a considerable increase of Alt a 1 concentration. Variations in the production of Alt a 1 protein, depend on the strain and extraction methods. These studies revealed no impact of the technical material from the workplaces on the production of Alt a 1 protein.

  17. Characterization and Pathogenicity of Alternaria burnsii from Seeds of Cucurbita maxima (Cucurbitaceae) in Bangladesh

    PubMed Central

    Paul, Narayan Chandra; Deng, Jian Xin

    2015-01-01

    In the course of survey of endophytic fungi from Bangladesh pumpkin seeds in 2011~2012, two strains (CNU111042 and CNU111043) with similar colony characteristics were isolated and characterized by their morphology and by molecular phylogenetic analysis of the internal transcribed spacer, glyceraldehydes-3-phosphate dehydrogenase (gpd), and Alternaria allergen a1 (Alt a1) sequences. Phylogenetic analysis of all three sequences and their combined dataset revealed that the fungus formed a subclade within the A. alternata clade, matching A. burnsi and showing differences with its other closely related Alternaria species, such as A. longipes, A. tomato, and A. tomaticola. Long ellipsoid, obclavate or ovoid beakless conidia, shorter and thinner conidial size (16~60 [90] × 6.5~14 [~16] µm) distinguish this fungus from other related species. These isolates showed more transverse septation (2~11) and less longitudinal septation (0~3) than did other related species. Moreover, the isolate did not produce any diffusible pigment on media. Therefore, our results reveal that the newly recorded fungus from a new host, Cucurbita maxima, is Alternaria burnsii Uppal, Patel & Kamat. PMID:26839497

  18. Sensitization to Alternaria and Cladosporium in patients with respiratory allergy and outdoor counts of mold spores in Ankara atmosphere, Turkey.

    PubMed

    Bavbek, Sevim; Erkekol, Ferda Oner; Ceter, Talip; Mungan, Dilşad; Ozer, Faruk; Pinar, Münevver; Misirligil, Zeynep

    2006-08-01

    Sensitization to Alternaria and Cladosporium has been reported to be 3% to 30% in European countries. However, in Turkey, there is limited data about the prevalence of sensitization to these molds and the intensity of the two mold spores in Ankara atmosphere. This study was designed to evaluate the sensitization to Alternaria and Cladosporium in patients with respiratory allergy in Ankara and also the concentration of the two molds in Ankara atmosphere. Allergic rhinitis and asthma patients living in Ankara were included in the study. Demographic and diagnostic data of the patients were recorded. A skin prick test with extracts supplied by three different laboratories was used to evaluate the sensitization to Alternaria and Cladosporium. Mold spores were measured using a Burkard 7-day recording volumetric spore trap in Ankara atmosphere during a year. Overall sensitization to the two molds was found to be 14.8%, and isolated Alternaria or Cladosporiumsensitization was 3%. Considering the positive reaction to at least one of the three suppliers, the sensitization rate was 11.9% and 8.1% for Alternaria and Cladosporium, respectively. Cochran's Q homogenization test demonstrated that the positive and negative reaction were not homogeneous among three laboratories. The total number of mold spores in Ankara atmosphere was 429,264 spores/m3 of which 75.5% and 6% were constituted by Cladosporium and Alternaria, respectively. The prevalence of Cladosporium and Alternaria sensitization in respiratory allergy patients is quite similar to European countries; however, our data indicate that commercial mold extracts should be standardized to establish the real sensitization rates. Additionally, considering the great numbers of these mold spores in Ankara atmosphere, long-term follow-up studies are needed to evaluate the relationship between the mold load and sensitization patterns.

  19. De novo genome assembly of Geosmithia morbida, the causal agent of thousand cankers disease

    PubMed Central

    Westbrook, Anthony; Woeste, Keith

    2016-01-01

    Geosmithia morbida is a filamentous ascomycete that causes thousand cankers disease in the eastern black walnut tree. This pathogen is commonly found in the western U.S.; however, recently the disease was also detected in several eastern states where the black walnut lumber industry is concentrated. G. morbida is one of two known phytopathogens within the genus Geosmithia, and it is vectored into the host tree via the walnut twig beetle. We present the first de novo draft genome of G. morbida. It is 26.5 Mbp in length and contains less than 1% repetitive elements. The genome possesses an estimated 6,273 genes, 277 of which are predicted to encode proteins with unknown functions. Approximately 31.5% of the proteins in G. morbida are homologous to proteins involved in pathogenicity, and 5.6% of the proteins contain signal peptides that indicate these proteins are secreted. Several studies have investigated the evolution of pathogenicity in pathogens of agricultural crops; forest fungal pathogens are often neglected because research efforts are focused on food crops. G. morbida is one of the few tree phytopathogens to be sequenced, assembled and annotated. The first draft genome of G. morbida serves as a valuable tool for comprehending the underlying molecular and evolutionary mechanisms behind pathogenesis within the Geosmithia genus. PMID:27168971

  20. The Xanthomonas axonopodis pv. citri flagellum is required for mature biofilm and canker development.

    PubMed

    Malamud, Florencia; Torres, Pablo S; Roeschlin, Roxana; Rigano, Luciano A; Enrique, Ramón; Bonomi, Hernán R; Castagnaro, Atilio P; Marano, María Rosa; Vojnov, Adrián A

    2011-03-01

    Xanthomonas axonopodis pv. citri (Xac) is the causative agent of citrus canker. This bacterium develops a characteristic biofilm on both biotic and abiotic surfaces. To evaluate the participation of the single flagellum of Xac in biofilm formation, mutants in the fliC (flagellin) and the flgE (hook) genes were generated. Swimming motility, assessed on 0.25 % agar plates, was markedly reduced in fliC and flgE mutants. However, the fliC and flgE mutants exhibited a flagellar-independent surface translocation on 0.5 % agar plates. Mutation of either the rpfF or the rpfC gene, which both encode proteins involved in cell-cell signalling mediated by diffusible signal factor (DSF), led to a reduction in both flagellar-dependent and flagellar-independent surface translocation, indicating a regulatory role for DSF in both types of motility. Confocal laser scanning microscopy of biofilms produced in static culture demonstrated that the flagellum is also involved in the formation of mushroom-shaped structures and water channels, and in the dispersion of biofilms. The presence of the flagellum was required for mature biofilm development on lemon leaf surfaces. The absence of flagellin produced a slight reduction in Xac pathogenicity and this reduction was more severe when the complete flagellum structure was absent.

  1. Description of Acinetobacter populi sp. nov. isolated from symptomatic bark of Populus x euramericana canker.

    PubMed

    Li, Yong; Chang, Jupu; Guo, Li-min; Wang, Hai-Ming; Xie, Shou-jiang; Piao, Chun-gen; He, Wei

    2015-12-01

    Five Gram-negative, non-motile, rod-shaped bacterial strains were isolated from cankers of Populus x euramericana collected from different locations in Puyang city, Henan Province, China. The five strains were characterized by nutritional and physiological testing and DNA sequence analysis. Haemolysis was not observed on agar media supplemented with sheep erythrocytes. The strains could be distinguished from members of most species of the genus Acinetobacter by their inability to assimilate L-arginine and benzoate. The five strains formed a single branch in phylogenetic trees based on 16S rRNA, gyrB and rpoB individual gene sequence analysis,indicating that they all belonged to a single taxon within the genus Acinetobacter. DNA-DNA hybridization results indicated that the five isolates represented to a single species that was separate from Acinetobacter puyangensis. On the basis of the phenotypic, genotypic and phylogenetic characteristics, the five strains are considered to represent a novel species of the genus Acinetobacter, for which the name Acinetobacter populi sp. nov. is proposed. The typestrain of A. populi sp. nov. is PBJ7T (CFCC 11170T=KCTC 42272T).

  2. Immunochemical quantitation of airborne short ragweed, Alternaria, antigen E, and Alt-I allergens: a two-year prospective study

    SciTech Connect

    Agarwal, M.K.; Swanson, M.C.; Reed, C.E.; Yunginger, J.W.

    1983-07-01

    We conducted a 2 yr prospective study to measure atmospheric short ragweed and Alternaria allergens by RAST inhibition analysis of eluates from filter sheets exposed in air samplers. In both years ragweed pollen and Alternaria spore counts, obtained with a rotoslide sampler, correlated significantly with immunochemically measured airborne ragweed and Alternaria allergenic activity. Airborne levels of the purified allergens AgE and Alt-I were successfully quantitated; these levels correlated closely with total airborne ragweed and Alternaria allergenic activities, respectively, and also with ragweed pollen and Alternaria spore counts. Eluates from filter sheets exposed during late summer and fall produced positive wheal-and-flare skin tests in patients with fall hay fever. In both years immunochemical measurements of allergenic activity due to airborne short ragweed correlated closely with mean symptom score indices in groups of short ragweed-sensitive individuals. Measurable levels of atmospheric ragweed allergenic activity were noted before and after the ragweed pollination season, and at these times we noted small increases in mean symptom score indices in the short ragweed-sensitive groups. Thus immunochemical analyses provide important information concerning levels of environmental allergens.

  3. Immunochemical quantitation of airborne short ragweed, Alternaria, antigen E, and Alt-I allergens: a two-year prospective study.

    PubMed

    Agarwal, M K; Swanson, M C; Reed, C E; Yunginger, J W

    1983-07-01

    We conducted a 2 yr prospective study to measure atmospheric short ragweed and Alternaria allergens by RAST inhibition analysis of eluates from filter sheets exposed in air samplers. In both years ragweed pollen and Alternaria spore counts, obtained with a rotoslide sampler, correlated significantly with immunochemically measured airborne ragweed and Alternaria allergenic activity. Airborne levels of the purified allergens AgE and Alt-I were successfully quantitated; these levels correlated closely with total airborne ragweed and Alternaria allergenic activities, respectively, and also with ragweed pollen and Alternaria spore counts. Eluates from filter sheets exposed during late summer and fall produced positive wheal-and-flare skin tests in patients with fall hay fever. In both years immunochemical measurements of allergenic activity due to airborne short ragweed correlated closely with mean symptom score indices in groups of short ragweed-sensitive individuals. Measurable levels of atmospheric ragweed allergenic activity were noted before and after the ragweed pollination season, and at these times we noted small increases in mean symptom score indices in the short ragweed-sensitive groups. Thus immunochemical analyses provide important information concerning levels of environmental allergens.

  4. Incorporating exposure to pitch canker disease to support management decisions of Pinus pinaster Ait. in the face of climate change.

    PubMed

    Serra-Varela, María Jesús; Alía, Ricardo; Pórtoles, Javier; Gonzalo, Julián; Soliño, Mario; Grivet, Delphine; Raposo, Rosa

    2017-01-01

    Climate change is gravely affecting forest ecosystems, resulting in large distribution shifts as well as in increasing infection diseases and biological invasions. Accordingly, forest management requires an evaluation of exposure to climate change that should integrate both its abiotic and biotic components. Here we address the implications of climate change in an emerging disease by analysing both the host species (Pinus pinaster, Maritime pine) and the pathogen's (Fusarium circinatum, pitch canker) environmental suitability i.e. estimating the host's risk of habitat loss and the disease`s future environmental range. We constrained our study area to the Spanish Iberian Peninsula, where accurate climate and pitch canker occurrence databases were available. While P. pinaster is widely distributed across the study area, the disease has only been detected in its north-central and north-western edges. We fitted species distribution models for the current distribution of the conifer and the disease. Then, these models were projected into nine Global Climate Models and two different climatic scenarios which totalled to 18 different future climate predictions representative of 2050. Based on the level of agreement among them, we created future suitability maps for the pine and for the disease independently, which were then used to assess exposure of current populations of P. pinaster to abiotic and biotic effects of climate change. Almost the entire distribution of P. pinaster in the Spanish Iberian Peninsula will be subjected to abiotic exposure likely to be driven by the predicted increase in drought events in the future. Furthermore, we detected a reduction in exposure to pitch canker that will be concentrated along the north-western edge of the study area. Setting up breeding programs is recommended in highly exposed and productive populations, while silvicultural methods and monitoring should be applied in those less productive, but still exposed, populations.

  5. Incorporating exposure to pitch canker disease to support management decisions of Pinus pinaster Ait. in the face of climate change

    PubMed Central

    Serra-Varela, María Jesús; Alía, Ricardo; Pórtoles, Javier; Gonzalo, Julián; Soliño, Mario; Grivet, Delphine; Raposo, Rosa

    2017-01-01

    Climate change is gravely affecting forest ecosystems, resulting in large distribution shifts as well as in increasing infection diseases and biological invasions. Accordingly, forest management requires an evaluation of exposure to climate change that should integrate both its abiotic and biotic components. Here we address the implications of climate change in an emerging disease by analysing both the host species (Pinus pinaster, Maritime pine) and the pathogen’s (Fusarium circinatum, pitch canker) environmental suitability i.e. estimating the host’s risk of habitat loss and the disease`s future environmental range. We constrained our study area to the Spanish Iberian Peninsula, where accurate climate and pitch canker occurrence databases were available. While P. pinaster is widely distributed across the study area, the disease has only been detected in its north-central and north-western edges. We fitted species distribution models for the current distribution of the conifer and the disease. Then, these models were projected into nine Global Climate Models and two different climatic scenarios which totalled to 18 different future climate predictions representative of 2050. Based on the level of agreement among them, we created future suitability maps for the pine and for the disease independently, which were then used to assess exposure of current populations of P. pinaster to abiotic and biotic effects of climate change. Almost the entire distribution of P. pinaster in the Spanish Iberian Peninsula will be subjected to abiotic exposure likely to be driven by the predicted increase in drought events in the future. Furthermore, we detected a reduction in exposure to pitch canker that will be concentrated along the north-western edge of the study area. Setting up breeding programs is recommended in highly exposed and productive populations, while silvicultural methods and monitoring should be applied in those less productive, but still exposed, populations

  6. A framework to gauge the epidemic potential of plant pathogens in environmental reservoirs: the example of kiwifruit canker.

    PubMed

    Bartoli, Claudia; Lamichhane, Jay Ram; Berge, Odile; Guilbaud, Caroline; Varvaro, Leonardo; Balestra, Giorgio M; Vinatzer, Boris A; Morris, Cindy E

    2015-02-01

    New economically important diseases on crops and forest trees emerge recurrently. An understanding of where new pathogenic lines come from and how they evolve is fundamental for the deployment of accurate surveillance methods. We used kiwifruit bacterial canker as a model to assess the importance of potential reservoirs of new pathogenic lineages. The current kiwifruit canker epidemic is at least the fourth outbreak of the disease on kiwifruit caused by Pseudomonas syringae in the mere 50 years in which this crop has been cultivated worldwide, with each outbreak being caused by different genetic lines of the bacterium. Here, we ask whether strains in natural (non-agricultural) environments could cause future epidemics of canker on kiwifruit. To answer this question, we evaluated the pathogenicity, endophytic colonization capacity and competitiveness on kiwifruit of P. syringae strains genetically similar to epidemic strains and originally isolated from aquatic and subalpine habitats. All environmental strains possessing an operon involved in the degradation of aromatic compounds via the catechol pathway grew endophytically and caused symptoms in kiwifruit vascular tissue. Environmental and epidemic strains showed a wide host range, revealing their potential as future pathogens of a variety of hosts. Environmental strains co-existed endophytically with CFBP 7286, an epidemic strain, and shared about 20 virulence genes, but were missing six virulence genes found in all epidemic strains. By identifying the specific gene content in genetic backgrounds similar to known epidemic strains, we developed criteria to assess the epidemic potential and to survey for such strains as a means of forecasting and managing disease emergence.

  7. DNA Polymorphisms and Biocontrol of Bacillus Antagonistic to Citrus Bacterial Canker with Indication of the Interference of Phyllosphere Biofilms

    PubMed Central

    Huang, Tzu-Pi; Tzeng, Dean Der-Syh; Wong, Amy C. L.; Chen, Chun-Han; Lu, Kuan-Min; Lee, Ya-Huei; Huang, Wen-Di; Hwang, Bing-Fang; Tzeng, Kuo-Ching

    2012-01-01

    Citrus bacterial canker caused by Xanthomonas axonopodis pv. citri is a devastating disease resulting in significant crop losses in various citrus cultivars worldwide. A biocontrol agent has not been recommended for this disease. To explore the potential of bacilli native to Taiwan to control this disease, Bacillus species with a broad spectrum of antagonistic activity against various phytopathogens were isolated from plant potting mixes, organic compost and the rhizosphere soil. Seven strains TKS1-1, OF3-16, SP4-17, HSP1, WG6-14, TLB7-7, and WP8-12 showing superior antagonistic activity were chosen for biopesticide development. The genetic identity based on 16S rDNA sequences indicated that all seven native strains were close relatives of the B. subtilis group and appeared to be discrete from the B. cereus group. DNA polymorphisms in strains WG6-14, SP4-17, TKS1-1, and WP8-12, as revealed by repetitive sequence-based PCR with the BOXA1R primers were similar to each other, but different from those of the respective Bacillus type strains. However, molecular typing of the strains using either tDNA-intergenic spacer regions or 16S–23S intergenic transcribed spacer regions was unable to differentiate the strains at the species level. Strains TKS1-1 and WG6-14 attenuated symptom development of citrus bacterial canker, which was found to be correlated with a reduction in colonization and biofilm formation by X. axonopodis pv. citri on leaf surfaces. The application of a Bacillus strain TKS1-1 endospore formulation to the leaf surfaces of citrus reduced the incidence of citrus bacterial canker and could prevent development of the disease. PMID:22848728

  8. Outbreak and Spread of Bacterial Canker of Kiwifruit Caused by Pseudomonas syringae pv. actinidiae Biovar 3 in Korea

    PubMed Central

    Kim, Gyoung Hee; Kim, Kwang-Hyung; Son, Kyeong In; Choi, Eu Ddeum; Lee, Young Sun; Jung, Jae Sung; Koh, Young Jin

    2016-01-01

    A bacterial pathogen, Pseudomonas syringae pv. actinidiae (Psa), is a causal agent of kiwifruit bacterial canker worldwide. Psa biovar 3 (Psa3) was first detected in 2011 at an orchard in Dodeok-myeon, Goheunggun, Jeonnam Province in Korea. In this study, we present the results of an epidemiological study regarding Psa3 occurrence on kiwifruit orchards in Korea for the period of 2013 to 2015. Since the first detection of Psa3 in 2011, there was no further case reported by 2013. However, Psa3 was rapidly spreading to 33 orchards in 2014; except for three orchards in Sacheonsi, Gyeongnam Province, most cases were reported in Jeju Island. Entering 2015, bacterial canker by Psa3 became a pandemic in Korea, spreading to 72 orchards in Jeju Island, Jeonnam, and Gyeongnam Provinces. Our epidemiological study indicated that the first Psa3 incidence in 2011 might result from an introduction of Psa3 through imported seedlings from China in 2006. Apart from this, it was estimated that most Psa3 outbreaks from 2014 to 2015 were caused by pollens imported from New Zealand and China for artificial pollination. Most kiwifruit cultivars growing in Korea were infected with Psa3; yellow-fleshed cultivars (Yellow-king, Hort16A, Enza-gold, Zecy-gold, and Haegeum), red-fleshed cultivars (Hongyang and Enza-Red), green-fleshed cultivars (Hayward and Daeheung), and even a kiwiberry (Skinny-green). However, susceptibility to canker differed among cultivars; yellow- and red-fleshed cultivars showed much more severe symptoms compared to the green-fleshed cultivars of kiwifruit and a kiwiberry. PMID:27904461

  9. Engineering canker-resistant plants through CRISPR/Cas9-targeted editing of the susceptibility gene CsLOB1 promoter in citrus.

    PubMed

    Peng, Aihong; Chen, Shanchun; Lei, Tiangang; Xu, Lanzhen; He, Yongrui; Wu, Liu; Yao, Lixiao; Zou, Xiuping

    2017-03-29

    Citrus canker, caused by Xanthomonas citri subsp. citri (Xcc), is severely damaging to the global citrus industry. Targeted editing of host disease-susceptibility genes represents an interesting and potentially durable alternative in plant breeding for resistance. Here, we report improvement of citrus canker resistance through CRISPR/Cas9-targeted modification of the susceptibility gene CsLOB1 promoter in citrus. Wanjincheng orange (Citrus sinensis Osbeck) harbors at least three copies of the CsLOB1(G) allele and one copy of the CsLOB1(-) allele. The promoter of both alleles contains the effector binding element (EBEPthA4 ), which is recognized by the main effector PthA4 of Xcc to activate CsLOB1 expression to promote citrus canker development. Five pCas9/CsLOB1sgRNA constructs were designed to modify the EBEPthA4 of the CsLOB1 promoter in Wanjincheng orange. Among these constructs, mutation rates were 11.5% to 64.7%. Homozygous mutants were generated directly from citrus explants. Sixteen lines that harbored EBEPthA4 modifications were identified from 38 mutant plants. Four mutation lines (S2-5, S2-6, S2-12 and S5-13), in which promoter editing disrupted CsLOB1 induction in response to Xcc infection, showed enhanced resistance to citrus canker compared with the wild type. No canker symptoms were observed in the S2-6 and S5-13 lines. Promoter editing of CsLOB1(G) alone was sufficient to enhance citrus canker resistance in Wanjincheng orange. Deletion of the entire EBEPthA4 sequence from both CsLOB1 alleles conferred a high degree of resistance to citrus canker. The results demonstrate that CRISPR/Cas9-mediated promoter editing of CsLOB1 is an efficient strategy for generation of canker-resistant citrus cultivars. This article is protected by copyright. All rights reserved.

  10. Exposure to Alternaria alternata in US homes is associated with asthma symptoms

    PubMed Central

    Salo, Päivi M.; Arbes, Samuel J.; Sever, Michelle; Jaramillo, Renee; Cohn, Richard D.; London, Stephanie J.; Zeldin, Darryl C.

    2007-01-01

    Background Exposure to the fungus Alternaria alternata is a risk factor for asthma. Few studies have examined Alternaria exposures in indoor environments. Objective We examined whether exposure to A alternata in US homes was associated with asthma-related outcomes. Methods The data for this study were collected as part of the National Survey of Lead and Allergens in Housing. This cross-sectional study surveyed a nationally representative sample of 831 housing units inhabited by 2456 individuals in 75 different locations throughout the United States. An interviewer-administered questionnaire obtained information on demographics, household characteristics, and occupants' health status. Exposure to A alternata was assessed by measuring concentrations of A alternata antigens in vacuumed dust samples using a polyclonal anti–A alternata antibody assay. Dust samples were collected from a bed, a sofa, or a chair, and from bedroom, living room, and kitchen floors. Results Lifetime prevalence of doctor-diagnosed asthma was 11.2%, and 6.9% of the study subjects reported active asthma symptoms in the past 12 months. The prevalence of current symptomatic asthma increased with increasing Alternaria concentrations in US homes; higher levels of A alternata antigens increased the odds of having asthma symptoms in the past year (relative to the lowest tertile, adjusted odds ratio was 1.52, 95% CI, 0.90−2.55 for the 2nd tertile; and 1.84, 95% CI, 1.18−2.85 for the 3rd tertile). Conclusion Exposure to A alternata in US homes is associated with active asthma symptoms. Clinical implications Measures that reduce indoor exposure to A alternata may help control asthma exacerbations. PMID:17030243

  11. Identification of putative TAL effector targets of the citrus canker pathogens shows functional convergence underlying disease development and defense response

    PubMed Central

    2014-01-01

    Background Transcriptional activator-like (TAL) effectors, formerly known as the AvrBs3/PthA protein family, are DNA-binding effectors broadly found in Xanthomonas spp. that transactivate host genes upon injection via the bacterial type three-secretion system. Biologically relevant targets of TAL effectors, i.e. host genes whose induction is vital to establish a compatible interaction, have been reported for xanthomonads that colonize rice and pepper; however, citrus genes modulated by the TAL effectors PthA“s” and PthC“s” of the citrus canker bacteria Xanthomonas citri (Xc) and Xanthomonas aurantifolii pathotype C (XaC), respectively, are poorly characterized. Of particular interest, XaC causes canker disease in its host lemon (Citrus aurantifolia), but triggers a defense response in sweet orange. Results Based on, 1) the TAL effector-DNA binding code, 2) gene expression data of Xc and XaC-infiltrated sweet orange leaves, and 3) citrus hypocotyls transformed with PthA2, PthA4 or PthC1, we have identified a collection of Citrus sinensis genes potentially targeted by Xc and XaC TAL effectors. Our results suggest that similar with other strains of Xanthomonas TAL effectors, PthA2 and PthA4, and PthC1 to some extent, functionally converge. In particular, towards induction of genes involved in the auxin and gibberellin synthesis and response, cell division, and defense response. We also present evidence indicating that the TAL effectors act as transcriptional repressors and that the best scoring predicted DNA targets of PthA“s” and PthC“s” in citrus promoters predominantly overlap with or localize near to TATA boxes of core promoters, supporting the idea that TAL effectors interact with the host basal transcriptional machinery to recruit the RNA pol II and start transcription. Conclusions The identification of PthA“s” and PthC“s” targets, such as the LOB (LATERAL ORGAN BOUNDARY) and CCNBS genes that we report here, is key for the understanding

  12. Rapid and sensitive detection of Citrus Bacterial Canker by loop-mediated isothermal amplification combined with simple visual evaluation methods

    PubMed Central

    2010-01-01

    Background Citrus Bacterial Canker (CBC) is a major, highly contagious disease of citrus plants present in many countries in Asia, Africa and America, but not in the Mediterranean area. There are three types of Citrus Bacterial Canker, named A, B, and C that have different genotypes and posses variation in host range within citrus species. The causative agent for type A CBC is Xanthomonas citri subsp. citri, while Xanthomonas fuscans subsp. aurantifolii, strain B causes type B CBC and Xanthomonas fuscans subsp. aurantifolii strain C causes CBC type C. The early and accurate identification of those bacteria is essential for the protection of the citrus industry. Detection methods based on bacterial isolation, antibodies or polymerase chain reaction (PCR) have been developed previously; however, these approaches may be time consuming, laborious and, in the case of PCR, it requires expensive laboratory equipment. Loop-mediated isothermal amplification (LAMP), which is a novel isothermal DNA amplification technique, is sensitive, specific, fast and requires no specialized laboratory equipment. Results A loop-mediated isothermal amplification assay for the diagnosis of Citrus Bacterial Canker (CBC-LAMP) was developed and evaluated. DNA samples were obtained from infected plants or cultured bacteria. A typical ladder-like pattern on gel electrophoresis was observed in all positive samples in contrast to the negative controls. In addition, amplification products were detected by visual inspection using SYBRGreen and using a lateral flow dipstick, eliminating the need for gel electrophoresis. The sensitivity and specificity of the assay were evaluated in different conditions and using several sample sources which included purified DNA, bacterium culture and infected plant tissue. The sensitivity of the CBC-LAMP was 10 fg of pure Xcc DNA, 5 CFU in culture samples and 18 CFU in samples of infected plant tissue. No cross reaction was observed with DNA of other phytopathogenic

  13. Mycotoxin production in liquid culture and on plants infected with Alternaria spp. isolated from rocket and cabbage.

    PubMed

    Siciliano, Ilenia; Ortu, Giuseppe; Gilardi, Giovanna; Gullino, Maria Lodovica; Garibaldi, Angelo

    2015-03-05

    Fungi belonging to the genus Alternaria are common pathogens of fruit and vegetables with some species able to produce secondary metabolites dangerous to human health. Twenty-eight Alternaria isolates from rocket and cabbage were investigated for their mycotoxin production. Five different Alternaria toxins were extracted from synthetic liquid media and from plant material (cabbage, cultivated rocket, cauliflower). A modified Czapek-Dox medium was used for the in vitro assay. Under these conditions, more than 80% of the isolates showed the ability to produce at least one mycotoxin, generally with higher levels for tenuazonic acid. However, the same isolates analyzed in vivo seemed to lose their ability to produce tenuazonic acid. For the other mycotoxins; alternariol, alternariol monomethyl ether, altenuene and tentoxin a good correlation between in vitro and in vivo production was observed. In vitro assay is a useful tool to predict the possible mycotoxin contamination under field and greenhouse conditions.

  14. Mycotoxin Production in Liquid Culture and on Plants Infected with Alternaria spp. Isolated from Rocket and Cabbage

    PubMed Central

    Siciliano, Ilenia; Ortu, Giuseppe; Gilardi, Giovanna; Gullino, Maria Lodovica; Garibaldi, Angelo

    2015-01-01

    Fungi belonging to the genus Alternaria are common pathogens of fruit and vegetables with some species able to produce secondary metabolites dangerous to human health. Twenty-eight Alternaria isolates from rocket and cabbage were investigated for their mycotoxin production. Five different Alternaria toxins were extracted from synthetic liquid media and from plant material (cabbage, cultivated rocket, cauliflower). A modified Czapek-Dox medium was used for the in vitro assay. Under these conditions, more than 80% of the isolates showed the ability to produce at least one mycotoxin, generally with higher levels for tenuazonic acid. However, the same isolates analyzed in vivo seemed to lose their ability to produce tenuazonic acid. For the other mycotoxins; alternariol, alternariol monomethyl ether, altenuene and tentoxin a good correlation between in vitro and in vivo production was observed. In vitro assay is a useful tool to predict the possible mycotoxin contamination under field and greenhouse conditions. PMID:25751147

  15. Proteomic analysis of Alternaria alternata (Fr.) Keissler responds to COS fumigation.

    PubMed

    Liu, Tao; Li, Li; Qu, Haixia; Zhan, Guoping; Liu, Bo; Wang, Yuejin

    2010-01-01

    Carbonyl sulfide (COS) is a new fumigant which has been a potential alternative to methyl bromide and phosphine in many applications. In this study, we investigated the fungitoxicity of COS towards the pathogen of pear black spot disease Alternaria alternata (Fr.) Keissler (A. alternata). Moreover, proteomic analysis and RT-PCR was performed and our results showed that during the fumigation, the regulation of 21 proteins in protein expression and mRNA accumulation levels is involved, which respond to growth inhibition caused by COS. These results provide new clues for the mechanism of the fungitoxicity of COS.

  16. Alternethanoxins A and B, polycyclic ethanones produced by Alternaria sonchi , potential mycoherbicides for Sonchus arvensis biocontrol.

    PubMed

    Evidente, Antonio; Punzo, Biancavaleria; Andolfi, Anna; Berestetskiy, Alexander; Motta, Andrea

    2009-08-12

    Alternaria sonchi is a fungal pathogen isolated from Sonchus arvensis and proposed as a biocontrol agent of this noxious perennial weed. Different phytotoxic metabolites were isolated from solid culture of the fungus. Two new polycyclic ethanones, named alternethanoxins A and B, were characterized using essentially spectroscopic and chemical methods. Tested by leaf disk-puncture assay on the fungal host plant and a number of nonhost plants, alternethanoxins A and B were shown to be phytotoxic, whereas they did not possess antimicrobial activity tested at 100 microg/disk. Hence, alternethanoxins A and B have potential as nonselective natural herbicides. Some structure-activity relationship observations were also made.

  17. Chemotactic signal transduction and phosphate metabolism as adaptive strategies during citrus canker induction by Xanthomonas citri.

    PubMed

    Moreira, Leandro Marcio; Facincani, Agda Paula; Ferreira, Cristiano Barbalho; Ferreira, Rafael Marine; Ferro, Maria Inês Tiraboshi; Gozzo, Fabio Cesar; de Oliveira, Julio Cezar Franco; Ferro, Jesus Aparecido; Soares, Márcia Regina

    2015-03-01

    The genome of Xanthomonas citri subsp. Citri strain 306 pathotype A (Xac) was completely sequenced more than 10 years; to date, few studies involving functional genomics Xac and its host compatible have been developed, specially related to adaptive events that allow the survival of Xac within the plant. Proteomic analysis of Xac showed that the processes of chemotactic signal transduction and phosphate metabolism are key adaptive strategies during the interaction of a pathogenic bacterium with its plant host. The results also indicate the importance of a group of proteins that may not be directly related to the classical virulence factors, but that are likely fundamental to the success of the initial stages of the infection, such as methyl-accepting chemotaxis protein (Mcp) and phosphate specific transport (Pst). Furthermore, the analysis of the mutant of the gene pstB which codifies to an ABC phosphate transporter subunit revealed a complete absence of citrus canker symptoms when inoculated in compatible hosts. We also conducted an in silico analysis which established the possible network of genes regulated by two-component systems PhoPQ and PhoBR (related to phosphate metabolism), and possible transcriptional factor binding site (TFBS) motifs of regulatory proteins PhoB and PhoP, detaching high degree of conservation of PhoB TFBS in 84 genes of Xac genome. This is the first time that chemotaxis signal transduction and phosphate metabolism were therefore indicated to be fundamental to the process of colonization of plant tissue during the induction of disease associated with Xanthomonas genus bacteria.

  18. Lateral organ boundaries 1 is a disease susceptibility gene for citrus bacterial canker disease.

    PubMed

    Hu, Yang; Zhang, Junli; Jia, Hongge; Sosso, Davide; Li, Ting; Frommer, Wolf B; Yang, Bing; White, Frank F; Wang, Nian; Jones, Jeffrey B

    2014-01-28

    Citrus bacterial canker (CBC) disease occurs worldwide and incurs considerable costs both from control measures and yield losses. Bacteria that cause CBC require one of six known type III transcription activator-like (TAL) effector genes for the characteristic pustule formation at the site of infection. Here, we show that Xanthomonas citri subspecies citri strain Xcc306, with the type III TAL effector gene pthA4 or with the distinct yet biologically equivalent gene pthAw from strain XccA(w), induces two host genes, CsLOB1 and CsSWEET1, in a TAL effector-dependent manner. CsLOB1 is a member of the Lateral Organ Boundaries (LOB) gene family of transcription factors, and CsSWEET1 is a homolog of the SWEET sugar transporter and rice disease susceptibility gene. Both TAL effectors drive expression of CsLOB1 and CsSWEET1 promoter reporter gene fusions when coexpressed in citrus or Nicotiana benthamiana. Artificially designed TAL effectors directed to sequences in the CsLOB1 promoter region, but not the CsSWEET1 promoter, promoted pustule formation and higher bacterial leaf populations. Three additional distinct TAL effector genes, pthA*, pthB, and pthC, also direct pustule formation and expression of CsLOB1. Unlike pthA4 and pthAw, pthB and pthC do not promote the expression of CsSWEET1. CsLOB1 expression was associated with the expression of genes associated with cell expansion. The results indicate that CBC-inciting species of Xanthomonas exploit a single host disease susceptibility gene by altering the expression of an otherwise developmentally regulated gene using any one of a diverse set of TAL effector genes in the pathogen populations.

  19. Lateral organ boundaries 1 is a disease susceptibility gene for citrus bacterial canker disease

    PubMed Central

    Hu, Yang; Zhang, Junli; Jia, Hongge; Sosso, Davide; Li, Ting; Frommer, Wolf B.; Yang, Bing; White, Frank F.; Wang, Nian; Jones, Jeffrey B.

    2014-01-01

    Citrus bacterial canker (CBC) disease occurs worldwide and incurs considerable costs both from control measures and yield losses. Bacteria that cause CBC require one of six known type III transcription activator-like (TAL) effector genes for the characteristic pustule formation at the site of infection. Here, we show that Xanthomonas citri subspecies citri strain Xcc306, with the type III TAL effector gene pthA4 or with the distinct yet biologically equivalent gene pthAw from strain XccAw, induces two host genes, CsLOB1 and CsSWEET1, in a TAL effector-dependent manner. CsLOB1 is a member of the Lateral Organ Boundaries (LOB) gene family of transcription factors, and CsSWEET1 is a homolog of the SWEET sugar transporter and rice disease susceptibility gene. Both TAL effectors drive expression of CsLOB1 and CsSWEET1 promoter reporter gene fusions when coexpressed in citrus or Nicotiana benthamiana. Artificially designed TAL effectors directed to sequences in the CsLOB1 promoter region, but not the CsSWEET1 promoter, promoted pustule formation and higher bacterial leaf populations. Three additional distinct TAL effector genes, pthA*, pthB, and pthC, also direct pustule formation and expression of CsLOB1. Unlike pthA4 and pthAw, pthB and pthC do not promote the expression of CsSWEET1. CsLOB1 expression was associated with the expression of genes associated with cell expansion. The results indicate that CBC-inciting species of Xanthomonas exploit a single host disease susceptibility gene by altering the expression of an otherwise developmentally regulated gene using any one of a diverse set of TAL effector genes in the pathogen populations. PMID:24474801

  20. Bayesian analysis for inference of an emerging epidemic: citrus canker in urban landscapes.

    PubMed

    Neri, Franco M; Cook, Alex R; Gibson, Gavin J; Gottwald, Tim R; Gilligan, Christopher A

    2014-04-01

    Outbreaks of infectious diseases require a rapid response from policy makers. The choice of an adequate level of response relies upon available knowledge of the spatial and temporal parameters governing pathogen spread, affecting, amongst others, the predicted severity of the epidemic. Yet, when a new pathogen is introduced into an alien environment, such information is often lacking or of no use, and epidemiological parameters must be estimated from the first observations of the epidemic. This poses a challenge to epidemiologists: how quickly can the parameters of an emerging disease be estimated? How soon can the future progress of the epidemic be reliably predicted? We investigate these issues using a unique, spatially and temporally resolved dataset for the invasion of a plant disease, Asiatic citrus canker in urban Miami. We use epidemiological models, Bayesian Markov-chain Monte Carlo, and advanced spatial statistical methods to analyse rates and extent of spread of the disease. A rich and complex epidemic behaviour is revealed. The spatial scale of spread is approximately constant over time and can be estimated rapidly with great precision (although the evidence for long-range transmission is inconclusive). In contrast, the rate of infection is characterised by strong monthly fluctuations that we associate with extreme weather events. Uninformed predictions from the early stages of the epidemic, assuming complete ignorance of the future environmental drivers, fail because of the unpredictable variability of the infection rate. Conversely, predictions improve dramatically if we assume prior knowledge of either the main environmental trend, or the main environmental events. A contrast emerges between the high detail attained by modelling in the spatiotemporal description of the epidemic and the bottleneck imposed on epidemic prediction by the limits of meteorological predictability. We argue that identifying such bottlenecks will be a fundamental step in

  1. Bayesian Analysis for Inference of an Emerging Epidemic: Citrus Canker in Urban Landscapes

    PubMed Central

    Neri, Franco M.; Cook, Alex R.; Gibson, Gavin J.; Gottwald, Tim R.; Gilligan, Christopher A.

    2014-01-01

    Outbreaks of infectious diseases require a rapid response from policy makers. The choice of an adequate level of response relies upon available knowledge of the spatial and temporal parameters governing pathogen spread, affecting, amongst others, the predicted severity of the epidemic. Yet, when a new pathogen is introduced into an alien environment, such information is often lacking or of no use, and epidemiological parameters must be estimated from the first observations of the epidemic. This poses a challenge to epidemiologists: how quickly can the parameters of an emerging disease be estimated? How soon can the future progress of the epidemic be reliably predicted? We investigate these issues using a unique, spatially and temporally resolved dataset for the invasion of a plant disease, Asiatic citrus canker in urban Miami. We use epidemiological models, Bayesian Markov-chain Monte Carlo, and advanced spatial statistical methods to analyse rates and extent of spread of the disease. A rich and complex epidemic behaviour is revealed. The spatial scale of spread is approximately constant over time and can be estimated rapidly with great precision (although the evidence for long-range transmission is inconclusive). In contrast, the rate of infection is characterised by strong monthly fluctuations that we associate with extreme weather events. Uninformed predictions from the early stages of the epidemic, assuming complete ignorance of the future environmental drivers, fail because of the unpredictable variability of the infection rate. Conversely, predictions improve dramatically if we assume prior knowledge of either the main environmental trend, or the main environmental events. A contrast emerges between the high detail attained by modelling in the spatiotemporal description of the epidemic and the bottleneck imposed on epidemic prediction by the limits of meteorological predictability. We argue that identifying such bottlenecks will be a fundamental step in

  2. STEM, STEM Education, STEMmania

    ERIC Educational Resources Information Center

    Sanders, Mark

    2009-01-01

    In this article, the author introduces integrative STEM (science, technology, engineering, and/or mathematics) education and discusses the importance of the program. The notion of integrative STEM education includes approaches that explore teaching and learning between/among any two or more of the STEM subject areas, and/or between a STEM subject…

  3. Recovery Plan for Phytophthora kernoviae Causing Bleeding Trunk Cankers, Leaf Blight and Stem Dieback in Trees and Shrubs

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Phytophthora kernoviae, a recently described species of Phytophthora, is an invasive pathogen of forest trees and shrubs such as beech (Fagus sylvatica) and rhododendron (Rhododendron ponticum) that has become established in woodlands and public gardens in Cornwall, United Kingdom. Although the ori...

  4. Biological Control of Alternaria Fruit Rot of Chili by Trichoderma Species under Field Conditions.

    PubMed

    Begum, Most Ferdousi; Rahman, M A; Alam, M Firoz

    2010-06-01

    Trichoderma strains were evaluated under field conditions to assay their efficacy in suppressing Alternaria fruit rot disease and promoting chili plant growth. The experiment was conducted at the Botanical Garden, Rajshahi University, Bangladesh from July 2006 to March 2007. Application of Trichoderma harzianum IMI 392432 significantly (p = 0.05) suppressed the disease compared to Alternaria tenuis (T2) treatment and improved both growth and yield. The treatment T4 (T. harzianum IMI-392432 + A. tenuis) was most effective in reducing disease percentage (72.27%) compared to A. tenuis (T1) treatment. The highest seed germination rate (85.56%) and the highest growth and yield (12.5 g/plant) was also recorded in the same treatment (T4), followed by T5 (T. harzianum IMI-392433 000000 + A. tenuis), T6 (T. harzianum IMI-392434 +A. tenuis), T2 (T. virens IMI-392430 + A. tenuis), and T3 (T. pseudokoningii IMI-392431 +A. tenuis) treatment, while single treatment with A. tenuis significantly decreased these values.

  5. Altering Conidial Dispersal of Alternaria solani by Modifying Microclimate in Tomato Crop Canopy

    PubMed Central

    Jambhulkar, Prashant Prakash; Jambhulkar, Nitiprasad; Meghwal, Madanlal; Ameta, Gauri Shankar

    2016-01-01

    Early blight of tomato caused by Alternaria solani, is responsible for severe yield losses in tomato. The conidia survive on soil surface and old dry lower leaves of the plant and spread when suitable climatic conditions are available. Macroclimatic study reveals that highest inoculum concentration of Alternaria spores appeared in May 2012 to 2013 and lowest concentration during January 2012 to 2013. High night temperature positively correlated and significantly (P < 0.01) involved in conidial spore dispersal and low relative humidity (RH) displayed significant (P < 0.05) but negative correlation with conidial dispersal. The objective of the study was to modify microclimatic conditions of tomato crop canopy which may hamper conidial dispersal and reduce disease severity. We evaluated effect of marigold intercropping and plastic mulching singly and in consortia on A. solani conidial density, tomato leaf damage and microclimatic parameters as compar to tomato alone (T). Tomato-marigold intercropping–plastic mulching treatment (T + M + P) showed 35–39% reduction in disease intensity as compared to tomato alone. When intercropped with tomato, marigold served as barrier to conidial movement and plastic mulching prevented evapotranspiration and reduced the canopy RH that resulted in less germination of A. solani spores. Marigold intercropping and plastic mulching served successfully as physical barrier against conidial dissemination to diminish significantly the tomato foliar damage produced by A. solani. PMID:27904457

  6. Altering Conidial Dispersal of Alternaria solani by Modifying Microclimate in Tomato Crop Canopy.

    PubMed

    Jambhulkar, Prashant Prakash; Jambhulkar, Nitiprasad; Meghwal, Madanlal; Ameta, Gauri Shankar

    2016-12-01

    Early blight of tomato caused by Alternaria solani, is responsible for severe yield losses in tomato. The conidia survive on soil surface and old dry lower leaves of the plant and spread when suitable climatic conditions are available. Macroclimatic study reveals that highest inoculum concentration of Alternaria spores appeared in May 2012 to 2013 and lowest concentration during January 2012 to 2013. High night temperature positively correlated and significantly (P < 0.01) involved in conidial spore dispersal and low relative humidity (RH) displayed significant (P < 0.05) but negative correlation with conidial dispersal. The objective of the study was to modify microclimatic conditions of tomato crop canopy which may hamper conidial dispersal and reduce disease severity. We evaluated effect of marigold intercropping and plastic mulching singly and in consortia on A. solani conidial density, tomato leaf damage and microclimatic parameters as compar to tomato alone (T). Tomato-marigold intercropping-plastic mulching treatment (T + M + P) showed 35-39% reduction in disease intensity as compared to tomato alone. When intercropped with tomato, marigold served as barrier to conidial movement and plastic mulching prevented evapotranspiration and reduced the canopy RH that resulted in less germination of A. solani spores. Marigold intercropping and plastic mulching served successfully as physical barrier against conidial dissemination to diminish significantly the tomato foliar damage produced by A. solani.

  7. Possibility of Alternaria toxins reduction by extrusion processing of whole wheat flour.

    PubMed

    Janić Hajnal, Elizabet; Čolović, Radmilo; Pezo, Lato; Orčić, Dejan; Vukmirović, Đuro; Mastilović, Jasna

    2016-12-15

    This study represents the first report about possibility of reduction of Alternaria toxins in wheat using the extrusion process. Effects of extrusion processing parameters - moisture content (w=16, 20, 24g/100g), feeding rate (q=15, 20, 25kg/h), and screw speed (v=300, 390, 480rpm), on reduction rate of tenuazonic acid (TeA), alternariol (AOH) and alternariol monomethyl ether (AME), in whole wheat flour were investigated. Temperature ranged between 111.1 and 160.8°C, while the absolute pressure was from 0.17 to 0.23MPa. The simultaneous influence of w and v was the most important for TeA reduction (p<0.05), while v and q were the most influential for AOH reduction (p<0.01). Level of AME reduction was mostly influenced by w and v (p<0.10). Optimal parameters for reduction of all three Alternaria toxins were as follows: w=24g/100g, q=25kg/h, v=390rpm, with a reduction of 65.6% for TeA, 87.9% for AOH and 94.5% for AME.

  8. Genome, Proteome and Structure of a T7-Like Bacteriophage of the Kiwifruit Canker Phytopathogen Pseudomonas syringae pv. actinidiae.

    PubMed

    Frampton, Rebekah A; Acedo, Elena Lopez; Young, Vivienne L; Chen, Danni; Tong, Brian; Taylor, Corinda; Easingwood, Richard A; Pitman, Andrew R; Kleffmann, Torsten; Bostina, Mihnea; Fineran, Peter C

    2015-06-24

    Pseudomonas syringae pv. actinidiae is an economically significant pathogen responsible for severe bacterial canker of kiwifruit (Actinidia sp.). Bacteriophages infecting this phytopathogen have potential as biocontrol agents as part of an integrated approach to the management of bacterial canker, and for use as molecular tools to study this bacterium. A variety of bacteriophages were previously isolated that infect P. syringae pv. actinidiae, and their basic properties were characterized to provide a framework for formulation of these phages as biocontrol agents. Here, we have examined in more detail φPsa17, a phage with the capacity to infect a broad range of P. syringae pv. actinidiae strains and the only member of the Podoviridae in this collection. Particle morphology was visualized using cryo-electron microscopy, the genome was sequenced, and its structural proteins were analysed using shotgun proteomics. These studies demonstrated that φPsa17 has a 40,525 bp genome, is a member of the T7likevirus genus and is closely related to the pseudomonad phages φPSA2 and gh-1. Eleven structural proteins (one scaffolding) were detected by proteomics and φPsa17 has a capsid of approximately 60 nm in diameter. No genes indicative of a lysogenic lifecycle were identified, suggesting the phage is obligately lytic. These features indicate that φPsa17 may be suitable for formulation as a biocontrol agent of P. syringae pv. actinidiae.

  9. Variables Associated with Severity of Bacterial Canker and Wilt Caused by Clavibacter michiganensis subsp. michiganensis in Tomato Greenhouses.

    PubMed

    Blank, L; Cohen, Y; Borenstein, M; Shulhani, R; Lofthouse, M; Sofer, M; Shtienberg, D

    2016-03-01

    Clavibacter michiganensis subsp. michiganensis, the causal agent of bacterial canker and wilt of tomato, is considered to be one of the most important bacterial pathogens worldwide. In the year 2000 there was an increase in the number of infected greenhouses and in the severity of the disease in Israel. As part of the effort to cope with the disease, a comprehensive survey was conducted. Scouts recorded disease severity monthly in 681 production units. At the end of the season the scouts met with the growers and together recorded relevant details about the crop and cultural practices employed. The results suggested an absence of anisotropy pattern in the study region. Global Moran's I analysis showed that disease severity had significant spatial autocorrelation. The strongest spatial autocorrelation occurred within a 1,500 m neighborhood, which is comparable to the distance between production units maintained by one grower (Farm). Next, we tested three groups of variables including or excluding the Farm as a variable. When the Farm was included the explained variation increased in all the studied models. Overall, results of this study demonstrate that the most influential factor on bacterial canker severity was the Farm. This variable probably encompasses variation in experience, differences in agricultural practices between growers, and the quality of implementation of management practices.

  10. Exogenous treatment with salicylic acid attenuates occurrence of citrus canker in susceptible navel orange (Citrus sinensis Osbeck).

    PubMed

    Wang, Yin; Liu, Ji-Hong

    2012-08-15

    Citrus canker caused by Xanthomonas axonopodis pv. citri (Xac) is a devastating bacterial disease threatening the citrus industry. Salicylic acid (SA) plays a key role in plant defense response to biotic stress, but information is scarce concerning the application of SA to enhancing Xac resistance. In the present research attempts were made to investigate how exogenous application of SA influenced canker disease outbreak in navel orange (Citrus sinensis). Exogenously applied SA at 0.25 mM significantly enhanced the endogenous free and bound SA, particularly the latter. Upon exposure to Xac, lower disease incidence rate and smaller lesion sites were observed in the samples pre-treated with SA, accompanied by repression of bacterial growth at the lesion sites. Concurrent with the augmented disease resistance, SA-treated leaves had higher H₂O₂ level and smaller stomata apertures before or after Xac infection when compared with their counterparts pre-treated with water (control). SA treatment elevated the activities of phenylalanine ammonia-lyase and β-1,3-glucanase, but only the latter was higher in the SA-treated samples after Xac infection. In addition, mRNA levels of two pathogenesis-related genes, CsCHI and CsPR4A, were higher in the SA-treated samples relative to the control. Taken together, our results strongly suggest that the exogenously applied SA has evoked a cascade of physiological and molecular events that function singly or in concert to confer resistance to Xac invasion.

  11. Wind speed effects on the quantity of Xanthomonas citri subsp. citri dispersed downwind from canopies of grapefruit trees infected with citrus canker

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The epidemic of citrus canker (Xanthomonas citri subsp. citri, Xcc) in Florida continues to expand since termination of the eradication program in 2006. Storms are known to be associated with disease spread, but little information exists on the interaction of fundamental physical and biological proc...

  12. Fusicoccum arbuti sp. nov. causing cankers on pacific madrone in western North America with notes on Fusicoccum dimidiatum, the correct name for Scytalidium dimidiatum and Nattrassia mangiferae.

    PubMed

    Farr, David F; Elliott, Marianne; Rossman, Amy Y; Edmonds, Robert L

    2005-01-01

    Pacific madrone (Arbutus menziesii) is a broadleaf evergreen tree native to western North America that has been in decline for the past 30 years. A fungus has been isolated and was verified as the cause of cankers on dying trees. It was determined to belong in the genus Fusicoccum, an asexual state of Botryosphaeria. This genus in both its sexual and asexual states commonly causes canker diseases of deciduous woody plants. Using morphological and molecular data the fungus causing cankers on Pacific madrone is characterized, described and illustrated as a new species of Fusicoccum, F. arbuti D.F. Farr & M. Elliott sp. nov. No sexual state is known for F. arbuti. Evidence from the literature, cultures and specimens suggests that F. arbuti, often mistakenly identified as Nattrassia mangiferae, has been causing madrone canker since at least 1968. Authentic isolates of Nattrassia mangiferae as the synanamorph Scytalidium dimidiatum were sequenced and determined to be different from Fusicoccum arbuti and to belong in Botryosphaeria/Fusicoccum. In addition to molecular sequence data, the morphology of the pycnidial and arthric conidial states of Nattrassia mangiferae/ Scytalidium dimidiatum resembles that of Fusicoccum. Therefore the correct name for Nattrassia mangiferae and its numerous synonyms (Dothiorella mangiferae, Torula dimidata, Scytilidium dimidiatum, Fusicoccum eucalypti, Hendersonula toruloidea, H. cypria, Exosporina fawcetii, H. agathidia, and S. lignicola) is Fusicoccum dimidiatum (Penz.) D.F. Farr, comb. nov.

  13. Characterization of Cytospora isolates from wood cankers of declining grapevine in North America, with the descriptions of two new Cytospora species

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Cytospora species are ubiquitous pathogens of woody plants, causing dieback and wood cankers in numerous perennial hosts, including agronomic crops (e.g., Prunus), timber trees (e.g., Eucalyptus), and riparian hosts (e.g., Salix). Cytospora chrysosperma, C. cincta, and C. leucostoma have been report...

  14. Processess involved in the dispersal of Xanthomonas citri pv. citri from canker-infected citrus canopies, and in the infection of citrus foliage

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus canker (Xanthomonas citri subsp. citri, Xcc) is now considered endemic in Florida, and epidemics result in yield loss and market penalties both in Florida and elsewhere, where the pathogen occurs and susceptible citrus is cultivated. The bacterium is dispersed in rain splash, and storms with...

  15. Processes involved in the dispersal of Xanthomonas citri pv. citri from canker-infectd citrus canopies, and in the infection of citrus foliage

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus canker (Xanthomonas citri subsp. citri, Xcc) is now considered endemic in Florida, and epidemics result in yield loss and market penalties both in Florida, and elsewhere where the pathogen occurs, and susceptible citrus is cultivated. The bacterium is dispersed in rain splash, and storms wit...

  16. Complete Genome Sequences of Six Copper-Resistant Xanthomonas citri pv. citri Strains Causing Asiatic Citrus Canker, Obtained Using Long-Read Technology

    PubMed Central

    Richard, Damien; Boyer, Claudine; Vernière, Christian; Canteros, Blanca I.; Lefeuvre, Pierre

    2017-01-01

    ABSTRACT The gammaproteobacterium Xanthomonas citri pv. citri causes Asiatic citrus canker. Pathotype A strains have a broad host range, which includes most commercial citrus species, and they cause important economic losses worldwide. Control often relies on frequent copper sprays. We present here the complete genomes of six X. citri pv. citri copper-resistant strains. PMID:28336584

  17. Development of consumer-friendly transgenic citrus plants with potential broad spectrum resistance to HLB, Citrus canker, Phytopthora and other exotic diseases

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The second year of this CRB funded project has started, which is focused on the development of citrus cultivars that exhibit disease resistance to multiple pathogens such as HLB, Phytophthora and citrus canker diseases. We are using precise genetic engineering to introduce into disease susceptible ...

  18. The epidemiological significance of post-packinghouse survival of Xanthomonas citri ssp. citri for dissemination of Asiatic citrus canker via infected fruit

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The risk of introduction of Xanthomonas citri spp. citri (Xcc) to new, unaffected citrus producing areas is a major concern for those citrus industries attempting to remain free of citrus canker. Citrus fruit, as a potential pathway for Xcc to enter and become established in these areas, is assumed...

  19. Two novel Fusarium species that cause canker disease of Sichuan pepper (Zanthoxylum bungeanum Maxim.) in northern China form a novel clade with F. torreyae

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Canker disease of prickly ash (Zanthoxylum bungeanum) has caused a decline in the production of this economically important spice in northern China in the past 25 y. To identify the etiological agent, 38 fungal isolates were recovered from symptomatic tissues from trees in five provinces in China. T...

  20. Stem Cells

    MedlinePlus

    Stem cells are cells with the potential to develop into many different types of cells in the body. ... the body. There are two main types of stem cells: embryonic stem cells and adult stem cells. Stem ...

  1. Characterization of Alternaria isolates from the infectoria species-group and a new taxon from Arrhenatherum, Pseudoalternaria arrhenatheria sp. nov

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The infectoria species-group within the genus Alternaria was originally conceived by Simmons in 1993 and was based upon common morphological characteristics that included the development of conidial chains with primary, secondary, and tertiary branching resulting in substantial three-dimensional com...

  2. Nonhost-specific phytotoxicity of the polyketide-derived toxin solanapyrone A produced by Ascochyta rabiei and Alternaria solani

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Solanapyrone A is a polyketide-derived metabolite produced by Ascochyta rabiei and Alternaria solani, which are the most destructive necrotrophic pathogens of chickpea and potato/tomato, respectively. They belong to the Order Pleosporales within the Class Dothideomycetes, but are phylogenetically di...

  3. Phytotoxin solanapyrone A produced by Ascochyta rabiei and Alternaria solani is nonessential for pathogenicity, but likely plays ecological roles

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Ascochyta rabiei and Alternaria solani, causal agents of chickpea and potato blights respectively, produce the same phytotoxin solanapyrone A (SolA).The toxicity of SolA to plants has been documented, but its role in pathogenicity has not been investigated. In this study, we generated solanapyrone-d...

  4. Inducible expression of Bs2 R gene from Capsicum chacoense in sweet orange (Citrus sinensis L. Osbeck) confers enhanced resistance to citrus canker disease.

    PubMed

    Sendín, Lorena Noelia; Orce, Ingrid Georgina; Gómez, Rocío Liliana; Enrique, Ramón; Grellet Bournonville, Carlos Froilán; Noguera, Aldo Sergio; Vojnov, Adrián Alberto; Marano, María Rosa; Castagnaro, Atilio Pedro; Filippone, María Paula

    2017-04-01

    Transgenic expression of the pepper Bs2 gene confers resistance to Xanthomonas campestris pv. vesicatoria (Xcv) pathogenic strains which contain the avrBs2 avirulence gene in susceptible pepper and tomato varieties. The avrBs2 gene is highly conserved among members of the Xanthomonas genus, and the avrBs2 of Xcv shares 96% homology with the avrBs2 of Xanthomonas citri subsp. citri (Xcc), the causal agent of citrus canker disease. A previous study showed that the transient expression of pepper Bs2 in lemon leaves reduced canker formation and induced plant defence mechanisms. In this work, the effect of the stable expression of Bs2 gene on citrus canker resistance was evaluated in transgenic plants of Citrus sinensis cv. Pineapple. Interestingly, Agrobacterium-mediated transformation of epicotyls was unsuccessful when a constitutive promoter (2× CaMV 35S) was used in the plasmid construction, but seven transgenic lines were obtained with a genetic construction harbouring Bs2 under the control of a pathogen-inducible promoter, from glutathione S-transferase gene from potato. A reduction of disease symptoms of up to 70% was observed in transgenic lines expressing Bs2 with respect to non-transformed control plants. This reduction was directly dependent on the Xcc avrBs2 gene since no effect was observed when a mutant strain of Xcc with a disruption in avrBs2 gene was used for inoculations. Additionally, a canker symptom reduction was correlated with levels of the Bs2 expression in transgenic plants, as assessed by real-time qPCR, and accompanied by the production of reactive oxygen species. These results indicate that the pepper Bs2 resistance gene is also functional in a family other than the Solanaceae, and could be considered for canker control.

  5. Responsiveness of different citrus genotypes to the Xanthomonas citri ssp. citri-derived pathogen-associated molecular pattern (PAMP) flg22 correlates with resistance to citrus canker.

    PubMed

    Shi, Qingchun; Febres, Vicente J; Jones, Jeffrey B; Moore, Gloria A

    2015-06-01

    The bacterial agent of citrus canker disease (Xanthomonas citri ssp. citri, Xcc) has caused tremendous economic losses to the citrus industry around the world. Pathogen-associated molecular pattern (PAMP)-triggered immunity (PTI) is important to plant immunity. In this study, we compared the defence responses of citrus canker-resistant and citrus canker-susceptible genotypes to the Xcc-derived PAMP flg22 (Xflg22) by analysing the expression of 20 citrus defence-associated genes. We showed that, in the most resistant genotype, 'Nagami' kumquat, there was significant induction of several defence genes (EDS1, NDR1, PBS1, RAR1, SGT1, PAL1, NPR2 and NPR3) as early as 6 h and up to 72 h after Xflg22 treatment. At the other end of the spectrum, highly susceptible 'Duncan' grapefruit showed no induction of the same defence genes, even 120 h after treatment. Citrus genotypes with partial levels of resistance showed intermediate levels of transcriptional reprogramming that correlated with their resistance level. Xflg22 also triggered a rapid oxidative burst in all genotypes which was higher and accompanied by the induction of PTI marker genes (WRKY22 and GST1) only in the more resistant genotypes. Pretreatment with Xflg22 prior to Xcc inoculation inhibited bacterial growth in kumquat, but not in grapefruit. A flagellin-deficient Xcc strain (XccΔfliC) showed greater growth increase relative to wild-type Xcc in kumquat than in grapefruit. Taken together, our results indicate that Xflg22 initiates strong PTI in canker-resistant genotypes, but not in susceptible ones, and that a robust induction of PTI is an important component of citrus resistance to canker.

  6. Use of a climatic rule and fuzzy sets to model geographic distribution of climatic risk for European canker (Neonectria galligena) of apple.

    PubMed

    Kim, Kwang Soo; Beresford, Robert M

    2012-02-01

    A rule-based model was developed to assess climatic risk of European canker (Neonectria galligena), which is a major disease of apple in some temperate zones. A descriptive rule was derived from published observations on climatic conditions favorable for European canker development. Fuzzy set theory was used to evaluate the descriptive rule quantitatively. The amount and frequency of rainfall and the average number of hours between 11 and 16°C/day were used as input variables whose values were matched with terms in the rule, e.g., 'high' or 'low'. The degree of a term, e.g., the state of being high or low, to a given input value was determined using a membership function that converts an input value to a number between 0 and 1. The rule was evaluated by combining the degree of the terms associated with monthly climate data. Monthly risk index values derived using the rule were combined for pairs of consecutive months over 12 months. The annual risk of European canker development was represented by the maximum risk index value for 2 months combined. The membership function parameters were adjusted iteratively to achieve a specified level of risk at Talca (Chile), Loughgall (Northern Ireland), East Malling (UK), and Sebastopol (USA), where European canker risk was known. The rule-based model was validated with data collected from Canada, Ecuador, Denmark, Germany, Norway, Poland, Sweden, the Netherlands, New Zealand, and the Pacific Northwest (USA), where European canker has been reported to occur. In these validation areas, the model's risk prediction agreed with reports of disease occurrence. The rule-based model also predicted high risk areas more reliably than the climate matching model, CLIMEX, which relies on correlations between the spatial distribution of a species and climatic conditions. The combination of a climatic rule and fuzzy sets could be used for other applications where prediction of the geographic distribution of organisms is required for

  7. Transformation of sweet orange [Citrus sinensis (L.) Osbeck] with pthA-nls for acquiring resistance to citrus canker disease.

    PubMed

    Yang, Li; Hu, Chunhua; Li, Na; Zhang, Jiayin; Yan, Jiawen; Deng, Ziniu

    2011-01-01

    The COOH terminal of pthA encoding three nuclear localizing signals (NLS) was amplified by polymerase chain reaction (PCR) from the plasmid of Xanthomonas axonopodis pv. citri, the pathogen of citrus canker disease. Then the sense and antisense strands of the nls were cloned into pBI121 vector. pthA-nls driven by the CaMV35 s promoter was transferred into sweet orange via Agrobacterium -mediated transformation. Successful integration was confirmed by PCR and Southern blotting, and 12 sense-nls (nls (+)) and 9 antisense-nls (nls (-)) transgenic clones were obtained. The expression of nls fragment was analyzed by RT-PCR, Real time q-PCR and Western blotting, in which the specific NLS protein was detected only in nls (+) transgenic clones. In an in vitro assay, when pin-puncture inoculation was performed with 2.5 × 10(7) cfu/ml of bacterial solution, the nls (+) transgenic clones showed no typical lesion development, while typical symptoms were observed in the wild types and the nls (-) transgenic clones. In vivo assay results indicated that the nls (+) transgenic clones showed less disease incidence, in comparison with the wild types and the nls (-) transgenic clones, when pin-puncture inoculation was performed with 10(4)-10(5) cfu/ml. The minimum disease incidence was 23.3% for 'Sucarri' sweet orange and 33.3% for 'Bingtang' sweet orange. When 10(4)-10(7) cfu/ml of pathogen was spray inoculated, the nls (+) transgenic clones did not show any symptom, and even the concentration raised to 10(9) cfu/ml, the disease incidence was 20-80%, while the wild types and the nls (-) transgenic clones had 100% disease development with whatever concentration of inoculum. Two transgenic clones were confirmed to be resistant to citrus canker disease in the repeated inoculation. The results suggested that the transformation of nls sense strands may offer an effective way to acquire resistance to citrus canker disease.

  8. [Production and properties of a beta-galactosidase preparation from Alternaria tenuis].

    PubMed

    Zagustina, N A; Tikhomirova, A S; Rafalovskaia, T Ia; Iangol', L M; Gracheva, A Iu

    1975-01-01

    Different methods of the preparation of fungal beta-galactosidase from the 72-hour culture of Alternaria tenuis were tested: lyophilization of the culture liquid, precipitation with ethanol, acetone, ammonium sulphate. Optimal results were obtained with precipitation by 1.5 acetone volume. Studies of the properties of fungal beta-galactosidase demonstrated that the preparation retained its activity during 22 month storage at 5 degrees C. The fungal preparation had pH optimum at a more acidic zone (4.2 versus 6.9), was active in a wider pH range 2.8-5.7 and 6.2-7.5), had a much higher temperature optimum (65 degrees and 30 degrees) and better thermostability as compared with the yeast preparation. Data on other properties of the preparation are presented.

  9. First Report of Leaf Spot Caused by Alternaria tenuissima on Black Chokeberry (Aronia melanocarpa) in Korea.

    PubMed

    Wee, Jung-In; Park, Jong-Han; Back, Chang-Gi; You, Young-Hyun; Chang, Taehyun

    2016-09-01

    In July 2015, diseased leaves of black chokeberry (Aronia melanocarpa) were observed in Danyang and Gochang, Korea. The symptoms appeared as circular or irregular brown leaf spots, from which Alternaria tenuissima was isolated. The isolates were cultured on potato dextrose agar, and their morphological characteristics were observed under a light microscope. The colonies were whitish to ash colored. The pathogenicity test on healthy black chokeberry leaves produced circular brown spots, in line with the original symptoms. Molecular analyses of the ITS, GPD, RPB2, and TEF genes were conducted to confirm the identity of the pathogen. The phylogeny of the multi-gene sequences indicated that the causal agent was A. tenuissima. This study is the first report of A. tenuissima leaf spot on black chokeberry (A. melanocarpa).

  10. First Report of Leaf Spot Caused by Alternaria tenuissima on Black Chokeberry (Aronia melanocarpa) in Korea

    PubMed Central

    Wee, Jung-In; Park, Jong-Han; Back, Chang-Gi; You, Young-Hyun

    2016-01-01

    In July 2015, diseased leaves of black chokeberry (Aronia melanocarpa) were observed in Danyang and Gochang, Korea. The symptoms appeared as circular or irregular brown leaf spots, from which Alternaria tenuissima was isolated. The isolates were cultured on potato dextrose agar, and their morphological characteristics were observed under a light microscope. The colonies were whitish to ash colored. The pathogenicity test on healthy black chokeberry leaves produced circular brown spots, in line with the original symptoms. Molecular analyses of the ITS, GPD, RPB2, and TEF genes were conducted to confirm the identity of the pathogen. The phylogeny of the multi-gene sequences indicated that the causal agent was A. tenuissima. This study is the first report of A. tenuissima leaf spot on black chokeberry (A. melanocarpa). PMID:27790070

  11. First Report of Foliar Blight on Dendropanax morbifera Caused by Alternaria panax.

    PubMed

    Deng, Jian Xin; Kim, Chang Sun; Oh, Eun Sung; Yu, Seung Hun

    2010-12-01

    Leaf spot and blight disease was observed on two-year-old seedlings of Dendropanax morbifera (Korean name: Hwangchil tree) during July of 2008 in Jindo Island, Korea. Symptoms included yellow-brown to dark brown irregularly enlarged spots frequently located along the veins of leaves. The lesions were often surrounded by chlorotic haloes. Severe leaf blight and subsequent defoliation occurred when conditions favored disease outbreak. The causal organism of the disease was identified as Alternaria panax based on morphological characteristics and sequence analysis of the internal transcribed spacer region of rDNA. A. panax isolates induced leaf spots and blight symptoms not only on D. morbifera but also on the other members of Araliaceae tested. This is the first report of foliar blight caused by A. panax on D. morbifera.

  12. Transcriptomics Analysis of Apple Leaves in Response to Alternaria alternata Apple Pathotype Infection

    PubMed Central

    Zhu, Longming; Ni, Weichen; Liu, Shuai; Cai, Binhua; Xing, Han; Wang, Sanhong

    2017-01-01

    Alternaria blotch disease of apple (Malus × domestica Borkh.), caused by the apple pathotype of Alternaria alternata, is one of the most serious fungal diseases to affect apples. To develop an understanding of how apples respond to A. alternata apple pathotype (AAAP) infection, we examined the host transcript accumulation over the period between 0 and 72 h post AAAP inoculation. Large-scale gene expression analysis was conducted of the compatible interaction between “Starking Delicious” apple cultivar and AAAP using RNA-Seq and digital gene expression (DGE) profiling methods. Our results show that a total of 9080 differentially expressed genes (DEGs) were detected (>two-fold and FDR < 0.001) by RNA-Seq. During the early phase of infection, 12 h post inoculation (HPI), AAAP exhibited limited fungal development and little change in the transcript accumulation status (950 DEGs). During the intermediate phase of infection, the period between 18 and 36 HPI, increased fungal development, active infection, and increased transcript accumulation were detected (4111 and 3838 DEGs detected at each time point, respectively). The majority of DEGs were detected by 72 HPI, suggesting that this is an important time point in the response of apples' AAAP infection. Subsequent gene ontology (GO) and pathway enrichment analyses showed that DEGs are predominately involved in biological processes and metabolic pathways; results showed that almost gene associated with photosynthesis, oxidation-reduction were down-regulated, while transcription factors (i.e., WRKY, MYB, NAC, and Hsf) and DEGs involved in cell wall modification, defense signaling, the synthesis of defense-related metabolites, including pathogenesis-related (PRs) genes and phenylpropanoid/cyanoamino acid /flavonoid biosynthesis, were activated during this process. Our study also suggested that the cell wall defensive vulnerability and the down-regulation of most PRs and HSP70s in “Starking Delicious” following AAAP

  13. Alternaria alternata allergens: Markers of exposure, phylogeny and risk of fungi-induced respiratory allergy.

    PubMed

    Gabriel, Marta F; Postigo, Idoia; Tomaz, Cândida T; Martínez, Jorge

    2016-01-01

    Alternaria alternata spores are considered a well-known biological contaminant and a very common potent aeroallergen source that is found in environmental samples. The most intense exposure to A. alternata allergens is likely to occur outdoors; however, Alternaria and other allergenic fungi can colonize in indoor environments and thereby increase the fungal aeroallergen exposure levels. A consequence of human exposure to fungal aeroallergens, sensitization to A. alternata, has been unequivocally associated with increased asthma severity. Among allergenic proteins described in this fungal specie, the major allergen, Alt a 1, has been reported as the main elicitor of airborne allergies in patients affected by a mold allergy and considered a marker of primary sensitization to A. alternata. Moreover, A. alternata sensitization seems to be a triggering factor in the development of poly-sensitization, most likely because of the capability of A. alternata to produce, in addition to Alt a 1, a broad and complex array of cross-reactive allergens that present homologs in several other allergenic sources. The study and understanding of A. alternata allergen information may be the key to explaining why sensitization to A. alternata is a risk factor for asthma and also why the severity of asthma is associated to this mold. Compared to other common environmental allergenic sources, such as pollens and dust mites, fungi are reported to be neglected and underestimated. The rise of the A. alternata allergy has enabled more research into the role of this fungal specie and its allergenic components in the induction of IgE-mediated respiratory diseases. Indeed, recent research on the identification and characterization of A. alternata allergens has allowed for the consideration of new perspectives in the categorization of allergenic molds, assessment of exposure and diagnosis of fungi-induced allergies.

  14. A nonribosomal peptide synthetase mediates siderophore production and virulence in the citrus fungal pathogen Alternaria alternata.

    PubMed

    Chen, Li-Hung; Lin, Ching-Hsuan; Chung, Kuang-Ren

    2013-06-01

    Alternaria species produce and excrete dimethyl coprogen siderophores to acquire iron. The Alternaria alternata gene AaNPS6, encoding a polypeptide analogous to fungal nonribosomal peptide synthetases, was found to be required for the production of siderophores and virulence on citrus. Siderophores purified from culture filtrates of the wild-type strain did not induce any phytotoxicity on the leaves of citrus. Fungal strains lacking AaNPS6 produced little or no detectable extracellular siderophores and displayed an increased sensitivity to H₂O₂, superoxide-generating compounds (KO₂ and menadione) and iron depletion. Δnps6 mutants were also defective for the production of melanin and conidia. The introduction of a wild-type AaNPS6 under the control of its endogenous promoter to a Δnps6 null mutant at least partially restored siderophore production and virulence to citrus, demonstrating a functional link between iron uptake and fungal pathogenesis. Elevated sensitivity to H₂O₂, seen for the Δnps6 null strain could be relieved by exogenous application of ferric iron. The expression of the AaNPS6 gene was highly up-regulated under low-iron conditions and apparently controlled by the redox-responsive yeast transcriptional regulator YAP1. Hence, the maintenance of iron homeostasis via siderophore-mediated iron uptake also plays an important role in resistance to toxic reactive oxygen species (ROS). Our results demonstrate further the critical role of ROS detoxification for the pathogenicity of A. alternata in citrus.

  15. Agrobacterium tumefaciens-mediated transformation of the causative agent of Valsa canker of apple tree Valsa mali var. mali.

    PubMed

    Hu, Yang; Dai, Qingqing; Liu, Yangyang; Yang, Zhe; Song, Na; Gao, Xiaoning; Voegele, Ralf Thomas; Kang, Zhensheng; Huang, Lili

    2014-06-01

    Valsa mali var. mali (Vmm), which is the causative agent of Valsa canker of apple tree, causes heavy damage to apple production in eastern Asia. In this article, we report Agrobacterium tumefaciens-mediated transformation (ATMT) of Vmm and expression of gfp (green fluorescent protein) in this fungus. The transformation system was optimized to a transformation efficiency of approximately 150 transformants/10(6) conidia, and a library containing over 4,000 transformants was generated. The tested transformants were mitotically stable. One hundred percent hph (hygromycin B phosphotransferase) integration into Vmm was identified by PCR and five single-copy integration of T-DNA was detected in the eighteen transformants by Southern blot. To our knowledge, this is the first report of ATMT of Vmm. Furthermore, this library has been used to identify genes involved in the virulence of the pathogen, and the transformation system may also be useful to the transformation of other species of the genus Valsa.

  16. Modifications of Xanthomonas axonopodis pv. citri lipopolysaccharide affect the basal response and the virulence process during citrus canker.

    PubMed

    Petrocelli, Silvana; Tondo, María Laura; Daurelio, Lucas D; Orellano, Elena G

    2012-01-01

    Xanthomonas axonopodis pv. citri (Xac) is the phytopathogen responsible for citrus canker, one of the most devastating citrus diseases in the world. A broad range of pathogens is recognized by plants through so-called pathogen-associated molecular patterns (PAMPs), which are highly conserved fragments of pathogenic molecules. In plant pathogenic bacteria, lipopolisaccharyde (LPS) is considered a virulence factor and it is being recognized as a PAMP. The study of the participation of Xac LPS in citrus canker establishment could help to understand the molecular bases of this disease. In the present work we investigated the role of Xac LPS in bacterial virulence and in basal defense during the interaction with host and non host plants. We analyzed physiological features of Xac mutants in LPS biosynthesis genes (wzt and rfb303) and the effect of these mutations on the interaction with orange and tobacco plants. Xac mutants showed an increased sensitivity to external stresses and differences in bacterial motilities, in vivo and in vitro adhesion and biofilm formation. Changes in the expression levels of the LPS biosynthesis genes were observed in a medium that mimics the plant environment. Xacwzt exhibited reduced virulence in host plants compared to Xac wild-type and Xacrfb303. However, both mutant strains produced a lower increase in the expression levels of host plant defense-related genes respect to the parental strain. In addition, Xac LPS mutants were not able to generate HR during the incompatible interaction with tobacco plants. Our findings indicate that the structural modifications of Xac LPS impinge on other physiological attributes and lead to a reduction in bacterial virulence. On the other hand, Xac LPS has a role in the activation of basal defense in host and non host plants.

  17. Air pollution by allergenic spores of the genus Alternaria in the air of central and eastern Europe.

    PubMed

    Kasprzyk, Idalia; Rodinkova, Victoria; Šaulienė, Ingrida; Ritenberga, Olga; Grinn-Gofron, Agnieszka; Nowak, Malgorzata; Sulborska, Aneta; Kaczmarek, Joanna; Weryszko-Chmielewska, Elzbieta; Bilous, Elena; Jedryczka, Malgorzata

    2015-06-01

    Spores of the genus Alternaria belong to one of the most prevailing constituents of the air in all regions of the world. They form infectious inoculum of numerous plant species as well as severe inhaled allergies. The aim of this study was to compare the biological pollution with Alternaria spores of the air of 12 cities located in central and eastern Europe. The experiment was done in 2010 and it covered the territory of Latvia (LV), Lithuania (LT), Poland (PL) and Ukraine (UA). The spores were counted using an identical method and standard equipment (7-day Lanzoni volumetric sampler) followed by extensive statistical calculations. The timing of the day of maximum concentration changed mainly along the N-S direction and had a positive correlation with latitude. The most important factor determining the increase in Alternaria spore concentration was the temperature, whereas other weather parameters were not related or of low significance. Regardless of geographical location, the first phase of the season (0-0.9 % of Alternaria spores in the air) was the longest (up to 60 days) and the last (97.5 to 99 %) was the shortest (22 days or less). The means of daily concentrations of Alternaria spores ranged from 11 spores m(-3) in Klaipeda (LT, Baltic Sea coast) to 187 in Poznan (west PL, agricultural plain). The threshold value of 80 spores m(-3) that triggers the first allergy symptoms was exceeded in 8 to 86 days (Vinnitsa, UA, temperate continental, forest-steppes region). There were considerable differences between the highest number of spores per cubic metre of air, varying from 139 in the north (Klaipeda, LT) to 2,295 in central west (Poznan, PL). The biological pollution by Alternaria spores in several places of central and eastern Europe was high; the number of days exceeding the threshold value of 300 spores m(-3) connected with serious health problems of atopic people ranged from 0 to 1 on the north (LV, LT) to 29 in central west (Poznan, PL).

  18. Antibacterial Activity of Cinnamaldehyde and Estragole Extracted from Plant Essential Oils against Pseudomonas syringae pv. actinidiae Causing Bacterial Canker Disease in Kiwifruit

    PubMed Central

    Song, Yu-Rim; Choi, Min-Seon; Choi, Geun-Won; Park, Il-Kwon; Oh, Chang-Sik

    2016-01-01

    Pseudomonas syringae pv. actinidiae (Psa) causes bacterial canker disease in kiwifruit. Antibacterial activity of plant essential oils (PEOs) originating from 49 plant species were tested against Psa by a vapor diffusion and a liquid culture assays. The five PEOs from Pimenta racemosa, P. dioica, Melaleuca linariifolia, M. cajuputii, and Cinnamomum cassia efficiently inhibited Psa growth by either assays. Among their major components, estragole, eugenol, and methyl eugenol showed significant antibacterial activity by only the liquid culture assay, while cinnamaldehyde exhibited antibacterial activity by both assays. The minimum inhibitory concentrations (MICs) of estragole and cinnamaldehyde by the liquid culture assay were 1,250 and 2,500 ppm, respectively. The MIC of cinnamaldehyde by the vapor diffusion assay was 5,000 ppm. Based on the formation of clear zones or the decrease of optical density caused by these compounds, they might kill the bacterial cells and this feature might be useful for managing the bacterial canker disease in kiwifruit. PMID:27493612

  19. Alternaria-Associated Fungus Ball of Orbit Nose and Paranasal Sinuses: Case Report of a Rare Clinical Entity.

    PubMed

    Pesic, Zoran; Otasevic, Suzana; Mihailovic, Dragan; Petrovic, Sladjana; Arsic-Arsenijevic, Valentina; Stojanov, Dragan; Petrovic, Milica

    2015-08-01

    Alternaria-associated fungus ball of maxillar, ethmoidal paranasal sinuses, nasal cavity and orbit with bone erosion is extremely rare. Till recently, only two cases of this infection in immune competitive patients have been reported. We are herein describing the case of immune-competent woman who suffered of nasal congestion for 10 years. Patient was treated for tumor-like lesion in right maxillar sinus, where propagation in right nose cavity, right ethmoidal cells and right orbita was present. The organism that was seen in surgical removal of fungal debris by histological study, in using mycological testing, was proven as Alternaria alternata. Combination of surgical intervention and treatment with itraconazole eradicated fungal infection, and the disease was not relapsed in follow-up period of 2 years.

  20. Importance of house dust mite and Alternaria allergens in childhood asthma: an epidemiological study in two climatic regions of Australia.

    PubMed

    Peat, J K; Tovey, E; Mellis, C M; Leeder, S R; Woolcock, A J

    1993-10-01

    The relation of house dust mite allergen levels to asthma and allergy was examined in two population samples of children aged 8-11 years in northern New South Wales. We studied 805 children in Lismore (a hot, humid, coastal region) and 770 in Moree/Narrabri (a hot, dry inland region). Respiratory symptoms were measured by questionnaire, bronchial hyperresponsiveness (BHR) by histamine inhalation test, and allergy by skin-prick tests. Current asthma was defined as the presence of both wheeze in last 12 months and BHR. Der p I levels were measured in dust from the bed and floors in the homes of 57 randomly selected children in the coastal region and of 74 inland children. Der p I levels were significantly higher by the coast (83.0 vs 11.2 microg/g, P < 0.001). House dust mite sensitivity was of similar prevalence in both regions (28.6 vs 26.4%, n.s.) but Alternaria sensitivity was higher inland (4.0 vs 15.2% P<0.001). Bronchial responsiveness was more severe in coastal children sensitized to house dust mites and in inland children who were sensitized to Alternaria. The adjusted odds ratios for current asthma in children sensitized to house dust mites were 21.3 (95% CI 10.5, 43.2) by the coast and 2.7 (95% CI 1.3, 5.4) inland, and in children sensitized to Alternaria were 3.4 (95% CI 1.3, 9.1) in the coastal region and 5.6 (95% CI 3.1, 10.1 inland. These studies suggest that high house dust mite allergen levels in a humid, subtropical region act to significantly increase bronchial responsiveness in sensitized children, and that Alternaria allergens have a similar but less potent action in a dry, rural region.

  1. Effect of climate change on Alternaria leaf spot of rocket salad and black spot of basil under controlled environment.

    PubMed

    Pugliese, M; Cogliati, E; Gullino, M L; Garibaldi, A

    2012-01-01

    Plant responses to elevated CO2 and temperature have been much studied in recent years, but effects of climate change on pathological responses are still largely unknown. The pathosystems rocket (Eruca vesicaria subsp. sativa)--Alternaria leaf spot (Alternaria japonica) and basil (Ocimum basilicum)--black spot (Colletotrichum gloeosporioides) were chosen as models to assess the potential impact of increased CO2 and temperature on disease incidence and severity under controlled environment. Potted plants were grown in phytotrons under 4 different simulated climatic conditions: (1) standard temperature (ranging from 18 degrees to 22 degrees C) and standard CO2 concentration (400 ppm); (2) standard temperature and elevated CO2 concentration (800 ppm); (3) elevated temperature (ranging from 22 degrees to 26 degrees C, 4 degrees C higher than standard) and standard CO2 concentration; (4) elevated temperature and CO2 concentration. Each plant was inoculated with a spore suspension containing 1 x 10(5) cfu/ml of the pathogen. Disease incidence and severity were assessed 14 days after inoculation. Increasing CO2 to 800 ppm showed a clear increment in the percentage of Alternaria leaf spot on rocket leaves compared to standard conditions. Basil plants grown at 800 ppm of CO2 showed increased black spot symptoms compared to 400 ppm. Disease incidence and severity were always influenced by the combination of rising CO2 and increased temperature, compared to standard conditions (400 ppm of CO2 - 22 degrees C). Considering the rising concentrations of CO2 and global temperature, we can assume that this could increase the severity of Alternaria japonica on rocket and Colletotrichum gloeosporioides on basil.

  2. Alternaria sp. MG1, a resveratrol-producing fungus: isolation, identification, and optimal cultivation conditions for resveratrol production.

    PubMed

    Shi, Junling; Zeng, Qin; Liu, Yanlin; Pan, Zhongli

    2012-07-01

    Due to its potential in preventing or slowing the occurrence of many diseases, resveratrol (3,5,4'-trihydroxystilbene) has attracted great research interest. The objective of this study was to identify microorganisms from selected plants that produce resveratrol and to optimize the conditions for resveratrol production. Endophytes from Merlot wine grapes (Vitis vinifera L. cv. Merlot), wild Vitis (Vitis quinquangularis Rehd.), and Japanese knotweed (Polygonum cuspidatum Siebold & Zucc.) were isolated, and their abilities to produce resveratrol were evaluated. A total of 65 isolates were obtained and 21 produced resveratrol (6-123 μg/L) in liquid culture. The resveratrol-producing isolates belonged to seven genera, Botryosphaeria, Penicillium, Cephalosporium, Aspergillus, Geotrichum, Mucor, and Alternaria. The resveratrol-producing capability decreased or was completely lost in most isolates after three rounds of subculture. It was found that only the strain Alternaria sp. MG1 (isolated from cob of Merlot using GA1 medium) had stable and high resveratrol-producing capability in all subcultures. During liquid cultivation of Alternaria sp. MG1 in potato dextrose medium, the synthesis of resveratrol began on the first day, increased to peak levels on day 7, and then decreased sharply thereafter. Cell growth increased during cultivation and reached a stable and high level of biomass after 5 days. The best fermentation conditions for resveratrol production in liquid cultures of Alternaria sp. MG1 were an inoculum size of 6 %, a medium volume of 125 mL in a 250-mL flask, a rotation speed of 101 rpm, and a temperature of 27 °C.

  3. Transcriptional analysis of the sweet orange interaction with the citrus canker pathogens Xanthomonas axonopodis pv. citri and Xanthomonas axonopodis pv. aurantifolii.

    PubMed

    Cernadas, Raúl Andrés; Camillo, Luciana Rodrigues; Benedetti, Celso Eduardo

    2008-09-01

    Xanthomonas axonopodis pv. citri (Xac) and Xanthomonas axonopodis pv. aurantifolii pathotype C (Xaa) are responsible for citrus canker disease; however, while Xac causes canker on all citrus varieties, Xaa is restricted to Mexican lime, and in sweet oranges it triggers a defence response. To gain insights into the differential pathogenicity exhibited by Xac and Xaa and to survey the early molecular events leading to canker development, a detailed transcriptional analysis of sweet orange plants infected with the pathogens was performed. Using differential display, suppressed subtractive hybridization and microarrays, we identified changes in transcript levels in approximately 2.0% of the approximately 32,000 citrus genes examined. Genes with altered expression in response to Xac/Xaa surveyed at 6 and 48 h post-infection (hpi) were associated with cell-wall modifications, cell division and expansion, vesicle trafficking, disease resistance, carbon and nitrogen metabolism, and responses to hormones auxin, gibberellin and ethylene. Most of the genes that were commonly modulated by Xac and Xaa were associated with basal defences triggered by pathogen-associated molecular patterns, including those involved in reactive oxygen species production and lignification. Significantly, we detected clear changes in the transcriptional profiles of defence, cell-wall, vesicle trafficking and cell growth-related genes in Xac-infected leaves between 6 and 48 hpi. This is consistent with the notion that Xac suppresses host defences early during infection and simultaneously changes the physiological status of the host cells, reprogramming them for division and growth. Notably, brefeldin A, an inhibitor of vesicle trafficking, retarded canker development. In contrast, Xaa triggered a mitogen-activated protein kinase signalling pathway involving WRKY and ethylene-responsive transcriptional factors known to activate downstream defence genes.

  4. Combination of UV-C treatment and Metschnikowia pulcherrimas for controlling Alternaria rot in postharvest winter jujube fruit.

    PubMed

    Guo, Dongqi; Zhu, Lixia; Hou, Xujie

    2015-01-01

    The potential of using antagonistic yeast Metschnikowia pulcherrimas alone or in combination with ultraviolet-C (UV-C) treatment for controlling Alternaria rot of winter jujube, and its effects on postharvest quality of fruit was investigated. The results showed that spore germination of Alternaria alternata was significantly inhibited by each of the 3 doses (1, 5, and 10 kJ m(-2) ) in vitro. In vivo, UV-C treatment (5 kJ m(-2) ) or antagonist yeast was capable of reducing the percentage of infected wounds and lesion diameter in artificially inoculated jujube fruits, however, in fruit treated with combination of UV-C treatment and M. pulcherrima, the percentage of infected wounds and lesion diameter was only 16.0% and 0.60 cm, respectively. The decay incidence on winter jujube fruits treated with the combination of UV-C treatment and M. pulcherrima was 23% after storage at 0 ± 1 °C for 45 d followed by 22 °C for 7 d. None of the treatments impaired quality parameters of jujube fruit. Thus, the combination of UV-C radiation and M. pulcherrima could be an alternative to synthetic fungicides for controlling postharvest Alternaria rot of winter jujube.

  5. Spotlight on the Underdogs—An Analysis of Underrepresented Alternaria Mycotoxins Formed Depending on Varying Substrate, Time and Temperature Conditions

    PubMed Central

    Zwickel, Theresa; Kahl, Sandra M.; Klaffke, Horst; Rychlik, Michael; Müller, Marina E. H.

    2016-01-01

    Alternaria (A.) is a genus of widespread fungi capable of producing numerous, possibly health-endangering Alternaria toxins (ATs), which are usually not the focus of attention. The formation of ATs depends on the species and complex interactions of various environmental factors and is not fully understood. In this study the influence of temperature (7 °C, 25 °C), substrate (rice, wheat kernels) and incubation time (4, 7, and 14 days) on the production of thirteen ATs and three sulfoconjugated ATs by three different Alternaria isolates from the species groups A. tenuissima and A. infectoria was determined. High-performance liquid chromatography coupled with tandem mass spectrometry was used for quantification. Under nearly all conditions, tenuazonic acid was the most extensively produced toxin. At 25 °C and with increasing incubation time all toxins were formed in high amounts by the two A. tenuissima strains on both substrates with comparable mycotoxin profiles. However, for some of the toxins, stagnation or a decrease in production was observed from day 7 to 14. As opposed to the A. tenuissima strains, the A. infectoria strain only produced low amounts of ATs, but high concentrations of stemphyltoxin III. The results provide an essential insight into the quantitative in vitro AT formation under different environmental conditions, potentially transferable to different field and storage conditions. PMID:27869760

  6. Variation in Host and Pathogen in the Neonectria/Malus Interaction; toward an Understanding of the Genetic Basis of Resistance to European Canker

    PubMed Central

    Gómez-Cortecero, Antonio; Saville, Robert J.; Scheper, Reiny W. A.; Bowen, Joanna K.; Agripino De Medeiros, Hugo; Kingsnorth, Jennifer; Xu, Xiangming; Harrison, Richard J.

    2016-01-01

    Apple canker caused by the phytopathogenic fungus Neonectria ditissima is an economically important disease, which has spread in recent years to almost all pome-producing regions of the world. N. ditissima is able to cross-infect a wide range of apple varieties and causes branch and trunk lesions, known as cankers. Most modern apple varieties are susceptible and in extreme cases suffer from high mortality (up to 50%) in the early phase of orchard establishment. There is no known race structure of the pathogen and the global level of genetic diversity of the pathogen population is unknown. Resistance breeding is underway in many global breeding programmes, but nevertheless, a total resistance to canker has not yet been demonstrated. Here we present preliminary data from a survey of the phylogenetic relationships between global isolates of N. ditissima which reveals only slight evidence for population structure. In addition we report the results of four rapid screening tests to assess the response to N. ditissima in different apple scion and rootstock varieties, which reveals abundant variation in resistance responses in both cultivar and rootstock material. Further seedling tests show that the segregation patterns of resistance and susceptibility vary widely between crosses. We discuss inconsistencies in test performance with field observations and discuss future research opportunities in this area. PMID:27695463

  7. D-PSA-K: A Model for Estimating the Accumulated Potential Damage on Kiwifruit Canes Caused by Bacterial Canker during the Growing and Overwintering Seasons

    PubMed Central

    Do, Ki Seok; Chung, Bong Nam; Joa, Jae Ho

    2016-01-01

    We developed a model, termed D-PSA-K, to estimate the accumulated potential damage on kiwifruit canes caused by bacterial canker during the growing and overwintering seasons. The model consisted of three parts including estimation of the amount of necrotic lesion in a non-frozen environment, the rate of necrosis increase in a freezing environment during the overwintering season, and the amount of necrotic lesion on kiwifruit canes caused by bacterial canker during the overwintering and growing seasons. We evaluated the model’s accuracy by comparing the observed maximum disease incidence on kiwifruit canes against the damage estimated using weather and disease data collected at Wando during 1994–1997 and at Seogwipo during 2014–2015. For the Hayward cultivar, D-PSA-K estimated the accumulated damage as approximately nine times the observed maximum disease incidence. For the Hort16A cultivar, the accumulated damage estimated by D-PSA-K was high when the observed disease incidence was high. D-PSA-K could assist kiwifruit growers in selecting optimal sites for kiwifruit cultivation and establishing improved production plans by predicting the loss in kiwifruit production due to bacterial canker, using past weather or future climate change data. PMID:27904460

  8. D-PSA-K: A Model for Estimating the Accumulated Potential Damage on Kiwifruit Canes Caused by Bacterial Canker during the Growing and Overwintering Seasons.

    PubMed

    Do, Ki Seok; Chung, Bong Nam; Joa, Jae Ho

    2016-12-01

    We developed a model, termed D-PSA-K, to estimate the accumulated potential damage on kiwifruit canes caused by bacterial canker during the growing and overwintering seasons. The model consisted of three parts including estimation of the amount of necrotic lesion in a non-frozen environment, the rate of necrosis increase in a freezing environment during the overwintering season, and the amount of necrotic lesion on kiwifruit canes caused by bacterial canker during the overwintering and growing seasons. We evaluated the model's accuracy by comparing the observed maximum disease incidence on kiwifruit canes against the damage estimated using weather and disease data collected at Wando during 1994-1997 and at Seogwipo during 2014-2015. For the Hayward cultivar, D-PSA-K estimated the accumulated damage as approximately nine times the observed maximum disease incidence. For the Hort16A cultivar, the accumulated damage estimated by D-PSA-K was high when the observed disease incidence was high. D-PSA-K could assist kiwifruit growers in selecting optimal sites for kiwifruit cultivation and establishing improved production plans by predicting the loss in kiwifruit production due to bacterial canker, using past weather or future climate change data.

  9. Botryosphaeriaceae species associated with dieback and canker disease of bay laurel in northern California with the description of Dothiorella californica sp. nov.

    PubMed

    Lawrence, Daniel P; Peduto Hand, Francesca; Gubler, W Douglas; Trouillas, Florent P

    2017-04-01

    Members of the Botryosphaeriaceae are cosmopolitan fungi that may exist as seemingly innocuous endophytes or as destructive pathogens of numerous woody hosts, including fruit and nut crops, urban ornamental trees and shrubs, and forest trees. Surveys of bay laurel in northern California have revealed symptoms of dieback and branch canker of unknown aetiology. The goals of this study were to identify and clarify the species of Botryosphaeriaceae associated with these symptoms and to confirm their pathogenicity. To understand the role of members of the Botryosphaeriaceae in the dieback and canker disease of bay laurel, 23 isolates were isolated from symptomatic wood. Phylogenetic analyses of ITS, translation elongation factor 1-α, and beta-tubulin revealed three species: Botryosphaeria dothidea, Neofusicoccum nonquaesitum, and the newly described and typified species Dothiorella californica sp. nov. When select isolates were inoculated to 2- to 3-year-old branches of Umbellularia californica in a natural forest, both B. dothidea and N. nonquaesitum were pathogenic with N. nonquaesitum producing the largest lesions at 12- and 18-months post inoculation, respectively, while Do. californica did not cause wood lesions significantly greater than the mock-inoculated controls. This study represents the first attempt to identify and test the pathogenicity of Botryosphaeriaceae species associated with dieback and canker disease of bay laurel in a northern California forest.

  10. Natural Occurrence of Alternaria Toxins in Wheat-Based Products and Their Dietary Exposure in China

    PubMed Central

    Zhao, Kai; Shao, Bing; Yang, Dajin; Li, Fengqin; Zhu, Jianghui

    2015-01-01

    A total of 181 wheat flour and 142 wheat-based foods including dried noodle, steamed bread and bread collected in China were analyzed for alternariol (AOH), alternariol monomethyl ether (AME), tentoxin (TEN) and tenuazonic acid (TeA) by ultra-performance liquid chromatography-electrospray ionization-tandem mass spectrometry. TeA was the predominant toxin found in 99.4% wheat flour samples at levels ranging from 1.76 μg/kg to 520 μg/kg. TEN was another Alternaria toxin frequently detected in wheat flour samples (97.2%) at levels between 2.72 μg/kg and 129 μg/kg. AOH and AME were detected in 11 (6.1%) samples at levels ranging from 16.0 μg/kg to 98.7 μg/kg (AOH) and in 165 (91.2%) samples with a range between 0.320 μg/kg and 61.8 μg/kg (AME). AOH was quantified at higher levels than AME with the ratio of AOH/AME ranging from 1.0 to 3.7. Significant linear regressions of correlation in toxin concentrations were observed between AOH and AME, AME and TeA, TEN and TeA, AOH+AME and TeA. At an average and 95th percentile, dietary exposure to AOH and AME in the Chinese general population and different age subgroups exceeded the relevant threshold value of toxicological concern (TTC), with the highest exposure found in children which deserves human health concern. TEN and TeA seem unlikely to be health concerns for the Chinese via wheat-based products but attention should be paid to synergistic or additive effects of TeA with AOH, AME, TEN and a further assessment will be performed once more data on toxicity-guided fractionation of the four toxins are available. It is necessary to conduct a systemic surveillance of Alternaria toxins in raw and processed foods in order to provide the scientific basis for making regulations on these toxins in China. PMID:26121047

  11. Population genetic analysis reveals cryptic sex in the phytopathogenic fungus Alternaria alternata

    PubMed Central

    Meng, Jing-Wen; Zhu, Wen; He, Meng-Han; Wu, E-Jiao; Duan, Guo-Hua; Xie, Ye-Kun; Jin, Yu-Jia; Yang, Li-Na; Shang, Li-Ping; Zhan, Jiasui

    2015-01-01

    Reproductive mode can impact population genetic dynamics and evolutionary landscape of plant pathogens as well as on disease epidemiology and management. In this study, we monitored the spatial dynamics and mating type idiomorphs in ~700 Alternaria alternata isolates sampled from the main potato production areas in China to infer the mating system of potato early blight. Consistent with the expectation of asexual species, identical genotypes were recovered from different locations separated by hundreds of kilometers of geographic distance and spanned across many years. However, high genotype diversity, equal MAT1-1 and MAT1-2 frequencies within and among populations, no genetic differentiation and phylogenetic association between two mating types, combined with random association amongst neutral markers in some field populations, suggested that sexual reproduction may also play an important role in the epidemics and evolution of the pathogen in at least half of the populations assayed despite the fact that no teleomorphs have been observed yet naturally or artificially. Our results indicated that A. alternata may adopt an epidemic mode of reproduction by combining many cycles of asexual propagation with fewer cycles of sexual reproduction, facilitating its adaptation to changing environments and making the disease management on potato fields even more difficult. PMID:26666175

  12. Anastomosis Is Required for Virulence of the Fungal Necrotroph Alternaria brassicicola▿

    PubMed Central

    Craven, Kelly D.; Vélëz, Heriberto; Cho, Yangrae; Lawrence, Christopher B.; Mitchell, Thomas K.

    2008-01-01

    A fungal mycelium is typically composed of radially extending hyphal filaments interconnected by bridges created through anastomoses. These bridges facilitate the dissemination of nutrients, water, and signaling molecules throughout the colony. In this study, we used targeted gene deletion and nitrate utilization mutants of the cruciferous pathogen Alternaria brassicicola and two closely related species to investigate hyphal fusion (anastomosis) and its role in the ability of fungi to cause disease. All eight of the A. brassicicola isolates tested, as well as A. mimicula and A. japonica, were capable of self-fusion, with two isolates of A. brassicicola being capable of non-self-fusion. Disruption of the anastomosis gene homolog (Aso1) in A. brassicicola resulted in both the loss of self-anastomosis and pathogenicity on cabbage. This finding, combined with our discovery that a previously described nonpathogenic A. brassicicola mutant defective for a mitogen-activated protein kinase gene (amk1) also lacked the capacity for self-anastomosis, suggests that self-anastomosis is associated with pathogenicity in A. brassicicola. PMID:18310356

  13. Host-selective toxins produced by the plant pathogenic fungus Alternaria alternata.

    PubMed

    Tsuge, Takashi; Harimoto, Yoshiaki; Akimitsu, Kazuya; Ohtani, Kouhei; Kodama, Motoichiro; Akagi, Yasunori; Egusa, Mayumi; Yamamoto, Mikihiro; Otani, Hiroshi

    2013-01-01

    Host-selective toxins (HSTs) produced by fungal plant pathogens are generally low-molecular-weight secondary metabolites with a diverse range of structures that function as effectors controlling pathogenicity or virulence in certain plant-pathogen interactions. There are now seven known diseases caused by Alternaria alternata in which HSTs are responsible for fungal pathogenesis. The pathogens have been defined as pathotypes of A. alternata because of morphological similarity but pathological differences. Chemical structures of HSTs from six pathotypes have been determined. The role of A. alternata HSTs in pathogenesis has been studied extensively, and discovery of the release of HSTs from germinating conidia prior to penetration aids in understanding the early participation of HSTs to induce susceptibility of host cells by suppressing their defence reactions. Many attempts have been made to find the target sites of A. alternata HSTs, and four cellular components, plasma membrane, mitochondrion, chloroplast and a metabolically important enzyme, have been identified as the primary sites of each HST action, leading to elucidation of the molecular mechanisms of HST sensitivity in host plants. Studies of the molecular genetics of HST production have identified supernumerary chromosomes encoding HST gene clusters and have provided new insights into the evolution of A. alternata pathotypes.

  14. Sensitivity to Alternaria alternata toxin in citrus because of altered mitochondrial RNA processing

    PubMed Central

    Ohtani, Kouhei; Yamamoto, Hiroyuki; Akimitsu, Kazuya

    2002-01-01

    Specificity in the interaction between rough lemon (Citrus jambhiri Lush.) and the fungal pathogen Alternaria alternata rough lemon pathotype is determined by a host-selective toxin, ACR-toxin. Mitochondria from rough lemon are sensitive to ACR-toxin whereas mitochondria from resistant plants, including other citrus species, are resistant. We have identified a C. jambhiri mitochondrial DNA sequence, designated ACRS (ACR-toxin sensitivity gene), that confers toxin sensitivity to Escherichia coli. ACRS is located in the group II intron of the mitochondrial tRNA-Ala and is translated into a SDS-resistant oligomeric protein in C. jambhiri mitochondria but is not translated in the toxin-insensitive mitochondria. ACRS is present in the mitochondrial genome of both toxin-sensitive and -insensitive citrus. However, in mitochondria of toxin-insensitive plants, the transcripts from ACRS are shorter than those in mitochondria of sensitive plants. These results demonstrate that sensitivity to ACR-toxin and hence specificity of the interaction between A. alternata rough lemon pathotype and C. jambhiri is due to differential posttranscriptional processing of a mitochondrial gene. PMID:11842194

  15. acetylcholinesterase inhibitory potential and insecticidal activity of an endophytic Alternaria sp. from Ricinus communis.

    PubMed

    Singh, Bahaderjeet; Thakur, Abhinay; Kaur, Sanehdeep; Chadha, B S; Kaur, Amarjeet

    2012-11-01

    Keeping in view the vast potential of endophytic fungi to produce bioactive molecules, this study aimed at isolating and screening endophytes for the production of acetylcholinesterase inhibitors. Fifty-four endophytic fungi were isolated from Ricinus communis and screened for their AChE inhibitory activity using Ellman's colorimetric assay method. Six isolates were found to possess AChE inhibitory activity with maximum inhibition of 78 % being evinced by culture Cas1 which was identified to be Alternaria sp. on the basis of molecular as well as microscopic methods. Optimization of inhibitor production was carried out using one factor at a time approach. Maximum production of inhibitor was obtained on potato dextrose broth after 10 days incubation. The IC(50) of the chloroform extract was observed to be 40 μg/ml. The extract was purified on silica gel and eluted stepwise with a gradient of chloroform/methanol. The insecticidal potential of the extract was evaluated by feeding the larvae of Spodoptera litura on diet containing varying concentrations of the extract. It was observed that with increase in the concentration of the extract, mortality of the larvae increased. The culture has the potential of being exploited in medicine as well as a biocontrol agent.

  16. Changes in concentration of Alternaria and Cladosporium spores during summer storms.

    PubMed

    Grinn-Gofroń, Agnieszka; Strzelczak, Agnieszka

    2013-09-01

    Fungal spores are known to cause allergic sensitization. Recent studies reported a strong association between asthma symptoms and thunderstorms that could be explained by an increase in airborne fungal spore concentrations. Just before and during thunderstorms the values of meteorological parameters rapidly change. Therefore, the goal of this study was to create a predictive model for hourly concentrations of atmospheric Alternaria and Cladosporium spores on days with summer storms in Szczecin (Poland) based on meteorological conditions. For this study we have chosen all days of June, July and August (2004-2009) with convective thunderstorms. There were statistically significant relationships between spore concentration and meteorological parameters: positive for air temperature and ozone content while negative for relative humidity. In general, before a thunderstorm, air temperature and ozone concentration increased, which was accompanied by a considerable increase in spore concentration. During and after a storm, relative humidity increased while both air temperature ozone concentration along with spore concentrations decreased. Artificial neural networks (ANN) were used to assess forecasting possibilities. Good performance of ANN models in this study suggest that it is possible to predict spore concentrations from meteorological variables 2 h in advance and, thus, warn people with spore-related asthma symptoms about the increasing abundance of airborne fungi on days with storms.

  17. Folic acid induces salicylic acid-dependent immunity in Arabidopsis and enhances susceptibility to Alternaria brassicicola.

    PubMed

    Wittek, Finni; Kanawati, Basem; Wenig, Marion; Hoffmann, Thomas; Franz-Oberdorf, Katrin; Schwab, Wilfried; Schmitt-Kopplin, Philippe; Vlot, A Corina

    2015-08-01

    Folates are essential for one-carbon transfer reactions in all organisms and contribute, for example, to de novo DNA synthesis. Here, we detected the folate precursors 7,8-dihydropteroate (DHP) and 4-amino-4-deoxychorismate (ADC) in extracts from Arabidopsis thaliana plants by Fourier transform ion cyclotron resonance-mass spectrometry. The accumulation of DHP, but not ADC, was induced after infection of plants with Pseudomonas syringae delivering the effector protein AvrRpm1. Application of folic acid or the DHP precursor 7,8-dihydroneopterin (DHN) enhanced resistance in Arabidopsis to P. syringae and elevated the transcript accumulation of the salicylic acid (SA) marker gene pathogenesis-related1 in both the treated and systemic untreated leaves. DHN- and folic acid-induced systemic resistance was dependent on SA biosynthesis and signalling. Similar to SA, folic acid application locally enhanced Arabidopsis susceptibility to the necrotrophic fungus Alternaria brassicicola. Together, the data associate the folic acid pathway with innate immunity in Arabidopsis, simultaneously activating local and systemic SA-dependent resistance to P. syringae and suppressing local resistance to A. brassicicola.

  18. Plasma-Based Degradation of Mycotoxins Produced by Fusarium, Aspergillus and Alternaria Species

    PubMed Central

    ten Bosch, Lars; Pfohl, Katharina; Avramidis, Georg; Wieneke, Stephan; Viöl, Wolfgang; Karlovsky, Petr

    2017-01-01

    The efficacy of cold atmospheric pressure plasma (CAPP) with ambient air as working gas for the degradation of selected mycotoxins was studied. Deoxynivalenol, zearalenone, enniatins, fumonisin B1, and T2 toxin produced by Fusarium spp., sterigmatocystin produced by Aspergillus spp. and AAL toxin produced by Alternaria alternata were used. The kinetics of the decay of mycotoxins exposed to plasma discharge was monitored. All pure mycotoxins exposed to CAPP were degraded almost completely within 60 s. Degradation rates varied with mycotoxin structure: fumonisin B1 and structurally related AAL toxin were degraded most rapidly while sterigmatocystin exhibited the highest resistance to degradation. As compared to pure compounds, the degradation rates of mycotoxins embedded in extracts of fungal cultures on rice were reduced to a varying extent. Our results show that CAPP efficiently degrades pure mycotoxins, the degradation rates vary with mycotoxin structure, and the presence of matrix slows down yet does not prevent the degradation. CAPP appears promising for the decontamination of food commodities with mycotoxins confined to or enriched on surfaces such as cereal grains. PMID:28287436

  19. A conditionally dispensable chromosome controls host-specific pathogenicity in the fungal plant pathogen Alternaria alternata.

    PubMed Central

    Hatta, Rieko; Ito, Kaoru; Hosaki, Yoshitsugu; Tanaka, Takayoshi; Tanaka, Aiko; Yamamoto, Mikihiro; Akimitsu, Kazuya; Tsuge, Takashi

    2002-01-01

    The filamentous fungus Alternaria alternata contains seven pathogenic variants (pathotypes), which produce host-specific toxins and cause diseases on different plants. Previously, the gene cluster involved in host-specific AK-toxin biosynthesis of the Japanese pear pathotype was isolated, and four genes, named AKT genes, were identified. The AKT homologs were also found in the strawberry and tangerine pathotypes, which produce AF-toxin and ACT-toxin, respectively. This result is consistent with the fact that the toxins of these pathotypes share a common 9,10-epoxy-8-hydroxy-9-methyl-decatrienoic acid structural moiety. In this study, three of the AKT homologs (AFT1-1, AFTR-1, and AFT3-1) were isolated on a single cosmid clone from strain NAF8 of the strawberry pathotype. In NAF8, all of the AKT homologs were present in multiple copies on a 1.05-Mb chromosome. Transformation-mediated targeting of AFT1-1 and AFT3-1 in NAF8 produced AF-toxin-minus, nonpathogenic mutants. All of the mutants lacked the 1.05-Mb chromosome encoding the AFT genes. This chromosome was not essential for saprophytic growth of this pathogen. Thus, we propose that a conditionally dispensable chromosome controls host-specific pathogenicity of this pathogen. PMID:12019223

  20. Zinc triggers signaling mechanisms and defense responses promoting resistance to Alternaria brassicicola in Arabidopsis thaliana.

    PubMed

    Martos, Soledad; Gallego, Berta; Cabot, Catalina; Llugany, Mercè; Barceló, Juan; Poschenrieder, Charlotte

    2016-08-01

    According to the elemental defense hypothesis the accumulation of trace elements by plants may substitute for organic defenses, while the joint effects hypothesis proposes that trace elements and organic defenses can have additive or synergistic effects against pathogens or herbivores. To evaluate these hypotheses the response of the pathosystem Alternaria brassicicola-Arabidopsis thaliana to control (2μM) and surplus (12μM) Zn was evaluated using the camalexin deficient mutant pad3-1 and mtp1-1, a mutant with impaired Zn vacuolar storage, along with the corresponding wildtypes. In vitro, a 50% inhibition of fungal growth was achieved by 440μM Zn. A. thaliana leaves could accumulate equivalent concentrations without harm. In fact, surplus Zn enhanced the resistance of A. thaliana to fungal attack in Columbia (Col-0), Wassilewskija (WS), and mtp1-1. However, surplus Zn was unable to protect pad3-1 demonstrating that Zn cannot substitute for camalexin, the main organic defense in A. thaliana. High, non phytotoxic leaf Zn concentrations enhanced the resistance to A. brassicicola of A. thaliana genotypes able to produce camalexin. This was mainly due to Zn-induced enhancement of the JA/ETH signaling pathway leading to enhanced PAD3 expression. These results support the joint effects hypothesis and highlight the importance of adequate Zn supply for reinforced pathogen resistance.

  1. Stress Response and Pathogenicity of the Necrotrophic Fungal Pathogen Alternaria alternata

    PubMed Central

    Chung, Kuang-Ren

    2012-01-01

    The production of host-selective toxins by the necrotrophic fungus Alternaria alternata is essential for the pathogenesis. A. alternata infection in citrus leaves induces rapid lipid peroxidation, accumulation of hydrogen peroxide (H2O2), and cell death. The mechanisms by which A. alternata avoids killing by reactive oxygen species (ROS) after invasion have begun to be elucidated. The ability to coordinate of signaling pathways is essential for the detoxification of cellular stresses induced by ROS and for pathogenicity in A. alternata. A low level of H2O2, produced by the NADPH oxidase (NOX) complex, modulates ROS resistance and triggers conidiation partially via regulating the redox-responsive regulators (YAP1 and SKN7) and the mitogen-activated protein (MAP) kinase (HOG1) mediated pathways, which subsequently regulate the genes required for the biosynthesis of siderophore, an iron-chelating compound. Siderophore-mediated iron acquisition plays a key role in ROS detoxification because of the requirement of iron for the activities of antioxidants (e.g., catalase and SOD). Fungal strains impaired for the ROS-detoxifying system severely reduce the virulence on susceptible citrus cultivars. This paper summarizes the current state of knowledge of signaling pathways associated with cellular responses to multidrugs, oxidative and osmotic stress, and fungicides, as well as the pathogenicity/virulence in the tangerine pathotype of A. alternata. PMID:24278721

  2. Role of mannitol metabolism in the pathogenicity of the necrotrophic fungus Alternaria brassicicola

    PubMed Central

    Calmes, Benoit; Guillemette, Thomas; Teyssier, Lény; Siegler, Benjamin; Pigné, Sandrine; Landreau, Anne; Iacomi, Béatrice; Lemoine, Rémi; Richomme, Pascal; Simoneau, Philippe

    2013-01-01

    In this study, the physiological functions of fungal mannitol metabolism in the pathogenicity and protection against environmental stresses were investigated in the necrotrophic fungus Alternaria brassicicola. Mannitol metabolism was examined during infection of Brassica oleracea leaves by sequential HPLC quantification of the major soluble carbohydrates and expression analysis of genes encoding two proteins of mannitol metabolism, i.e., a mannitol dehydrogenase (AbMdh), and a mannitol-1-phosphate dehydrogenase (AbMpd). Knockout mutants deficient for AbMdh or AbMpd and a double mutant lacking both enzyme activities were constructed. Their capacity to cope with various oxidative and drought stresses and their pathogenic behavior were evaluated. Metabolic and gene expression profiling indicated an increase in mannitol production during plant infection. Depending on the mutants, distinct pathogenic processes, such as leaf and silique colonization, sporulation, survival on seeds, were impaired by comparison to the wild-type. This pathogenic alteration could be partly explained by the differential susceptibilities of mutants to oxidative and drought stresses. These results highlight the importance of mannitol metabolism with respect to the ability of A. brassicicola to efficiently accomplish key steps of its pathogenic life cycle. PMID:23717316

  3. Arabidopsis Responds to Alternaria alternata Volatiles by Triggering Plastid Phosphoglucose Isomerase-Independent Mechanisms1[OPEN

    PubMed Central

    Sánchez-López, Ángela María; Bahaji, Abdellatif; De Diego, Nuria; Baslam, Marouane; Li, Jun; Almagro, Goizeder; García-Gómez, Pablo; Ricarte-Bermejo, Adriana; Novák, Ondřej; Spíchal, Lukáš; Ciordia, Sergio; Mena, María Carmen

    2016-01-01

    Volatile compounds (VCs) emitted by phylogenetically diverse microorganisms (including plant pathogens and microbes that do not normally interact mutualistically with plants) promote photosynthesis, growth, and the accumulation of high levels of starch in leaves through cytokinin (CK)-regulated processes. In Arabidopsis (Arabidopsis thaliana) plants not exposed to VCs, plastidic phosphoglucose isomerase (pPGI) acts as an important determinant of photosynthesis and growth, likely as a consequence of its involvement in the synthesis of plastidic CKs in roots. Moreover, this enzyme plays an important role in connecting the Calvin-Benson cycle with the starch biosynthetic pathway in leaves. To elucidate the mechanisms involved in the responses of plants to microbial VCs and to investigate the extent of pPGI involvement, we characterized pPGI-null pgi1-2 Arabidopsis plants cultured in the presence or absence of VCs emitted by Alternaria alternata. We found that volatile emissions from this fungal phytopathogen promote growth, photosynthesis, and the accumulation of plastidic CKs in pgi1-2 leaves. Notably, the mesophyll cells of pgi1-2 leaves accumulated exceptionally high levels of starch following VC exposure. Proteomic analyses revealed that VCs promote global changes in the expression of proteins involved in photosynthesis, starch metabolism, and growth that can account for the observed responses in pgi1-2 plants. The overall data show that Arabidopsis plants can respond to VCs emitted by phytopathogenic microorganisms by triggering pPGI-independent mechanisms. PMID:27663407

  4. [Effects of Alternaria tenuis nees on Trifolium repens L. under Cu stress].

    PubMed

    Chu, Ling; Shao, Deng-hui; Jin, Song; Wu, Xue-feng

    2007-11-01

    With pot culture, this paper studied the effects of Alternaria tenuis Nees inoculation on the eco-physiological indices of Trifolium repens L. leaf under Cu stress. The results showed that in the control (not inoculated with the pathogen), the contents of photosynthetic pigments (chlorophyll a, b, a + b and carotenoid) and soluble protein in T. repens leaves decreased markedly with increasing Cu concentration (0-3000 mg x kg(-1)). The enhancement of cellular membrane lipids peroxidation with the increase of Cu concentration led to a rapid accumulation of malondialdehyde (MDA), the damage on cellular membrane structure, and an increase of electric conductivity. The balance of active oxygen metabolism systems was broken, SOD and CAT activities decreased, while POD activity increased. After inoculation with A. tenuis, the damages of Cu on plant membrane systems and active oxygen metabolism systems aggravated, the contents of photosynthetic pigments and soluble protein and the activities of SOD and CAT decreased to different degree, while the electric conductivity, MDA content and POD activity increased markedly, compared with the control.

  5. Genomic and transcriptomic analyses of the tangerine pathotype of Alternaria alternata in response to oxidative stress

    PubMed Central

    Wang, Mingshuang; Sun, Xuepeng; Yu, Dongliang; Xu, Jianping; Chung, Kuangren; Li, Hongye

    2016-01-01

    The tangerine pathotype of Alternaria alternata produces the A. citri toxin (ACT) and is the causal agent of citrus brown spot that results in significant yield losses worldwide. Both the production of ACT and the ability to detoxify reactive oxygen species (ROS) are required for A. alternata pathogenicity in citrus. In this study, we report the 34.41 Mb genome sequence of strain Z7 of the tangerine pathotype of A. alternata. The host selective ACT gene cluster in strain Z7 was identified, which included 25 genes with 19 of them not reported previously. Of these, 10 genes were present only in the tangerine pathotype, representing the most likely candidate genes for this pathotype specialization. A transcriptome analysis of the global effects of H2O2 on gene expression revealed 1108 up-regulated and 498 down-regulated genes. Expressions of those genes encoding catalase, peroxiredoxin, thioredoxin and glutathione were highly induced. Genes encoding several protein families including kinases, transcription factors, transporters, cytochrome P450, ubiquitin and heat shock proteins were found associated with adaptation to oxidative stress. Our data not only revealed the molecular basis of ACT biosynthesis but also provided new insights into the potential pathways that the phytopathogen A. alternata copes with oxidative stress. PMID:27582273

  6. Activation of the Nrf2-ARE pathway by the Alternaria alternata mycotoxins altertoxin I and II.

    PubMed

    Jarolim, Katharina; Del Favero, Giorgia; Pahlke, Gudrun; Dostal, Victoria; Zimmermann, Kristin; Heiss, Elke; Ellmer, Doris; Stark, Timo D; Hofmann, Thomas; Marko, Doris

    2017-01-01

    The mycotoxins altertoxin I and II (ATX I and II) are secondary metabolites produced by Alternaria alternata fungi and may occur as food and feed contaminants, especially after long storage periods. Although the toxic potential of altertoxins has been previously investigated, little is known about the pathways that play a role in their intracellular metabolism. In order to identify potential targets of ATX I and ATX II, the two toxins were tested for interaction with the nuclear factor erythroid-derived 2-like 2/antioxidant response element (Nrf2/ARE) pathway in mammalian cells. This pathway can be activated by various stressors resulting in the expression of enzymes important for metabolism and detoxification. In the present study, only ATX II triggered a concentration-dependent increase in Nrf2-ARE-dependent luciferase expression. Consistently, confocal microscopy revealed an ATX II-induced increase in Nrf2 signal in HT29 intestinal cells. In agreement with these data, ATX II induced the transcription of γ-glutamate cysteine ligase, the key enzyme in catalyzing GSH synthesis of the cells and which is regulated by Nrf2. Further investigations demonstrated that ATX II induced a concentration-dependent depletion of the cellular GSH levels after short incubation time (3 h) and an increase after longer incubation time (24 h). In conclusion, it was demonstrated that ATX II can interact at several levels of the Nrf2-ARE pathway in mammalian cells and that ATX I does not share the same mechanism of action.

  7. Candidate Resistant Genes of Sand Pear (Pyrus pyrifolia Nakai) to Alternaria alternata Revealed by Transcriptome Sequencing.

    PubMed

    Yang, Xiaoping; Hu, Hongju; Yu, Dazhao; Sun, Zhonghai; He, Xiujuan; Zhang, Jingguo; Chen, Qiliang; Tian, Rui; Fan, Jing

    2015-01-01

    Pear black spot (PBS) disease, which is caused by Alternaria alternata (Aa), is one of the most serious diseases affecting sand pear (Pyrus pyrifolia Nakai) cultivation worldwide. To investigate the defense mechanisms of sand pear in response to Aa, the transcriptome of a sand pear germplasm with differential resistance to Aa was analyzed using Illumina paired-end sequencing. Four libraries derived from PBS-resistant and PBS-susceptible sand pear leaves were characterized through inoculation or mock-inoculation. In total, 20.5 Gbp of sequence data and 101,632,565 reads were generated, representing 44717 genes. Approximately 66% of the genes or sequenced reads could be aligned to the pear reference genome. A large number (5213) of differentially expressed genes related to PBS resistance were obtained; 34 microsatellites were detected in these genes, and 28 genes were found to be closely related to PBS resistance. Using a transcriptome analysis in response to PBS inoculation and comparison analysis to the PHI database, 4 genes (Pbr039001, Pbr001627, Pbr025080 and Pbr023112) were considered to be promising candidates for sand pear resistance to PBS. This study provides insight into changes in the transcriptome of sand pear in response to PBS infection, and the findings have improved our understanding of the resistance mechanism of sand pear to PBS and will facilitate future gene discovery and functional genome studies of sand pear.

  8. Potent Selective Inhibition of Monoamine Oxidase A by Alternariol Monomethyl Ether Isolated from Alternaria brassicae.

    PubMed

    Lee, Hyun Woo; Kim, Yeon Ji; Nam, Sang-Jip; Kim, Hoon

    2017-02-28

    Alternariol monomethyl ether (AME), a dibenzopyrone derivative, was isolated from Alternaria brassicae along with altertoxin II (ATX-II). The compounds were tested for the inhibitory activity of monoamine oxidase (MAO), which catalyzes neurotransmitting monoamines. AME was found to be a highly potent and selective inhibitor of human MAO-A with an IC50 value of 1.71 µM; however, it was found to be ineffective for MAO-B inhibition. ATX-II was not effective for the inhibition of either MAO-A or MAO-B. The inhibition of MAO-A using AME was apparently instantaneous. MAO-A activity was almost completely recovered after the dilution of the inhibited enzyme with an excess amount of AME, suggesting AME is a reversible inhibitor. AME showed mixed inhibition for MAO-A in Lineweaver-Burk plots with a Ki value of 0.34 µM. The findings of this study suggest that microbial metabolites and dibenzopyrone could be potent MAO inhibitors. In addition, AME could be a useful lead compound for developing reversible MAO-A inhibitors to treat depression, Parkinson's disease, and Alzheimer's disease.

  9. Recent developments in stable isotope dilution assays in mycotoxin analysis with special regard to Alternaria toxins.

    PubMed

    Asam, Stefan; Rychlik, Michael

    2015-10-01

    Stable isotope dilution assays (SIDAs) are becoming ever commoner in mycotoxin analysis, and the number of synthesized or commercially available isotopically labelled compounds has greatly increased in the 7 years since our last review dealing with this topic. Thus, this review is conceived as an update for new applications or improvements of SIDAs for compounds discussed earlier, but the main focus is on newly introduced labelled substances and the development of SIDAs for, for example, fusarin C, moniliformin or the enniatins. Mycotoxin research has concentrated on the emerging group of Alternaria toxins in recent years, and a series of SIDAs have been developed, including ones for tenuazonic acid, alternariol, altertoxins and tentoxin that are discussed in detail in this review. Information about synthetic routes, isotopic purity and mass-spectrometric characterization of labelled compounds is given, as well as about the development and validation of SIDAs and their application to foods, feeds or biological samples. As the number of commercially available labelled standards is increasing continuously, a general tendency for the use of analytical methods based on liquid chromatography coupled with mass spectrometry capable of identifying a series of mycotoxins simultaneously ("multimethods") and using one or more labelled internal standards can be observed. An overview of these applications is given, thus demonstrating that SIDAs are increasingly being used in routine analysis.

  10. Green Chemistry Approach for the Synthesis of Gold Nanoparticles Using the Fungus Alternaria sp.

    PubMed

    Dhanasekar, Naresh Niranjan; Rahul, Ganga Ravindran; Narayanan, Kannan Badri; Raman, Gurusamy; Sakthivel, Natarajan

    2015-07-01

    The synthesis of gold nanoparticles has gained tremendous attention owing to their immense applications in the field of biomedical sciences. Although several chemical procedures are used for the synthesis of nanoparticles, the release of toxic and hazardous by-products restricts their use in biomedical applications. In the present investigation, gold nanoparticles were synthesized biologically using the culture filtrate of the filamentous fungus Alternaria sp. The culture filtrate of the fungus was exposed to three different concentrations of chloroaurate ions. In all cases, the gold ions were reduced to Au(0), leading to the formation of stable gold nanoparticles of variable sizes and shapes. UV-Vis spectroscopy analysis confirmed the formation of nanoparticles by reduction of Au(3+) to Au(0). TEM analysis revealed the presence of spherical, rod, square, pentagonal, and hexagonal morphologies for 1 mM chloroaurate solution. However, quasi-spherical and spherical nanoparticles/heart-like morphologies with size range of about 7-13 and 15-18 nm were observed for lower molar concentrations of 0.3 and 0.5 mM gold chloride solution, respectively. The XRD spectrum revealed the face-centered cubic crystals of synthesized gold nanoparticles. FT-IR spectroscopy analysis confirmed the presence of aromatic primary amines, and the additional SPR bands at 290 and 230 nm further suggested that the presence of amino acids such as tryptophan/tyrosine or phenylalanine acts as the capping agent on the synthesized mycogenic gold nanoparticles.

  11. Triterpenoids from the stems of Schisandra grandiflora and their biological activity.

    PubMed

    Shi, Wei; Liu, Han-Wei; Guo, Xin; Hou, Lan; Gao, Jin-Ming

    2016-07-01

    One new ursane-type triterpenoid (1), named granditriol, along with 14 known compounds (2-15), was isolated from the organic extracts of Schisandra grandiflora stems. The structure of the new compound was elucidated by extensive spectroscopic methods as 28-norursa-12,17,19,21-tetraen-2α,3α,23-triol. These isolates were evaluated for anti-phytopathogenic fungi activity and cytotoxicity against human cancer cell line (HepG2). Asiatic acid (8) and 2α,3α,19α-trihydroxyurs-12-en-28-oic acid (9) inhibited the growth of two plant pathogens, Alternaria alternata and Alternaria solani. In addition, compounds 12, 15, and 11 displayed notable anti-proliferative activity against HepG2 cells. Compound 1 is the first report of 28-nortriterpenoid from the Schisandraceae family. All these were obtained from this plant for the first time.

  12. Comparative ultrastructure of nonwounded Mexican lime and Yuzu leaves infected with the citrus canker bacterium Xanthomonas citri pv. citri.

    PubMed

    Lee, In Jung; Kim, Ki Woo; Hyun, Jae Wook; Lee, Yong Hoon; Park, Eun Woo

    2009-07-01

    Ultrastructural aspects of citrus canker development were investigated in nonwounded leaves of citrus species by transmission electron microscopy (TEM). A susceptible species Mexican lime and a resistant species Yuzu were spray-inoculated with a virulent strain of Xanthomonas citri pv. citri. Initial symptoms occurred on Mexican lime approximately 9 days after inoculation, whereas they appeared on Yuzu mostly 11 days after inoculation. In Mexican lime leaves, the bacterial invasion was usually accompanied by host cell wall dissolution and cellular disruption. Fibrillar materials from degenerated cell walls were usually found in intercellular spaces. Damaged host cells with necrotic cytoplasm showed the localized separation of plasma membrane from the cell wall. Bacterial multiplication and electron-transparent capsule-like structures around bacteria were commonly observed. Meanwhile, cell wall protuberances were prominent outside host cell walls in response to bacterial invasion in Yuzu leaves. Occlusion of intercellular spaces was also formed by the fusion of two or more individual cell wall protuberances originated from two adjacent host cells. Papillae-like materials accumulated locally within host cells in close proximity to bacteria. Some bacteria were found to be undergoing degeneration in xylem vessels. Also, the shrunken, inactive bacteria were surrounded by electron-translucent fibrillar materials in intercellular spaces, implying bacterial immobilization. These cellular responses are thought to be the consequences of defense responses of Yuzu leaves to invading bacteria. In both citrus species, X. citri pv. citri contained polyphosphate bodies showing electron-dense and elliptical structures in cytoplasm.

  13. Genetic Diversity and Pathogenicity of Xanthomonas axonopodis Strains Inducing Citrus Canker Disease in Iran and South Korea.

    PubMed

    Khodakaramian, G; Swings, J

    2011-06-01

    For the first time in 1989 citrus bacterial canker disease has seen on Citrusaurantiifolia in southern Iran. A total of 43 strains from affected citrus trees, ten strains from South Korea and representative from all known five pathotypes of Xanthomonas axonopodis pathogenic on citrus trees were used in this study. Isolated strains from Iran were indistinguishable by phenotypic, FAMEs, and SDS-PAGE analyses but showed different host range. First group were pathogenic on all tested citrus seedlings including C. aurantiifolia, C. limettioides, C. limon, C. jambhiri, Poncirus trifoliata X C. paradisi, C. aurantium, C. paradise, C. medica, P. trifoliate, C. grandis, C. sinensis, C. reticulate and C. sinensis X P. trifoliate. Pathogenicity of the second group were limited to C. aurantiifolia, C. limettioides, C. limon, C. jambhiri, P. trifoliata X C. paradis, and C. aurantium. Among the strains studied by AFLP fingerprinting six clusters were found. These clusters were: (1) strains of pathotype C; (2) strains of pathotypes B and D; (3) strains of pathotype A together with the main group of the Iranian strains; (4) strains isolated from Korea; (5) strains of pathotype E; and (6) seven strains from Iran which made a completely separate cluster. Strains from pathotypes B and D could not be differentiated by AFLP. The tested Iranian strains belongs to the two different groups and strains from Korea grouped as a subcluster from main cluster of Iranian strains belong to the pathotype A.

  14. Candidate effector proteins of the necrotrophic apple canker pathogen Valsa mali can suppress BAX-induced PCD

    PubMed Central

    Li, Zhengpeng; Yin, Zhiyuan; Fan, Yanyun; Xu, Ming; Kang, Zhensheng; Huang, Lili

    2015-01-01

    Canker caused by the Ascomycete Valsa mali is the most destructive disease of apple in Eastern Asia, resulting in yield losses of up to 100%. This necrotrophic fungus induces severe necrosis on apple, eventually leading to the death of the whole tree. Identification of necrosis inducing factors may help to unravel the molecular bases for colonization of apple trees by V. mali. As a first step toward this goal, we identified and characterized the V. mali repertoire of candidate effector proteins (CEPs). In total, 193 secreted proteins with no known function were predicted from genomic data, of which 101 were V. mali-specific. Compared to non-CEPs predicted for the V. mali secretome, CEPs have shorter sequence length and a higher content of cysteine residues. Based on transient over-expression in Nicotiana benthamiana performed for 70 randomly selected CEPs, seven V. mali Effector Proteins (VmEPs) were shown to significantly suppress BAX-induced PCD. Furthermore, targeted deletion of VmEP1 resulted in a significant reduction of virulence. These results suggest that V. mali expresses secreted proteins that can suppress PCD usually associated with effector-triggered immunity (ETI). ETI in turn may play an important role in the V. mali–apple interaction. The ability of V. mali to suppress plant ETI sheds a new light onto the interaction of a necrotrophic fungus with its host plant. PMID:26284095

  15. Genetic differentiation and spatial structure of Geosmithia morbida, the causal agent of thousand cankers disease in black walnut (Juglans nigra).

    PubMed

    Hadziabdic, Denita; Vito, Lisa M; Windham, Mark T; Pscheidt, Jay W; Trigiano, Robert N; Kolarik, Miroslav

    2014-05-01

    The main objectives of this study were to evaluate genetic composition of Geosmithia morbida populations in the native range of black walnut and provide a better understanding regarding demography of the pathogen. The fungus G. morbida, and the walnut twig beetle, Pityophthorus juglandis, have been associated with a disease complex of black walnut (Juglans nigra) known as thousand cankers disease (TCD). The disease is manifested as branch dieback and canopy loss, eventually resulting in tree death. In 2010, the disease was detected in black walnut in Tennessee, and subsequently in Virginia and Pennsylvania in 2011 and North Carolina in 2012. These were the first incidences of TCD east of Colorado, where the disease has been established for more than a decade on indigenous walnut species. A genetic diversity and population structure study of 62 G. morbida isolates from Tennessee, Pennsylvania, North Carolina and Oregon was completed using 15 polymorphic microsatellite loci. The results revealed high haploid genetic diversity among seven G. morbida populations with evidence of gene flow, and significant differentiation among two identified genetic clusters. There was a significant correlation between geographic and genetic distance. Understanding the genetic composition and demography of G. morbida can provide valuable insight into recognizing factors affecting the persistence and spread of an invasive pathogen, disease progression, and future infestation predictions. Overall, these data support the hypotheses of two separate, highly diverse pathogen introductions into the native range of black walnut.

  16. The structure of a Xanthomonas general stress protein involved in citrus canker reveals its flavin-binding property.

    PubMed

    Hilario, Eduardo; Li, Yang; Niks, Dimitri; Fan, Li

    2012-07-01

    Xanthomonas citri pv. citri (Xac) causes citrus canker and affects citrus agriculture worldwide. Functional genetic analysis has indicated that a putative general stress protein (XacGSP) encoded by the Xac2369 gene is involved in the bacterial infection. In this report, the crystal structure of XacGSP was determined to 2.5 Å resolution. There are four XacGSP molecules in the crystal asymmetric unit. Each XacGSP monomer folds into a six-stranded antiparallel β-barrel flanked by five α-helices. A C-terminal extension protrudes from the sixth β-strand of the β-barrel and pairs with its counterpart from another monomer to form a bridge between the two subunits of an XacGSP dimer. Two XacGSP dimers cross over each other to form a tetramer; the β-barrels from one dimer contact the β-barrels of the other, while the two bridges are distant from each other and do not make contacts. The three-dimensional structure of the XacGSP monomer is very similar to those of pyridoxine 5-phosphate oxidases, a group of enzymes that use flavin mononucleotide (FMN) as a cofactor. Consistent with this, purified XacGSP protein binds to both FMN and flavin adenine dinucleotide (FAD), suggesting that XacGSP may help the bacteria to react against the oxidative stress induced by the defense mechanisms of the plant.

  17. Bacterial canker on kiwifruit in Italy: anatomical changes in the wood and in the primary infection sites.

    PubMed

    Renzi, Marsilio; Copini, Paul; Taddei, Anna R; Rossetti, Antonio; Gallipoli, Lorenzo; Mazzaglia, Angelo; Balestra, Giorgio M

    2012-09-01

    The bacterial canker of kiwifruit caused by Pseudomonas syringae pv. actinidiae is a severe threat to kiwifruit production worldwide. Many aspects of P. syringae pv. actinidiae biology and epidemiology still require in-depth investigation. The infection by and spread of P. syringae pv. actinidiae in xylem and phloem was investigated by carrying out artificial inoculation experiments with histological and dendrochronological analyses of naturally diseased plants in Italy. We found that the bacterium can infect host plants by entering natural openings and lesions. In naturally infected kiwifruit plants, P. syringae pv. actinidiae is present in the lenticels as well as in the dead phloem tissue beneath the lenticels, surrounded by a lesion in the periderm which appears to indicate the importance of lenticels to kiwifruit infection. Biofilm formation was observed outside and inside plants. In cases of advanced stages of P. syringae pv. actinidiae infection, neuroses of the phloem occur, which are followed by cambial dieback and most likely by infection of the xylem. Anatomical changes in wood such as reduced ring width, a drastic reduction in vessel size, and the presence of tyloses were observed within several infected sites. In the field, these changes occur only a year after the first leaf symptoms are observed suggesting a significant time lapse between primary and secondary symptoms. It was possible to study the temporal development of P. syringae pv. actinidiae-induced cambial dieback by applying dendrochronology methods which revealed that cambial dieback occurs only during the growing season.

  18. Isolation and characterization of the grain mold fungi Cochliobolus and Alternaria spp. from sorghum using semiselective media and DNA sequence analyses.

    PubMed

    Funnell-Harris, Deanna L; Prom, Louis K; Pedersen, Jeffrey F

    2013-02-01

    Mold diseases, caused by fungal complexes including Alternaria, Cochliobolus, and Fusarium species, limit sorghum grain production. Media were tested by plating Fusarium thapsinum, Alternaria sp., and Curvularia lunata, individually and competitively. Dichloran chloramphenicol rose bengal (DRBC) and modified V8 juice (ModV8) agars, found to be useful, were compared with commonly used agar media, dichloran chloramphenicol peptone (DCPA) and pentachloronitrobenzene (PCNB). Radial growth, starting with mycelia or single-conidia and hyphal tips, demonstrated an effect of media. For isolation of grain fungi, DRBC and ModV8 were similar or superior to DCPA and PCNB. When seedlings were inoculated with conidia of C. lunata, Alternaria sp., F. thapsinum, or mixtures, the percentage of root infection ranged from 28% to 77%. For mixed inoculations, shoot weights, lesion lengths, and percentage of root infections were similar to F. thapsinum inoculations; most colonies recovered from roots were F. thapsinum. For Alternaria grain isolates, 5 morphological types, including Alternaria alternata, were distinguished by colony morphologies and conidial dimensions. Sequence analysis using a portion of the endo-polygalacturonase gene was able to further distinguish isolates. Cochliobolus isolates were identified morphologically as C. lunata, Curvularia sorghina, and Bipolaris sorghicola. Multiple molecular genotypes were apparent from rRNA internal transcribed spacer region sequences from Cochliobolus grain isolates.

  19. Comparison of methodologies for conidia production by Alternaria alternata from citrus

    PubMed Central

    Carvalho, Daniel D. C.; Alves, Eduardo; Batista, Tereza R. S.; Camargos, Renato B.; Lopes, Eloísa A. G. L.

    2008-01-01

    Conidia production is a problem in the study of Alternaria alternata from citrus. Thus, this study aimed to compare existing methodologies for conidial production of A. alternata isolated from Ponkan tangerine (2 isolates), Cravo lemon (1 isolate), Pêra orange (2 isolates) and Murcott tangor (1 isolate). The methodologies used were conidia production with 12 and 24 hours under white fluorescent light, evaluation with 24 and 48 hours after applying fungal mycelium stress technique, cold stress followed by injury of mycelium and evaluation with 24 hours, using healthy vegetable tissue and the use of black fluorescent near ultraviolet (NUV) lamp. Satisfactory result was obtained with A. alternata isolate from Murcott tangor, with the production of 2.8 × 105 conidia mL-1, when fungal mycelium was stressed (Petri dish with 66.66% of fungi growth) and subsequently 24 h of growth. The use of white light (24 h) and black fluorescent NUV lamp also induced expressive conidia production by one isolate of Ponkan tangerine, which produced 17.2 × 105 and 10.1 × 105conidia mL-1 and another of Murcott tangor, which produced 13.9 × 105 and 10.1 × 105 conidia mL-1, respectively. The remaining methodologies analyzed in this study were not able to induce conidia production in satisfactory quantity. The use of both mycelium stress technique and white light (24 h) and black fluorescent NUV lamp allowed the production of enough quantities of conidia to be used in vitro (detection of fungitoxic substances) and in vivo (pathogenicity test) assays, respectively. PMID:24031309

  20. Transcription Factor Amr1 Induces Melanin Biosynthesis and Suppresses Virulence in Alternaria brassicicola

    SciTech Connect

    Cho, Yangrae; Srivastava, Akhil; Ohm, Robin A.; Lawrence, Christopher B.; Wang, Koon-Hui; Grigoriev, Igor V.; Marahatta, Sharadchandra P.

    2012-05-01

    Alternaria brassicicola is a successful saprophyte and necrotrophic plant pathogen. Several A. brassicicola genes have been characterized as affecting pathogenesis of Brassica species. To study regulatory mechanisms of pathogenesis, we mined 421 genes in silico encoding putative transcription factors in a machine-annotated, draft genome sequence of A. brassicicola. In this study, targeted gene disruption mutants for 117 of the transcription factor genes were produced and screened. Three of these genes were associated with pathogenesis. Disruption mutants of one gene (AbPacC) were nonpathogenic and another gene (AbVf8) caused lesions less than half the diameter of wild-type lesions. Unexpectedly, mutants of the third gene, Amr1, caused lesions with a two-fold larger diameter than the wild type and complementation mutants. Amr1 is a homolog of Cmr1, a transcription factor that regulates melanin biosynthesis in several fungi. We created gene deletion mutants of ?amr1 and characterized their phenotypes. The ?amr1 mutants used pectin as a carbon source more efficiently than the wild type, were melanin-deficient, and more sensitive to UV light and glucanase digestion. The AMR1 protein was localized in the nuclei of hyphae and in highly melanized conidia during the late stage of plant pathogenesis. RNA-seq analysis revealed that three genes in the melanin biosynthesis pathway, along with the deleted Amr1 gene, were expressed at low levels in the mutants. In contrast, many hydrolytic enzyme-coding genes were expressed at higher levels in the mutants than in the wild type during pathogenesis. The results of this study suggested that a gene important for survival in nature negatively affected virulence, probably by a less efficient use of plant cell-wall materials. We speculate that the functions of the Amr1 gene are important to the success of A. brassicicola as a competitive saprophyte and plant parasite.

  1. Arabidopsis Cell Death in Compatible and Incompatible Interactions with Alternaria brassicicola

    PubMed Central

    Su’udi, Mukhamad; Kim, Min Gab; Park, Sang-Ryeol; Hwang, Duk-Ju; Bae, Shin-Chul; Ahn, Il-Pyung

    2011-01-01

    Two strains of necrotrophic Alternaria brassicicola, Ab40857 and Ab42464, are virulent on Korean cabbage and several wild types of Arabidopsis thaliana. Interaction between Ab42464 and Col-0 was compatible, whereas interaction between Ab40857 and Col-0 was incompatible. The loss of defense, no death (dnd) 1 function abrogated the compatibility between Ab42464 and Col-0, and the accelerated cell death (acd) 2 mutation attenuated the Col-0’s resistance against Ab40857. These two fungal strains induced PR1 transcription in Col-0. Ab40857 accelerated transcription of PDF1.2, THI2.1, CAT, and POX by 12 h compared to those challenged with Ab42464. More abundant cell death was observed in Col-0 infected with Ab42464, however, callose deposition was evident in the incompatible interaction. Remarkably, Ab40857-infected areas of acd2-2 underwent rampant cell death and Ab42464 triggered callose production in dnd1-1. Furthermore, the incompatibility between Ab40857 and Col-0 was nullified by the coronatine- insensitive 1 (coi1) and phytoalexin-deficient 3 (pad3) mutations but not by nonexpresser of PR genes (npr1) and pad4. Ab40857 induced abundant cell death in pad3. Taken together, cell death during the early infection stage is a key determinant that discriminates between a compatible interaction and an incompatible one, and the resistance within Col-0 against Ab40857 is dependent on a defensesignaling pathway mediated by jasmonic acid and PAD3. PMID:21688205

  2. Transcription Factor Amr1 Induces Melanin Biosynthesis and Suppresses Virulence in Alternaria brassicicola

    PubMed Central

    Cho, Yangrae; Srivastava, Akhil; Ohm, Robin A.; Lawrence, Christopher B.; Wang, Koon-Hui; Grigoriev, Igor V.; Marahatta, Sharadchandra P.

    2012-01-01

    Alternaria brassicicola is a successful saprophyte and necrotrophic plant pathogen. Several A. brassicicola genes have been characterized as affecting pathogenesis of Brassica species. To study regulatory mechanisms of pathogenesis, we mined 421 genes in silico encoding putative transcription factors in a machine-annotated, draft genome sequence of A. brassicicola. In this study, targeted gene disruption mutants for 117 of the transcription factor genes were produced and screened. Three of these genes were associated with pathogenesis. Disruption mutants of one gene (AbPacC) were nonpathogenic and another gene (AbVf8) caused lesions less than half the diameter of wild-type lesions. Unexpectedly, mutants of the third gene, Amr1, caused lesions with a two-fold larger diameter than the wild type and complementation mutants. Amr1 is a homolog of Cmr1, a transcription factor that regulates melanin biosynthesis in several fungi. We created gene deletion mutants of Δamr1 and characterized their phenotypes. The Δamr1 mutants used pectin as a carbon source more efficiently than the wild type, were melanin-deficient, and more sensitive to UV light and glucanase digestion. The AMR1 protein was localized in the nuclei of hyphae and in highly melanized conidia during the late stage of plant pathogenesis. RNA-seq analysis revealed that three genes in the melanin biosynthesis pathway, along with the deleted Amr1 gene, were expressed at low levels in the mutants. In contrast, many hydrolytic enzyme-coding genes were expressed at higher levels in the mutants than in the wild type during pathogenesis. The results of this study suggested that a gene important for survival in nature negatively affected virulence, probably by a less efficient use of plant cell-wall materials. We speculate that the functions of the Amr1 gene are important to the success of A. brassicicola as a competitive saprophyte and plant parasite. PMID:23133370

  3. Contribution of peroxisomes to secondary metabolism and pathogenicity in the fungal plant pathogen Alternaria alternata.

    PubMed

    Imazaki, Ai; Tanaka, Aiko; Harimoto, Yoshiaki; Yamamoto, Mikihiro; Akimitsu, Kazuya; Park, Pyoyun; Tsuge, Takashi

    2010-05-01

    The filamentous fungus Alternaria alternata includes seven pathogenic variants (pathotypes) which produce different host-selective toxins and cause diseases on different plants. The Japanese pear pathotype produces the host-selective AK-toxin, an epoxy-decatrienoic acid ester, and causes black spot of Japanese pear. Previously, we identified four genes, AKT1, AKT2, AKT3, and AKTR, involved in AK toxin biosynthesis. AKT1, AKT2, and AKT3 encode enzyme proteins with peroxisomal targeting signal type 1 (PTS1)-like tripeptides, SKI, SKL, and PKL, respectively, at the C-terminal ends. In this study, we verified the peroxisome localization of Akt1, Akt2, and Akt3 by using strains expressing N-terminal green fluorescent protein (GFP)-tagged versions of the proteins. To assess the role of peroxisome function in AK-toxin production, we isolated AaPEX6, which encodes a peroxin protein essential for peroxisome biogenesis, from the Japanese pear pathotype and made AaPEX6 disruption-containing transformants from a GFP-Akt1-expressing strain. The DeltaAaPEX6 mutant strains did not grow on fatty acid media because of a defect in fatty acid beta oxidation. The import of GFP-Akt1 into peroxisomes was impaired in the DeltaAaPEX6 mutant strains. These strains completely lost AK toxin production and pathogenicity on susceptible pear leaves. These data show that peroxisomes are essential for AK-toxin biosynthesis. The DeltaAaPEX6 mutant strains showed a marked reduction in the ability to cause lesions on leaves of a resistant pear cultivar with defense responses compromised by heat shock. This result suggests that peroxisome function is also required for plant invasion and tissue colonization in A. alternata. We also observed that mutation of AaPEX6 caused a marked reduction of conidiation.

  4. Elevated Atmospheric Carbon Dioxide Concentrations Amplify Alternaria alternata Sporulation and Total Antigen Production

    PubMed Central

    Wolf, Julie; O’Neill, Nichole R.; Rogers, Christine A.; Muilenberg, Michael L.; Ziska, Lewis H.

    2010-01-01

    Background Although the effect of elevated carbon dioxide (CO2) concentration on pollen production has been established in some plant species, impacts on fungal sporulation and antigen production have not been elucidated. Objective Our purpose was to examine the effects of rising atmospheric CO2 concentrations on the quantity and quality of fungal spores produced on timothy (Phleum pratense) leaves. Methods Timothy plants were grown at four CO2 concentrations (300, 400, 500, and 600 μmol/mol). Leaves were used as growth substrate for Alternaria alternata and Cladosporium phlei. The spore abundance produced by both fungi, as well as the size (microscopy) and antigenic protein content (ELISA) of A. alternata, were quantified. Results Leaf carbon-to-nitrogen ratio was greater at 500 and 600 μmol/mol, and leaf biomass was greater at 600 μmol/mol than at the lower CO2 concentrations. Leaf carbon-to-nitrogen ratio was positively correlated with A. alternata spore production per gram of leaf but negatively correlated with antigenic protein content per spore. At 500 and 600 μmol/mol CO2 concentrations, A. alternata produced nearly three times the number of spores and more than twice the total antigenic protein per plant than at lower concentrations. C. phlei spore production was positively correlated with leaf carbon-to-nitrogen ratio, but overall spore production was much lower than in A. alternata, and total per-plant production did not vary among CO2 concentrations. Conclusions Elevated CO2 concentrations often increase plant leaf biomass and carbon-to-nitrogen ratio. Here we demonstrate for the first time that these leaf changes are associated with increased spore production by A. alternata, a ubiquitous allergenic fungus. This response may contribute to the increasing prevalence of allergies and asthma. PMID:20462828

  5. Pseudomonas syringae pv. actinidiae (PSA) isolates from recent bacterial canker of kiwifruit outbreaks belong to the same genetic lineage.

    PubMed

    Mazzaglia, Angelo; Studholme, David J; Taratufolo, Maria C; Cai, Rongman; Almeida, Nalvo F; Goodman, Tokia; Guttman, David S; Vinatzer, Boris A; Balestra, Giorgio M

    2012-01-01

    Intercontinental spread of emerging plant diseases is one of the most serious threats to world agriculture. One emerging disease is bacterial canker of kiwi fruit (Actinidia deliciosa and A. chinensis) caused by Pseudomonas syringae pv. actinidiae (PSA). The disease first occurred in China and Japan in the 1980s and in Korea and Italy in the 1990s. A more severe form of the disease broke out in Italy in 2008 and in additional countries in 2010 and 2011 threatening the viability of the global kiwi fruit industry. To start investigating the source and routes of international transmission of PSA, genomes of strains from China (the country of origin of the genus Actinidia), Japan, Korea, Italy and Portugal have been sequenced. Strains from China, Italy, and Portugal have been found to belong to the same clonal lineage with only 6 single nucleotide polymorphisms (SNPs) in 3,453,192 bp and one genomic island distinguishing the Chinese strains from the European strains. Not more than two SNPs distinguish each of the Italian and Portuguese strains from each other. The Japanese and Korean strains belong to a separate genetic lineage as previously reported. Analysis of additional European isolates and of New Zealand isolates exploiting genome-derived markers showed that these strains belong to the same lineage as the Italian and Chinese strains. Interestingly, the analyzed New Zealand strains are identical to European strains at the tested SNP loci but test positive for the genomic island present in the sequenced Chinese strains and negative for the genomic island present in the European strains. Results are interpreted in regard to the possible direction of movement of the pathogen between countries and suggest a possible Chinese origin of the European and New Zealand outbreaks.

  6. Identification of Circular RNAs in Kiwifruit and Their Species-Specific Response to Bacterial Canker Pathogen Invasion

    PubMed Central

    Wang, Zupeng; Liu, Yifei; Li, Dawei; Li, Li; Zhang, Qiong; Wang, Shuaibin; Huang, Hongwen

    2017-01-01

    Research studies have recently focused on circle RNAs (circRNAs) in relation to their regulatory functions in animals. However, the systematic identification of circRNAs in plants, especially non-model plants, is limited. In addition, raw report on the prediction of the potential role of circRNAs in plant response to pathogen invasion is currently available. We conducted the systematic identification of circRNAs from four materials originating from three species belonging to genus Actinidia under different situations using ribosomal RNA (rRNA) depleted RNA-Seq data. A total of 3,582 circRNAs were identified in Actinidia, of which 64.01, 21.44, and 14.55% were intergenic circRNAs, exonic circRNAs, and intronic circRNAs, respectively. Tissue-specific expression of circRNAs was observed in kiwifruit, and a species-specific response was detected when infected with Pseudomonas syringae pv. actinidiae (Psa), which is the causative agent of kiwifruit bacterial canker disease. Furthermore, we found that both exonic and intronic circRNAs were significantly positively correlated to parent protein-coding genes, and intronic circRNAs are a class of highly remarkable regulators the parent genes comparing to that of exonic circRNAs. Expression and weighted gene co-expression network analysis (WGCNA) identified a set of circRNAs that were closely associated with plant defense response. The findings of the presents study suggest that circRNAs exhibit tissue- and species-specific expression, as well as play an important role in plant immune response.

  7. Population Structure of Geosmithia morbida, the Causal Agent of Thousand Cankers Disease of Walnut Trees in the United States

    PubMed Central

    Graves, Andrew D.; Hartel, Colleen; Pscheidt, Jay W.; Tonos, Jadelys; Broders, Kirk; Cranshaw, Whitney; Seybold, Steven J.; Tisserat, Ned

    2014-01-01

    The ascomycete Geosmithia morbida and the walnut twig beetle Pityophthorus juglandis are associated with thousand cankers disease of Juglans (walnut) and Pterocarya (wingnut). The disease was first reported in the western United States (USA) on several Juglans species, but has been found more recently in the eastern USA in the native range of the highly susceptible Juglans nigra. We performed a comprehensive population genetic study of 209 G. morbida isolates collected from Juglans and Pterocarya from 17 geographic regions distributed across 12 U.S. states. The study was based on sequence typing of 27 single nucleotide polymorphisms from three genomic regions and genotyping with ten microsatellite primer pairs. Using multilocus sequence-typing data, 197 G. morbida isolates were placed into one of 57 haplotypes. In some instances, multiple haplotypes were recovered from isolates collected on the same tree. Twenty-four of the haplotypes (42%) were recovered from more than one isolate; the two most frequently occurring haplotypes (H02 and H03) represented 36% of all isolates. These two haplotypes were abundant in California, but were not recovered from Arizona or New Mexico. G. morbida population structure was best explained by four genetically distinct groups that clustered into three geographic regions. Most of the haplotypes isolated from the native range of J. major (Arizona and New Mexico) were found in those states only or present in distinct genetic clusters. There was no evidence of sexual reproduction or genetic recombination in any population. The scattered distribution of the genetic clusters indicated that G. morbida was likely disseminated to different regions at several times and from several sources. The large number of haplotypes observed and the genetic complexity of G. morbida indicate that it evolved in association with at least one Juglans spp. and the walnut twig beetle long before the first reports of the disease. PMID:25393300

  8. Population structure of Geosmithia morbida, the causal agent of thousand cankers disease of walnut trees in the United States.

    PubMed

    Zerillo, Marcelo M; Ibarra Caballero, Jorge; Woeste, Keith; Graves, Andrew D; Hartel, Colleen; Pscheidt, Jay W; Tonos, Jadelys; Broders, Kirk; Cranshaw, Whitney; Seybold, Steven J; Tisserat, Ned

    2014-01-01

    The ascomycete Geosmithia morbida and the walnut twig beetle Pityophthorus juglandis are associated with thousand cankers disease of Juglans (walnut) and Pterocarya (wingnut). The disease was first reported in the western United States (USA) on several Juglans species, but has been found more recently in the eastern USA in the native range of the highly susceptible Juglans nigra. We performed a comprehensive population genetic study of 209 G. morbida isolates collected from Juglans and Pterocarya from 17 geographic regions distributed across 12 U.S. states. The study was based on sequence typing of 27 single nucleotide polymorphisms from three genomic regions and genotyping with ten microsatellite primer pairs. Using multilocus sequence-typing data, 197 G. morbida isolates were placed into one of 57 haplotypes. In some instances, multiple haplotypes were recovered from isolates collected on the same tree. Twenty-four of the haplotypes (42%) were recovered from more than one isolate; the two most frequently occurring haplotypes (H02 and H03) represented 36% of all isolates. These two haplotypes were abundant in California, but were not recovered from Arizona or New Mexico. G. morbida population structure was best explained by four genetically distinct groups that clustered into three geographic regions. Most of the haplotypes isolated from the native range of J. major (Arizona and New Mexico) were found in those states only or present in distinct genetic clusters. There was no evidence of sexual reproduction or genetic recombination in any population. The scattered distribution of the genetic clusters indicated that G. morbida was likely disseminated to different regions at several times and from several sources. The large number of haplotypes observed and the genetic complexity of G. morbida indicate that it evolved in association with at least one Juglans spp. and the walnut twig beetle long before the first reports of the disease.

  9. Phylogeography of the Walnut Twig Beetle, Pityophthorus juglandis, the Vector of Thousand Cankers Disease in North American Walnut Trees

    PubMed Central

    Rugman-Jones, Paul F.; Seybold, Steven J.; Graves, Andrew D.; Stouthamer, Richard

    2015-01-01

    Thousand cankers disease (TCD) of walnut trees (Juglans spp.) results from aggressive feeding in the phloem by the walnut twig beetle (WTB), Pityophthorus juglandis, accompanied by inoculation of its galleries with a pathogenic fungus, Geosmithia morbida. In 1960, WTB was only known from four U.S. counties (in Arizona, California, and New Mexico), but the species has now (2014) invaded over 115 counties, representing much of the western USA, and at least six states in the eastern USA. The eastern expansion places TCD in direct proximity to highly valuable (> $500 billion) native timber stands of eastern black walnut, Juglans nigra. Using mitochondrial DNA sequences, from nearly 1100 individuals, we examined variation among 77 samples of WTB populations across its extended range in the USA, revealing high levels of polymorphism and evidence of two divergent lineages. The highest level of genetic diversity for the different lineages was found in the neighboring Madrean Sky Island and Western New Mexico regions, respectively. Despite their proximity, there was little evidence of mixing between these regions, with only a single migrant detected among 179 beetles tested. Indeed, geographic overlap of the two lineages was only common in parts of Colorado and Utah. Just two haplotypes, from the same lineage, predominated over the vast majority of the recently expanded range. Tests for Wolbachia proved negative suggesting it plays no role in "driving" the spread of particular haplotypes, or in maintaining deep levels of intraspecific divergence in WTB. Genotyping of ribosomal RNA corroborated the mitochondrial lineages, but also revealed evidence of hybridization between them. Hybridization was particularly prevalent in the sympatric areas, also apparent in all invaded areas, but absent from the most haplotype-rich area of each mitochondrial lineage. Hypotheses about the specific status of WTB, its recent expansion, and potential evolutionary origins of TCD are discussed

  10. Pseudomonas syringae pv. actinidiae (PSA) Isolates from Recent Bacterial Canker of Kiwifruit Outbreaks Belong to the Same Genetic Lineage

    PubMed Central

    Taratufolo, Maria C.; Cai, Rongman; Almeida, Nalvo F.; Goodman, Tokia; Guttman, David S.; Vinatzer, Boris A.; Balestra, Giorgio M.

    2012-01-01

    Intercontinental spread of emerging plant diseases is one of the most serious threats to world agriculture. One emerging disease is bacterial canker of kiwi fruit (Actinidia deliciosa and A. chinensis) caused by Pseudomonas syringae pv. actinidiae (PSA). The disease first occurred in China and Japan in the 1980s and in Korea and Italy in the 1990s. A more severe form of the disease broke out in Italy in 2008 and in additional countries in 2010 and 2011 threatening the viability of the global kiwi fruit industry. To start investigating the source and routes of international transmission of PSA, genomes of strains from China (the country of origin of the genus Actinidia), Japan, Korea, Italy and Portugal have been sequenced. Strains from China, Italy, and Portugal have been found to belong to the same clonal lineage with only 6 single nucleotide polymorphisms (SNPs) in 3,453,192 bp and one genomic island distinguishing the Chinese strains from the European strains. Not more than two SNPs distinguish each of the Italian and Portuguese strains from each other. The Japanese and Korean strains belong to a separate genetic lineage as previously reported. Analysis of additional European isolates and of New Zealand isolates exploiting genome-derived markers showed that these strains belong to the same lineage as the Italian and Chinese strains. Interestingly, the analyzed New Zealand strains are identical to European strains at the tested SNP loci but test positive for the genomic island present in the sequenced Chinese strains and negative for the genomic island present in the European strains. Results are interpreted in regard to the possible direction of movement of the pathogen between countries and suggest a possible Chinese origin of the European and New Zealand outbreaks. PMID:22590555

  11. Phylogeography of the walnut twig beetle, Pityophthorus juglandis, the vector of thousand cankers disease in North American walnut trees.

    PubMed

    Rugman-Jones, Paul F; Seybold, Steven J; Graves, Andrew D; Stouthamer, Richard

    2015-01-01

    Thousand cankers disease (TCD) of walnut trees (Juglans spp.) results from aggressive feeding in the phloem by the walnut twig beetle (WTB), Pityophthorus juglandis, accompanied by inoculation of its galleries with a pathogenic fungus, Geosmithia morbida. In 1960, WTB was only known from four U.S. counties (in Arizona, California, and New Mexico), but the species has now (2014) invaded over 115 counties, representing much of the western USA, and at least six states in the eastern USA. The eastern expansion places TCD in direct proximity to highly valuable (> $500 billion) native timber stands of eastern black walnut, Juglans nigra. Using mitochondrial DNA sequences, from nearly 1100 individuals, we examined variation among 77 samples of WTB populations across its extended range in the USA, revealing high levels of polymorphism and evidence of two divergent lineages. The highest level of genetic diversity for the different lineages was found in the neighboring Madrean Sky Island and Western New Mexico regions, respectively. Despite their proximity, there was little evidence of mixing between these regions, with only a single migrant detected among 179 beetles tested. Indeed, geographic overlap of the two lineages was only common in parts of Colorado and Utah. Just two haplotypes, from the same lineage, predominated over the vast majority of the recently expanded range. Tests for Wolbachia proved negative suggesting it plays no role in "driving" the spread of particular haplotypes, or in maintaining deep levels of intraspecific divergence in WTB. Genotyping of ribosomal RNA corroborated the mitochondrial lineages, but also revealed evidence of hybridization between them. Hybridization was particularly prevalent in the sympatric areas, also apparent in all invaded areas, but absent from the most haplotype-rich area of each mitochondrial lineage. Hypotheses about the specific status of WTB, its recent expansion, and potential evolutionary origins of TCD are discussed.

  12. Types of Stem Cells

    MedlinePlus

    ... Stem Cell Glossary Search Toggle Nav Types of Stem Cells Stem cells are the foundation from which all ... Learn About Stem Cells > Types of Stem Cells Stem cells Stem cells are the foundation for every organ ...

  13. Sesquiterpene emissions from Alternaria alternata and Fusarium oxysporum: Effects of age, nutrient availability, and co-cultivation

    PubMed Central

    Weikl, Fabian; Ghirardo, Andrea; Schnitzler, Jörg-Peter; Pritsch, Karin

    2016-01-01

    Alternaria alternata is one of the most studied fungi to date because of its impact on human life – from plant pathogenicity to allergenicity. However, its sesquiterpene emissions have not been systematically explored. Alternaria regularly co-occurs with Fusarium fungi, which are common plant pathogens, on withering plants. We analyzed the diversity and determined the absolute quantities of volatile organic compounds (VOCs) in the headspace above mycelial cultures of A. alternata and Fusarium oxysporum under different conditions (nutrient rich and poor, single cultures and co-cultivation) and at different mycelial ages. Using stir bar sorptive extraction and gas chromatography–mass spectrometry, we observed A. alternata to strongly emit sesquiterpenes, particularly during the early growth stages, while emissions from F. oxysporum consistently remained comparatively low. The emission profile characterizing A. alternata comprised over 20 sesquiterpenes with few effects from nutrient quality and age on the overall emission profile. Co-cultivation with F. oxysporum resulted in reduced amounts of VOCs emitted from A. alternata although its profile remained similar. Both fungi showed distinct emission profiles, rendering them suitable biomarkers for growth-detection of their phylotype in ambient air. The study highlights the importance of thorough and quantitative evaluations of fungal emissions of volatile infochemicals such as sesquiterpenes. PMID:26915756

  14. Genetic Architecture of Resistance to Alternaria brassicae in Arabidopsis thaliana: QTL Mapping Reveals Two Major Resistance-Conferring Loci

    PubMed Central

    Rajarammohan, Sivasubramanian; Kumar, Amarendra; Gupta, Vibha; Pental, Deepak; Pradhan, Akshay K.; Kaur, Jagreet

    2017-01-01

    Alternaria brassicae, a necrotrophic fungal pathogen, causes Alternaria blight, one of the most important diseases of oleiferous Brassica crops. The current study utilized Arabidopsis as a model to decipher the genetic architecture of defense against A. brassicae. Significant phenotypic variation that was largely genetically determined was observed among Arabidopsis accessions in response to pathogen challenge. Three biparental mapping populations were developed from three resistant accessions viz. CIBC-5, Ei-2, and Cvi-0 and two susceptible accessions – Gre-0 and Zdr-1 (commonly crossed to CIBC-5 and Ei-2). A total of six quantitative trait locus (QTLs) governing resistance to A. brassicae were identified, five of which were population-specific while one QTL was common between all the three mapping populations. Interestingly, the common QTL had varying phenotypic contributions in different populations, which can be attributed to the genetic background of the parental accessions. The presence of both common and population-specific QTLs indicate that resistance to A. brassicae is quantitative, and that different genes may mediate resistance to the pathogen in different accessions. Two of the QTLs had moderate-to-large effects, one of which explained nearly 50% of the variation. The large effect QTLs may therefore contain genes that could play a significant role in conferring resistance even in heterologous hosts. PMID:28286515

  15. Involvement of an extracellular fungus laccase in the flavonoid metabolism in Citrus fruits inoculated with Alternaria alternata.

    PubMed

    Díaz, Licinio; Del Río, José Antonio; Pérez-Gilabert, Manuela; Ortuño, Ana

    2015-04-01

    Fungi of the genus Alternaria are responsible for substantial pre-harvest losses in Citrus. In this study a degradative metabolism of flavonoids (flavanones, flavones and polymethoxyflavones) was observed when 'Fortune' mandarin, Citrus limon and Citrus paradisi, fruits were inoculated with Alternaria alternata, a pre-harvest pathogenic fungus. Associated to this flavonic metabolism the de novo synthesis of the phytoalexin scoparone was detected. This metabolism of flavonoids is caused by an extracellular fungus laccase. The kinetic characterisation of this enzyme revealed that the activity was induced by Citrus flavonoids and was dependent on flavonoid concentrations. The enzyme exhibited a Km of 1.9 mM using ABTS as substrate with an optimum pH of 3.5 in citrate buffer 100 mM. The enzyme is active between 15 and 45 °C, the optimum temperature being around 35 °C, although 50% of the initial activity is lost after 45 min at 35 °C. The A. alternata laccase was inhibited by 0.5 mM l-cysteine and by caffeic acid. Study of the substrate specificity of this enzyme revealed that Citrus flavonoids are substrates of A. alternata laccase. These results suggest that the laccase enzyme could be involved in the pathogenesis of A. alternata in Citrus.

  16. Resistance to citrus canker induced by a variant of Xanthomonas citri ssp. citri is associated with a hypersensitive cell death response involving autophagy-associated vacuolar processes.

    PubMed

    Roeschlin, Roxana A; Favaro, María A; Chiesa, María A; Alemano, Sergio; Vojnov, Adrián A; Castagnaro, Atilio P; Filippone, María P; Gmitter, Frederick G; Gadea, José; Marano, María R

    2016-09-20

    Xanthomonas citri ssp. citri (X. citri) is the causal agent of Asiatic citrus canker, a disease that seriously affects most commercially important Citrus species worldwide. We have identified previously a natural variant, X. citri A(T) , that triggers a host-specific defence response in Citrus limon. However, the mechanisms involved in this canker disease resistance are unknown. In this work, the defence response induced by X. citri A(T) was assessed by transcriptomic, physiological and ultrastructural analyses, and the effects on bacterial biofilm formation were monitored in parallel. We show that X. citri A(T) triggers a hypersensitive response associated with the interference of biofilm development and arrest of bacterial growth in C. limon. This plant response involves an extensive transcriptional reprogramming, setting in motion cell wall reinforcement, the oxidative burst and the accumulation of salicylic acid (SA) and phenolic compounds. Ultrastructural analyses revealed subcellular changes involving the activation of autophagy-associated vacuolar processes. Our findings show the activation of SA-dependent defence in response to X. citri A(T) and suggest a coordinated regulation between the SA and flavonoid pathways, which is associated with autophagy mechanisms that control pathogen invasion in C. limon. Furthermore, this defence response protects C. limon plants from disease on subsequent challenges by pathogenic X. citri. This knowledge will allow the rational exploitation of the plant immune system as a biotechnological approach for the management of the disease.

  17. Evidence for the role of synchronicity between host phenology and pathogen activity in the distribution of sudden oak death canker disease.

    PubMed

    Dodd, Richard S; Hüberli, Daniel; Mayer, Wasima; Harnik, Tamar Y; Afzal-Rafli, Zara; Garbelotto, Matteo

    2008-07-01

    Variations in synchronicity between colonization rate by the pathogen and host phenology may account for unexplained spatial distribution of canker disease. The hypothesis that synchronous pathogenicity and host development are necessary for incidence of sudden oak death disease was tested by correlating seasonal variations in host cambial phenology and response to inoculation with Phytophthora ramorum. Response to infection was estimated by inoculating branch cuttings from coast live oak (Quercus agrifolia) trees at nine dates through a full annual cycle in 2003-2004. Host phenology was estimated from measurements of bud burst and cambial activity in spring 2006. Lesions were largest in the spring soon after the cambium resumed activity. A moderate genetic component to lesion size was detected. Variation among trees in date of largest lesions correlated with variation in timing of bud burst and cambial phenology. The data support the hypothesis that active host cambial tissue is a necessary requisite for successful infection with the pathogen that causes sudden oak death canker disease. Genetic variation in host phenology will buffer coast live oak against epidemics of this disease.

  18. Expression of ß-1,3-glucanase and ß-1,4-glucanase in two potato cultivars following challenge by the fungal pathogen Alternaria solani

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Early blight of potato, caused by Alternaria solani, is a ubiquitous disease in many countries around the world. We have previously found that variation in resistance phenotypes exist between two different Iranian cultivars of potato. Cultivar ‘Diamond’ is more resistant to multiple isolates of A. s...

  19. Functional analyses of the Diels-Alderase gene sol5 of Ascochyta rabiei and Alternaria solani indicate that the Solanapyrone phytotoxins are not required for pathogenicity

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Ascochyta rabiei and Alternaria solani, the causal agents of Ascochyta blight of chickpea (Cicer arietinum) and early blight of potato (Solanum tuberosum), respectively, produce a set of phytotoxic compounds incuding solanapyrones A, B, and C. Although both the phytotoxicity of solanopyrones and the...

  20. Occurrence of Cladosporium spp. and Alternaria spp. spores in Western, Northern and Central-Eastern Poland in 2004-2006 and relation to some meteorological factors

    NASA Astrophysics Data System (ADS)

    Grinn-Gofroń, Agnieszka; Rapiejko, Piotr

    2009-08-01

    The concentration of airborne spores of Cladosporium spp. and Alternaria spp. has been investigated at three monitoring stations situated along the west-north and central-east transect in Poland (Szczecin, Olsztyn, Warszawa,) i.e. from a height of 100 m to 149 m above sea level. The aerobiological monitoring of fungal spores was performed by means of three Lanzoni volumetric spore traps. Cladosporium spp. spores were dominant at all the stations. The highest Cladosporium spp. and Alternaria spp. numbers of spores were observed at all the cities in July and August. Statistically significant correlations have been found between the Cladosporium spp. and Alternaria spp. concentration in the air and the mean air temperature, amount of precipitation, air pressure and relative air humidity. The spore count of Cladosporium spp. and Alternaria spp. is determined by the diversity of local flora and weather conditions, especially by the air temperature. The identification of factors, which influence and shape spore concentrations, may significantly improve the current methods of allergy prevention.

  1. Stem cells.

    PubMed

    Behr, Björn; Ko, Sae Hee; Wong, Victor W; Gurtner, Geoffrey C; Longaker, Michael T

    2010-10-01

    Stem cells are self-renewing cells capable of differentiating into multiple cell lines and are classified according to their origin and their ability to differentiate. Enormous potential exists in use of stem cells for regenerative medicine. To produce effective stem cell-based treatments for a range of diseases, an improved understanding of stem cell biology and better control over stem cell fate are necessary. In addition, the barriers to clinical translation, such as potential oncologic properties of stem cells, need to be addressed. With renewed government support and continued refinement of current stem cell methodologies, the future of stem cell research is exciting and promises to provide novel reconstructive options for patients and surgeons limited by traditional paradigms.

  2. STEM Sell

    ERIC Educational Resources Information Center

    Pantic, Zorica

    2007-01-01

    Between 1994 and 2003, employment in science, technology, engineering and math (STEM) fields grew by a remarkable 23 percent, compared with 17 percent in non-STEM fields, according to federal data. The Bureau of Labor Statistics predicts continued strong growth in STEM job openings through 2014, with emphasis on life sciences, environmental…

  3. Alternaria toxin-induced resistance against rose aphids and olfactory response of aphids to toxin-induced volatiles of rose plants.

    PubMed

    Yang, Fa-zhong; Li, Li; Yang, Bin

    2012-02-01

    The search for active toxins for managing weeds or plant diseases is believed to be a promising avenue of investigation. However, the effects of Alternaria toxins on insects have just begun to be investigated. Bioactivities of toxins from four strains of Alternaria alternata on Rosa chinensis and rose aphid Macrosiphum rosivorum were tested in the present study. At a concentration of 50.0 μg/ml, the crude extract (toxin) of strain 7484 was found not to be harmful to rose plants with excised leaf-puncture method (P≥0.079), and rose plants showed enhanced resistance to rose aphids when this Alternaria toxin was sprayed on the plants (P≤0.001). However, this toxin caused no detrimental effects on aphids in insecticidal bioassay at a concentration of 10.0 to 160.0 μg/ml (P≥0.096). Therefore, the Alternaria toxin had significantly induced the resistance of rose plants against rose aphids, demonstrating that the resistance mechanism triggered by the Alternaria toxin in the rose plant may also be used by the plant to defend itself against insects. Further bioassays aimed to discover the olfactory responses of aphids to the toxin-induced volatiles of host plants. The aphids were significantly more attracted to both volatiles emitted and collected from control rose plants than to both volatiles emitted and collected from the toxin-treated rose plants (P≤0.014). This result showed that the toxin-induced resistance related to the volatile changes of host plants.

  4. Bioactive sulfur-containing sulochrin dimers and other metabolites from an Alternaria sp. isolate from a Hawaiian soil sample.

    PubMed

    Cai, Shengxin; King, Jarrod B; Du, Lin; Powell, Douglas R; Cichewicz, Robert H

    2014-10-24

    Polluxochrin (1) and dioschrin (2), two new dimers of sulochrin linked by thioether bonds, were purified from an Alternaria sp. isolate obtained from a Hawaiian soil sample. The structures of the two metabolites were established by NMR, mass spectrometry data, and X-ray analysis. Metabolite 1 was determined to be susceptible to intramolecular cyclization under aqueous conditions, resulting in the generation of 2 as well as another dimeric compound, castochrin (3). An additional nine new metabolites were also obtained, including four new pyrenochaetic acid derivatives (8-11), one new asterric acid analogue (13), and four new secalonic acid analogues (14-17). Bioassay analysis of these compounds revealed 1-3 displayed antimicrobial and weak cytotoxic activities.

  5. Characterization and temperature-dependent quantification of heat shock protein 60 of the immunogenic fungus Alternaria alternata.

    PubMed

    Buzina, Walter; Raggam, Renhard B; Paulitsch, Astrid; Heiling, Bettina; Marth, Egon

    2008-09-01

    Heat shock proteins or chaperones are found in mitochondrial and cytosolic compartments of cells. They are responsible for the correct folding of proteins and are up-regulated in reaction to various stressors. In addition, when released or presented on the surface of cells, they may play an important role in inflammatory and immunomodulating processes. To identify and characterize hsp60 in the common environmental mold Alternaria alternata, the fungus was cultivated and incubated at different temperatures to induce a possible heat shock response. Fully automated RNA extraction was followed by quantitative real-time RT-PCR targeting A. alternata specific Hsp60 mRNA and subsequent sequencing of the amplicon. While Hsp60 mRNA was constitutively expressed in all samples tested, a temperature-dependent expression of Hsp60 mRNA was observed. Sequencing revealed an identity of more than 85% to other fungal hsp60, indicating the existence of this protein in A. alternata.

  6. Specific Antibodies for the Detection of Alternaria Allergens and the Identification of Cross-Reactive Antigens in Other Fungi

    PubMed Central

    Twaroch, Teresa E.; Curin, Mirela; Sterflinger, Katja; Focke-Tejkl, Margit; Swoboda, Ines; Valenta, Rudolf

    2017-01-01

    Background The mould Alternaria alternata is an important source of respiratory allergens. A. alternata extracts show great variations regarding allergenic potency. The aim of this study was to generate antibody probes specific for important Alternaria allergens and to use them to study allergen expression, depending on different culture conditions, as well as to search for cross-reactive allergens in other mould species. Methods Synthetic peptides from antigenic regions of A. alternata allergens (Alt a 1, Alt a 2, Alt a 3, Alt a 6 and Alt a 8) were used to raise highly specific rabbit antibodies. These antibodies and IgE from allergic patients were used to detect allergens by immunoblotting in extracts of 4 A. alternata strains grown under varying culturing conditions, in commercial skin-prick extracts and in closely (Cladosporium herbarum and Aureobasidium pullulans) or distantly related (Aspergillus niger and Penicillium chrysogenum) mould species. Results There was a wide variation of expression of the individual A. Alternata allergens, depending on the strain and culture conditions, but the antibody probes allowed us to distinguish strains and culture conditions with low and high allergen expression. In the commercial skin-prick solutions, varying levels of Alt a 1 were found, but no other allergens were detectable. Alt a 1 was identified as species-specific A. Alternata allergen, whereas Alt a 3, 6- and Alt a 8-cross-reactive antigens were found in C. herbarum and/or A. pullulans. Conclusions and Clinical Relevance Peptide-specific antibodies are useful to analyze diagnostic and therapeutic mould extracts, to study the presence of A. Alternata allergens in biological samples and to search for cross-reactive allergens in other mould species. PMID:27780168

  7. Partial Resistance of Carrot to Alternaria dauci Correlates with In Vitro Cultured Carrot Cell Resistance to Fungal Exudates

    PubMed Central

    Voisine, Linda; Gatto, Julia; Hélesbeux, Jean-Jacques; Séraphin, Denis; Peña-Rodriguez, Luis M.; Richomme, Pascal; Boedo, Cora; Yovanopoulos, Claire; Gyomlai, Melvina; Briard, Mathilde; Simoneau, Philippe; Poupard, Pascal; Berruyer, Romain

    2014-01-01

    Although different mechanisms have been proposed in the recent years, plant pathogen partial resistance is still poorly understood. Components of the chemical warfare, including the production of plant defense compounds and plant resistance to pathogen-produced toxins, are likely to play a role. Toxins are indeed recognized as important determinants of pathogenicity in necrotrophic fungi. Partial resistance based on quantitative resistance loci and linked to a pathogen-produced toxin has never been fully described. We tested this hypothesis using the Alternaria dauci – carrot pathosystem. Alternaria dauci, causing carrot leaf blight, is a necrotrophic fungus known to produce zinniol, a compound described as a non-host selective toxin. Embryogenic cellular cultures from carrot genotypes varying in resistance against A. dauci were confronted with zinniol at different concentrations or to fungal exudates (raw, organic or aqueous extracts). The plant response was analyzed through the measurement of cytoplasmic esterase activity, as a marker of cell viability, and the differentiation of somatic embryos in cellular cultures. A differential response to toxicity was demonstrated between susceptible and partially resistant genotypes, with a good correlation noted between the resistance to the fungus at the whole plant level and resistance at the cellular level to fungal exudates from raw and organic extracts. No toxic reaction of embryogenic cultures was observed after treatment with the aqueous extract or zinniol used at physiological concentration. Moreover, we did not detect zinniol in toxic fungal extracts by UHPLC analysis. These results suggest that strong phytotoxic compounds are present in the organic extract and remain to be characterized. Our results clearly show that carrot tolerance to A. dauci toxins is one component of its partial resistance. PMID:24983469

  8. Simultaneous analysis of Alternaria toxins and citrinin in tomato: an optimised method using liquid chromatography-tandem mass spectrometry

    PubMed Central

    Tölgyesi, Ádám; Stroka, Joerg; Tamosiunas, Vytautas; Zwickel, Theresa

    2015-01-01

    Alternaria toxins and citrinin are mycotoxins produced by fungi growing on different raw materials and agricultural commodities. Maximum levels of these toxins in foods are currently under consideration by the European Commission as a risk management measure. In this study, a new quantitative method is described for the determination of five Alternaria toxins and citrinin in tomato and tomato juice samples based on LC-MS/MS detection. Samples were extracted with pure methanol, followed by a derivatisation step with 2,4-dinitrophenylhydrazine to improve the determination of tenuazonic acid and to decrease the wide polarity difference between the compounds of interest. Samples were purified on hydrophilic-modified styrene polymer solid-phase extraction cartridges. High-performance liquid chromatographic columns packed with different core–shell materials were tested for the separation of toxins and a C-18 phase was in the final method applied to achieve sufficient separation of all relevant analytes. A key element of this approach was to prove successful transferability of the method to three different triple quadrupole mass spectrometers. A full single laboratory method validation was performed on two LC-MS/MS systems and performance characteristics met the predefined requirements. Moreover, the method was used in an international proficiency test and the satisfactory z-scores obtained (−0.1 to 0.8 in tomato juice samples) demonstrated the reliability of the approach described. The method will be validated in an inter-laboratory collaborative study and if the criteria for method precision are met, the method will be proposed as a new Work Item to the European Committee for Standardisation. PMID:26212568

  9. Apoptosis: A Functional Paradigm for Programmed Plant Cell Death Induced by a Host-Selective Phytotoxin and Invoked during Development.

    PubMed Central

    Wang, H; Li, J; Bostock, RM; Gilchrist, DG

    1996-01-01

    The host-selective AAL toxins secreted by Alternaria alternata f sp lycopersici are primary chemical determinants in the Alternaria stem canker disease of tomato. The AAL toxins are members of a new class of sphinganine analog mycotoxins that cause cell death in both animals and plants. Here, we report detection of stereotypic hallmarks of apoptosis during cell death induced by these toxins in tomato. DNA ladders were observed during cell death in toxin-treated tomato protoplasts and leaflets. The intensity of the DNA ladders was enhanced by Ca2+ and inhibited by Zn2+. The progressive delineation of fragmented DNA into distinct bodies, coincident with the appearance of DNA ladders, also was observed during death of toxin-treated tomato protoplasts. In situ analysis of cells dying during development in both onion root caps and tomato leaf tracheary elements revealed DNA fragmentation localized to the dying cells as well as the additional formation of apoptotic-like bodies in sloughing root cap cells. We conclude that the fundamental elements of apoptosis, as characterized in animals, are conserved in plants. The apoptotic process may be expressed during some developmental transitions and is the functional process by which symptomatic lesions are formed in the Alternaria stem canker disease of tomato. Sphinganine analog mycotoxins may be used to characterize further signaling pathways leading to apoptosis in plants. PMID:12239387

  10. Identification of Pseudomonas syringae pv. actinidiae strains causing bacterial canker of kiwifruit in the Anhui Province of China, and determination of their streptomycin sensitivities.

    PubMed

    Yang, X; Yi, X-K; Chen, Y; Zhang, A-F; Zhang, J-Y; Gao, Z-H; Qi, Y-J; Xu, Y-L

    2015-07-27

    Bacterial canker, caused by Pseudomonas syringae pv. actinidiae, is one of the most severe diseases of kiwifruit. It has become an international pandemic and threatens the sustainable development of kiwifruit production in all main kiwi-growing regions worldwide. Streptomycin has been the major bactericide for the control of kiwifruit canker, especially in Anhui Province, one of the main kiwifruit production regions in China. However, until now, no studies on the baseline sensitivity to streptomycin of field isolates of P. syringae pv. actinidiae from China have been available. During 2012-2013, a total of 102 single-colony P. syringae pv. actinidiae strains were isolated: 36, 12, 13, 26, and 15 strains from Yuexi, Jinzhai, Huoshan, Qianshan, and Taihu counties, respectively. All strains were confirmed by production of a 280-bp fragment using the specific primers PsaF1/R2 upon polymerase chain reaction amplification, followed by an assay for confirmation of pathogenicity to fulfill Koch's postulates. In this study, the streptomycin sensitivity of the 102 isolated strains was determined. The half-maximal effective concentration values for inhibition of growth by streptomycin were 0.03-0.42 μg/mL (average 0.12 ± 0.06 μg/mL). The baseline sensitivity curve was unimodal, representing range-of-variation factors of 14.0. No resistant subpopulation was identified among the strains used in the study. Thus, these sensitivity data could be used as a baseline for monitoring the shift in sensitivity of P. syringae pv. actinidiae populations to streptomycin in Anhui Province. Continuous resistance monitoring should be carried out, as streptomycin is an at-risk bactericide agent.

  11. A filamentous hemagglutinin-like protein of Xanthomonas axonopodis pv. citri, the phytopathogen responsible for citrus canker, is involved in bacterial virulence.

    PubMed

    Gottig, Natalia; Garavaglia, Betiana S; Garofalo, Cecilia G; Orellano, Elena G; Ottado, Jorgelina

    2009-01-01

    Xanthomonas axonopodis pv. citri, the phytopathogen responsible for citrus canker has a number of protein secretion systems and among them, at least one type V protein secretion system belonging to the two-partner secretion pathway. This system is mainly associated to the translocation of large proteins such as adhesins to the outer membrane of several pathogens. Xanthomonas axonopodis pv. citri possess a filamentous hemagglutinin-like protein in close vicinity to its putative transporter protein, XacFhaB and XacFhaC, respectively. Expression analysis indicated that XacFhaB was induced in planta during plant-pathogen interaction. By mutation analysis of XacFhaB and XacFhaC genes we determined that XacFhaB is involved in virulence both in epiphytic and wound inoculations, displaying more dispersed and fewer canker lesions. Unexpectedly, the XacFhaC mutant in the transporter protein produced an intermediate virulence phenotype resembling wild type infection, suggesting that XacFhaB could be secreted by another partner different from XacFhaC. Moreover, XacFhaB mutants showed a general lack of adhesion and were affected in leaf surface attachment and biofilm formation. In agreement with the in planta phenotype, adhesin lacking cells moved faster in swarming plates. Since no hyperflagellation phenotype was observed in this bacteria, the faster movement may be attributed to the lack of cell-to-cell aggregation. Moreover, XacFhaB mutants secreted more exopolysaccharide that in turn may facilitate its motility. Our results suggest that this hemagglutinin-like protein is required for tissue colonization being mainly involved in surface attachment and biofilm formation, and that plant tissue attachment and cell-to-cell aggregation are dependent on the coordinated action of adhesin molecules and exopolysaccharides.

  12. The dual nature of trehalose in citrus canker disease: a virulence factor for Xanthomonas citri subsp. citri and a trigger for plant defence responses

    PubMed Central

    Piazza, Ainelén; Zimaro, Tamara; Garavaglia, Betiana S.; Ficarra, Florencia A.; Thomas, Ludivine; Marondedze, Claudius; Feil, Regina; Lunn, John E.; Gehring, Chris; Ottado, Jorgelina; Gottig, Natalia

    2015-01-01

    Xanthomonas citri subsp. citri (Xcc) is a bacterial pathogen that causes citrus canker in susceptible Citrus spp. The Xcc genome contains genes encoding enzymes from three separate pathways of trehalose biosynthesis. Expression of genes encoding trehalose-6-phosphate synthase (otsA) and trehalose phosphatase (otsB) was highly induced during canker development, suggesting that the two-step pathway of trehalose biosynthesis via trehalose-6-phosphate has a function in pathogenesis. This pathway was eliminated from the bacterium by deletion of the otsA gene. The resulting XccΔotsA mutant produced less trehalose than the wild-type strain, was less resistant to salt and oxidative stresses, and was less able to colonize plant tissues. Gene expression and proteomic analyses of infected leaves showed that infection with XccΔotsA triggered only weak defence responses in the plant compared with infection with Xcc, and had less impact on the host plant’s metabolism than the wild-type strain. These results suggested that trehalose of bacterial origin, synthesized via the otsA–otsB pathway, in Xcc, plays a role in modifying the host plant’s metabolism to its own advantage but is also perceived by the plant as a sign of pathogen attack. Thus, trehalose biosynthesis has both positive and negative consequences for Xcc. On the one hand, it enables this bacterial pathogen to survive in the inhospitable environment of the leaf surface before infection and exploit the host plant’s resources after infection, but on the other hand, it is a tell-tale sign of the pathogen’s presence that triggers the plant to defend itself against infection. PMID:25770587

  13. The dual nature of trehalose in citrus canker disease: a virulence factor for Xanthomonas citri subsp. citri and a trigger for plant defence responses.

    PubMed

    Piazza, Ainelén; Zimaro, Tamara; Garavaglia, Betiana S; Ficarra, Florencia A; Thomas, Ludivine; Marondedze, Claudius; Feil, Regina; Lunn, John E; Gehring, Chris; Ottado, Jorgelina; Gottig, Natalia

    2015-05-01

    Xanthomonas citri subsp. citri (Xcc) is a bacterial pathogen that causes citrus canker in susceptible Citrus spp. The Xcc genome contains genes encoding enzymes from three separate pathways of trehalose biosynthesis. Expression of genes encoding trehalose-6-phosphate synthase (otsA) and trehalose phosphatase (otsB) was highly induced during canker development, suggesting that the two-step pathway of trehalose biosynthesis via trehalose-6-phosphate has a function in pathogenesis. This pathway was eliminated from the bacterium by deletion of the otsA gene. The resulting XccΔotsA mutant produced less trehalose than the wild-type strain, was less resistant to salt and oxidative stresses, and was less able to colonize plant tissues. Gene expression and proteomic analyses of infected leaves showed that infection with XccΔotsA triggered only weak defence responses in the plant compared with infection with Xcc, and had less impact on the host plant's metabolism than the wild-type strain. These results suggested that trehalose of bacterial origin, synthesized via the otsA-otsB pathway, in Xcc, plays a role in modifying the host plant's metabolism to its own advantage but is also perceived by the plant as a sign of pathogen attack. Thus, trehalose biosynthesis has both positive and negative consequences for Xcc. On the one hand, it enables this bacterial pathogen to survive in the inhospitable environment of the leaf surface before infection and exploit the host plant's resources after infection, but on the other hand, it is a tell-tale sign of the pathogen's presence that triggers the plant to defend itself against infection.

  14. Ectopic expression of MdSPDS1 in sweet orange (Citrus sinensis Osbeck) reduces canker susceptibility: involvement of H2O2 production and transcriptional alteration

    PubMed Central

    2011-01-01

    Background Enormous work has shown that polyamines are involved in a variety of physiological processes, but information is scarce on the potential of modifying disease response through genetic transformation of a polyamine biosynthetic gene. Results In the present work, an apple spermidine synthase gene (MdSPDS1) was introduced into sweet orange (Citrus sinensis Osbeck 'Anliucheng') via Agrobacterium-mediated transformation of embryogenic calluses. Two transgenic lines (TG4 and TG9) varied in the transgene expression and cellular endogenous polyamine contents. Pinprick inoculation demonstrated that the transgenic lines were less susceptible to Xanthomonas axonopodis pv. citri (Xac), the causal agent of citrus canker, than the wild type plants (WT). In addition, our data showed that upon Xac attack TG9 had significantly higher free spermine (Spm) and polyamine oxidase (PAO) activity when compared with the WT, concurrent with an apparent hypersensitive response and the accumulation of more H2O2. Pretreatment of TG9 leaves with guazatine acetate, an inhibitor of PAO, repressed PAO activity and reduced H2O2 accumulation, leading to more conspicuous disease symptoms than the controls when both were challenged with Xac. Moreover, mRNA levels of most of the defense-related genes involved in synthesis of pathogenesis-related protein and jasmonic acid were upregulated in TG9 than in the WT regardless of Xac infection. Conclusion Our results demonstrated that overexpression of the MdSPDS1 gene prominently lowered the sensitivity of the transgenic plants to canker. This may be, at least partially, correlated with the generation of more H2O2 due to increased production of polyamines and enhanced PAO-mediated catabolism, triggering hypersensitive response or activation of defense-related genes. PMID:21439092

  15. Canker Sores (For Parents)

    MedlinePlus

    ... open sores that have a white or yellowish coating and a red "halo" around them. They tend ... take you to a site outside of KidsHealth's control. About KidsHealth Nemours.org Reading BrightStart! Contact Us ...

  16. Canker Sores: Treatment

    MedlinePlus

    ... small part of the underlying problem. A general rule of thumb recommended by health care professionals is that any ulcer that lasts for more than 10 to 14 days should be evaluated by a dentist and biopsied ...

  17. STEM Education

    PubMed Central

    Xie, Yu; Fang, Michael; Shauman, Kimberlee

    2015-01-01

    Improving science, technology, engineering, and mathematics (STEM) education, especially for traditionally disadvantaged groups, is widely recognized as pivotal to the U.S.’s long-term economic growth and security. In this article, we review and discuss current research on STEM education in the U.S., drawing on recent research in sociology and related fields. The reviewed literature shows that different social factors affect the two major components of STEM education attainment: (1) attainment of education in general, and (2) attainment of STEM education relative to non-STEM education conditional on educational attainment. Cognitive and social psychological characteristics matter for both major components, as do structural influences at the neighborhood, school, and broader cultural levels. However, while commonly used measures of socioeconomic status (SES) predict the attainment of general education, social psychological factors are more important influences on participation and achievement in STEM versus non-STEM education. Domestically, disparities by family SES, race, and gender persist in STEM education. Internationally, American students lag behind those in some countries with less economic resources. Explanations for group disparities within the U.S. and the mediocre international ranking of US student performance require more research, a task that is best accomplished through interdisciplinary approaches. PMID:26778893

  18. STEM Education.

    PubMed

    Xie, Yu; Fang, Michael; Shauman, Kimberlee

    2015-08-01

    Improving science, technology, engineering, and mathematics (STEM) education, especially for traditionally disadvantaged groups, is widely recognized as pivotal to the U.S.'s long-term economic growth and security. In this article, we review and discuss current research on STEM education in the U.S., drawing on recent research in sociology and related fields. The reviewed literature shows that different social factors affect the two major components of STEM education attainment: (1) attainment of education in general, and (2) attainment of STEM education relative to non-STEM education conditional on educational attainment. Cognitive and social psychological characteristics matter for both major components, as do structural influences at the neighborhood, school, and broader cultural levels. However, while commonly used measures of socioeconomic status (SES) predict the attainment of general education, social psychological factors are more important influences on participation and achievement in STEM versus non-STEM education. Domestically, disparities by family SES, race, and gender persist in STEM education. Internationally, American students lag behind those in some countries with less economic resources. Explanations for group disparities within the U.S. and the mediocre international ranking of US student performance require more research, a task that is best accomplished through interdisciplinary approaches.

  19. Determination of Alternaria mycotoxins in wine and juice using ionic liquid modified countercurrent chromatography as a pretreatment method followed by high-performance liquid chromatography.

    PubMed

    Fan, Chen; Cao, Xueli; Liu, Man; Wang, Wei

    2016-03-04

    Alternariol (AOH), alternariol monomethyl ether (AME), and tenuazonic acid (TeA) are some of the main Alternaria mycotoxins that can be found as contaminants in food materials. The objective of this study was to develop a pretreatment method with countercurrent chromatography (CCC) for enrichment and cleanup of trace Alternaria mycotoxins in food samples prior to high-performance liquid chromatography (HPLC) analysis. An Analytical CCC instrument with a column volume 22.5mL was used, and a two-phase solvent system composed of ethyl acetate and water modified with 6% [HOOMIM][Cl] in mass to volume ratio was selected. Under the optimized CCC operation conditions, trace amounts of AOH, AME, and TeA in large volume of liquid sample were efficiently extracted and enriched in the stationary phase, and then eluted out just by reversing the stationary phase as mobile phase in the opposite flowing direction tail-to-head. The enrichment and elution strategies are unique and can be fulfilled online with high enrichment factors (87-114) and high recoveries (81.14-110.94%). The method has been successively applied to the determination of Alternaria mycotoxins in real apple juice and wine samples with the limits of detection (LOD) in the range of 0.03-0.14μgL(-1). Totally 12 wine samples and 15 apple juice samples from the local market were analyzed. The detection rate of AOH and AME in both kinds of the samples were more than 50%, while TeA was found in relatively high level of 1.75-49.61μgL(-1) in some of the apple juice samples. The proposed method is simple, rapid, and sensitive and could also be used for the analysis and monitoring of Alternaria mycotoxin in other food samples.

  20. Ex Vivo Application of Secreted Metabolites Produced by Soil-Inhabiting Bacillus spp. Efficiently Controls Foliar Diseases Caused by Alternaria spp.

    PubMed Central

    El-Sayed, Ashraf S. A.; Patel, Jaimin S.; Green, Kari B.; Ali, Mohammad; Brennan, Mary; Norman, David

    2015-01-01

    Bacterial biological control agents (BCAs) are largely used as live products to control plant pathogens. However, due to variable environmental and ecological factors, live BCAs usually fail to produce desirable results against foliar pathogens. In this study, we investigated the potential of cell-free culture filtrates of 12 different bacterial BCAs isolated from flower beds for controlling foliar diseases caused by Alternaria spp. In vitro studies showed that culture filtrates from two isolates belonging to Bacillus subtilis and Bacillus amyloliquefaciens displayed strong efficacy and potencies against Alternaria spp. The antimicrobial activity of the culture filtrate of these two biological control agents was effective over a wider range of pH (3.0 to 9.0) and was not affected by autoclaving or proteolysis. Comparative liquid chromatography-mass spectrometry (LC-MS) analyses showed that a complex mixture of cyclic lipopeptides, primarily of the fengycin A and fengycin B families, was significantly higher in these two BCAs than inactive Bacillus spp. Interaction studies with mixtures of culture filtrates of these two species revealed additive activity, suggesting that they produce similar products, which was confirmed by LC-tandem MS analyses. In in planta pre- and postinoculation trials, foliar application of culture filtrates of B. subtilis reduced lesion sizes and lesion frequencies caused by Alternaria alternata by 68 to 81%. Taken together, our studies suggest that instead of live bacteria, culture filtrates of B. subtilis and B. amyloliquefaciens can be applied either individually or in combination for controlling foliar diseases caused by Alternaria species. PMID:26519395

  1. A 10-year study of Alternaria and Cladosporium in two Polish cities (Szczecin and Cracow) and relationship with the meteorological parameters.

    PubMed

    Grinn-Gofroń, Agnieszka; Strzelczak, Agnieszka; Stępalska, Danuta; Myszkowska, Dorota

    Alternaria and Cladosporium spores belong to the most frequent and allergenic particles in bioaerosol in the temperate climate. The investigation of Alternaria and Cladosporium spore concentrations was performed in two cities in Poland, Szczecin and Cracow, in 2004-2013. The meteorological parameters taken to assess their impact on fungal spores were average, maximum and minimum temperature, relative humidity and average wind velocity. In order to reveal whether changes in dynamics of spore seasons are driven by meteorological conditions, ordination methods were applied. Canonical correspondence analysis was used to explore redundancy among the predictors (meteorological parameters). Prior to ordination analyses, the data were log(x)-transformed. Concentrations of Alternaria and Cladosporium spores were significantly higher in Szczecin comparing to Cracow, but it was also observed the decreasing trend in the spore concentrations in Szczecin. As regards temperature, it was higher in Cracow and was still increasing in the studied years. Relative humidity and wind velocity were significantly lower in Cracow. In Szczecin meteorological conditions did not explain changes in spore season characteristics (insignificant redundancy analysis models), while in Cracow's redundancy analysis models indicated that spore season parameters were in over 40 % determined by meteorological conditions, mainly air temperature and wind velocity. If they increase, the peak value, total number of spores and their average concentrations in a season will also increase.

  2. The Arabidopsis thaliana-Alternaria brassicicola pathosystem: A model interaction for investigating seed transmission of necrotrophic fungi

    PubMed Central

    2012-01-01

    Background Seed transmission constitutes a major component of the parasitic cycle for several fungal pathogens. However, very little is known concerning fungal or plant genetic factors that impact seed transmission and mechanisms underlying this key biological trait have yet to be clarified. Such lack of available data could be probably explained by the absence of suitable model pathosystem to study plant-fungus interactions during the plant reproductive phase. Results Here we report on setting up a new pathosystem that could facilitate the study of fungal seed transmission. Reproductive organs of Arabidopsis thaliana were inoculated with Alternaria brassicicola conidia. Parameters (floral vs fruit route, seed collection date, plant and silique developmental stages) that could influence the seed transmission efficiency were tested to define optimal seed infection conditions. Microscopic observations revealed that the fungus penetrates siliques through cellular junctions, replum and stomata, and into seed coats either directly or through cracks. The ability of the osmosensitive fungal mutant nik1Δ3 to transmit to A. thaliana seeds was analyzed. A significant decrease in seed transmission rate was observed compared to the wild-type parental strain, confirming that a functional osmoregulation pathway is required for efficient seed transmission of the fungus. Similarly, to test the role of flavonoids in seed coat protection against pathogens, a transparent testa Arabidopsis mutant (tt4-1) not producing any flavonoid was used as host plant. Unexpectedly, tt4-1 seeds were infected to a significantly lower extent than wild-type seeds, possibly due to over-accumulation of other antimicrobial metabolites. Conclusions The Arabidopsis thaliana-Alternaria brassicicola pathosystem, that have been widely used to study plant-pathogen interactions during the vegetative phase, also proved to constitute a suitable model pathosystem for detailed analysis of plant

  3. Identification of Novel Source of Resistance and Differential Response of Allium Genotypes to Purple Blotch Pathogen, Alternaria porri (Ellis) Ciferri

    PubMed Central

    Nanda, Satyabrata; Chand, Subodh Kumar; Mandal, Purander; Tripathy, Pradyumna; Joshi, Raj Kumar

    2016-01-01

    Purple blotch, caused by Alternaria porri (Ellis) Cifferi, is a serious disease incurring heavy yield losses in the bulb and seed crop of onion and garlic worldwide. There is an immediate need for identification of effective resistance sources for use in host resistance breeding. A total of 43 Allium genotypes were screened for purple blotch resistance under field conditions. Allium cepa accession ‘CBT-Ac77’ and cultivar ‘Arka Kalyan’ were observed to be highly resistant. In vitro inoculation of a selected set of genotypes with A. porri, revealed that 7 days after inoculation was suitable to observe the disease severity. In vitro screening of 43 genotypes for resistance to A. porri revealed two resistant lines. An additional 14 genotypes showed consistent moderate resistance in the field as well as in vitro evaluations. Among the related Allium species, A. schoenoprasum and A. roylei showed the least disease index and can be used for interspecific hybridization with cultivated onion. Differential reaction analysis of three A. porri isolates (Apo-Chiplima, Apn-Nasik, Apg-Guntur) in 43 genotypes revealed significant variation among the evaluated Allium species (P = 0.001). All together, the present study suggest that, the newly identified resistance sources can be used as potential donors for ongoing purple blotch resistance breeding program in India. PMID:27904458

  4. Conidiogenic effects of mannose-binding lectins isolated from cotyledons of red kidney bean (Phaseolus vulgaris) on Alternaria alternata.

    PubMed

    Alizadeh, Hossein; Leung, David W M; Cole, Anthony L J

    2011-01-01

    Effect of proteinaceous extracts from red kidney bean cotyledons on mycelium of Alternaria alternata growing on potato dextrose agar (PDA) plates was investigated. Unexpectedly, conidia formation was induced in response to applied crude extracts. A PDA disc method was developed to quantify conidia formed. A purified fraction retaining conidiation inducing effect (CIE) was obtained following several protein purification procedures including the last step of eluting bound proteins from an Affi-gel blue gel column. Based on MALDI (matrix assisted laser desorption/ionization) mass spectrometric analysis, a previously identified mannose-binding lectin (MBL) called PvFRIL (Phaseolus vulgaris fetal liver tyrosine kinase 3-receptor interacting lectin) was present in this conidiation inducing fraction. The PvFRIL was subsequently purified using a single step mannose-agarose affinity column chromatography. When the lectin was applied exogenously to A. alternata, increased conidiation resulted. The conidia produced in response to the MBL were similar to those induced by other methods and their germ tubes were longer after 12 h growth than those induced under white light. To our knowledge this is the first report of exogenous application of a PvFRIL or another purified protein from a plant inducing conidia formation in a fungus.

  5. TeA is a key virulence factor for Alternaria alternata (Fr.) Keissler infection of its host.

    PubMed

    Kang, Ye; Feng, Hongwei; Zhang, Jingxu; Chen, Shiguo; Valverde, Bernal E; Qiang, Sheng

    2017-03-04

    A toxin-deficient mutant strain, HP001 mutant of Alternaria alternata, whose mycelium is unable to infect its host, produces little tenuazonic acid (TeA) toxin. How TeA plays a role in initiating host infection by A. alternata remains unclear. In this research we use Imaging-PAM based on chlorophyll fluorescence parameters and transmission electron microscopy to explore the role of TeA toxin during the infection process of A. alternata. Photosystem II damage began even before wild type mycelium infected the leaves of its host, croftonweed (Ageratina adenophora). Compared with the wild type, HP001 mutant produces morphologically different colonies, hyphae with thinner cell walls, has higher reactive oxygen species (ROS) content and lower peroxidase activity, and fails to form appressoria on the host surface. Adding TeA toxin allows the mutant to partially recover these characters and more closely resemble the wild type. Additionally, we found that the mutant is able to elicit disease symptoms when its mycelium is placed on leaves whose epidermis has been manually removed, which indicates that TeA may be determinant in the fungus recognition of its plant host. Lack of TeA toxin appears responsible for the loss of pathogenicity of the HP001 mutant. As a key virulence factor, TeA toxin not only damages the host plant but also is involved in maintaining ROS content, host recognition, inducing appressoria to infect the host and for allowing completion of the infection process.

  6. Resistance to oxidative stress via regulating siderophore-mediated iron acquisition by the citrus fungal pathogen Alternaria alternata.

    PubMed

    Chen, Li-Hung; Yang, Siwy Ling; Chung, Kuang-Ren

    2014-05-01

    The ability of the necrotrophic fungus Alternaria alternata to detoxify reactive oxygen species (ROS) is crucial for pathogenesis to citrus. We report regulation of siderophore-mediated iron acquisition and ROS resistance by the NADPH oxidase (NOX), the redox activating yes-associated protein 1 (YAP1) regulator, and the high-osmolarity glycerol 1 (HOG1) mitogen-activated protein kinase (MAPK). The A. alternata nonribosomal peptide synthetase (NPS6) is essential for the biosynthesis of siderophores, contributing to iron uptake under low-iron conditions. Fungal strains impaired for NOX, YAP1, HOG1 or NPS6 all display increased sensitivity to ROS. Exogenous addition of iron at least partially rescues ROS sensitivity seen for NPS6, YAP1, HOG1, and NOX mutants. Importantly, expression of the NPS6 gene and biosynthesis of siderophores are regulated by NOX, YAP1 and HOG1, supporting a functional link among these regulatory pathways. Although iron fully rescues H2O2 sensitivity seen in mutants impaired for the response regulator SKN7, neither expression of NPS6 nor biosynthesis of siderophores is controlled by SKN7. Our results indicate that the acquisition of environmental iron has profound effects on ROS detoxification.

  7. Decolorization and biodegradation of congo red dye by a novel white rot fungus Alternaria alternata CMERI F6.

    PubMed

    Chakraborty, Samayita; Basak, Bikram; Dutta, Subhasish; Bhunia, Biswanath; Dey, Apurba

    2013-11-01

    A novel white rot fungus Alternaria alternata CMERI F6 decolorized 99.99% of 600 mg/L congo red within 48 h in yeast extract-glucose medium at 25 °C, pH 5 and 150 rpm. Physicochemical parameters like carbon and nitrogen sources, temperature, pH and aeration were optimized to develop faster decolorization process. Dye decolorization rate was maximal (20.21 mg/L h) at 25 °C, pH 5, 150 rpm and 800 mg/L dye, giving 78% final decolorization efficiency. Scanning electron microscopy and X-ray Diffraction analysis revealed that the fungus become amorphous after dye adsorption. HPLC and FTIR analysis of the extracted metabolites suggested that the decolorization occurred through biosorption and biodegradation. Thermogravimetric analysis (TGA), differential thermal analysis (DTA) and acid-alkali and 70% ethanol treatment revealed the efficient dye retention capability of the fungus. The foregoing results justify the applicability of the strain in removal of congo red from textile wastewaters and their safe disposal.

  8. Phosphoproteome profiles of the phytopathogenic fungi Alternaria brassicicola and Botrytis cinerea during exponential growth in axenic cultures.

    PubMed

    Davanture, Marlène; Dumur, Jérôme; Bataillé-Simoneau, Nelly; Campion, Claire; Valot, Benoît; Zivy, Michel; Simoneau, Philippe; Fillinger, Sabine

    2014-07-01

    This study describes the gel-free phosphoproteomic analysis of the phytopathogenic fungi Alternaria brassicicola and Botrytis cinerea grown in vitro under nonlimiting conditions. Using a combination of strong cation exchange and IMAC prior to LC-MS, we identified over 1350 phosphopeptides per fungus representing over 800 phosphoproteins. The preferred phosphorylation sites were found on serine (>80%) and threonine (>15%), whereas phosphorylated tyrosine residues were found at less than 1% in A. brassicicola and at a slightly higher ratio in B. cinerea (1.5%). Biological processes represented principally among the phoshoproteins were those involved in response and transduction of stimuli as well as in regulation of cellular and metabolic processes. Most known elements of signal transduction were found in the datasets of both fungi. This study also revealed unexpected phosphorylation sites in histidine kinases, a category overrepresented in filamentous ascomycetes compared to yeast. The data have been deposited to the ProteomeXchange database with identifier PXD000817 (http://proteomecentral.proteomexchange.org/dataset/PXD000817).

  9. LaeA and VeA are involved in growth morphology, asexual development, and mycotoxin production in Alternaria alternata.

    PubMed

    Estiarte, N; Lawrence, C B; Sanchis, V; Ramos, A J; Crespo-Sempere, A

    2016-12-05

    Alternaria alternata is a common filamentous fungus that contaminates various fruits, grains and vegetables causing important economic losses to farmers and the food industry. A. alternata is a mycotoxigenic mould, which may jeopardize human and animal health. Two of the most common A. alternata mycotoxins found in food and feed are alternariol and alternariol monomethyl ether. In this study we examined the role of LaeA and VeA, two regulatory proteins belonging to the velvet family, which have been described to be involved in several functions in many fungi including secondary metabolism. We found that deletion of laeA and veA genes, respectively, greatly reduced sporulation and strongly compromised mycotoxin production, both in vitro or during pathogenesis of tomato fruits. We have also studied how the loss of laeA and veA may affect expression of genes related to alternariol and alternariol monomethyl ether biosynthesis (pksJ and altR), and to melanin biosynthesis (cmrA, pksA).

  10. Zinc hyperaccumulation substitutes for defense failures beyond salicylate and jasmonate signaling pathways of Alternaria brassicicola attack in Noccaea caerulescens.

    PubMed

    Gallego, Berta; Martos, Soledad; Cabot, Catalina; Barceló, Juan; Poschenrieder, Charlotte

    2017-04-01

    The hypothesis of metal defense as a substitute for a defective biotic stress signaling system in metal hyperaccumulators was tested using the pathosystem Alternaria brassicicola-Noccaea caerulescens under low (2 µM), medium (12 µM) and high (102 µM) Zn supply. Regardless the Zn supply, N. caerulescens responded to fungal attack with the activation of both HMA4 coding for a Zn transporter, and biotic stress signaling pathways. Salicylate, jasmonate, abscisic acid and indoleacetic acid concentrations, as well as biotic stress marker genes (PDF1.2, CHIB, LOX2, PR1 and BGL2) were activated 24 h upon inoculation. Based on the activation of defense genes 24 h after the inoculation an incompatible fungal-plant interaction could be predicted. Nonetheless, in the longer term (7 days) no effective protection against A. brassicicola was achieved in plants exposed to low and medium Zn supply. After 1 week the biotic stress markers were even further increased in these plants, and this compatible interaction was apparently not caused by a failure in the signaling of the fungal attack, but due to the lack of specificity in the type of the activated defense mechanisms. Only plants receiving high Zn exhibited an incompatible fungal interaction. High Zn accumulation in these plants, possibly in cooperation with high glucosinolate concentrations, substituted for the ineffective defense system and the interaction turned into incompatible. In a threshold-type response, these joint effects efficiently hampered fungal spread and, consequently decreased the biotic stress signaling.

  11. Inhibitors of BRD4 Protein from a Marine-Derived Fungus Alternaria sp. NH-F6

    PubMed Central

    Ding, Hui; Zhang, Dashan; Zhou, Biao; Ma, Zhongjun

    2017-01-01

    Bromodomains (BRD) are readers of the epigenetic code that regulate gene transcription through their recognition of acetyl-lysine modified histone tails. Recently, bromodomain-containing proteins such as BRD4 have been demonstrated to be druggable through the discovery of potent inhibitors. These protein–protein interaction inhibitors have the potential to modulate multiple diseases by their profound anti-inflammatory and antiproliferative effects. In order to explore new BRD4 inhibitors as well as lead compounds for the development of new drugs, the secondary metabolites of Alternaria sp. NH-F6, a fungus isolated from deep-sea sediment samples, were analyzed systematically. Five new compounds including two new perylenequinones (1–2), one new alternaric acid (3), 2-(N-vinylacetamide)-4-hydroxymethyl-3-ene-butyrolactone (4), one new cerebroside (5), together with 19 known compounds (6–24) were isolated from the ethyl acetate extracts of this strain. Their structures were elucidated using nuclear magnetic resonance (NMR) and high resolution electrospray ionization mass spectrometry (HR-ESI-MS) analyses. Finally, all these compounds were evaluated for their inhibitory activity against BRD4 protein, and compound 2 exhibited a potent inhibition rate of 88.1% at a concentration of 10 µM. This research provides a new BRD4 inhibitor which may possess potential antitumoral, antiviral, or anti-inflammatory pharmaceutical values. PMID:28300771

  12. Characterization of the programmed cell death induced by metabolic products of Alternaria alternata in tobacco BY-2 cells.

    PubMed

    Cheng, Dan-Dan; Jia, Yu-Jiao; Gao, Hui-Yuan; Zhang, Li-Tao; Zhang, Zi-Shan; Xue, Zhong-Cai; Meng, Qing-Wei

    2011-02-01

    Alternaria alternata has received considerable attention in current literature and most of the studies are focused on its pathogenic effects on plant chloroplasts, but little is known about the characteristics of programmed cell death (PCD) induced by metabolic products (MP) of A. alternata, the effects of the MP on mitochondrial respiration and its relation to PCD. The purpose of this study was to explore the mechanism of MP-induced PCD in non-green tobacco BY-2 cells and to explore the role of mitochondrial inhibitory processes in the PCD of tobacco BY-2 cells. MP treatment led to significant cell death that was proven to be PCD by the concurrent cytoplasm shrinkage, chromatin condensation and DNA laddering observed in the cells. Moreover, MP treatment resulted in the overproduction of reactive oxygen species (ROS), rapid ATP depletion and a respiratory decline in the tobacco BY-2 cells. It was concluded that the direct inhibition of the mitochondrial electron transport chain (ETC), alternative pathway (AOX) capacity and catalase (CAT) activity by the MP might be the main contributors to the MP-induced ROS burst observed in tobacco BY-2 cells. The addition of adenosine together with the MP significantly inhibited ATP depletion without preventing PCD; however, when the cells were treated with the MP plus CAT, ROS overproduction was blocked and PCD did not occur. The data presented here demonstrate that the ROS burst played an important role in MP-induced PCD in the tobacco BY-2 cells.

  13. Evaluation of Models for Alternaria Brown Spot of Mandarin Under Mediterranean Conditions by Partial Receiver Operating Characteristic Curve Analysis.

    PubMed

    Bassimba, Daniel D M; Mira, Jose L; Vicent, Antonio

    2017-04-12

    Alternaria brown spot (ABS) is a serious fungal disease of mandarin in the Mediterranean Basin. Due to the rigorous fruit quality standards, models for ABS should avoid false negatives. Experiments were conducted with susceptible 'Fortune' and 'Nova' inoculated at different temperatures and leaf wetness durations, including interrupted periods. Effects of temperature and time elapsed after inoculation were also studied. Disease incidence data were fitted to generalized additive models and a generic infection model. Exposure of trap plants in affected orchards was used for model evaluation, including the Alter-Rater and a simple rule system (SRS). The predictive ability of the models was analyzed using the partial area under the receiver operating characteristic curve in the high-sensitivity range between 0.9 and 1. Postinoculation temperature had a significant effect on disease incidence, with maximum symptom expression after 30 h on Fortune and 60 h on Nova. ABS incidence did not increase after a leaf wetness interruption of 1 h on Nova and 2 h on Fortune. All the models evaluated had high false-positive rates on Fortune. Only the SRS showed a substantial strength of agreement in Nova, with a true-positive rate of 0.93 and false-positive rate of 0.16.

  14. In vitro antifungal activity of four chemotypes of Lippia alba (Verbenaceae) essential oils against Alternaria solani (Pleosporeaceae) isolates.

    PubMed

    Tomazoni, Elisa Z; Pansera, Márcia R; Pauletti, Gabriel F; Moura, Sidnei; Ribeiro, Rute T S; Schwambach, Joséli

    2016-05-31

    Several volatile natural compounds produced by plant secondary metabolism have been proven to present antimicrobial action, enabling their use in phytopathogen control. They also present low environmental impact when compared to conventional pesticides. Essential oils contain these compounds and can be found in several plant species, such as Lippia alba (Mill.) N.E. Brown (Verbenaceae). Essential oils of four chemotypes of L. alba, characterized by their major compounds, namely camphor, citral, linalool and camphor/1,8-cineole, were tested against the phytopathogen Alternaria solani Sorauer (Pleosporaceae), which causes early blight on tomatoes and is responsible for great economic losses regarding production. Essential oils antifungal action was tested in vitro using potato dextrose agar medium with essential oil concentrations at 0.1, 0.5, 1.0, 1.5 and 2.0 µL mL-1. The chemotype that had the best performance was citral, showing significant inhibition compared to the others, starting at the 0.5 µL mL-1 concentration. The essential oil belonging to the linalool chemotype was efficient starting at the 1.5 µL mL-1 concentration. Conversely, the camphor chemotype did not show any action against the phytopathogen. Moreover, the essential oils had no remarkable effect on tomato germination and growth. In conclusion, these essential oils presented fungicidal action against A. solani.

  15. Identification of a Polyketide Synthase Required for Alternariol (AOH) and Alternariol-9-Methyl Ether (AME) Formation in Alternaria alternata

    PubMed Central

    Saha, Debjani; Fetzner, Ramona; Burkhardt, Britta; Podlech, Joachim; Metzler, Manfred; Dang, Ha; Lawrence, Christopher; Fischer, Reinhard

    2012-01-01

    Alternaria alternata produces more than 60 secondary metabolites, among which alternariol (AOH) and alternariol-9-methyl ether (AME) are important mycotoxins. Whereas the toxicology of these two polyketide-based compounds has been studied, nothing is known about the genetics of their biosynthesis. One of the postulated core enzymes in the biosynthesis of AOH and AME is polyketide synthase (PKS). In a draft genome sequence of A. alternata we identified 10 putative PKS-encoding genes. The timing of the expression of two PKS genes, pksJ and pksH, correlated with the production of AOH and AME. The PksJ and PksH proteins are predicted to be 2222 and 2821 amino acids in length, respectively. They are both iterative type I reducing polyketide synthases. PksJ harbors a peroxisomal targeting sequence at the C-terminus, suggesting that the biosynthesis occurs at least partly in these organelles. In the vicinity of pksJ we found a transcriptional regulator, altR, involved in pksJ induction and a putative methyl transferase, possibly responsible for AME formation. Downregulation of pksJ and altR caused a large decrease of alternariol formation, suggesting that PksJ is the polyketide synthase required for the postulated Claisen condensations during the biosynthesis. No other enzymes appeared to be required. PksH downregulation affected pksJ expression and thus caused an indirect effect on AOH production. PMID:22792370

  16. Identification of Novel Source of Resistance and Differential Response of Allium Genotypes to Purple Blotch Pathogen, Alternaria porri (Ellis) Ciferri.

    PubMed

    Nanda, Satyabrata; Chand, Subodh Kumar; Mandal, Purander; Tripathy, Pradyumna; Joshi, Raj Kumar

    2016-12-01

    Purple blotch, caused by Alternaria porri (Ellis) Cifferi, is a serious disease incurring heavy yield losses in the bulb and seed crop of onion and garlic worldwide. There is an immediate need for identification of effective resistance sources for use in host resistance breeding. A total of 43 Allium genotypes were screened for purple blotch resistance under field conditions. Allium cepa accession 'CBT-Ac77' and cultivar 'Arka Kalyan' were observed to be highly resistant. In vitro inoculation of a selected set of genotypes with A. porri, revealed that 7 days after inoculation was suitable to observe the disease severity. In vitro screening of 43 genotypes for resistance to A. porri revealed two resistant lines. An additional 14 genotypes showed consistent moderate resistance in the field as well as in vitro evaluations. Among the related Allium species, A. schoenoprasum and A. roylei showed the least disease index and can be used for interspecific hybridization with cultivated onion. Differential reaction analysis of three A. porri isolates (Apo-Chiplima, Apn-Nasik, Apg-Guntur) in 43 genotypes revealed significant variation among the evaluated Allium species (P = 0.001). All together, the present study suggest that, the newly identified resistance sources can be used as potential donors for ongoing purple blotch resistance breeding program in India.

  17. Spatial and temporal distribution of Alternaria spores in the Iberian Peninsula atmosphere, and meteorological relationships: 1993-2009

    NASA Astrophysics Data System (ADS)

    Aira, María-Jesús; Rodríguez-Rajo, Francisco-Javier; Fernández-González, María; Seijo, Carmen; Elvira-Rendueles, Belén; Abreu, Ilda; Gutiérrez-Bustillo, Montserrat; Pérez-Sánchez, Elena; Oliveira, Manuela; Recio, Marta; Tormo, Rafael; Morales, Julia

    2013-03-01

    This paper provides an updated of airborne Alternaria spore spatial and temporal distribution patterns in the Iberian Peninsula, using a common non-viable volumetric sampling method. The highest mean annual spore counts were recorded in Sevilla (39,418 spores), Mérida (33,744) and Málaga (12,947), while other sampling stations never exceeded 5,000. The same cities also recorded the highest mean daily spore counts (Sevilla 109 spores m-3; Mérida 53 spores m-3 and Málaga 35 spores m-3) and the highest number of days on which counts exceeded the threshold levels required to trigger allergy symptoms (Sevilla 38 % and Mérida 30 % of days). Analysis of annual spore distribution patterns revealed either one or two peaks, depending on the location and prevailing climate of sampling stations. For all stations, average temperature was the weather parameter displaying the strongest positive correlation with airborne spore counts, whilst negative correlations were found for rainfall and relative humidity.

  18. Selection and differentiation of Bacillus spp. Antagonistic to Fusarium oxysporum f.sp. lycopersici and Alternaria solani infecting Tomato.

    PubMed

    Shanmugam, Veerubommu; Atri, Kamini; Gupta, Samriti; Kanoujia, Nandina; Naruka, Digvijay Singh

    2011-03-01

    Antagonistic Bacillus spp. displaying in vitro production of siderophore, chitinase, and β-1,3-glucanase were identified from dual culture assays. In independent greenhouse studies, seed bacterization and soil application of Bacillus atrophaeus S2BC-2 challenge inoculated with Fusarium oxysporum f.sp. lycopersici (FOL) and Alternaria solani (AS) recorded low percent disease index of 25.3 and 28.7, respectively, over nonbacterised pathogen control (44.3 and 56.4). The low disease incidence corroborated with tomato growth promotion with high vigor index (8,041.2) and fresh plant weight (82.5 g) on challenge inoculation with FOL. Analysis of root and leaf samples in rhizobacterial treatment challenged with FOL and AS revealed maximum induction of chitinase (1.9 and 1.7 U/mg of protein, respectively) and β-1,3-glucanase (23.5 and 19.2 U/mg of protein, respectively). In native gel activity assays, the rhizobacterial treatment on challenge inoculation strongly expressed three high intensity PO isoforms along with one low intensity isoform. In studies on genetic diversity of the Bacillus strains by repetitive extragenomic palindromic-polymerase chain reaction (REP-PCR) and amplified rDNA restriction analysis (ARDRA) patterns, ARDRA was more highly discriminant than REP-PCR and allowed grouping of the strains and differentiation of the antagonistic strains from other isolates.

  19. Content of the Alternaria mycotoxin tenuazonic acid in food commodities determined by a stable isotope dilution assay.

    PubMed

    Asam, Stefan; Lichtenegger, Martina; Liu, Yang; Rychlik, Michael

    2012-02-01

    The Alternaria mycotoxin tenuazonic acid (TA) was quantified in fruit juices (n = 50), cereals (n = 12) and spices (n = 38) using a recently developed stable isotope dilution assay (SIDA). [(13) C6,(15) N]-TA was used as the internal standard. Method validation revealed low limits of detection (LODs) of 0.15 μg/kg (fruit juices), 1.0 μg/kg (cereals) and 17 μg/kg (spices). The respective limits of quantitation were about three times higher. Recovery was about 100% for all matrices. The precision (relative standard deviation of replicate analyses of naturally contaminated samples) was 4.2% (grape juice; 1.7 μg/kg), 3.5% (whole wheat flour; 36 μg/kg) and 0.9% (curry powder; 215 μg/kg). The median content of TA in the analyzed samples was 1.8 μg/kg (fruit juices), 16 μg/kg (cereals) and 500 μg/kg (spices). Positive samples amounted to 86% (fruit juices), 92% (cereals) and 87% (spices).

  20. Stem Cell Basics

    MedlinePlus

    ... Tips Info Center Research Topics Federal Policy Glossary Stem Cell Information General Information Clinical Trials Funding Information Current ... Basics » Stem Cell Basics I. Back to top Stem Cell Basics I. Introduction: What are stem cells, and ...

  1. Inhibitory activity of Indian spice plant Cinnamomum zeylanicum extracts against Alternaria solani and Curvularia lunata, the pathogenic dematiaceous moulds

    PubMed Central

    Mishra, Ajay K; Mishra, Amita; Kehri, HK; Sharma, Bechan; Pandey, Abhay K

    2009-01-01

    Background Dematiaceous moulds are pathogenic microorganisms and act as etiological agents of mycoses with different degrees of severity in humans and animals. These moulds also cause loss of food crops and storage food products. The information regarding antimicrobial efficacy of the plant preparations on these moulds is scanty. The present study reveals phytochemical characterization and the effect of bark and leaf extracts of Indian spice plant, Cinnamomum zeylanicum (Cz), against the growth of two species of dematiaceous moulds, Alternaria solani and Curvularia lunata. Methods Cz bark and leaf samples were sequentially extracted in different solvents using Soxhlet apparatus. Phytochemical analyses of extracts were done as per standard protocols. The antifungal bioassay of extracts was done by hanging drop technique. The inhibition of fungal spore germination was monitored under influence of three different concentrations of extracts. Results The lowest test concentration (50 μg/ml) of extracts of Cz bark prepared into acetone and that of Cz leaf into petroleum ether and ethanol exhibited complete inhibition (100%) of spore germination in both the moulds. At 100 μg/ml concentration all the extracts showed about 50 to 100% inhibition. However, the treatment of the spores of the two fungal species with highest concentration (500 μg/ml) of bark and leaf extracts in all the solvents showed 100% fungicidal activity as it completely arrested the germination of spores. Relatively lower activity of aqueous extracts at 50 and 100 μg/ml concentrations suggests that the antifungal ingredients present in Cz bark and leaf are more soluble in organic solvents than water. Conclusion The results demonstrated that the Cz bark and leaves contain certain fungicidal constituents exhibiting potential antimould activity against A. solani and C. lunata. PMID:19267932

  2. [Peculiarities of linear growth of the melanin-containing fungi Cladosporium sphaerospermum Penz. and Alternaria alternata (Fr.) Keissler].

    PubMed

    Vember, V V; Zhdanova, N N

    2001-01-01

    Results on determination of radial (linear) growth rate of melanin-containing fungi Cladosporium sphaerospermum Penz. and Alternaria alternata (Fr.) Keissler concerning place of their isolation and ecologic peculiarities are presented in the paper. Essential differences, as to all studied indices of the strains of C. sphaerospermum strains, isolated from premices of the ChNPP 4th unit from the control strains of this species have been established. Only strain 34 of C. sphaerospermum, isolated from the cable driving region of the "Shelter" object, which did not differ from the control strains as to all the studied indices, made the exception. The radial growth rate was 2-4 times as low and variability amplitude 10-30 times as high in strains 60 and 5-1 of C. sphaerospermum, (which suffered high radiation load), as in the control strains of this species. The radial growth rate of C. sphaerospermum studied strains was an order lower as a whole than that of A. alternata strains. All the above said evidence for implementation of K-type vital strategy by C. sphaerospermum species and in this connection one can suppose its active growth on the surface of walls and ferroconcrete structures of the ChNPP 4th unit. Absence of differences, as to the studied radial growth indices, between A. alternata strains isolated from the ChNPP 4th unit premices and control strains of the species can evidence for the existence of A. alternata species under the conditions of high radioactivity in the surviving state.

  3. Biopriming of Infected Carrot Seed with an Antagonist, Clonostachys rosea, Selected for Control of Seedborne Alternaria spp.

    PubMed

    Jensen, Birgit; Knudsen, Inge M B; Madsen, Mette; Jensen, Dan Funck

    2004-06-01

    ABSTRACT An ecological approach was used to select fungal antagonists effective against the seedborne pathogens Alternaria dauci and A. radicina on carrot. Twenty-five and 105 isolates originating from cereal and carrot habitats were screened against the pathogens in planta, respectively. Irrespective of isolate origin, fungal isolates belonging to Clonostachys rosea controlled pre- and postemergence death caused by A. dauci and A. radicina as effectively as the fungicide iprodione. Isolate IK726 of C. rosea was used in biopriming a seed lot with 29% A. radicina and 11% A. dauci (highly infected), and a seed lot with 4% A. radicina and 7% A. dauci (low infection). Seeds were primed with water alone (hydropriming) or with addition of C. rosea IK726 (biopriming). The occurrence of A. radicina and A. dauci increased twofold and fivefold, respectively, during 14 days hydropriming, irrespective of the initial infection level. On highly infected seed, biopriming reduced the incidence of A. radicina to <2.3% and that of A. dauci to <4.8% while the level of both pathogens was <0.5% on bioprimed seed with a low initial infection rate. In sand stand establishment tests, hydroprimed seeds had a lower healthy seedling stand than nonprimed seeds, mainly due to a high degree of postemergence seedling death. In contrast, biopriming resulted in a seedling stand that was better than that of both nonprimed and hydroprimed seeds. C. rosea IK726 multiplied fivefold to eightfold, and microscopic observations using C. rosea IK726 transformed with a green fluorescent protein (GFP) reporter gene showed that seeds were covered with a fine web of sporulating mycelium of C. rosea. The positive effect of biopriming on healthy seedling stand remained after 5 months of storage at 4 degrees C and IK726 survived at high numbers on these seed. In this study, we demonstrated that bio-priming with the biocontrol strain C. rosea IK726 facilitates priming of infected seeds without risking adverse

  4. Hyperspectral remote sensing for advanced detection of early blight (Alternaria solani) disease in potato (Solanum tuberosum) plants

    NASA Astrophysics Data System (ADS)

    Atherton, Daniel

    Early detection of disease and insect infestation within crops and precise application of pesticides can help reduce potential production losses, reduce environmental risk, and reduce the cost of farming. The goal of this study was the advanced detection of early blight (Alternaria solani) in potato (Solanum tuberosum) plants using hyperspectral remote sensing data captured with a handheld spectroradiometer. Hyperspectral reflectance spectra were captured 10 times over five weeks from plants grown to the vegetative and tuber bulking growth stages. The spectra were analyzed using principal component analysis (PCA), spectral change (ratio) analysis, partial least squares (PLS), cluster analysis, and vegetative indices. PCA successfully distinguished more heavily diseased plants from healthy and minimally diseased plants using two principal components. Spectral change (ratio) analysis provided wavelengths (490-510, 640, 665-670, 690, 740-750, and 935 nm) most sensitive to early blight infection followed by ANOVA results indicating a highly significant difference (p < 0.0001) between disease rating group means. In the majority of the experiments, comparisons of diseased plants with healthy plants using Fisher's LSD revealed more heavily diseased plants were significantly different from healthy plants. PLS analysis demonstrated the feasibility of detecting early blight infected plants, finding four optimal factors for raw spectra with the predictor variation explained ranging from 93.4% to 94.6% and the response variation explained ranging from 42.7% to 64.7%. Cluster analysis successfully distinguished healthy plants from all diseased plants except for the most mildly diseased plants, showing clustering analysis was an effective method for detection of early blight. Analysis of the reflectance spectra using the simple ratio (SR) and the normalized difference vegetative index (NDVI) was effective at differentiating all diseased plants from healthy plants, except for the

  5. RNA-Seq derived identification of differential transcription in the chrysanthemum leaf following inoculation with Alternaria tenuissima

    PubMed Central

    2014-01-01

    Background A major production constraint on the important ornamental species chrysanthemum is black spot which is caused by the necrotrophic fungus Alternaria tenuissima. The molecular basis of host resistance to A. tenuissima has not been studied as yet in any detail. Here, high throughput sequencing was taken to characterize the transcriptomic response of the chrysanthemum leaf to A. tenuissima inoculation. Results The transcriptomic data was acquired using RNA-Seq technology, based on the Illumina HiSeq™ 2000 platform. Four different libraries derived from two sets of leaves harvested from either inoculated or mock-inoculated plants were characterized. Over seven million clean reads were generated from each library, each corresponding to a coverage of >350,000 nt. About 70% of the reads could be mapped to a set of chrysanthemum unigenes. Read frequency was used as a measure of transcript abundance and therefore as an identifier of differential transcription in the four libraries. The differentially transcribed genes identified were involved in photosynthesis, pathogen recognition, reactive oxygen species generation, cell wall modification and phytohormone signalling; in addition, a number of varied transcription factors were identified. A selection of 23 of the genes was transcription-profiled using quantitative RT-PCR to validate the RNA-Seq output. Conclusions A substantial body of chrysanthemum transcriptomic sequence was generated, which led to a number of insights into the molecular basis of the host response to A. tenuissima infection. Although most of the differentially transcribed genes were up-regulated by the presence of the pathogen, those involved in photosynthesis were down-regulated. PMID:24387266

  6. Inducible and constitutive expression of an elicitor gene Hrip1 from Alternaria tenuissima enhances stress tolerance in Arabidopsis.

    PubMed

    Peng, Xue-Cong; Qiu, De-Wen; Zeng, Hong-Mei; Guo, Li-Hua; Yang, Xiu-Fen; Liu, Zheng

    2015-02-01

    Hrip1 is a novel hypersensitive response-inducing protein secreted by Alternaria tenuissima that activates defense responses and systemic acquired resistance in tobacco. This study investigates the role that Hrip1 plays in responses to abiotic and biotic stress using transgenic Arabidopsis thaliana expressing the Hrip1 gene under the control of the stress-inducible rd29A promoter or constitutive cauliflower mosaic virus 35S promoter. Bioassays showed that inducible Hrip1 expression in rd29A∷Hrip1 transgenic lines had a significantly higher effect on plant height, silique length, plant dry weight, seed germination and root length under salt and drought stress compared to expression in 35S∷Hrip1 lines and wild type plants. The level of enhancement of resistance to Botrytis cinerea by the 35S∷Hrip1 lines was higher than in the rd29A∷Hrip1 lines. Moreover, stress-related gene expression in the transgenic Arabidopsis lines was significantly increased by 200 mM NaCl and 200 mM mannitol treatments, and defense genes in the jasmonic acid and ethylene signaling pathway were significantly up-regulated after Botrytis inoculation in the Hrip1 transgenic plants. Furthermore, the activity of some antioxidant enzymes, such as peroxidase and catalase increased after salt and drought stress and Botrytis infection. These results suggested that the Hrip1 protein contributes to abiotic and biotic resistance in transgenic Arabidopsis and may be used as a useful gene for resistance breeding in crops. Although the constitutive expression of Hrip1 is suitable for biotic resistance, inducible Hrip1 expression is more responsive for abiotic resistance.

  7. Characterisation of Alternaria alternata manganese-dependent superoxide dismutase, a cross-reactive allergen homologue to Asp f 6.

    PubMed

    Gabriel, Marta F; Postigo, Idoia; Gutiérrez-Rodríguez, Antonio; Suñén, Ester; Guisantes, Jorge; Tomaz, Cândida T; Martínez, Jorge

    2015-07-01

    It is well known that Alternaria alternata presents a significant level of allergenic cross-reactivity with several other phylogenetically related and non-related allergenic moulds. To improve the molecular diagnosis, the identification and characterisation of all clinically relevant allergens, including both species-specific and cross-reacting proteins, is required. In this study we report the molecular and immunological characterisation of the A. alternata manganese-dependent superoxide dismutase (Alt a MnSOD) and its cross-reactivity with Asp f 6, a diagnostic marker allergen in allergic bronchopulmonary aspergillosis (ABPA). The cDNA coding for Alt a MnSOD sequence was isolated by RACE and PCR. Alt a MnSOD is a protein of 191 amino acids that presented significant homology and potential cross-reactive epitopes with Asp f 6. The recombinant protein was produced in Escherichia coli and the immunoreactivity was evaluated in patient sera. Immunoblotting analyses showed that seven of sixty-one A. alternata-sensitised patient sera and two ABPA patient sera reacted with the recombinant Alt a MnSOD. The native counterpart contained in both A. alternata and Aspergillus fumigatus extracts inhibited IgE binding to the recombinant molecule. The allergen was named Alt a 14 by the official Allergen nomenclature subcommittee. Thus, Alt a 14 is a relevant allergen in A. alternata sensitisation that may be used to improve diagnostic procedures. Evidence of cross-reactivity between Asp f 6 and Alt a 14-recognition by ABPA patient sera suggest the existence of an Alt a 14-mediated mechanism that, similar to Asp f 6, may be related to the pathogenesis of ABPA.

  8. Systemic jasmonic acid modulation in mycorrhizal tomato plants and its role in induced resistance against Alternaria alternata.

    PubMed

    Nair, A; Kolet, S P; Thulasiram, H V; Bhargava, S

    2015-05-01

    Tomato plants colonised with the arbuscular mycorrhizal (AM) fungus Glomus fasciculatum show systemic induced resistance to the foliar pathogen Alternaria alternata, as observed in interactions of other AM-colonised plants with a range of pathogens. The role of jasmonic (JA) and salicylic (SA) acid in expression of this mycorrhiza-induced resistance (MIR) against A. alternata was studied by measuring: (i) activity of enzymes reported to be involved in their biosynthesis, namely lipoxygenase (LOX) and phenylammonia lyase (PAL); and (ii) levels of methyl jasmonate (MeJA) and SA. Transcript abundance of some defence genes associated with JA and SA response pathways were also studied. Both LOX and PAL activity increased twofold in response to pathogen application to control plants. AM-colonised plants had three-fold higher LOX activity compared to control plants, but unlike controls, this did not increase further in response to pathogen application. Higher LOX activity in AM-colonised plants correlated with four-fold higher MeJA in leaves of AM-colonised plants compared to controls. Treatment of plants with the JA biosynthesis inhibitor salicylhydroxamic acid (SHAM) led to 50% lower MeJA in both control and AM-colonised plants and correlated with increased susceptibility to A. alternata, suggesting a causal role for JA in expression of MIR against the pathogen. Genes involved in JA biosynthesis (OPR3) and response (COI1) showed six- and 42-fold higher expression, respectively, in leaves of AM-colonised plants compared to controls. AM-colonised plants also showed increased expression of the SA response gene PR1 and that of the wound-inducible polypeptide prosystemin. Our results suggest that the systemic increase in JA in response to AM colonisation plays a key role in expression of MIR against A. alternata.

  9. Alternaria toxin-induced resistance in rose plants against rose aphid (Macrosiphum rosivorum): effect of tenuazonic acid.

    PubMed

    Yang, Fa-zhong; Yang, Bin; Li, Bei-bei; Xiao, Chun

    2015-04-01

    Many different types of toxins are produced by the fungus, Alternaria alternata (Fr.) Keissler. Little is known, however, regarding the influence of these toxins on insects. In this study, we investigated the toxin-induced inhibitory effects of the toxin produced by A. alternata on the rose aphid, Macrosiphum rosivorum, when the toxin was applied to leaves of the rose, Rosa chinensis. The results demonstrated that the purified crude toxin was non-harmful to rose plants and rose aphids, but had an intensive inhibitory effect on the multiplication of aphids. The inhibitory index against rose aphids reached 87.99% when rose plants were sprayed with the toxin solution at a low concentration. Further results from bioassays with aphids and high performance liquid chromatography (HPLC) analyses demonstrated that tenuazonic acid (TeA) was one of the most important resistance-related active components in the crude toxin. The content of TeA was 0.1199% in the crude toxin under the HPLC method. Similar to the crude toxin, the inhibitory index of pure TeA reached 83.60% 15 d after the rose plants were sprayed with pure TeA solution at the lower concentration of 0.060 μg/ml, while the contents of residual TeA on the surface and in the inner portion of the rose plants were only 0.04 and 0.00 ng/g fresh weight of TeA-treated rose twigs, respectively, 7 d after the treatment. Our results show that TeA, an active component in the A. alternata toxin, can induce the indirect plant-mediated responses in rose plants to intensively enhance the plant's resistances against rose aphids, and the results are very helpful to understand the plant-mediated interaction between fungi and insects on their shared host plants.

  10. Fungal-specific transcription factor AbPf2 activates pathogenicity in Alternaria brassicicola

    SciTech Connect

    Cho, Yangrae; Ohm, Robin A.; Grigoriev, Igor V.; Srivastava, Akhil

    2013-05-24

    Alternaria brassicicola is a successful saprophyte and necrotrophic plant pathogen. To identify molecular determinants of pathogenicity, we created non-pathogenic mutants of a transcription factor-encoding gene, AbPf2. The frequency and timing of germination and appressorium formation on host plants were similar between the non-pathogenic abpf2 mutants and wild-type A. brassicicola. The mutants were also similar in vitro to wild-type A. brassicicola in terms of vegetative growth, conidium production, and responses to a phytoalexin, reactive oxygen species and osmolites. The hyphae of the mutants grew slowly but did not cause disease symptoms on the surface of host plants. Transcripts of the AbPf2 gene increased exponentially soon after wild-type conidia contacted their host plants . A small amount of AbPf2 protein, as monitored using GFP fusions, was present in young, mature conidia. The protein level decreased during saprophytic growth, but increased and was located primarily in fungal nuclei during pathogenesis. Levels of the proteins and transcripts sharply decreased following colonization of host tissues beyond the initial infection site. When expression of the transcription factor was induced in the wild-type during early pathogenesis, 106 fungal genes were also induced in the wild-type but not in the abpf2 mutants. Notably, 33 of the 106 genes encoded secreted proteins, including eight putative effector proteins. Plants inoculated with abpf2 mutants expressed higher levels of genes associated with photosynthesis, the pentose phosphate pathway and primary metabolism, but lower levels of defense-related genes. Our results suggest that AbPf2 is an important regulator of pathogenesis, but does not affect other cellular processes in A. brassicicola.

  11. Phenylacetaldehyde O-methyloxime: a volatile compound produced by grapefruit leaves infected with the citrus canker pathogen, Xanthomonas axonopodis pv. citri.

    PubMed

    Zhang, Aijun; Hartung, John S

    2005-06-29

    An aldehyde oxime O-methyl ether, phenylacetaldehyde O-methyloxime, was detected using solid-phase microextraction (SPME) and gas chromatography-mass spectrometry (GC-MS) in the headspace above grapefruit leaves infected with Xanthomonas axonopodis pv. citri, the causal agent of citrus bacterial canker disease (CBCD). This disease is a major phytosanitary concern, and an eradication campaign against it is currently underway in Florida. Phenylacetaldehyde O-methyloxime has been reported to be produced by other plants and fragrant flowers, but it was not observed in the headspace above uninfected grapefruit leaves, the pathogenic bacterium X. axonopodis pv. citri itself, or grapefruit leaves infected with another closely related bacterial pathogen, X. axonopodis pv. citrumelo, which causes citrus bacterial spot, a disease of no phytosanitary significance. It was also not detected from CBCD infected fruits, including orange, lemon, grapefruit, and lime. We conclude that phenylacetaldehyde O-methyloxime may potentially be used to identify CBCD infestations. However, more intensive studies will be required to fully evaluate the potential of phenylacetaldehyde O-methyloxime as a diagnostic compound for CBCD. Using SPME and GC-MS to measure phenylacetaldehyde O-methyloxime may provide an easy and feasible tool to complement current methods used to detect X. axonopodis pv. citri in environmental samples.

  12. The filamentous phage XacF1 causes loss of virulence in Xanthomonas axonopodis pv. citri, the causative agent of citrus canker disease.

    PubMed

    Ahmad, Abdelmonim Ali; Askora, Ahmed; Kawasaki, Takeru; Fujie, Makoto; Yamada, Takashi

    2014-01-01

    In this study, filamentous phage XacF1, which can infect Xanthomonas axonopodis pv. citri (Xac) strains, was isolated and characterized. Electron microscopy showed that XacF1 is a member of the family Inoviridae and is about 600 nm long. The genome of XacF1 is 7325 nucleotides in size, containing 13 predicted open reading frames (ORFs), some of which showed significant homology to Ff-like phage proteins such as ORF1 (pII), ORF2 (pV), ORF6 (pIII), and ORF8 (pVI). XacF1 showed a relatively wide host range, infecting seven out of 11 strains tested in this study. Frequently, XacF1 was found to be integrated into the genome of Xac strains. This integration occurred at the host dif site (attB) and was mediated by the host XerC/D recombination system. The attP sequence was identical to that of Xanthomonas phage Cf1c. Interestingly, infection by XacF1 phage caused several physiological changes to the bacterial host cells, including lower levels of extracellular polysaccharide production, reduced motility, slower growth rate, and a dramatic reduction in virulence. In particular, the reduction in virulence suggested possible utilization of XacF1 as a biological control agent against citrus canker disease.

  13. Corticicoccus populi gen. nov., sp. nov., a member of the family Staphylococcaceae, isolated from symptomatic bark of Populus × euramericana canker.

    PubMed

    Li, Yong; Wang, Sheng-Kun; Xue, Han; Chang, Ju-Pu; Guo, Li-Min; Yang, Xu-Qi

    2016-10-24

    Two gram-stain positive, aerobic, non-motile, bacterial strains were isolated from symptomatic bark tissue of Populus × euramericana canker. The isolates were able to grow between 10 and 37°C, pH 6 to 10 and 0-4% (w/v) NaCl, with optimal growth at 28-30°C, pH 7.0-8.0 and 2% (w/v) NaCl The strains were found to be oxidase positive and catalase negative. The menaquinone of the 26D10-3-4T was found to be MK-7 and the peptidoglycan type A3α based on L-Lys-Gly3-?Ala. The polar lipid profiles of 26D10-3-4T showed diphosphatidylglycerol, phosphatidylglycerol, two unidentified phospholipids and three unidentified glycolipids; the major fatty acids found were anteiso-C15:0, iso-C15:0, iso-C17:0 and anteiso-C17:0. The DNA G+C content was 38.2 mol%. The two novel isolates shared the highest 16S rRNA gene sequence similarity with Salinicoccus qingdaonensis ZXM223T (95.0%). Based on phenotypic and genotypic characteristics, these two strains represent a novel species of a novel genus of the family Staphylococcaceae; the name Corticicoccus populi gen. nov., sp. nov. is proposed. The type strain is 26D10-3-4T (=CFCC 12725T =KCTC 33575T).

  14. Populibacterium corticicola gen. nov., sp. nov., a member of the family Jonesiaceae, isolated from symptomatic bark of Populus × euramericana canker.

    PubMed

    Li, Yong; Guo, Min-Wei; Wang, Laifa; Wang, Xi-Zhuo; Lin, Cai-Li

    2016-09-01

    Four Gram-stain-positive, aerobic, motile bacterial strains were isolated from the bark tissue of Populus × euramericana canker. Growth occurred between 10 and 37 °C and at pH 6-10, with optimal growth at 28-30 °C and pH 7.0-8.0. Growth occurred at 0-3 % (w/v) salinity. The strains were positive for oxidase and catalase activity. The major fatty acids were anteiso-C15 : 0 and C16 : 0. The phospholipid profiles contained diphosphatidylglycerol, phosphatidylinositol, phosphatidylglycerol, two phospholipids and five glycolipids. The peptidoglycan type was A4α, which is based on l-Lys-d-Ser-d-Asp. The DNA G+C content was 58.5 mol%. Based on 16S rRNA gene sequence analysis, as well as physiological and biochemical characteristics, the strains are considered to represent a novel species of a new genus in the family Jonesiaceae. The name proposed is Populibacterium corticicola gen. nov., sp. nov. The type strain of Populibacterium corticicola is 2D-4T (=CFCC 11886T=KCTC 33576T).

  15. Study on Interactions between the Major Apple Valsa Canker Pathogen Valsa mali and Its Biocontrol Agent Saccharothrix yanglingensis Hhs.015 Using RT-qPCR

    PubMed Central

    Fan, Dongying; Li, Yanfang; Zhao, Lingyun; Li, Zhengpeng; Huang, Lili; Yan, Xia

    2016-01-01

    The mechanism of biocontrol agent Saccharothrix yanglingensis Hhs.015 action against Valsa mali, a major apple Valsa canker pathogen, was examined using a novel, sensitive (minimum detection limit 100 pg/μL) and reliably RT-qPCR technique. Prior to lesion formation, total concentration of V. mali in the bark showed a significant decrease (p<0.05) after 24 h of Hhs.015 treatment. This was more pronounced at 48 and 96 h post treatment. After lesion formation, levels of V. mali remained constant at the boundary between infected and uninfected bark tissues, although the relative expansion rate of the lesion was significantly reduced (p<0.05). Gene expression levels of endo-polygalacturonase, a marker for fungal pathogenicity, were sharply reduced while host induced resistance callose synthase levels increased significantly (p<0.05) at the boundary bark at 9 d after Hhs.015 treatment. The results showed that biocontrol agent Hhs.015 prevented infection of V. mali by inhibiting pathogen growth, down-regulating pathogenicity factor expression and inducing a high level of host resistance. PMID:27611855

  16. Composition of tobaccos from countries with high and low incidences of lung cancer. I. Selenium, polonium-210, Alternaria, tar, and nicotine

    SciTech Connect

    Bogden, J.D.; Kemp, F.W.; Buse, M.; Thind, I.S.; Louria, D.B.; Forgacs, J.; Llanos, G.; Terrones, I.M.

    1981-01-01

    Tobaccos from countries with high and low incidences of lung cancer were analyzed. Tobacco concentrations of polonium-210 were similar in cigarettes from high- and low-incidence countries, as were levels of cigarette smoke tar and nicotine. Tobaccos from low-incidence countries had significantly lower Alternaria spore counts. Mean selenium concentrations of tobaccos from the high-incidence countries (0.16 +- 0.05 ..mu..g/g) were significantly lower than those of tobaccos from the low-incidence countries (0.49 +- 0.22 ..mu..g/g).

  17. STEM Thinking!

    ERIC Educational Resources Information Center

    Reeve, Edward M.

    2015-01-01

    Science, Technology, Engineering, and Mathematics (STEM) is a term seen almost daily in the news. In 2009, President Obama launched the Educate to Innovate initiative to move American students from the middle to the top of the pack in science and math achievement over the next decade (The White House, n.d.). Learning about the attributes of STEM…

  18. Why STEM?

    ERIC Educational Resources Information Center

    Mitts, Charles R.

    2016-01-01

    The International Technology and Engineering Educators Association (ITEEA) defines STEM as a new transdisciplinary subject in schools that integrates the disciplines of science, technology, engineering, and mathematics into a single course of study. There are three major problems with this definition: There is no consensus in support of the ITEEA…

  19. Transcriptome Analysis Reveals the Genetic Basis of the Resveratrol Biosynthesis Pathway in an Endophytic Fungus (Alternaria sp. MG1) Isolated from Vitis vinifera

    PubMed Central

    Che, Jinxin; Shi, Junling; Gao, Zhenhong; Zhang, Yan

    2016-01-01

    Alternaria sp. MG1, an endophytic fungus previously isolated from Merlot grape, produces resveratrol from glucose, showing similar metabolic flux to the phenylpropanoid biosynthesis pathway, currently found solely in plants. In order to identify the resveratrol biosynthesis pathway in this strain at the gene level, de novo transcriptome sequencing was conducted using Illumina paired-end sequencing. A total of 22,954,434 high-quality reads were assembled into contigs and 18,570 unigenes were identified. Among these unigenes, 14,153 were annotated in the NCBI non-redundant protein database and 5341 were annotated in the Swiss-Prot database. After KEGG mapping, 2701 unigenes were mapped onto 115 pathways. Eighty-four unigenes were annotated in major pathways from glucose to resveratrol, coding 20 enzymes for glycolysis, 10 for phenylalanine biosynthesis, 4 for phenylpropanoid biosynthesis, and 4 for stilbenoid biosynthesis. Chalcone synthase was identified for resveratrol biosynthesis in this strain, due to the absence of stilbene synthase. All the identified enzymes indicated a reasonable biosynthesis pathway from glucose to resveratrol via glycolysis, phenylalanine biosynthesis, phenylpropanoid biosynthesis, and stilbenoid pathways. These results provide essential evidence for the occurrence of resveratrol biosynthesis in Alternaria sp. MG1 at the gene level, facilitating further elucidation of the molecular mechanisms involved in this strain's secondary metabolism. PMID:27588016

  20. Validated UPLC-MS/MS Methods To Quantitate Free and Conjugated Alternaria Toxins in Commercially Available Tomato Products and Fruit and Vegetable Juices in Belgium.

    PubMed

    Walravens, Jeroen; Mikula, Hannes; Rychlik, Michael; Asam, Stefan; Devos, Tom; Njumbe Ediage, Emmanuel; Diana Di Mavungu, José; Jacxsens, Liesbeth; Van Landschoot, Anita; Vanhaecke, Lynn; De Saeger, Sarah

    2016-06-22

    Ultraperformance liquid chromatography tandem mass spectrometry and Quick, Easy, Cheap, Effective, Rugged, and Safe based analytical methodologies to quantitate both free (alternariol (1), alternariol monomethyl ether (2), tenuazonic acid (3), tentoxin (4), altenuene (5), altertoxin-I (6)) and conjugated (sulfates and glucosides of 1 and 2) Alternaria toxins in fruit and vegetable juices and tomato products were developed and validated. Acceptable limits of quantitation (0.7-5.7 μg/kg), repeatability (RSDr < 15.7%), reproducibility (RSDR < 17.9%), and apparent recovery (87.0-110.6%) were obtained for all analytes in all matrices investigated. 129 commercial foodstuffs were analyzed, and 3 was detected in 100% of tomato product samples (Alternaria toxins (sulfates of 1 and 2) were repeatedly detected. A deterministic dietary exposure assessment revealed the possible risk for human health related to the presence of 1 and 2 in tomato based foodstuffs, whereas 3 is unlikely to be of human health concern.

  1. Serine protease identification (in vitro) and molecular structure predictions (in silico) from a phytopathogenic fungus, Alternaria solani.

    PubMed

    Chandrasekaran, Murugesan; Chandrasekar, Raman; Sa, Tongmin; Sathiyabama, Muthukrishnan

    2014-07-01

    Serine proteases are involved in an enormous number of biological processes. The present study aims at characterizing three-dimensional (3D) molecular architecture of serine proteases from early blight pathogen, Alternaria solani that are hypothesized to be markers of phytopathogenicity. A serine protease was purified to homogeneity and MALDI-TOF-MS/MS analysis revealed that protease produced by A. solani belongs to alkaline serine proteases (AsP). AsP is made up of 403 amino acid residues with molecular weight of 42.1 kDa (Isoelectric point - 6.51) and its molecular formula was C1859 H2930 N516 O595 S4 . AsP structure model was built based on its comparative homology with serine protease using the program, MODELER. AsP had 16 β-sheets and 10 α-helices, with Ser(350) (G347-G357), Asp(158) (D158-H169), and His(193) (H193-G203) in separate turn/coil structures. Biological metal binding region situated near 6th-helix and His(193) residue is responsible for metal binding site. Also, calcium ion (Ca(2+)) is coordinated by the carboxyl groups of Lys(84), Ile(85), Lys(86), Asp(87), Phe(88), Ala(89), Ala(90) (K84-A90) for first Ca(2+) binding site and carbonyl oxygen atom of Lys(244), Gly(245), Arg(246), Thr(247), Lys(248), Lys(249), and Ala(250) (K244-A250), for second Ca(2+) binding site. Moreover, Ramachandran plot analysis of protein residues falling into most favored secondary structures were determined (83.3%). The predicted molecular 3D structural model was further verified using PROCHECK, ERRAT, and VADAR servers to confirm the geometry and stereo-chemical parameters of the molecular structural design. The functional analysis of AsP 3D molecular structure predictions familiar in the current study may provide a new perspective in the understanding and identification of antifungal protease inhibitor designing.

  2. Development and Validation of an Infection Risk Model for Bacterial Canker of Kiwifruit, Using a Multiplication and Dispersal Concept for Forecasting Bacterial Diseases.

    PubMed

    Beresford, R M; Tyson, J L; Henshall, W R

    2017-02-01

    A weather-based disease prediction model for bacterial canker of kiwifruit (known worldwide as Psa; Pseudomonas syringae pv. actinidiae biovar 3) was developed using a new mechanistic scheme for bacterial disease forecasters, the multiplication and dispersal concept. Bacterial multiplication is estimated from a temperature function, the M index, accumulated from hourly air temperature over 3 days for hours when the leaf canopy is wet. Rainfall provides free water to move inoculum to infection sites, and the daily risk indicator, the R index, is the 3-day accumulation of the M index output on days with total rainfall >1 mm; otherwise, R is zero. The model was field-tested using potted kiwifruit trap plants exposed for discrete periods in infected kiwifruit orchards to identify when leaf infection occurred. In a 9-week study during spring, the R index predicted leaf-spot intensity with high accuracy (R(2) = 93%) and, in an 82-week seasonal accuracy study, prediction of infection incidence was most accurate from spring to late summer and lower during other times. To implement the risk model for the New Zealand kiwifruit industry, a modified risk index, R', used relative humidity (RH) >81% instead of wetness, so that 2- and 6-day weather forecasts of RH could be used. Risk index values were affected by the shape of the temperature function and an alternative 'low temperature' function for the M index was identified that could be used in climates in which high temperatures are known to limit Psa development during some parts of the year. This study has shown how infection risk for bacterial diseases can be conceptualized as separate processes for temperature-dependent bacterial multiplication and rain-dependent dispersal and infection. This concept has potentially wide application for bacterial disease prediction in the same way that the infection monocycle concept has had for fungal disease prediction.

  3. Stem Cell Sciences plc.

    PubMed

    Daniels, Sebnem

    2006-09-01

    Stem Cell Sciences' core objective is to develop safe and effective stem cell-based therapies for currently incurable diseases. In order to achieve this goal, Stem Cell Sciences recognizes the need for multiple technologies and a globally integrated stem cell initiative. The key challenges for the successful application of stem cells in the clinic is the need for a reproducible supply of pure, fully characterized stem cells that have been grown in suitable conditions for use in the clinic.

  4. Development and Validation of a LC-ESI-MS/MS Method for the Determination of Alternaria Toxins Alternariol, Alternariol Methyl-Ether and Tentoxin in Tomato and Tomato-Based Products

    PubMed Central

    Rodríguez-Carrasco, Yelko; Mañes, Jordi; Berrada, Houda; Juan, Cristina

    2016-01-01

    Alternaria species are capable of producing several secondary toxic metabolites in infected plants and in agricultural commodities, which play important roles in food safety. Alternaria alternata turn out to be the most frequent fungal species invading tomatoes. Alternariol (AOH), alternariol monomethyl ether (AME), and tentoxin (TEN) are some of the main Alternaria mycotoxins that can be found as contaminants in food. In this work, an analytical method based on liquid chromatography (LC) tandem mass spectrometry (MS/MS) detection for the simultaneous quantification of AOH, AME, and TEN in tomato and tomato-based products was developed. Mycotoxin analysis was performed by dispersive liquid-liquid microextraction (DLLME) combined with LC-ESI-MS/MS. Careful optimization of the MS/MS parameters was performed with an LC/MS system with the ESI interface in the positive ion mode. Mycotoxins were efficiently extracted from sample extract into a droplet of chloroform (100 µL) by DLLME technique using acetonitrile as a disperser solvent. Method validation following the Commission Decision No. 2002/657/EC was carried out by using tomato juice as a blank matrix. Limits of detection and quantitation were, respectively, in the range 0.7 and 3.5 ng/g. Recovery rates were above 80%. Relative standard deviations of repeatability (RSDr) and intermediate reproducibility (RSDR) were ≤ 9% and ≤ 15%, respectively, at levels of 25 and 50 ng/g. Five out of 30 analyzed samples resulted positive to at least one Alternaria toxin investigated. AOH was the most common Alternaria toxin found, but at levels close to LOQ (average content: 3.75 ng/g). PMID:27845716

  5. Stem Cell Information: Glossary

    MedlinePlus

    ... cells (skeletal stem cells) Cell-based therapies Cell culture Cell division Chromosome Clone Cloning Cord blood stem cells Culture medium Differentiation Directed differentiation DNA Ectoderm Embryo Embryoid ...

  6. Identification of a Novel Alternaria alternata Strain Able to Hyperparasitize Puccinia striiformis f. sp. tritici, the Causal Agent of Wheat Stripe Rust.

    PubMed

    Zheng, Li; Zhao, Jie; Liang, Xiaofei; Zhan, Gangming; Jiang, Shuchang; Kang, Zhensheng

    2017-01-01

    The obligate bitrophic fungus Puccinia striiformis f. sp. tritici (Pst) causes stripe (yellow) rust on wheat worldwide. Here, we report a novel fungal strain able to hyperparasitize Pst. The strain was isolated from gray-colored rust pustules, and was identified as Alternaria alternata (Fr.: Fr.) keissler based on a combination of morphological characteristics and multi-locus (ITS, GAPDH, and RPB2) phylogeny. Upon artificial inoculation, the hyperparasite reduced the production and viability of urediniospores, and produced a typical gray-colored rust pustule symptom. Scanning electron microscopy demonstrated that the strain could efficiently penetrate and colonize Pst urediniospores. This study first demonstrates that A. alternata could parasitize Pst and indicates its potential application in the biological control of wheat stripe rust disease.

  7. Modification of primary and secondary metabolism of potato plants by nitrogen application differentially affects resistance to Phytophthora infestans and Alternaria solani.

    PubMed

    Mittelstrass, K; Treutter, D; Plessl, M; Heller, W; Elstner, E F; Heiser, I

    2006-09-01

    Potato plants ( SOLANUM TUBEROSUM L. cv. Indira) were grown at two levels of N supply in the greenhouse. Plants supplied with 0.8 g N per plant (high N variant) showed significantly increased biomass as compared to plants without additional N fertilisation (low N variant). C/N ratio was lower and protein content was higher in leaves of the high N variant. The concentration of chlorogenic acids and flavonols was significantly lower in leaves from the high N variant. Whereas resistance to ALTERNARIA SOLANI increased when plants were supplied with additional nitrogen, these plants were more susceptible to PHYTOPHTHORA INFESTANS. After infection with both pathogens, we found a strong induction of p-coumaroylnoradrenaline and p-coumaroyloctopamine, which are identified for the first time in potato leaves and are discussed as resistance factors of other solanaceous plants.

  8. Identification of a Novel Alternaria alternata Strain Able to Hyperparasitize Puccinia striiformis f. sp. tritici, the Causal Agent of Wheat Stripe Rust

    PubMed Central

    Zheng, Li; Zhao, Jie; Liang, Xiaofei; Zhan, Gangming; Jiang, Shuchang; Kang, Zhensheng

    2017-01-01

    The obligate bitrophic fungus Puccinia striiformis f. sp. tritici (Pst) causes stripe (yellow) rust on wheat worldwide. Here, we report a novel fungal strain able to hyperparasitize Pst. The strain was isolated from gray-colored rust pustules, and was identified as Alternaria alternata (Fr.: Fr.) keissler based on a combination of morphological characteristics and multi-locus (ITS, GAPDH, and RPB2) phylogeny. Upon artificial inoculation, the hyperparasite reduced the production and viability of urediniospores, and produced a typical gray-colored rust pustule symptom. Scanning electron microscopy demonstrated that the strain could efficiently penetrate and colonize Pst urediniospores. This study first demonstrates that A. alternata could parasitize Pst and indicates its potential application in the biological control of wheat stripe rust disease. PMID:28197134

  9. Toxin production by Alternaria alternata in tomatoes and apples stored under various conditions and quantitation of the toxins by high-performance liquid chromatography.

    PubMed

    Ozcelik, S; Ozcelik, N; Beuchat, L R

    1990-12-01

    Alternaria alternata strain 8442-3 was inoculated into tomatoes (Lycopersicon esculentum Mill.) and Red Delicious cultivar apples (Malus domestica Borkh.). Half of the lots of each fruit were shrink-wrapped in high-density polyethylene film. Wrapped and unwrapped fruits were incubated under darkness at 4, 15 and 25 degrees C for up to 5 weeks. A high-performance liquid chromatography method was developed to quantitate tenuazonic acid (TeA), alternariol (AOH), alternariol methyl ether (AME), and altenuene (ALT). Shrink-wrapping retarded, but did not completely inhibit growth in tomatoes for 3-7 days. Concentrations of up to 120.6 mg of AOH and 63.7 mg of AME per 100 g of tissue were produced in unwrapped tomatoes stored at 15 degrees C for 4 weeks; 19.0 mg of ALT per 100 g of tomato tissue was produced after 3 weeks at 25 degrees C. AOH, AME and ALT were also produced in unwrapped tomatoes stored at 4 degrees C; however, no TeA was detected in decayed tomatoes, regardless of type of wrapping or storage temperature. Shrink-wrapping resulted in decreased production of AOH, AME, and ALT. Alternaria toxins were not detected in apples stored at 4 and 15 degrees C. The highest concentration of AOH produced (48.8 mg per 100 g of tissue) was in unwrapped apples stored at 25 degrees C for 2 weeks; 12.3 mg per 100 g of tissue of shrink-wrapped apples was detected after 5 weeks of storage at 25 degrees C, while ALT reached 5.7 mg per 100 g after 4 weeks. TeA was not detected in apples infected with A. alternata.

  10. Rotatable stem and lock

    DOEpatents

    Deveney, Joseph E.; Sanderson, Stephen N.

    1984-01-01

    A valve stem and lock include a housing surrounding a valve stem, a solenoid affixed to an interior wall of the housing, an armature affixed to the valve stem and a locking device for coupling the armature to the housing body. When the solenoid is energized, the solenoid moves away from the housing body, permitting rotation of the valve stem.

  11. Rotatable stem and lock

    DOEpatents

    Deveney, J.E.; Sanderson, S.N.

    1981-10-27

    A valve stem and lock is disclosed which includes a housing surrounding a valve stem, a solenoid affixed to an interior wall of the housing, an armature affixed to the valve stem and a locking device for coupling the armature to the housing body. When the solenoid is energized, the solenoid moves away from the housing body, permitting rotation of the valve stem.

  12. [Pancreatic cancer stem cell].

    PubMed

    Hamada, Shin; Masamune, Atsushi; Shimosegawa, Tooru

    2015-05-01

    Prognosis of pancreatic cancer remains dismal due to the resistance against conventional therapies. Metastasis and massive invasion toward surrounding organs hamper radical resection. Small part of entire cancer cells reveal resistance against chemotherapy or radiotherapy, increased tumorigenicity and migratory phenotype. These cells are called as cancer stem cells, as a counter part of normal stem cells. In pancreatic cancer, several cancer stem cell markers have been identified, which enabled detailed characterization of pancreatic cancer stem cells. Recent researches clarified that conventional chemotherapy itself could increase cancer cells with stem cell-phenotype, suggesting the necessity of cancer stem cell-targeting therapy. Based on these observations, pancreatic cancer stem cell-targeting therapies have been tested, which effectively eliminated cancer stem cell fraction and attenuated cancer progression in experimental models. Clinical efficacy of these therapies need to be evaluated, and cancer stem cell-targeting therapy will contribute to improve the prognosis of pancreatic cancer.

  13. Origin of the Outbreak in France of Pseudomonas syringae pv. actinidiae Biovar 3, the Causal Agent of Bacterial Canker of Kiwifruit, Revealed by a Multilocus Variable-Number Tandem-Repeat Analysis

    PubMed Central

    Cunty, A.; Cesbron, S.; Poliakoff, F.; Jacques, M.-A.

    2015-01-01

    The first outbreaks of bacterial canker of kiwifruit caused by Pseudomonas syringae pv. actinidiae biovar 3 were detected in France in 2010. P. syringae pv. actinidiae causes leaf spots, dieback, and canker that sometimes lead to the death of the vine. P. syringae pv. actinidifoliorum, which is pathogenic on kiwi as well, causes only leaf spots. In order to conduct an epidemiological study to track the spread of the epidemics of these two pathogens in France, we developed a multilocus variable-number tandem-repeat (VNTR) analysis (MLVA). MLVA was conducted on 340 strains of P. syringae pv. actinidiae biovar 3 isolated in Chile, China, France, Italy, and New Zealand and on 39 strains of P. syringae pv. actinidifoliorum isolated in Australia, France, and New Zealand. Eleven polymorphic VNTR loci were identified in the genomes of P. syringae pv. actinidiae biovar 3 ICMP 18744 and of P. syringae pv. actinidifoliorum ICMP 18807. MLVA enabled the structuring of P. syringae pv. actinidiae biovar 3 and P. syringae pv. actinidifoliorum strains in 55 and 16 haplotypes, respectively. MLVA and discriminant analysis of principal components revealed that strains isolated in Chile, China, and New Zealand are genetically distinct from P. syringae pv. actinidiae strains isolated in France and in Italy, which appear to be closely related at the genetic level. In contrast, no structuring was observed for P. syringae pv. actinidifoliorum. We developed an MLVA scheme to explore the diversity within P. syringae pv. actinidiae biovar 3 and to trace the dispersal routes of epidemic P. syringae pv. actinidiae biovar 3 in Europe. We suggest using this MLVA scheme to trace the dispersal routes of P. syringae pv. actinidiae at a global level. PMID:26209667

  14. Origin of the Outbreak in France of Pseudomonas syringae pv. actinidiae Biovar 3, the Causal Agent of Bacterial Canker of Kiwifruit, Revealed by a Multilocus Variable-Number Tandem-Repeat Analysis.

    PubMed

    Cunty, A; Cesbron, S; Poliakoff, F; Jacques, M-A; Manceau, C

    2015-10-01

    The first outbreaks of bacterial canker of kiwifruit caused by Pseudomonas syringae pv. actinidiae biovar 3 were detected in France in 2010. P. syringae pv. actinidiae causes leaf spots, dieback, and canker that sometimes lead to the death of the vine. P. syringae pv. actinidifoliorum, which is pathogenic on kiwi as well, causes only leaf spots. In order to conduct an epidemiological study to track the spread of the epidemics of these two pathogens in France, we developed a multilocus variable-number tandem-repeat (VNTR) analysis (MLVA). MLVA was conducted on 340 strains of P. syringae pv. actinidiae biovar 3 isolated in Chile, China, France, Italy, and New Zealand and on 39 strains of P. syringae pv. actinidifoliorum isolated in Australia, France, and New Zealand. Eleven polymorphic VNTR loci were identified in the genomes of P. syringae pv. actinidiae biovar 3 ICMP 18744 and of P. syringae pv. actinidifoliorum ICMP 18807. MLVA enabled the structuring of P. syringae pv. actinidiae biovar 3 and P. syringae pv. actinidifoliorum strains in 55 and 16 haplotypes, respectively. MLVA and discriminant analysis of principal components revealed that strains isolated in Chile, China, and New Zealand are genetically distinct from P. syringae pv. actinidiae strains isolated in France and in Italy, which appear to be closely related at the genetic level. In contrast, no structuring was observed for P. syringae pv. actinidifoliorum. We developed an MLVA scheme to explore the diversity within P. syringae pv. actinidiae biovar 3 and to trace the dispersal routes of epidemic P. syringae pv. actinidiae biovar 3 in Europe. We suggest using this MLVA scheme to trace the dispersal routes of P. syringae pv. actinidiae at a global level.

  15. Stem Cell Transplant

    MedlinePlus

    ... transplant is a procedure that infuses healthy blood stem cells into your body to replace your damaged or ... A bone marrow transplant is also called a stem cell transplant. A bone marrow transplant may be necessary ...

  16. STEM Club Participation and STEM Schooling Outcomes

    ERIC Educational Resources Information Center

    Gottfried, Michael A.; Williams, Darryl N.

    2013-01-01

    To develop a more robust understanding of the relationship between non-formal, school-based STEM activities and students' success and persistence in STEM fields, this study evaluates how math club participation influences math GPA and how science club participation influences science GPA. Additionally, this study evaluates how math or science club…

  17. Understanding STEM: Current Perceptions

    ERIC Educational Resources Information Center

    Brown, Ryan; Brown, Joshua; Reardon, Kristin; Merrill, Chris

    2011-01-01

    In many ways, the push for STEM (science, technology, engineering, and mathematics) education appears to have grown from a concern for the low number of future professionals to fill STEM jobs and careers and economic and educational competitiveness. The proponents of STEM education believe that by increasing math and science requirements in…

  18. FTIR spectroscopic evaluation of changes in the cellular biochemical composition of the phytopathogenic fungus Alternaria alternata induced by extracts of some Greek medicinal and aromatic plants

    NASA Astrophysics Data System (ADS)

    Skotti, Efstathia; Kountouri, Sophia; Bouchagier, Pavlos; Tsitsigiannis, Dimitrios I.; Polissiou, Moschos; Tarantilis, Petros A.

    2014-06-01

    In this study, the biological activity of aquatic extracts of selected Greek medicinal and aromatic plants to the phytopathogenic fungus Alternaria alternata was investigated. Lamiaceae species (Hyssopus officinalis L., Melissa officinalis L., Origanum dictamnus L., Origanum vulgare L. and Salvia officinalis L.) were found to enhance significantly the mycelium growth whereas Crocus sativus appears to inhibit it slightly. M. officinalis and S. officinalis caused the highest stimulation in mycelium growth (+97%) and conidia production (+65%) respectively. In order to further investigate the bioactivity of plant extracts to A. alternata, we employed Fourier Transform Infrared Spectroscopy (FTIR). Differences of original spectra were assigned mainly to amides of proteins. The second derivative transformation of spectra revealed changes in spectral regions corresponding to absorptions of the major cellular constituents such as cell membrane and proteins. Principal component analysis of the second derivative transformed spectra confirmed that fatty acids of the cell membranes, amides of proteins and polysaccharides of the cell wall had the major contribution to data variation. FTIR band area ratios were found to correlate with fungal mycelium growth.

  19. A Zinc-Finger-Family Transcription Factor, AbVf19, Is Required for the Induction of a Gene Subset Important for Virulence in Alternaria brassicicola

    SciTech Connect

    Srivastava, Akhil; Ohm, Robin A.; Oxiles, Lindsay; Brooks, Fred; Lawrence, Christopher B.; Grigoriev, Igor V.; Cho, Yangrae

    2011-10-26

    Alternaria brassicicola is a successful saprophyte and necrotrophic plant pathogen with a broad host range within the family Brassicaceae. It produces secondary metabolites that marginally affect virulence. Cell wall degrading enzymes (CDWE) have been considered important for pathogenesis but none of them individually have been identified as significant virulence factors in A. brassicicola. In this study, knockout mutants of a gene, AbVf19, were created and produced considerably smaller lesions than the wild type on inoculated host plants. The presence of tandem zinc-finger domains in the predicted amino acid sequence and nuclear localization of AbVf19- reporter protein suggested that it was a transcription factor. Gene expression comparisons using RNA-seq identified 74 genes being downregulated in the mutant during a late stage of infection. Among the 74 downregulated genes, 28 were putative CWDE genes. These were hydrolytic enzyme genes that composed a small fraction of genes within each family of cellulases, pectinases, cutinases, and proteinases. The mutants grew slower than the wild type on an axenic medium with pectin as a major carbon source. This study demonstrated the existence and the importance of a transcription factor that regulates a suite of genes that are important for decomposing and utilizing plant material during the late stage of plant infection.

  20. The glutathione peroxidase-mediated reactive oxygen species resistance, fungicide sensitivity and cell wall construction in the citrus fungal pathogen Alternaria alternata.

    PubMed

    Yang, Siwy Ling; Yu, Pei-Ling; Chung, Kuang-Ren

    2016-03-01

    The ability to detoxify reactive oxygen species (ROS) is critical for pathogenicity in the necrotrophic fungus Alternaria alternata. We report a glutathione peroxidase 3 (AaGPx3) involved in the complex signalling network that is essential for the detoxification of cellular stresses induced by ROS and for A. alternata pathogenesis in citrus. AaGPx3 deletion mutants displayed increased sensitivity to H2 O2 and many ROS-generating compounds. AaGPx3 is required for correct fungal development as the AaGPx3 mutant strains showed a severe reduction in conidiation. AaGPx3 mutants accumulated higher chitin content than the wild-type and were less sensitive to the cell wall-targeting compounds calcofluor white and Congo red, as well as the fungicides fludioxonil and vinclozolin, suggesting a role of the glutathione systems in fungal cell wall construction. Virulence assays revealed that AaGPx3 is required for full virulence. The expression of AaGPx3 was downregulated in fungal strains carrying defective NADPH oxidase (Nox) or the oxidative stress responsive regulators YAP1 and HOG1, all implicated in ROS resistance. These results further support the important role of ROS detoxification during A. alternata pathogenesis in citrus. Overall, our study provides genetic evidence to define the central role of AaGPx3 in the biological and pathological functions of A. alternata.

  1. Analysis of alternariol and alternariol monomethyl ether on flavedo and albedo tissues of tangerines (Citrus reticulata) with symptoms of alternaria brown spot.

    PubMed

    Magnani, Rodrigo F; De Souza, Gezimar D; Rodrigues-Filho, Edson

    2007-06-27

    A method was developed for the quantification of alternariol and alternariol monomethyl ether on tangerines with and without symptoms of Alternaria brown spot disease. The method employs solid-phase extraction for cleanup, followed by high-performance liquid chromatography coupled with tandem mass spectrometry (HPLC-MS/MS) for detection. This method was validated on flavedo (exocarp or epicarp, exterior yellow peel) and on albedo tissue (mesocarp, interior white peel). An excellent linearity over a range of 0.50-20.0 mg/kg was achieved, with r2 >or= 0.997. The limits of detection (LOD) and quantification (LOQ) were fewer than 0.13 and 0.50 microg/kg, respectively. The relative standard deviations (RSDs) were

  2. FTIR spectroscopic evaluation of changes in the cellular biochemical composition of the phytopathogenic fungus Alternaria alternata induced by extracts of some Greek medicinal and aromatic plants.

    PubMed

    Skotti, Efstathia; Kountouri, Sophia; Bouchagier, Pavlos; Tsitsigiannis, Dimitrios I; Polissiou, Moschos; Tarantilis, Petros A

    2014-06-05

    In this study, the biological activity of aquatic extracts of selected Greek medicinal and aromatic plants to the phytopathogenic fungus Alternaria alternata was investigated. Lamiaceae species (Hyssopus officinalis L., Melissa officinalis L., Origanum dictamnus L., Origanum vulgare L. and Salvia officinalis L.) were found to enhance significantly the mycelium growth whereas Crocus sativus appears to inhibit it slightly. M. officinalis and S. officinalis caused the highest stimulation in mycelium growth (+97%) and conidia production (+65%) respectively. In order to further investigate the bioactivity of plant extracts to A. alternata, we employed Fourier Transform Infrared Spectroscopy (FTIR). Differences of original spectra were assigned mainly to amides of proteins. The second derivative transformation of spectra revealed changes in spectral regions corresponding to absorptions of the major cellular constituents such as cell membrane and proteins. Principal component analysis of the second derivative transformed spectra confirmed that fatty acids of the cell membranes, amides of proteins and polysaccharides of the cell wall had the major contribution to data variation. FTIR band area ratios were found to correlate with fungal mycelium growth.

  3. Fine Mapping for Identification of Citrus Alternaria Brown Spot Candidate Resistance Genes and Development of New SNP Markers for Marker-Assisted Selection

    PubMed Central

    Cuenca, Jose; Aleza, Pablo; Garcia-Lor, Andres; Ollitrault, Patrick; Navarro, Luis

    2016-01-01

    Alternaria brown spot (ABS) is a serious disease affecting susceptible citrus genotypes, which is a strong concern regarding citrus breeding programs. Resistance is conferred by a recessive locus (ABSr) previously located by our group within a 3.3 Mb genome region near the centromere in chromosome III. This work addresses fine-linkage mapping of this region for identifying candidate resistance genes and develops new molecular markers for ABS-resistance effective marker-assisted selection (MAS). Markers closely linked to ABSr locus were used for fine mapping using a 268-segregating diploid progeny derived from a heterozygous susceptible × resistant cross. Fine mapping limited the genomic region containing the ABSr resistance gene to 366 kb, flanked by markers at 0.4 and 0.7 cM. This region contains nine genes related to pathogen resistance. Among them, eight are resistance (R) gene homologs, with two of them harboring a serine/threonine protein kinase domain. These two genes along with a gene encoding a S-adenosyl-L-methionine-dependent-methyltransferase protein, should be considered as strong candidates for ABS-resistance. Moreover, the closest SNP was genotyped in 40 citrus varieties, revealing very high association with the resistant/susceptible phenotype. This new marker is currently used in our citrus breeding program for ABS-resistant parent and cultivar selection, at diploid, triploid and tetraploid level. PMID:28066498

  4. A Major Facilitator Superfamily Transporter-Mediated Resistance to Oxidative Stress and Fungicides Requires Yap1, Skn7, and MAP Kinases in the Citrus Fungal Pathogen Alternaria alternata.

    PubMed

    Chen, Li-Hung; Tsai, Hsieh-Chin; Yu, Pei-Ling; Chung, Kuang-Ren

    2017-01-01

    Major Facilitator Superfamily (MFS) transporters play an important role in multidrug resistance in fungi. We report an AaMFS19 gene encoding a MFS transporter required for cellular resistance to oxidative stress and fungicides in the phytopathogenic fungus Alternaria alternata. AaMFS19, containing 12 transmembrane domains, displays activity toward a broad range of substrates. Fungal mutants lacking AaMFS19 display profound hypersensitivities to cumyl hydroperoxide, potassium superoxide, many singlet oxygen-generating compounds (eosin Y, rose Bengal, hematoporphyrin, methylene blue, and cercosporin), and the cell wall biosynthesis inhibitor, Congo red. AaMFS19 mutants also increase sensitivity to copper ions, clotrimazole, fludioxonil, and kocide fungicides, 2-chloro-5-hydroxypyridine (CHP), and 2,3,5-triiodobenzoic acid (TIBA). AaMFS19 mutants induce smaller necrotic lesions on leaves of a susceptible citrus cultivar. All observed phenotypes in the mutant are restored by introducing and expressing a wild-type copy of AaMFS19. The wild-type strain of A. alternata treated with either CHP or TIBA reduces radial growth and formation and germination of conidia, increases hyphal branching, and results in decreased expression of the AaMFS19 gene. The expression of AaMFS19 is regulated by the Yap1 transcription activator, the Hog1 and Fus3 mitogen-activated protein (MAP) kinases, the 'two component' histidine kinase, and the Skn7 response regulator. Our results demonstrate that A. alternata confers resistance to different chemicals via a membrane-bound MFS transporter.

  5. Antifungal activity of food additives in vitro and as ingredients of hydroxypropyl methylcellulose-lipid edible coatings against Botrytis cinerea and Alternaria alternata on cherry tomato fruit.

    PubMed

    Fagundes, Cristiane; Pérez-Gago, María B; Monteiro, Alcilene R; Palou, Lluís

    2013-09-16

    The antifungal activity of food additives or 'generally recognized as safe' (GRAS) compounds was tested in vitro against Botrytis cinerea and Alternaria alternata. Radial mycelial growth of each pathogen was measured in PDA Petri dishes amended with food preservatives at 0.2, 1.0, or 2.0% (v/v) after 3, 5, and 7 days of incubation at 25 °C. Selected additives and concentrations were tested as antifungal ingredients of hydroxypropyl methylcellulose (HPMC)-lipid edible coatings. The curative activity of stable coatings was tested in in vivo experiments. Cherry tomatoes were artificially inoculated with the pathogens, coated by immersion about 24 h later, and incubated at 20 °C and 90% RH. Disease incidence and severity (lesion diameter) were determined after 6, 10, and 15 days of incubation and the 'area under the disease progress stairs' (AUDPS) was calculated. In general, HPMC-lipid antifungal coatings controlled black spot caused by A. alternata more effectively than gray mold caused by B. cinerea. Overall, the best results for reduction of gray mold on cherry tomato fruit were obtained with coatings containing 2.0% of potassium carbonate, ammonium phosphate, potassium bicarbonate, or ammonium carbonate, while 2.0% sodium methylparaben, sodium ethylparaben, and sodium propylparaben were the best ingredients for coatings against black rot.

  6. Examination of fungi in domestic interiors by using factor analysis: Correlations and associations with home factors. [Cladosporium, Alternaria, Epicoccum, Aureobasidium, Aspergillus; Penicillium

    SciTech Connect

    Su, H.J.; Rotnitzky, A.; Spengler, J.D. ); Burge, H.A. )

    1992-01-01

    Factor analysis was utilized to investigate correlations among airborne microorganisms collected with Andersen samplers from homes in Topeka, Kans., during the winter of 1987 to 1988. The factors derived were used to relate microbial concentrations with categorical, questionnaire-derived descriptions of housing conditions. This approach successfully identified groups of common aboveground decay fungi including Cladosporium, Alternaria, Epicoccum, and Aureobasidium spp. The common soil fungi Aspergillus and Penicillium spp. were also separated as a group. These previously known ecological groupings were confirmed with air sampling data by a quantitative evaluation technique. The above ground decay fungi sampled indoors in winter were present at relatively high concentrations in homes with gas stoves for cooking, suggesting a possible association between these fungi and increased humidity from the combustion process. Elevated concentrations of the soil fungi were significantly associated with the dirt floor, crawl-space type of basement. Elevated concentrations of water-requiring fungi, such as Fusarium spp., were shown to be associated with water collection in domestic interiors. Also, elevated mean concentrations for the group of fungi including Cladosporium, Epicoccum, Aureobasidium, and yeast spp. were found to be associated with symptoms reported on a health questionnaire. This finding was consistent with the authors previous study of associations between respiratory health and airborne microorganisms by univariate logistic regression analysis.

  7. Assessment of genetic diversity and distribution of endophytic fungal communities of Alternaria solani isolates associated with the dominant Karanja plants in Sanganer Region of Rajasthan.

    PubMed

    Tiwari, Kartikeya; Chittora, Manish

    2013-12-01

    Higher plants are ubiquitously colonized with fungal endophytes that often lack readily detectable structures. Current study examines the distribution of endophytic fungal communities within Karanja plants and diversity of novel fungal endophyte Alternaria solani isolates collected from different locations of Sanganer region of Rajasthan. Results confirmed that A. solani is a major fungal endophyte consortium associated with Karanja plants. PCR Amplified fragments using random amplified polymorphic DNA (RAPD) primers were subjected to unweighted pair group method analysis (UPGMA), which clearly distinguished twelve ecologically diverse A. solani isolates. A total of 58 RAPD loci were amplified, out of which 35 (60.34%) were polymorphic and 23 were monomorphic (39.66%) in nature. These polymorphic loci were identified with an average of 2.92 bands per primer. The efficacy of RAPD markers proved as an efficient marker system with respect to detection of polymorphism and number of loci scored and can be used for the identification of a particular isolates, thereby defining core collections and strengthening their exploitation in acquiring novel products produced by them.

  8. Comparative Transcriptomic Analysis Reveals That Ethylene/H2O2-Mediated Hypersensitive Response and Programmed Cell Death Determine the Compatible Interaction of Sand Pear and Alternaria alternata

    PubMed Central

    Wang, Hong; Lin, Jing; Chang, Youhong; Jiang, Cai-Zhong

    2017-01-01

    A major restriction on sand pear (Pyrus pyrifolia) production is black spot disease caused by the necrotrophic fungus Alternaria alternata. However, the pear response mechanism to A. alternata is unknown at the molecular level. Here, host responses of a resistant cultivar Cuiguan (CG) and a susceptible cultivar Sucui1 (SC1) to A. alternata infection were investigated. We found that the primary necrotic lesion formed at 1 dpi and the expansion of lesions was aggressive in SC1. Data from transcriptomic profiles using RNA-Seq technology identified a large number of differentially expressed genes (DEGs) between CG and SC1 in the early phase of A. alternata infection. K-mean cluster and Mapman analysis revealed that genes involved in ethylene (ET) biosynthesis and ET signaling pathway, such as ACS, ACOs, and ERFs, and in hypersensitive response (HR) and programmed cell death (PCD) were significantly enriched and up-regulated in the susceptible cultivar SC1. Conversely, genes involved in response to hydrogen peroxide and superoxide were differentially up-regulated in the resistant cultivar CG after inoculation with the fungus. Furthermore, ET levels were highly accumulated in SC1, but not in CG. Higher activities of detoxifying enzymes such as catalases were detected in CG. Our results demonstrate that the ET-/H2O2-mediated PCD and detoxifying processes play a vital role in the interaction of pear and A. alternata. PMID:28261248

  9. Genome-wide identification and expression analysis of the apple ASR gene family in response to Alternaria alternata f. sp. mali.

    PubMed

    Huang, Kaihui; Zhong, Yan; Li, Yingjun; Zheng, Dan; Cheng, Zong-Ming

    2016-10-01

    The ABA/water stress/ripening-induced (ASR) gene family exists universally in higher plants, and many ASR genes are up-regulated during periods of environmental stress and fruit ripening. Although a considerable amount of research has been performed investigating ASR gene response to abiotic stresses, relatively little is known about their roles in response to biotic stresses. In this report, we identified five ASR genes in apple (Malus × domestica) and explored their phylogenetic relationship, duplication events, and selective pressure. Five apple ASR genes (Md-ASR) were divided into two clades based on phylogenetic analysis. Species-specific duplication was detected in M. domestica ASR genes. Leaves of 'Golden delicious' and 'Starking' were infected with Alternaria alternata f. sp. mali, which causes apple blotch disease, and examined for the expression of the ASR genes in lesion areas during the first 72 h after inoculation. Md-ASR genes showed different expression patterns at different sampling times in 'Golden delicious' and 'Starking'. The activities of stress-related enzymes, peroxidase (POD), superoxide dismutase (SOD), catalase (CAT), phenylalanine ammonia lyase (PAL), and polyphenoloxidase (PPO), and the content of malondialdehyde (MDA) were also measured in different stages of disease development in two cultivars. The ASR gene expression patterns and theses physiological indexes for disease resistance suggested that Md-ASR genes are involved in biotic stress responses in apple.

  10. Nail stem cells.

    PubMed

    Sellheyer, Klaus

    2013-03-01

    Our knowledge on stem cells of the hair follicle has increased exponentially after the bulge was characterized as the stem cell niche two decades ago. In contrast, little is known about stem cells in the nail unit. Whereas hair follicles are plentiful and easy to access, the human body has only twenty nails and they are rarely biopsied. Therefore, examining fetal material offers unique advantages. In the following mini-review, our current knowledge on nail stem cells is summarized and analogies to the hair follicle stem cells are drawn.

  11. Liver cancer stem cells.

    PubMed

    Sell, Stewart; Leffert, Hyam L

    2008-06-10

    In an effort to review the evidence that liver cancer stem cells exist, two fundamental questions must be addressed. First, do hepatocellular carcinomas (HCC) arise from liver stem cells? Second, do HCCs contain cells that possess properties of cancer stem cells? For many years the finding of preneoplastic nodules in the liver during experimental induction of HCCs by chemicals was interpreted to support the hypothesis that HCC arose by dedifferentiation of mature liver cells. More recently, recognition of the role of small oval cells in the carcinogenic process led to a new hypothesis that HCC arises by maturation arrest of liver stem cells. Analysis of the cells in HCC supports the presence of cells with stem-cell properties (ie, immortality, transplantability, and resistance to therapy). However, definitive markers for these putative cancer stem cells have not yet been found and a liver cancer stem cell has not been isolated.

  12. Plant stem cell niches.

    PubMed

    Aichinger, Ernst; Kornet, Noortje; Friedrich, Thomas; Laux, Thomas

    2012-01-01

    Multicellular organisms possess pluripotent stem cells to form new organs, replenish the daily loss of cells, or regenerate organs after injury. Stem cells are maintained in specific environments, the stem cell niches, that provide signals to block differentiation. In plants, stem cell niches are situated in the shoot, root, and vascular meristems-self-perpetuating units of organ formation. Plants' lifelong activity-which, as in the case of trees, can extend over more than a thousand years-requires that a robust regulatory network keep the balance between pluripotent stem cells and differentiating descendants. In this review, we focus on current models in plant stem cell research elaborated during the past two decades, mainly in the model plant Arabidopsis thaliana. We address the roles of mobile signals on transcriptional modules involved in balancing cell fates. In addition, we discuss shared features of and differences between the distinct stem cell niches of Arabidopsis.

  13. Investigating Preservice STEM Teacher Conceptions of STEM Education

    ERIC Educational Resources Information Center

    Radloff, Jeff; Guzey, Selcen

    2016-01-01

    Surrounding the national emphasis on improving STEM education, effective STEM educators are required. Connected, yet often overlooked, is the need for effective preservice STEM teaching instruction for incoming educators. At a basic level, preservice STEM teacher education should include STEM content, pedagogy, and conceptualization. However, the…

  14. Global Collaborative STEM Education

    NASA Astrophysics Data System (ADS)

    Meabh Kelly, Susan; Smith, Walter

    2016-04-01

    Global Collaborative STEM Education, as the name suggests, simultaneously supports two sets of knowledge and skills. The first set is STEM -- science, technology, engineering and math. The other set of content knowledge and skills is that of global collaboration. Successful global partnerships require awareness of one's own culture, the biases embedded within that culture, as well as developing awareness of the collaborators' culture. Workforce skills fostered include open-mindedness, perseverance when faced with obstacles, and resourceful use of technological "bridges" to facilitate and sustain communication. In respect for the 2016 GIFT Workshop focus, Global Collaborative STEM Education projects dedicated to astronomy research will be presented. The projects represent different benchmarks within the Global Collaborative STEM Education continuum, culminating in an astronomy research experience that fully reflects how the global STEM workforce collaborates. To facilitate wider engagement in Global Collaborative STEM Education, project summaries, classroom resources and contact information for established international collaborative astronomy research projects will be disseminated.

  15. Characterization and Comparison of the CPK Gene Family in the Apple (Malus × domestica) and Other Rosaceae Species and Its Response to Alternaria alternata Infection

    PubMed Central

    Wei, Menghan; Wang, Sanhong; Dong, Hui; Cai, Binhua; Tao, Jianmin

    2016-01-01

    As one of the Ca2+ sensors, calcium-dependent protein kinase (CPK) plays vital roles in immune and stress signaling, growth and development, and hormone responses, etc. Recently, the whole genome of apple (Malus × domestica), pear (Pyrus communis), peach (Prunus persica), plum (Prunus mume) and strawberry (Fragaria vesca) in Rosaceae family has been fully sequenced. However, little is known about the CPK gene family in these Rosaceae species. In this study, 123 CPK genes were identified from five Rosaceae species, including 37 apple CPKs, 37 pear CPKs, 17 peach CPKs, 16 strawberry CPKs, and 16 plum CPKs. Based on the phylogenetic tree topology and structural characteristics, we divided the CPK gene family into 4 distinct subfamilies: Group I, II, III, and IV. Whole-genome duplication (WGD) or segmental duplication played vital roles in the expansion of the CPK in these Rosaceae species. Most of segmental duplication pairs in peach and plum may have arisen from the γ triplication (~140 million years ago [MYA]), while in apple genome, many duplicated genes may have been derived from a recent WGD (30~45 MYA). Purifying selection also played a critical role in the function evolution of CPK family genes. Expression of apple CPK genes in response to apple pathotype of Alternaria alternata was verified by analysis of quantitative real-time RT-PCR (qPCR). Expression data demonstrated that CPK genes in apple might have evolved independently in different biological contexts. The analysis of evolution history and expression profile laid a foundation for further examining the function and complexity of the CPK gene family in Rosaceae. PMID:27186637

  16. Characterization and Comparison of the CPK Gene Family in the Apple (Malus × domestica) and Other Rosaceae Species and Its Response to Alternaria alternata Infection.

    PubMed

    Wei, Menghan; Wang, Sanhong; Dong, Hui; Cai, Binhua; Tao, Jianmin

    2016-01-01

    As one of the Ca2+ sensors, calcium-dependent protein kinase (CPK) plays vital roles in immune and stress signaling, growth and development, and hormone responses, etc. Recently, the whole genome of apple (Malus × domestica), pear (Pyrus communis), peach (Prunus persica), plum (Prunus mume) and strawberry (Fragaria vesca) in Rosaceae family has been fully sequenced. However, little is known about the CPK gene family in these Rosaceae species. In this study, 123 CPK genes were identified from five Rosaceae species, including 37 apple CPKs, 37 pear CPKs, 17 peach CPKs, 16 strawberry CPKs, and 16 plum CPKs. Based on the phylogenetic tree topology and structural characteristics, we divided the CPK gene family into 4 distinct subfamilies: Group I, II, III, and IV. Whole-genome duplication (WGD) or segmental duplication played vital roles in the expansion of the CPK in these Rosaceae species. Most of segmental duplication pairs in peach and plum may have arisen from the γ triplication (~140 million years ago [MYA]), while in apple genome, many duplicated genes may have been derived from a recent WGD (30~45 MYA). Purifying selection also played a critical role in the function evolution of CPK family genes. Expression of apple CPK genes in response to apple pathotype of Alternaria alternata was verified by analysis of quantitative real-time RT-PCR (qPCR). Expression data demonstrated that CPK genes in apple might have evolved independently in different biological contexts. The analysis of evolution history and expression profile laid a foundation for further examining the function and complexity of the CPK gene family in Rosaceae.

  17. Biocontrol Potential of Streptomyces hydrogenans Strain DH16 toward Alternaria brassicicola to Control Damping Off and Black Leaf Spot of Raphanus sativus

    PubMed Central

    Manhas, Rajesh K.; Kaur, Talwinder

    2016-01-01

    Biocontrol agents and their bioactive metabolites provide one of the best alternatives to decrease the use of chemical pesticides. In light of this, the present investigation reports the biocontrol potential of Streptomyces hydrogenans DH16 and its metabolites towards Alternaria brassicicola, causal agent of black leaf spot and damping off of seedlings of crucifers. In vitro antibiosis of strain against pathogen revealed complete suppression of mycelial growth of pathogen, grown in potato dextrose broth supplemented with culture supernatant (20% v/v) of S. hydrogenans DH16. Microscopic examination of the fungal growth showed severe morphological abnormalities in the mycelium caused by antifungal metabolites. In vivo studies showed the efficacy of streptomycete cells and culture supernatant as seed dressings to control damping off of Raphanus sativus seedlings. Treatment of pathogen infested seeds with culture supernatant (10%) and streptomycete cells significantly improved seed germination (75–80%) and vigor index (1167–1538). Furthermore, potential of cells and culture supernatant as foliar treatment to control black leaf spot was also evaluated. Clearly visible symptoms of disease were observed in the control plants with 66.81% disease incidence and retarded growth of root system. However, disease incidence reduced to 6.78 and 1.47% in plants treated with antagonist and its metabolites, respectively. Additionally, treatment of seeds and plants with streptomycete stimulated various growth traits of plants over uninoculated control plants in the absence of pathogen challenge. These results indicate that S. hydrogenans and its culture metabolites can be developed as biofungicides as seed dressings to control seed borne pathogens, and as sprays to control black leaf spot of crucifers. PMID:28018402

  18. Enhanced resistance to Phytophthora infestans and Alternaria solani in leaves and tubers, respectively, of potato plants with decreased activity of the plastidic ATP/ADP transporter.

    PubMed

    Conrath, Uwe; Linke, Christoph; Jeblick, Wolfgang; Geigenberger, Peter; Quick, W Paul; Neuhaus, H Ekkehard

    2003-05-01

    Recently, it has been reported that tubers of transgenic potato ( Solanum tuberosum L.) plants with decreased activity of the plastidic ATP/ADP transporter (AATP1) contain less starch, despite having an increased glucose level [P. Geigenberger et al. (2001) Plant Physiol 125:1667-1678]. The metabolic alterations correlated with enhanced resistance to the bacterium Erwinia carotovora. Here it is shown that transgenic potato tubers, possessing less starch yet increased glucose levels due to the expression of a cytoplasm-localized yeast invertase, exhibit drastic susceptibility to E. carotovora. In addition, it is demonstrated that AATP1 anti-sense tubers show an increased capacity to ward off the pathogenic fungus Alternaria solani. In contrast to AATP1 anti-sense tubers, the corresponding leaf tissue does not show changes in carbohydrate accumulation. However, upon elicitor treatment, AATP1 anti-sense leaves possess an increased capacity to release H(2)O(2) and activate various defence-related genes, reactions that are associated with substantially delayed appearance of disease symptoms caused by Phytophthora infestans. Grafting experiments between AATP1 anti-sense plants and wild-type plants indicate the presence of a signal that is generated in AATP1 rootstocks and primes wild-type scions for potentiated activation of cellular defence responses in leaves. Together, the results suggest that (i) the enhanced pathogen tolerance of AATP1 anti-sense tubers is not due to "high sugar resistance", (ii) the increased disease resistance of AATP1 anti-sense tubers is effective against different types of pathogen and (iii) a systemic signal induced by antisensing the plastidic ATP/ADP transporter in potato tubers confers increased resistance to pathogens.

  19. Induction of protection against the necrotrophic pathogens Phytophthora citrophthora and Alternaria solani in Lycopersicon esculentum Mill. by a novel synthetic glycoside combined with amines.

    PubMed

    Flors, Víctor; Miralles, M Carmen; González-Bosch, Carmen; Carda, Miguel; García-Agustín, Pilar

    2003-04-01

    1,2,3,4-tetra-O-acetyl-6-ethyladipate-beta- D-glucopyranose (TOGE), a glycoside derivative of adipic acid monoethyl ester and 1,2,3,4-tetra-O-acetyl-beta- D-glucopyranose, was synthesized and the resistance-inducing activity shown by TOGE-1 (TOGE plus furfurylamine) and TOGE-2 (TOGE plus 1,3-diaminepropane) was assayed. TOGE-1 and TOGE-2 protected tomato plants against two different fungal pathogens Phytophthora citrophthora and Alternaria solani. Foliar treatments at very low concentrations (2.5 mg l(-1) TOGE, 0.5 mg l(-1) amine) clearly reduced the disease incidence for both pathogens. TOGE-2 application was the most effective on intact plants as well as on detached leaves, reducing fungal growth by more than 46% with respect to control plants. On the other hand TOGE-1 treatment reduced fungal advance by 21%. These results demonstrate a high protective effect against fungal infections for both chemicals. A possible direct antimicrobial effect was discounted due to the weak activity observed in vitro against these pathogens at the low concentrations used in plants. TOGE-2 treatment clearly activates resistance against both pathogens and improves the protective effect previously shown by FGA mixture (adipic acid monoethyl ester, 1,2,3,4-tetra-O-acetyl-beta- D-glucopyranose and furfurylamine) [V. Flors et al. (2001) J Agric Food Chem 49:2569-2575]. The obtained results indicate that TOGE-1 and TOGE-2 act as resistance inducers. Although their mode of action is still unknown, pre-challenge studies have demonstrated the induction of the phenylpropanoid pathway and antioxidant activities. Both chemicals have demonstrated a beneficial effect on plant development, increasing chlorophyll and protein contents, photosynthetic rate and water-use efficiency. The improvement of plant growth and development produced by these treatments suggests crop tolerance to these chemicals, although effective formulations that are safe to humans must be developed before this technology can

  20. Chilling stress--the key predisposing factor for causing Alternaria alternata infection and leading to cotton (Gossypium hirsutum L.) leaf senescence.

    PubMed

    Zhao, Jingqing; Li, Sha; Jiang, Tengfei; Liu, Zhi; Zhang, Wenwei; Jian, Guiliang; Qi, Fangjun

    2012-01-01

    Leaf senescence plays a vital role in nutrient recycling and overall capacity to assimilate carbon dioxide. Cotton premature leaf senescence, often accompanied with unexpected short-term low temperature, has been occurring with an increasing frequency in many cotton-growing areas and causes serious reduction in yield and quality of cotton. The key factors for causing and promoting cotton premature leaf senescence are still unclear. In this case, the relationship between the pre-chilling stress and Alternaria alternata infection for causing cotton leaf senescence was investigated under precisely controlled laboratory conditions with four to five leaves stage cotton plants. The results showed short-term chilling stress could cause a certain degree of physiological impairment to cotton leaves, which could be recovered to normal levels in 2-4 days when the chilling stresses were removed. When these chilling stress injured leaves were further inoculated with A. alternata, the pronounced appearance and development of leaf spot disease, and eventually the pronounced symptoms of leaf senescence, occurred on these cotton leaves. The onset of cotton leaf senescence at this condition was also reflected in various physiological indexes such as irreversible increase in malondialdehyde (MDA) content and electrolyte leakage, irreversible decrease in soluble protein content and chlorophyll content, and irreversible damage in leaves' photosynthesis ability. The presented results demonstrated that chilling stress acted as the key predisposing factor for causing A. alternata infection and leading to cotton leaf senescence. It could be expected that the understanding of the key factors causing and promoting cotton leaf senescence would be helpful for taking appropriate management steps to prevent cotton premature leaf senescence.

  1. Site-directed mutagenesis of α-L-rhamnosidase from Alternaria sp. L1 to enhance synthesis yield of reverse hydrolysis based on rational design.

    PubMed

    Xu, Li; Liu, Xiaohong; Yin, Zhenhao; Liu, Qian; Lu, Lili; Xiao, Min

    2016-12-01

    The α-L-rhamnosidase catalyzes the hydrolytic release of rhamnose from polysaccharides and glycosides and is widely used due to its applications in a variety of industrial processes. Our previous work reported that a wild-type α-L-rhamnosidase (RhaL1) from Alternaria sp. L1 could synthesize rhamnose-containing chemicals (RCCs) though reverse hydrolysis reaction with inexpensive rhamnose as glycosyl donor. To enhance the yield of reverse hydrolysis reaction and to determine the amino acid residues essential for the catalytic activity of RhaL1, site-directed mutagenesis of 11 residues was performed in this study. Through rationally designed mutations, the critical amino acid residues which may form direct or solvent-mediated hydrogen bonds with donor rhamnose (Asp(252), Asp(257), Asp(264), Glu(530), Arg(548), His(553), and Trp(555)) and may form the hydrophobic pocket in stabilizing donor (Trp(261), Tyr(302), Tyr(316), and Trp(369)) in active-site of RhaL1 were analyzed, and three positive mutants (W261Y, Y302F, and Y316F) with improved product yield stood out. From the three positive variants, mutant W261Y accelerated the reverse hydrolysis with a prominent increase (43.7 %) in relative yield compared to the wild-type enzyme. Based on the 3D structural modeling, we supposed that the improved yield of mutant W261Y is due to the adjustment of the spatial position of the putative catalytic acid residue Asp(257). Mutant W261Y also exhibited a shift in the pH-activity profile in hydrolysis reaction, indicating that introducing of a polar residue in the active site cavity may affect the catalysis behavior of the enzyme.

  2. Isolation, characterization and molecular three-dimensional structural predictions of metalloprotease from a phytopathogenic fungus, Alternaria solani (Ell. and Mart.) Sor.

    PubMed

    Chandrasekaran, Murugesan; Chandrasekar, Raman; Chun, Se-Chul; Sathiyabama, Muthukrishnan

    2016-08-01

    The present study aims at isolation, identification, characterization and prediction of three-dimensional molecular architecture of a proteolytic enzyme from the early blight pathogen, Alternaria solani which are hypothesized to be a marker of phytopathogenicity. Maximum enzyme production by A. solani was observed in Czapex's Dox broth amended with 2% (w/v) casein than other inducer amendments. Results indicate that the enzyme remained highly active in a pH range of 7.0-10.0 and a temperature range of 45-50°C. The enzyme was strongly inhibited by EDTA, whereas phenylmethylsulfonyl fluoride and monovalent cations (Na(+), K(+)) had little effect. Metal ions such as MgSO4, CaCl2, KCl at 10 mM concentration showed a stimulatory effect (>85%) on protease activity. Matrix-assisted laser desorption and ionization time of flight/mass spectrometry analysis of partially purified enzyme revealed the presence of protease belonging to a keratinolytic protein (metalloprotease) of exopeptidase nature. Putative A. solani keratinolytic enzyme (AsK) is made up of 216 amino acid residues with molecular weight (MW) 24.5 kDa, having a molecular formula of C1094H1704N290O342S4. Ramachandran plot analysis of the protein residues falling into the most favored secondary structures was observed at 84.2%. The major protein structural blocks, 2-β-sheets, and 9-α-helices have a greater tendency to be conserved during the evolutionary process than do mere sequences of amino acids. Besides, AsK, model prediction showed the presence of a Zinc atom at helix regions (Helix 3, 6, 7: His(57), His(130), His(169), and Cys(123)). Thus, it can be concluded that the major proteinases of AsK are divalent cation-requiring metalloproteinases and make them potential targets of protease inhibitors designing.

  3. A Major Facilitator Superfamily Transporter-Mediated Resistance to Oxidative Stress and Fungicides Requires Yap1, Skn7, and MAP Kinases in the Citrus Fungal Pathogen Alternaria alternata

    PubMed Central

    Chen, Li-Hung; Tsai, Hsieh-Chin; Yu, Pei-Ling

    2017-01-01

    Major Facilitator Superfamily (MFS) transporters play an important role in multidrug resistance in fungi. We report an AaMFS19 gene encoding a MFS transporter required for cellular resistance to oxidative stress and fungicides in the phytopathogenic fungus Alternaria alternata. AaMFS19, containing 12 transmembrane domains, displays activity toward a broad range of substrates. Fungal mutants lacking AaMFS19 display profound hypersensitivities to cumyl hydroperoxide, potassium superoxide, many singlet oxygen-generating compounds (eosin Y, rose Bengal, hematoporphyrin, methylene blue, and cercosporin), and the cell wall biosynthesis inhibitor, Congo red. AaMFS19 mutants also increase sensitivity to copper ions, clotrimazole, fludioxonil, and kocide fungicides, 2-chloro-5-hydroxypyridine (CHP), and 2,3,5-triiodobenzoic acid (TIBA). AaMFS19 mutants induce smaller necrotic lesions on leaves of a susceptible citrus cultivar. All observed phenotypes in the mutant are restored by introducing and expressing a wild-type copy of AaMFS19. The wild-type strain of A. alternata treated with either CHP or TIBA reduces radial growth and formation and germination of conidia, increases hyphal branching, and results in decreased expression of the AaMFS19 gene. The expression of AaMFS19 is regulated by the Yap1 transcription activator, the Hog1 and Fus3 mitogen-activated protein (MAP) kinases, the ‘two component’ histidine kinase, and the Skn7 response regulator. Our results demonstrate that A. alternata confers resistance to different chemicals via a membrane-bound MFS transporter. PMID:28060864

  4. Precise determination of the Alternaria mycotoxins alternariol and alternariol monomethyl ether in cereal, fruit and vegetable products using stable isotope dilution assays.

    PubMed

    Asam, Stefan; Konitzer, Katharina; Rychlik, Michael

    2011-02-01

    Cereal, fruit and vegetable products were analyzed for contamination with the Alternaria mycotoxins alternariol (AOH) and alternariol monomethyl ether (AME) using stable isotope dilution assays (SIDAs). Both toxins were practically not detected in cereals and cereal products: AOH-one out of 13 samples at a content of 4.1 μg/kg; AME-two out of 13 samples at contents ranging between 0.2 and 0.6 μg/kg. However, if cereals for animal nutrition were analyzed, much higher values were found: AOH-five out of six samples (13-250 μg/kg); AME-six out of six samples (3-100 μg/kg). This finding may pose a potential problem concerning animal health. AOH and AME were frequently detected in vegetable products: AOH-5 out of 10 samples (2.6-25 μg/kg); AME-6 out of 10 samples (0.1-5 μg/kg). Tomato products were affected, especially. The highest content of AOH (25 μg/kg) and AME (5 μg/kg) were found in triple concentrated tomato paste. Special wines like "Trockenbeerenauslese" or "Spätlese" (affected by noble rot in the vineyard) contained AOH (4/6 samples; 1.2-4.9 μg/kg) and AME (4/6 samples; 0.1-0.3 μg/kg), but the values did not exceed the values of both toxins that were found generally in wines.

  5. Stress and stem cells.

    PubMed

    Tower, John

    2012-01-01

    The unique properties and functions of stem cells make them particularly susceptible to stresses and also lead to their regulation by stress. Stem cell division must respond to the demand to replenish cells during normal tissue turnover as well as in response to damage. Oxidative stress, mechanical stress, growth factors, and cytokines signal stem cell division and differentiation. Many of the conserved pathways regulating stem cell self-renewal and differentiation are also stress-response pathways. The long life span and division potential of stem cells create a propensity for transformation (cancer) and specific stress responses such as apoptosis and senescence act as antitumor mechanisms. Quiescence regulated by CDK inhibitors and a hypoxic niche regulated by FOXO transcription factor function to reduce stress for several types of stem cells to facilitate long-term maintenance. Aging is a particularly relevant stress for stem cells, because repeated demands on stem cell function over the life span can have cumulative cell-autonomous effects including epigenetic dysregulation, mutations, and telomere erosion. In addition, aging of the organism impairs function of the stem cell niche and systemic signals, including chronic inflammation and oxidative stress.

  6. Stem Cell Transplants (For Teens)

    MedlinePlus

    ... Loss Surgery? A Week of Healthy Breakfasts Shyness Stem Cell Transplants KidsHealth > For Teens > Stem Cell Transplants Print ... Does it Take to Recover? Coping What Are Stem Cells? As you probably remember from biology class, every ...

  7. Stem Cell Transplants (For Teens)

    MedlinePlus

    ... Loss Surgery? A Week of Healthy Breakfasts Shyness Stem Cell Transplants KidsHealth > For Teens > Stem Cell Transplants A ... Does it Take to Recover? Coping What Are Stem Cells? As you probably remember from biology class, every ...

  8. Investigating Preservice STEM Teacher Conceptions of STEM Education

    NASA Astrophysics Data System (ADS)

    Radloff, Jeff; Guzey, Selcen

    2016-10-01

    Surrounding the national emphasis on improving STEM education, effective STEM educators are required. Connected, yet often overlooked, is the need for effective preservice STEM teaching instruction for incoming educators. At a basic level, preservice STEM teacher education should include STEM content, pedagogy, and conceptualization. However, the literature suggests no leading conception of STEM education, and little is known about how preservice STEM teachers are conceptualizing STEM education. In order to explore preservice STEM teacher conceptions of STEM education, preservice teachers at a large, Midwestern research university were given an open-ended survey eliciting both textual and visual responses. Here, we report and discuss the results of employing this instrument in relation with the current STEM conceptualization literature.

  9. Myeloproliferative neoplasm stem cells.

    PubMed

    Mead, Adam J; Mullally, Ann

    2017-03-23

    Myeloproliferative neoplasms (MPNs) arise in the hematopoietic stem cell (HSC) compartment as a result of the acquisition of somatic mutations in a single HSC that provides a selective advantage to mutant HSC over normal HSC and promotes myeloid differentiation to engender a myeloproliferative phenotype. This population of somatically mutated HSC, which initiates and sustains MPNs, is termed MPN stem cells. In >95% of cases, mutations that drive the development of an MPN phenotype occur in a mutually exclusive manner in 1 of 3 genes: JAK2, CALR, or MPL The thrombopoietin receptor, MPL, is the key cytokine receptor in MPN development, and these mutations all activate MPL-JAK-STAT signaling in MPN stem cells. Despite common biological features, MPNs display diverse disease phenotypes as a result of both constitutional and acquired factors that influence MPN stem cells, and likely also as a result of heterogeneity in the HSC in which MPN-initiating mutations arise. As the MPN clone expands, it exerts cell-extrinsic effects on components of the bone marrow niche that can favor the survival and expansion of MPN stem cells over normal HSC, further sustaining and driving malignant hematopoiesis. Although developed as targeted therapies for MPNs, current JAK2 inhibitors do not preferentially target MPN stem cells, and as a result, rarely induce molecular remissions in MPN patients. As the understanding of the molecular mechanisms underlying the clonal dominance of MPN stem cells advances, this will help facilitate the development of therapies that preferentially target MPN stem cells over normal HSC.

  10. Learn About Stem Cells

    MedlinePlus

    ... develops and ages, the number and type of stem cells changes. Totipotent cells are no longer present after dividing into the cells that generate the placenta and umbilical cord. Pluripotent cells ... organs and tissues. The stem cells that stay in your body throughout your ...

  11. Designing for STEM Integration

    ERIC Educational Resources Information Center

    Berland, Leema K.

    2013-01-01

    We are increasingly seeing an emphasis on STEM integration in high school classrooms such that students will learn and apply relevant math and science content while simultaneously developing engineering habits of mind. However, research in both science education and engineering education suggests that this goal of truly integrating STEM is rife…

  12. STEM School Discourse Patterns

    ERIC Educational Resources Information Center

    Tofel-Grehl, Colby; Callahan, Carolyn M.

    2016-01-01

    Analysis of discursive practices in science classrooms within STEM schools may provide meaningful information about the nature of these classrooms and, potentially, their uniqueness. Full descriptions of current practice can serve as a foundation for exploring the differences in instructional norms within STEM specialized schools and across…

  13. Teaching STEM by Design

    ERIC Educational Resources Information Center

    Billiar, Kristen; Hubelbank, Jeanne; Oliva, Thomas; Camesano, Terri

    2014-01-01

    Developing innovative science, technology, engineering and mathematics (STEM) curricula that elicit student excitement for learning is a continuous challenge for K-12 STEM teachers. Generating these lessons while meeting conflicting pedagogical objectives and constraints of time, content, and cost from various parties is truly a challenging task…

  14. Making STEM Connections

    ERIC Educational Resources Information Center

    Stump, Sheryl L.; Bryan, Joel A.; McConnell, Tom J.

    2016-01-01

    Integrated approaches to education in science, technology, engineering, and mathematics (STEM), especially those set in the context of real-world situations, can motivate and deepen students' learning of the STEM subjects (National Academy of Engineering and National Research Council 2014). This article describes two integrated investigations used…

  15. Colorectal cancer stem cells.

    PubMed

    Salama, Paul; Platell, Cameron

    2009-10-01

    Somatic stem cells reside at the base of the crypts throughout the colonic mucosa. These cells are essential for the normal regeneration of the colonic epithelium. The stem cells reside within a special 'niche' comprised of intestinal sub-epithelial myofibroblasts that tightly control their function. It has been postulated that mutations within these adult colonic stem cells may induce neoplastic changes. Such cells can then dissociate from the epithelium and travel into the mesenchyme and thus form invasive cancers. This theory is based on the observation that within a colon cancer, less than 1% of the neoplastic cells have the ability to regenerate the tumour. It is this group of cells that exhibits characteristics of colonic stem cells. Although anti-neoplastic agents can induce remissions by inhibiting cell division, the stem cells appear to be remarkably resistant to both standard chemotherapy and radiotherapy. These stem cells may therefore persist after treatment and form the nucleus for cancer recurrence. Hence, future treatment modalities should focus specifically on controlling the cancer stem cells. In this review, we discuss the biology of normal and malignant colonic stem cells.

  16. Bringing STEM to Life

    ERIC Educational Resources Information Center

    Berkeihiser, Mike; Ray, Dori

    2013-01-01

    The interdisciplinary approach that science, technology, engineering and mathematics (STEM) projects inspire in both teachers and students "brings to light a larger picture that promotes real-world scientific applications, which has in turn been shown to increase undergraduate persistence in STEM." The high school students have been…

  17. STEM Careers Ambassadors

    ERIC Educational Resources Information Center

    Eaton, Denise

    2011-01-01

    This article is designed to help teachers feel more confident in their work with STEM Ambassadors to further enhance enrichment activities. Skills shortages in Science, Technology, Engineering, Maths (STEM) and the Built Environment are well documented, and will continue to be an issue whether people are in a period of recession or recovery. The…

  18. Lock For Valve Stem

    NASA Technical Reports Server (NTRS)

    Burley, Richard K.; Guirguis, Kamal S.

    1991-01-01

    Simple, cheap device locks valve stem so its setting cannot be changed by unauthorized people. Device covers valve stem; cover locked in place with standard padlock. Valve lock made of PVC pipe and packing band. Shears, drill or punch, and forming rod only tools needed.

  19. Expanding STEM Education | Poster

    Cancer.gov

    Editor’s note: This article is written as a reflection on experiential STEM education by a student who completed her Werner H. Kirsten internship in June 2015. Here, she advocates for incorporating hands-on experience into STEM curricula. If the only way for high school students to learn science, technology, engineering, and mathematics (STEM) is through textbooks, then count me out. But how then do you get students to learn STEM outside of the classroom? The focus of this article is to advocate for high school STEM education through experiential learning. Tom Freston, one of the founders and the chief executive officer (CEO) of MTV Productions, said in an interview in Men’s Journal that “innovation is taking two things that already exist and...

  20. Intraoperative Stem Cell Therapy

    PubMed Central

    Coelho, Mónica Beato; Cabral, Joaquim M.S.; Karp, Jeffrey M.

    2013-01-01

    Stem cells hold significant promise for regeneration of tissue defects and disease-modifying therapies. Although numerous promising stem cell approaches are advancing in clinical trials, intraoperative stem cell therapies offer more immediate hope by integrating an autologous cell source with a well-established surgical intervention in a single procedure. Herein, the major developments in intraoperative stem cell approaches, from in vivo models to clinical studies, are reviewed, and the potential regenerative mechanisms and the roles of different cell populations in the regeneration process are discussed. Although intraoperative stem cell therapies have been shown to be safe and effective for several indications, there are still critical challenges to be tackled prior to adoption into the standard surgical armamentarium. PMID:22809140

  1. Horizontal gene transfer and gene dosage drives adaptation to wood colonization in a tree pathogen

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Some of the most damaging tree pathogens can attack woody stems, causing lesions (cankers) that may be lethal. To identify the genomic determinants of wood colonization leading to canker formation, we sequenced the genomes of the poplar canker pathogen, Mycosphaerella populorum, and the closely rela...

  2. [On plant stem cells and animal stem cells].

    PubMed

    You, Yun; Jiang, Chao; Huang, Lu-Qi

    2014-01-01

    A comparison of plant and animal stem cells can highlight core aspects of stem-cell biology. In both kingdoms, stem cells are defined by their clonogenic properties and are maintained by intercellular signals. The signaling molecules are different in plants and animals stem cell niches, but the roles of argonaute and polycomb group proteins suggest that there are some molecular similarities.

  3. Genetically based location from triploid populations and gene ontology of a 3.3-mb genome region linked to Alternaria brown spot resistance in citrus reveal clusters of resistance genes.

    PubMed

    Cuenca, José; Aleza, Pablo; Vicent, Antonio; Brunel, Dominique; Ollitrault, Patrick; Navarro, Luis

    2013-01-01

    Genetic analysis of phenotypical traits and marker-trait association in polyploid species is generally considered as a challenge. In the present work, different approaches were combined taking advantage of the particular genetic structures of 2n gametes resulting from second division restitution (SDR) to map a genome region linked to Alternaria brown spot (ABS) resistance in triploid citrus progeny. ABS in citrus is a serious disease caused by the tangerine pathotype of the fungus Alternaria alternata. This pathogen produces ACT-toxin, which induces necrotic lesions on fruit and young leaves, defoliation and fruit drop in susceptible genotypes. It is a strong concern for triploid breeding programs aiming to produce seedless mandarin cultivars. The monolocus dominant inheritance of susceptibility, proposed on the basis of diploid population studies, was corroborated in triploid progeny. Bulk segregant analysis coupled with genome scan using a large set of genetically mapped SNP markers and targeted genetic mapping by half tetrad analysis, using SSR and SNP markers, allowed locating a 3.3 Mb genomic region linked to ABS resistance near the centromere of chromosome III. Clusters of resistance genes were identified by gene ontology analysis of this genomic region. Some of these genes are good candidates to control the dominant susceptibility to the ACT-toxin. SSR and SNP markers were developed for efficient early marker-assisted selection of ABS resistant hybrids.

  4. Genetically Based Location from Triploid Populations and Gene Ontology of a 3.3-Mb Genome Region Linked to Alternaria Brown Spot Resistance in Citrus Reveal Clusters of Resistance Genes

    PubMed Central

    Cuenca, José; Aleza, Pablo; Vicent, Antonio; Brunel, Dominique; Ollitrault, Patrick; Navarro, Luis

    2013-01-01

    Genetic analysis of phenotypical traits and marker-trait association in polyploid species is generally considered as a challenge. In the present work, different approaches were combined taking advantage of the particular genetic structures of 2n gametes resulting from second division restitution (SDR) to map a genome region linked to Alternaria brown spot (ABS) resistance in triploid citrus progeny. ABS in citrus is a serious disease caused by the tangerine pathotype of the fungus Alternaria alternata. This pathogen produces ACT-toxin, which induces necrotic lesions on fruit and young leaves, defoliation and fruit drop in susceptible genotypes. It is a strong concern for triploid breeding programs aiming to produce seedless mandarin cultivars. The monolocus dominant inheritance of susceptibility, proposed on the basis of diploid population studies, was corroborated in triploid progeny. Bulk segregant analysis coupled with genome scan using a large set of genetically mapped SNP markers and targeted genetic mapping by half tetrad analysis, using SSR and SNP markers, allowed locating a 3.3 Mb genomic region linked to ABS resistance near the centromere of chromosome III. Clusters of resistance genes were identified by gene ontology analysis of this genomic region. Some of these genes are good candidates to control the dominant susceptibility to the ACT-toxin. SSR and SNP markers were developed for efficient early marker-assisted selection of ABS resistant hybrids. PMID:24116149

  5. Eco-friendly PEG-based controlled release nano-formulations of Mancozeb: Synthesis and bioefficacy evaluation against phytopathogenic fungi Alternaria solani and Sclerotium rolfsii.

    PubMed

    Majumder, Sujan; Shakil, Najam A; Kumar, Jitendra; Banerjee, Tirthankar; Sinha, Parimal; Singh, Braj B; Garg, Parul

    2016-12-01

    Controlled release (CR) nano-formulations of Mancozeb (manganese-zinc double salt of N,N-bisdithiocarbamic acid), a protective fungicide, have been prepared using laboratory-synthesized poly(ethylene glycols) (PEGs)-based functionalized amphiphilic copolymers without using any surfactants or external additives. The release kinetics of the developed Mancozeb CR formulations were studied and compared with that of commercially available 42% suspension concentrate and 75% wettable powder. Maximum amount of Mancozeb was released on 42nd day for PEG-600 and octyl chain, PEG-1000 and octyl chain, and PEG-600 and hexadecyl chain, on 35th day for PEG-1000 and hexadecyl chain, on 28th day for PEG-1500 and octyl chain, PEG-2000 and octyl chain, PEG-1500 and hexadecyl chain, and PEG-2000 and hexadecyl chain in comparison to both commercial formulations (15th day). The diffusion exponent (n value) of Mancozeb in water ranged from 0.42 to 0.62 in tested formulations. The half-release (t1/2) values ranged from 17.35 to 35.14 days, and the period of optimum availability of Mancozeb ranged from 18.54 to 35.42 days. Further, the in vitro bioefficacy evaluation of developed formulations was done against plant pathogenic fungi Alternaria solani and Sclerotium rolfsii by poison food technique. Effective dose for 50% inhibition in mgL(-1) (ED50) values of developed formulations varied from 1.31 to 2.79 mg L(-1) for A. solani, and 1.60 to 3.14 mg L(-1) for S. rolfsii. The present methodology is simple, economical, and eco-friendly for the development of environment-friendly CR formulations of Mancozeb. These formulations can be used to optimize the release of Mancozeb to achieve disease control for the desired period depending upon the matrix of the polymer used. Importantly, the maximum amount of active ingredient remains available for a reasonable period after application. In addition, the developed CR formulations were found to be suitable for fungicidal applications, allowing use

  6. Comparative Evaluation of Biochemical Changes in Tomato (Lycopersicon esculentum Mill.) Infected by Alternaria alternata and Its Toxic Metabolites (TeA, AOH, and AME)

    PubMed Central

    Meena, Mukesh; Zehra, Andleeb; Dubey, Manish K.; Aamir, Mohd; Gupta, Vijai K.; Upadhyay, Ram S.

    2016-01-01

    In the present study, we have evaluated the comparative biochemical defense response generated against Alternaria alternata and its purified toxins viz. alternariol (AOH), alternariol monomethyl ether (AME), and tenuazonic acid (TeA). The necrotic lesions developed due to treatment with toxins were almost similar as those produced by the pathogen, indicating the crucial role of these toxins in plant pathogenesis. An oxidative burst reaction characterized by the rapid and transient production of a large amount of reactive oxygen species (ROS) occurs following the pathogen infection/toxin exposure. The maximum concentration of hydrogen peroxide (H2O2) produced was reported in the pathogen infected samples (22.2-fold) at 24 h post inoculation followed by TeA (18.2-fold), AOH (15.9-fold), and AME (14.1-fold) in treated tissues. 3,3′- Diaminobenzidine staining predicted the possible sites of H2O2 accumulation while the extent of cell death was measured by Evans blue dye. The extent of lipid peroxidation and malondialdehyde (MDA) content was higher (15.8-fold) at 48 h in the sample of inoculated leaves of the pathogen when compared to control. The cellular damages were observed as increased MDA content and reduced chlorophyll. The activities of antioxidative defense enzymes increased in both the pathogen infected as well as toxin treated samples. Superoxide dismutase (SOD) activity was 5.9-fold higher at 24 h post inoculation in leaves followed by TeA (5.0-fold), AOH (4.1-fold) and AME (2.3-fold) treated leaves than control. Catalase (CAT) activity was found to be increased upto 48 h post inoculation and maximum in the pathogen challenged samples followed by other toxins. The native PAGE results showed the variations in the intensities of isozyme (SOD and CAT) bands in the pathogen infected and toxin treated samples. Ascorbate peroxidase (APx) and glutathione reductase (GR) activities followed the similar trend to scavenge the excess H2O2. The reduction in CAT

  7. Comparative Evaluation of Biochemical Changes in Tomato (Lycopersicon esculentum Mill.) Infected by Alternaria alternata and Its Toxic Metabolites (TeA, AOH, and AME).

    PubMed

    Meena, Mukesh; Zehra, Andleeb; Dubey, Manish K; Aamir, Mohd; Gupta, Vijai K; Upadhyay, Ram S

    2016-01-01

    In the present study, we have evaluated the comparative biochemical defense response generated against Alternaria alternata and its purified toxins viz. alternariol (AOH), alternariol monomethyl ether (AME), and tenuazonic acid (TeA). The necrotic lesions developed due to treatment with toxins were almost similar as those produced by the pathogen, indicating the crucial role of these toxins in plant pathogenesis. An oxidative burst reaction characterized by the rapid and transient production of a large amount of reactive oxygen species (ROS) occurs following the pathogen infection/toxin exposure. The maximum concentration of hydrogen peroxide (H2O2) produced was reported in the pathogen infected samples (22.2-fold) at 24 h post inoculation followed by TeA (18.2-fold), AOH (15.9-fold), and AME (14.1-fold) in treated tissues. 3,3'- Diaminobenzidine staining predicted the possible sites of H2O2 accumulation while the extent of cell death was measured by Evans blue dye. The extent of lipid peroxidation and malondialdehyde (MDA) content was higher (15.8-fold) at 48 h in the sample of inoculated leaves of the pathogen when compared to control. The cellular damages were observed as increased MDA content and reduced chlorophyll. The activities of antioxidative defense enzymes increased in both the pathogen infected as well as toxin treated samples. Superoxide dismutase (SOD) activity was 5.9-fold higher at 24 h post inoculation in leaves followed by TeA (5.0-fold), AOH (4.1-fold) and AME (2.3-fold) treated leaves than control. Catalase (CAT) activity was found to be increased upto 48 h post inoculation and maximum in the pathogen challenged samples followed by other toxins. The native PAGE results showed the variations in the intensities of isozyme (SOD and CAT) bands in the pathogen infected and toxin treated samples. Ascorbate peroxidase (APx) and glutathione reductase (GR) activities followed the similar trend to scavenge the excess H2O2. The reduction in CAT activities

  8. Nuclear receptor regulation of stemness and stem cell differentiation

    PubMed Central

    Jeong, Yangsik

    2009-01-01

    Stem cells include a diverse number of toti-, pluri-, and multi-potent cells that play important roles in cellular genesis and differentiation, tissue development, and organogenesis. Genetic regulation involving various transcription factors results in the self-renewal and differentiation properties of stem cells. The nuclear receptor (NR) superfamily is composed of 48 ligand-activated transcription factors involved in diverse physiological functions such as metabolism, development, and reproduction. Increasing evidence shows that certain NRs function in regulating stemness or differentiation of embryonic stem (ES) cells and tissue-specific adult stem cells. Here, we review the role of the NR superfamily in various aspects of stem cell biology, including their regulation of stemness, forward- and trans-differentiation events; reprogramming of terminally differentiated cells; and interspecies differences. These studies provide insights into the therapeutic potential of the NR superfamily in stem cell therapy and in treating stem cell-associated diseases (e.g., cancer stem cell). PMID:19696553

  9. Innovation and STEM Schools

    ERIC Educational Resources Information Center

    Roberts, Julia Link

    2015-01-01

    How do schools with a focus on science, technology, engineering, and mathematics (STEM) fit in with state goals to increase innovation and to boost the economy? This article briefly discusses how educators can encourage creativity and innovation.

  10. Hematopoietic Stem Cells Therapies.

    PubMed

    Chivu-Economescu, Mihaela; Rubach, Martin

    2017-01-01

    Stem cell-based therapies are recognized as a new way to treat various diseases and injuries, with a wide range of health benefits. The goal is to heal or replace diseased or destroyed organs or body parts with healthy new cells provided by stem cell transplantation. The current practical form of stem cell therapy is the hematopoietic stem cells transplant applied for the treatment of hematological disorders. There are over 2100 clinical studies in progress concerning hematopoietic stem cell therapies. All of them are using hematopoietic stem cells to treat various diseases like: cancers, leukemia, lymphoma, cardiac failure, neural disorders, auto-immune diseases, immunodeficiency, metabolic or genetic disorders. Several challenges are to be addressed prior to developing and applying large scale cell therapies: 1) to explain and control the mechanisms of differentiation and development toward a specific cell type needed to treat the disease, 2) to obtain a sufficient number of desired cell type for transplantation, 3) to overcome the immune rejection and 4) to show that transplanted cells fulfill their normal functions in vivo after transplants.

  11. Ribosomal DNA sequence divergence and group I introns within the Leucostoma species L. cinctum, L. persoonii, and L. parapersoonii sp. nov., ascomycetes that cause Cytospora canker of fruit trees.

    PubMed

    Adams, Gerard C; Surve-Iyer, Rupa S; Iezzoni, Amy F

    2002-01-01

    Leucostoma species that are the causal agents of Cytospora canker of stone and pome fruit trees were studied in detail. DNA sequence of the internal transcribed spacer regions and the 5.8S of the nuclear ribosomal DNA operon (ITS rDNA) supplied sufficient characters to assess the phylogenetic relationships among species of Leucostoma, Valsa, Valsella, and related anamorphs in Cytospora. Parsimony analysis of the aligned sequence divided Cytospora isolates from fruit trees into clades that generally agreed with the morphological species concepts, and with some of the phenetic groupings (PG 1-6) identified previously by isozyme analysis and cultural characteristics. Phylogenetic analysis inferred that isolates of L. persoonii formed two well-resolved clades distinct from isolates of L. cinctum. Phylogenetic analysis of the ITS rDNA, isozyme analysis, and cultural characteristics supported the inference that L. persoonii groups PG 2 and PG 3 were populations of a new species apparently more genetically different from L. persoonii PG 1 than from isolates representative of L. massariana, L. niveum, L. translucens, and Valsella melastoma. The new species, L. parapersoonii, was described. A diverse collection of isolates of L. cinctum, L. persoonii, and L. parapersoonii were examined for genetic variation using restriction fragment length polymorphism (RFLP) analysis of the ITS rDNA and the five prime end of the large subunit of the rDNA (LSU rDNA). HinfI and HpaII endonucleases were each useful in dividing the Leucostoma isolates into RFLP profiles corresponding to the isozyme phenetic groups, PG 1-6. RFLP analysis was more effective than isozyme analysis in uncovering variation among isolates of L. persoonii PG 1, but less effective within L. cinctum populations. Isolates representative of seven of the L. persoonii formae speciales proposed by G. Défago in 1935 were found to be genetically diverse isolates of PG 1. Two large insertions, 415 and 309 nucleotides long, in

  12. Chemotherapy targeting cancer stem cells

    PubMed Central

    Liu, Haiguang; Lv, Lin; Yang, Kai

    2015-01-01

    Conventional chemotherapy is the main treatment for cancer and benefits patients in the form of decreased relapse and metastasis and longer overall survival. However, as the target therapy drugs and delivery systems are not wholly precise, it also results in quite a few side effects, and is less efficient in many cancers due to the spared cancer stem cells, which are considered the reason for chemotherapy resistance, relapse, and metastasis. Conventional chemotherapy limitations and the cancer stem cell hypothesis inspired our search for a novel chemotherapy targeting cancer stem cells. In this review, we summarize cancer stem cell enrichment methods, the search for new efficient drugs, and the delivery of drugs targeting cancer stem cells. We also discuss cancer stem cell hierarchy complexity and the corresponding combination therapy for both cancer stem and non-stem cells. Learning from cancer stem cells may reveal novel strategies for chemotherapy in the future. PMID:26045975

  13. Valve stem and packing assembly

    DOEpatents

    Wordin, John J.

    1991-01-01

    A valve stem and packing assembly is provided in which a rotatable valve stem includes a first tractrix surface for sliding contact with a stem packing and also includes a second tractrix surface for sliding contact with a bonnet. Force is applied by means of a spring, gland flange, and gland on the stem packing so the stem packing seals to the valve stem and bonnet. This configuration serves to create and maintain a reliable seal between the stem packing and the valve stem. The bonnet includes a second complementary tractrix surface for contacting the second sliding tractrix surface, the combination serving as a journal bearing for the entire valve stem and packing assembly. The journal bearing so configured is known as a Schiele's pivot. The Schiele's pivot also serves to maintain proper alignment of the valve stem with respect to the bonnet. Vertical wear between the surfaces of the Schiele's pivot is uniform at all points of contact between the second sliding tractrix surface and the second complementary tractrix surface of a bonnet. The valve stem is connected to a valve plug by means of a slip joint. The valve is opened and closed by rotating the valve stem. The slip joint compensates for wear on the Schiele's pivot and on the valve plug. A ledge is provided on the valve bonnet for the retaining nut to bear against. The ledge prevents overtightening of the retaining nut and the resulting excessive friction between stem and stem packing.

  14. Valve stem and packing assembly

    DOEpatents

    Wordin, J.J.

    1991-09-03

    A valve stem and packing assembly is provided in which a rotatable valve stem includes a first tractrix surface for sliding contact with a stem packing and also includes a second tractrix surface for sliding contact with a bonnet. Force is applied by means of a spring, gland flange, and gland on the stem packing so the stem packing seals to the valve stem and bonnet. This configuration serves to create and maintain a reliable seal between the stem packing and the valve stem. The bonnet includes a second complementary tractrix surface for contacting the second sliding tractrix surface, the combination serving as a journal bearing for the entire valve stem and packing assembly. The journal bearing so configured is known as a Schiele's pivot. The Schiele's pivot also serves to maintain proper alignment of the valve stem with respect to the bonnet. Vertical wear between the surfaces of the Schiele's pivot is uniform at all points of contact between the second sliding tractrix surface and the second complementary tractrix surface of a bonnet. The valve stem is connected to a valve plug by means of a slip joint. The valve is opened and closed by rotating the valve stem. The slip joint compensates for wear on the Schiele's pivot and on the valve plug. A ledge is provided on the valve bonnet for the retaining nut to bear against. The ledge prevents over tightening of the retaining nut and the resulting excessive friction between stem and stem packing. 2 figures.

  15. Identification of QTLs for early blight ( Alternaria solani) resistance in tomato using backcross populations of a Lycopersicon esculentum x L. hirsutum cross.

    PubMed

    Foolad, R.; Zhang, P.; Khan, A. A.; Niño-Liu, D.; Lin, Y.

    2002-05-01

    Most commercial cultivars of tomato, Lycopersicon esculentum Mill., are susceptible to early blight (EB), a devastating fungal ( Alternaria solani Sorauer) disease of tomato in the northern and eastern parts of the U.S. and elsewhere in the world. The disease causes plant defoliation, which reduces yield and fruit quality, and contributes to significant crop loss. Sources of resistance have been identified within related wild species of tomato. The purpose of this study was to identify and validate quantitative trait loci (QTLs) for EB resistance in backcross populations of a cross between a susceptible tomato breeding line (NC84173; maternal and recurrent parent) and a resistant Lycopersicon hirsutum Humb. and Bonpl. accession (PI126445). Sixteen hundred BC(1) plants were grown to maturity in a field in 1998. Plants that were self-incompatible, indeterminant in growth habit, and/or extremely late in maturity, were discarded in order to eliminate confounding effects of these factors on disease evaluation, QTL mapping, and future breeding research. The remaining 145 plants (referred to as the BC(1) population) were genotyped for 141 restriction fragment length polymorphism (RFLP) markers and 23 resistance gene analogs (RGAs), and a genetic linkage map was constructed. BC(1) plants were evaluated for disease symptoms throughout the season, and the area under the disease progress curve (AUDPC) and the final percent defoliation (disease severity) were determined for each plant. BC(1) plants were self-pollinated and produced BC(1)S(1) seed. The BC(1)S(1) population, consisting of 145 BC(1)S(1) families, was grown and evaluated for disease symptoms in replicated field trials in two subsequent years (1999 and 2000) and AUDPC and/or final percent defoliation were determined for each family in each year. Two QTL mapping approaches, simple interval mapping (SIM) and composite interval mapping (CIM), were used to identify QTLs for EB resistance in the BC(1) and BC(1)S(1

  16. Fifth Annual Stem Cell Summit.

    PubMed

    Knowlton, Daniel

    2010-04-01

    The Fifth Annual Stem Cell Summit, held in New York, included topics covering new commercial developments in the research field of stem cell-based therapies. This conference report highlights selected presentations on embryonic and adult stem cells, stem cell-based therapies for the treatment of orthopedic and cardiovascular indications and inflammatory diseases, as well as technologies for processing and storing stem cells. Investigational therapies discussed include placental expanded (PLX) cells (Pluristem Therapeutics Inc), StemEx (Gamida-Teva Joint Venture/Teva Pharmaceutical Industries Ltd) and remestemcel-L (Osiris Therapeutics Inc/Genzyme Corp/JCR Pharmaceuticals Co Ltd/ Mochida Pharmaceutical Co Ltd).

  17. 7. VIEW OF UPRIGHT OUTLET GATE, WHEEL STEM AND STEM ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    7. VIEW OF UPRIGHT OUTLET GATE, WHEEL STEM AND STEM GUIDE (14' DIAMETER CIRCULAR CALCO CAST IRON SLIDE GATE), LOOKING SOUTHEAST - High Mountain Dams in Bonneville Unit, Fire Lake Dam, Wasatch National Forest, Kamas, Summit County, UT

  18. 5. VIEW OF INCLINED OUTLET GATE WHEEL, STEM AND STEM ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    5. VIEW OF INCLINED OUTLET GATE WHEEL, STEM AND STEM GUIDE, (12' DIAMETER HARDESTY MODEL 112 CIRCULAR GATE), LOOKING NORTHEAST - High Mountain Dams in Bonneville Unit, Island Lake Dam, Wasatch National Forest, Kamas, Summit County, UT

  19. 5. VIEW SHOWING INCLINED OUTLET GATE WHEEL, STEM AND STEM ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    5. VIEW SHOWING INCLINED OUTLET GATE WHEEL, STEM AND STEM GUIDE (28' WIDE HARDESTY CAST IRON SLIDE HEADGATE), LOOKING NORTHEAST - High Mountain Dams in Bonneville Unit, Duck Lake Dam, Wasatch National Forest, Kamas, Summit County, UT

  20. 5. VIEW OF UPRIGHT OUTLET GATE WHEEL, STEM AND STEM ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    5. VIEW OF UPRIGHT OUTLET GATE WHEEL, STEM AND STEM GUIDE, LOOKING NORTHWEST - High Mountain Dams in Upalco Unit, Drift Lake Dam, Ashley National Forest, 11.4 miles Northwest of Swift Creek Campground, Mountain Home, Duchesne County, UT

  1. 6. VIEW SHOWING INCLINED OUTLET GATE WHEEL, STEM AND STEM ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    6. VIEW SHOWING INCLINED OUTLET GATE WHEEL, STEM AND STEM GUIDE (18' HARDESTY GATE), LOOKING SOUTHEAST - High Mountain Dams in Bonneville Unit, Long Lake Dam, Wasatch National Forest, Kamas, Summit County, UT

  2. 5. VIEW OF UPRIGHT OUTLET GATE, STEM, STEM GUIDE AND ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    5. VIEW OF UPRIGHT OUTLET GATE, STEM, STEM GUIDE AND WHEEL (10' HARDESTY CAST IRON VERTICAL LIFT GATE), LOOKING WEST - High Mountain Dams in Bonneville Unit, Weir Lake Dam, Wasatch National Forest, Kamas, Summit County, UT

  3. 7. VIEW OF INCLINED OUTLET GATE WHEEL, STEM AND STEM ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    7. VIEW OF INCLINED OUTLET GATE WHEEL, STEM AND STEM GUIDE (15' HARDESTY MODEL 115 GATE), LOOKING NORTHWEST - High Mountain Dams in Bonneville Unit, Marjorie Lake Dam, Wasatch National Forest, Kamas, Summit County, UT

  4. 4. VIEW OF INCLINED OUTLET GATE, STEM, STEM GUIDE AND ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    4. VIEW OF INCLINED OUTLET GATE, STEM, STEM GUIDE AND WHEEL (10' HARDESTY VERTICAL LIFT GATE), LOOKING NORTHWEST - High Mountain Dams in Bonneville Unit, Pot Lake Dam, Wasatch National Forest, Kamas, Summit County, UT

  5. 5. VIEW OF UPRIGHT OUTLET GATE WHEEL, STEM AND STEM ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    5. VIEW OF UPRIGHT OUTLET GATE WHEEL, STEM AND STEM GUIDE (HARDESTY CAST IRON RECTANGULAR SLIDE GATE), LOOKING SOUTHWEST - High Mountain Dams in Bonneville Unit, Lost Lake Dam, Kamas, Summit County, UT

  6. Dental pulp stem cells

    PubMed Central

    Ashri, Nahid Y.; Ajlan, Sumaiah A.; Aldahmash, Abdullah M.

    2015-01-01

    Inflammatory periodontal disease is a major cause of loss of tooth-supporting structures. Novel approaches for regeneration of periodontal apparatus is an area of intensive research. Periodontal tissue engineering implies the use of appropriate regenerative cells, delivered through a suitable scaffold, and guided through signaling molecules. Dental pulp stem cells have been used in an increasing number of studies in dental tissue engineering. Those cells show mesenchymal (stromal) stem cell-like properties including self-renewal and multilineage differentiation potentials, aside from their relative accessibility and pleasant handling properties. The purpose of this article is to review the biological principles of periodontal tissue engineering, along with the challenges facing the development of a consistent and clinically relevant tissue regeneration platform. This article includes an updated review on dental pulp stem cells and their applications in periodontal regeneration, in combination with different scaffolds and growth factors. PMID:26620980

  7. Reversing breast cancer stem cell into breast somatic stem cell.

    PubMed

    Wijaya, L; Agustina, D; Lizandi, A O; Kartawinata, M M; Sandra, F

    2011-02-01

    Stem cells have an important role in cell biology, allowing tissues to be renewed by freshly created cells throughout their lifetime. The specific micro-environment of stem cells is called stem cell niche; this environment influences the development of stem cells from quiescence through stages of differentiation. Recent advance researches have improved the understanding of the cellular and molecular components of the micro-environment--or niche--that regulates stem cells. We point out an important trend to the study of niche activity in breast cancers. Breast cancer has long been known to conserve a heterogeneous population of cells. While the majority of cells that make up tumors are destined to differentiate and eventually stop dividing, only minority populations of cells, termed cancer stem cell, possess extensive self renewal capability. These cancer stem cells possess characteristics of both stem cells and cancer cells. Breast cancer stem cells reversal to breast somatic stem cells offer a new therapy, that not only can stop the spread of breast cancer cells, but also can differentiate breast cancer stem cells into normal breast somatic stem cells. These can replace damaged breast tissue. Nevertheless, the complexity of realizing this therapy approach needs further research.

  8. Development and validation of an ultra-high-performance liquid chromatography tandem mass spectrometric method for the simultaneous determination of free and conjugated Alternaria toxins in cereal-based foodstuffs.

    PubMed

    Walravens, Jeroen; Mikula, Hannes; Rychlik, Michael; Asam, Stefan; Ediage, Emmanuel Njumbe; Di Mavungu, José Diana; Van Landschoot, Anita; Vanhaecke, Lynn; De Saeger, Sarah

    2014-10-31

    A UPLC-ESI(+/-)-MS/MS method for the simultaneous determination of free (alternariol, alternariol monomethyl ether, altenuene, tenuazonic acid, tentoxin, altertoxin-I) and conjugated (sulfates and glucosides of alternariol and alternariol monomethyl ether) Alternaria toxins in cereals and cereal products (rice, oat flakes and barley) was developed. Optimization of the sample preparation and extraction methodology was achieved through experimental design, using full factorial design for extraction solvent composition optimization and fractional factorial design to identify the critical factors in the sample preparation protocol, which were in turn subjected to optimization. Final extracts were analysed using an Waters Acquity UPLC system coupled to a Quattro Premier XE mass spectrometer equipped with an electrospray interface operated in both positive and negative ionization mode. Chromatographic separation was achieved using an Acquity UPLC HSS T3 column, and the applied gradient elution programme allowed for the simultaneous determination of 10 Alternaria toxins in a one-step chromatographic run with a total run time of only 7min. Subsequently, the method, applying isotopically labelled internal standards ([(2)H4]-alternariol monomethyl ether and [(13)C6,(15)N]-tenuazonic acid), was validated for several parameters such as linearity, apparent recovery, limit of detection, limit of quantification, precision, measurement uncertainty and specificity (in agreement with the criteria mentioned in Commission Regulation No. 401/2006/EC and Commission Decision No. 2002/657/EC). During validation, quality of the bioanalytical data was improved by counteracting the observed heteroscedasticity through the application of weighted least squares linear regression (WLSLR). Finally, 24 commercially available cereal-based foodstuffs were subjected to analysis, revealing the presence of tenuazonic acid in both rice and oat flake samples (

  9. Stem cells and healthy aging.

    PubMed

    Goodell, Margaret A; Rando, Thomas A

    2015-12-04

    Research into stem cells and aging aims to understand how stem cells maintain tissue health, what mechanisms ultimately lead to decline in stem cell function with age, and how the regenerative capacity of somatic stem cells can be enhanced to promote healthy aging. Here, we explore the effects of aging on stem cells in different tissues. Recent research has focused on the ways that genetic mutations, epigenetic changes, and the extrinsic environmental milieu influence stem cell functionality over time. We describe each of these three factors, the ways in which they interact, and how these interactions decrease stem cell health over time. We are optimistic that a better understanding of these changes will uncover potential strategies to enhance stem cell function and increase tissue resiliency into old age.

  10. Multipotent Stem Cell and Reproduction.

    PubMed

    Khanlarkhani, Neda; Baazm, Maryam; Mohammadzadeh, Farzaneh; Najafi, Atefeh; Mehdinejadiani, Shayesteh; Sobhani, Aligholi

    2016-01-01

    Stem cells are self-renewing and undifferentiated cell types that can be differentiate into functional cells. Stem cells can be classified into two main types based on their source of origin: Embryonic and Adult stem cells. Stem cells also classified based on the range of differentiation potentials into Totipotent, Pluripotent, Multipotent, and Unipotent. Multipotent stem cells have the ability to differentiate into all cell types within one particular lineage. There are plentiful advantages and usages for multipotent stem cells. Multipotent Stem cells act as a significant key in procedure of development, tissue repair, and protection. The accessibility and adaptability of these amazing cells create them a great therapeutic choice for different part of medical approaches, and it becomes interesting topic in the scientific researches to found obvious method for the most advantageous use of MSC-based therapies. Recent studies in the field of stem cell biology have provided new perspectives and opportunities for the treatment of infertility disorders.

  11. Stem Cells and Female Reproduction

    PubMed Central

    Du, Hongling; Taylor, Hugh S.

    2011-01-01

    Several recent findings in stem cell biology have resulted in new opportunities for the treatment of reproductive disease. Endometrial regeneration can be driven by bone marrow derived stem cells. This finding has potential implications for the treatment of uterine disorders. It also supports a new theory for the etiology of endometriosis. The ovaries have been shown to contain stem cells that form oocytes in adults and can be cultured in vitro to develop mature oocytes. Stem cells from the fetus have been demonstrated to lead to microchimerism in the mother and implicated in several maternal diseases. Additionally the placenta may be another source of hematopoietic stem cell. Finally endometrial derived stem cells have been demonstrated to differentiate into non-reproductive tissues. While we are just beginning to understand stem cells and many key questions remain, the potential advantages of stem cells in reproductive biology and medicine are apparent. PMID:19208782

  12. Stem Cell Organoid Engineering

    PubMed Central

    Yin, Xiaolei; Mead, Benjamin E.; Safaee, Helia; Langer, Robert; Karp, Jeffrey M.; Levy, Oren

    2016-01-01

    Organoid systems leverage the self-organizing properties of stem cells to create diverse multi-cellular tissue proxies. Most organoid models only represent single or partial components of a tissue, and it is often difficult to control the cell type, organization, and cell-cell/cell-matrix interactions within these systems. Herein, we discuss basic approaches to generate stem cell-based organoids, their advantages and limitations, and how bioengineering strategies can be used to steer the cell composition and their 3D organization within organoids to further enhance their utility in research and therapies. PMID:26748754

  13. Engineering Stem Cell Organoids.

    PubMed

    Yin, Xiaolei; Mead, Benjamin E; Safaee, Helia; Langer, Robert; Karp, Jeffrey M; Levy, Oren

    2016-01-07

    Organoid systems leverage the self-organizing properties of stem cells to create diverse multi-cellular tissue proxies. Most organoid models only represent single or partial components of a tissue, and it is often difficult to control the cell type, organization, and cell-cell/cell-matrix interactions within these systems. Herein, we discuss basic approaches to generate stem cell-based organoids, their advantages and limitations, and how bioengineering strategies can be used to steer the cell composition and their 3D organization within organoids to further enhance their utility in research and therapies.

  14. Cloning of Mammary Stem Cells

    DTIC Science & Technology

    2001-11-01

    these parity-induced cells do represent a totipotent mammary stem cell population per se, but these cells might support stem cell maintenance as... Stem Cells PRINCIPAL INVESTIGATOR: Dr. Kay-Uwe Wagner CONTRACTING ORGANIZATION: University of Nebraska Medical Center Omaha, Nebraska 68198-6810 REPORT...Mammary Stem Cells DAMD17-00-1-0641 6. AUTHOR(S) Dr. Kay-Uwe Wagner 7. PERFORMING ORGANIZATION NAME(S) AND ADDRESS(ES) 8. PERFORMING ORGANIZATION REPORT

  15. Laser biomodulation on stem cells

    NASA Astrophysics Data System (ADS)

    Liu, Timon C.; Duan, Rui; Li, Yan; Li, Xue-Feng; Tan, Li-Ling; Liu, Songhao

    2001-08-01

    Stem cells are views from the perspectives of their function, evolution, development, and cause. Counterintuitively, most stem cells may arise late in development, to act principally in tissue renewal, thus ensuring an organisms long-term survival. Surprisingly, recent reports suggest that tissue-specific adult stem cells have the potential to contribute to replenishment of multiple adult tissues. Stem cells are currently in the news for two reasons: the successful cultivation of human embryonic stem cell lines and reports that adult stem cells can differentiate into developmentally unrelated cell types, such as nerve cells into blood cells. The spotlight on stem cells has revealed gaps in our knowledge that must be filled if we are to take advantage of their full potential for treating devastating degenerative diseases such as Parkinsons's disease and muscular dystrophy. We need to know more about the intrinsic controls that keep stem cells as stem cells or direct them along particular differentiation pathways. Such intrinsic regulators are, in turn, sensitive to the influences of the microenvironment, or niche, where stem cells normally reside. Both intrinsic and extrinsic signals regular stem cell fate and some of these signals have now been identified. Vacek et al and Wang et al have studied the effect of low intensity laser on the haemopoietic stem cells in vitro. There experiments show there is indeed the effect of low intensity laser on the haemopoietic stem cells in vitro, and the present effect is the promotion of haemopoietic stem cells proliferation. In other words, low intensity laser irradiation can act as an extrinsic signal regulating stem cell fate. In this paper, we study how low intensity laser can be used to regulate stem cell fate from the viewpoint of collective phototransduction.

  16. Stem Cell Transplants (For Parents)

    MedlinePlus

    ... Old Feeding Your 1- to 2-Year-Old Stem Cell Transplants KidsHealth > For Parents > Stem Cell Transplants A A A What's in this article? ... Recovery Coping en español Trasplantes de células madre Stem cells are cells in the body that have the ...

  17. Information on Stem Cell Research

    MedlinePlus

    ... Home » Current Research » Focus on Research Focus on Stem Cell Research Stem cells possess the unique ability to differentiate into many ... they also retain the ability to produce more stem cells, a process termed self-renewal. There are multiple ...

  18. STEM--Beyond the Acronym

    ERIC Educational Resources Information Center

    Vasquez, Jo Anne

    2015-01-01

    When most educators think of STEM education, they think of fully integrated projects seamlessly combining all four disciplines--science, technology, engineering, and mathematics. Although such transdisciplinary STEM units are ideal, writes Vasquez, they are not the only way to give students valuable STEM experiences. She gives examples of two…

  19. A Problem with STEM

    ERIC Educational Resources Information Center

    Marder, Michael

    2013-01-01

    Striking differences between physics and biology have important implications for interdisciplinary science, technology, engineering, and mathematics (STEM) education. The author is a physicist with interdisciplinary connections. The research group in which he works, the Center for Nonlinear Dynamics at the University of Texas at Austin, is…

  20. "Excellence" in STEM Education

    ERIC Educational Resources Information Center

    Clark, Aaron C.

    2012-01-01

    So what does it take to achieve excellence in STEM education? That is the title of the author's presentation delivered at International Technology and Engineering Educators Association's (ITEEA's) FTEE "Spirit of Excellence" Breakfast on March 16, 2012, in Long Beach, California. In preparation for this presentation, the author went back and read…

  1. STEM Sense and Nonsense

    ERIC Educational Resources Information Center

    Charette, Robert N.

    2015-01-01

    If you can believe the daily flood of mass media stories, journal articles, and white papers, the United States is facing a STEM worker crisis. Business leaders and politicians warn that the nation is falling hopelessly behind in the global economic race because our students are unprepared for and uninterested in science, technology, engineering,…

  2. STEMMING the Gap

    ERIC Educational Resources Information Center

    Kahler, Jim; Valentine, Nancy

    2011-01-01

    America has a gap when it comes to youth pursuing science and technology careers. In an effort to improve the knowledge and application of science, technology, engineering, and math (STEM), after-school programs can work in conjunction with formal in-school curriculum to improve science education. One organization that actively addresses this…

  3. STEM Comes to Preschool

    ERIC Educational Resources Information Center

    Moomaw, Sally; Davis, Jaumall A.

    2010-01-01

    Math and science and the related technology and engineering are natural pairings. These four disciplines form the acronym STEM (Science, Technology, Engineering, and Math) and can be readily combined into an integrated curriculum for early childhood classrooms. Many educators believe that children learn best when disciplines are interconnected. An…

  4. Advancing Diversity in STEM

    ERIC Educational Resources Information Center

    Hill, Paul L.; Shaw, Rose A.; Taylor, Jan R.; Hallar, Brittan L.

    2011-01-01

    Although progress has been made, greater efforts are needed to promote faculty diversity at the college and university levels, especially in STEM fields. Thus, it is important to elucidate best practices both for increasing awareness of diversity issues pertaining to higher education and for implementing change. This article focuses on the…

  5. Helping STEM Take Root

    ERIC Educational Resources Information Center

    Schachter, Ron

    2011-01-01

    STEM--shorthand for "science, technology, engineering, and mathematics"--has been part of the school improvement discussion for more than a decade, as educational leaders and policy makers have underscored the importance of these areas in an internationally competitive, 21st-century economy. But building and implementing programs that…

  6. Helping STEM Take Root

    ERIC Educational Resources Information Center

    Schachter, Ron

    2011-01-01

    STEM--the catchy shorthand for "science, technology, engineering and mathematics"--has been part of the school improvement discussion for more than a decade, as educational leaders and policy makers have underscored the importance of these areas in preparing students for an internationally competitive, 21st-century economy. But while the…

  7. Stem cell mobilization.

    PubMed

    Cottler-Fox, Michele H; Lapidot, Tsvee; Petit, Isabelle; Kollet, Orit; DiPersio, John F; Link, Dan; Devine, Steven

    2003-01-01

    Successful blood and marrow transplant (BMT), both autologous and allogeneic, requires the infusion of a sufficient number of hematopoietic progenitor/stem cells (HPCs) capable of homing to the marrow cavity and regenerating a full array of hematopoietic cell lineages in a timely fashion. At present, the most commonly used surrogate marker for HPCs is the cell surface marker CD34, identified in the clinical laboratory by flow cytometry. Clinical studies have shown that infusion of at least 2 x 10(6) CD34(+) cells/kg recipient body weight results in reliable engraftment as measured by recovery of adequate neutrophil and platelet counts approximately 14 days after transplant. Recruitment of HPCs from the marrow into the blood is termed mobilization, or, more commonly, stem cell mobilization. In Section I, Dr. Tsvee Lapidot and colleagues review the wide range of factors influencing stem cell mobilization. Our current understanding focuses on chemokines, proteolytic enzymes, adhesion molecules, cytokines and stromal cell-stem cell interactions. On the basis of this understanding, new approaches to mobilization have been designed and are now starting to undergo clinical testing. In Section II, Dr. Michele Cottler-Fox describes factors predicting the ability to mobilize the older patient with myeloma. In addition, clinical approaches to improving collection by individualizing the timing of apheresis and adjusting the volume of blood processed to achieve a desired product are discussed. Key to this process is the daily enumeration of blood CD34(+) cells. Newer methods of enumerating and mobilizing autologous blood HPCs are discussed. In Section III, Dr. John DiPersio and colleagues provide data on clinical results of mobilizing allogeneic donors with G-CSF, GM-CSF and the combination of both as relates to the number and type of cells collected by apheresis. Newer methods of stem cell mobilization as well as the relationship of graft composition on immune reconstitution

  8. Inflammation and cancer stem cells.

    PubMed

    Shigdar, Sarah; Li, Yong; Bhattacharya, Santanu; O'Connor, Michael; Pu, Chunwen; Lin, Jia; Wang, Tao; Xiang, Dongxi; Kong, Lingxue; Wei, Ming Q; Zhu, Yimin; Zhou, Shufeng; Duan, Wei

    2014-04-10

    Cancer stem cells are becoming recognised as being responsible for metastasis and treatment resistance. The complex cellular and molecular network that regulates cancer stem cells and the role that inflammation plays in cancer progression are slowly being elucidated. Cytokines, secreted by tumour associated immune cells, activate the necessary pathways required by cancer stem cells to facilitate cancer stem cells progressing through the epithelial-mesenchymal transition and migrating to distant sites. Once in situ, these cancer stem cells can secrete their own attractants, thus providing an environment whereby these cells can continue to propagate the tumour in a secondary niche.

  9. Stem Cells in Mammalian Gonads.

    PubMed

    Wu, Ji; Ding, Xinbao; Wang, Jian

    Stem cells have great value in clinical application because of their ability to self-renew and their potential to differentiate into many different cell types. Mammalian gonads, including testes for males and ovaries for females, are composed of germline and somatic cells. In male mammals, spermatogonial stem cells maintain spermatogenesis which occurs continuously in adult testis. Likewise, a growing body of evidence demonstrated that female germline stem cells could be found in mammalian ovaries. Meanwhile, prior studies have shown that somatic stem cells exist in both testes and ovaries. In this chapter, we focus on mammalian gonad stem cells and discuss their characteristics as well as differentiation potentials.

  10. Gravitropism in Leafy Dicot Stems

    NASA Technical Reports Server (NTRS)

    Salisbury, F. B.

    1985-01-01

    A polarizing research microscope with rotating stage and associated camera equipment were ordered, and techniques of fixation and preparation of specimens were perfected for studying possible changes in orientation of cellulose microfibrils in cell walls of gravistimulated dicot stems. Acid ethephon solutions or acid without ethephon caused elongation of stem tissues where they were applied; stems bent away from the side of application. Acid solutions applied to the bottom of horizontal stems greatly delayed bending. Research in tissue sensitivity changes during gravitropic bending of soybean hypocotyls while immersed in auxin and in castor bean stems is also reported.

  11. Stem cells and transplant arteriosclerosis.

    PubMed

    Xu, Qingbo

    2008-05-09

    Stem cells can differentiate into a variety of cells to replace dead cells or to repair damaged tissues. Recent evidence indicates that stem cells are involved in the pathogenesis of transplant arteriosclerosis, an alloimmune initiated vascular stenosis that often results in transplant organ failure. Although the pathogenesis of transplant arteriosclerosis is not yet fully understood, recent developments in stem cell research have suggested novel mechanisms of vascular remodeling in allografts. For example, stem cells derived from the recipient may repair damaged endothelial cells of arteries in transplant organs. Further evidence suggests that stem cells or endothelial progenitor cells may be released from both bone marrow and non-bone marrow tissues. Vascular stem cells appear to replenish cells that died in donor vessels. Concomitantly, stem/progenitor cells may also accumulate in the intima, where they differentiate into smooth muscle cells. However, several issues concerning the contribution of stem cells to the pathogenesis of transplant arteriosclerosis are controversial, eg, whether bone marrow-derived stem cells can differentiate into smooth muscle cells that form neointimal lesions of the vessel wall. This review summarizes recent research on the role of stem cells in transplant arteriosclerosis, discusses the mechanisms of stem cell homing and differentiation into mature endothelial and smooth muscle cells, and highlights the controversial issues in the field.

  12. Materials as stem cell regulators

    PubMed Central

    Murphy, William L.; McDevitt, Todd C.; Engler, Adam J.

    2014-01-01

    The stem cell/material interface is a complex, dynamic microenvironment in which the cell and the material cooperatively dictate one another's fate: the cell by remodelling its surroundings, and the material through its inherent properties (such as adhesivity, stiffness, nanostructure or degradability). Stem cells in contact with materials are able to sense their properties, integrate cues via signal propagation and ultimately translate parallel signalling information into cell fate decisions. However, discovering the mechanisms by which stem cells respond to inherent material characteristics is challenging because of the highly complex, multicomponent signalling milieu present in the stem cell environment. In this Review, we discuss recent evidence that shows that inherent material properties may be engineered to dictate stem cell fate decisions, and overview a subset of the operative signal transduction mechanisms that have begun to emerge. Further developments in stem cell engineering and mechanotransduction are poised to have substantial implications for stem cell biology and regenerative medicine. PMID:24845994

  13. Neural stem cells: an overview.

    PubMed

    Parati, E A; Pozzi, S; Ottolina, A; Onofrj, M; Bez, A; Pagano, S F

    2004-01-01

    Multipotent stem cells are present in the majority of mammalian tissues where they are a renewable source of specialized cells. According to the several biological portions from which multipotent stem cells can be derived, they are characterized as a) embryonic stem cells (ESCs) isolated from the pluripotent inner-cell mass of the pre-implantation blastocyste-stage embryo; b) multipotent fetal stem cells (FSCs) from aborted fetuses; and c) adult stem cells (ASCs) localized in small zones of several organs known as "niche" where a subset of tissue cells and extracellular substrates can indefinitely house one or more stem cells and control their self-renewal and progeny production in vivo. ECSs have an high self-renewing capacity, plasticity and pluripotency over the years. Pluripotency is a property that makes a stem cell able to give rise to all cell type found in the embryo and adult animals.

  14. Materials as stem cell regulators

    NASA Astrophysics Data System (ADS)

    Murphy, William L.; McDevitt, Todd C.; Engler, Adam J.

    2014-06-01

    The stem cell/material interface is a complex, dynamic microenvironment in which the cell and the material cooperatively dictate one another's fate: the cell by remodelling its surroundings, and the material through its inherent properties (such as adhesivity, stiffness, nanostructure or degradability). Stem cells in contact with materials are able to sense their properties, integrate cues via signal propagation and ultimately translate parallel signalling information into cell fate decisions. However, discovering the mechanisms by which stem cells respond to inherent material characteristics is challenging because of the highly complex, multicomponent signalling milieu present in the stem cell environment. In this Review, we discuss recent evidence that shows that inherent material properties may be engineered to dictate stem cell fate decisions, and overview a subset of the operative signal transduction mechanisms that have begun to emerge. Further developments in stem cell engineering and mechanotransduction are poised to have substantial implications for stem cell biology and regenerative medicine.

  15. [Stem cell colloquy: conclusion].

    PubMed

    Tubiana, Maurice

    2002-10-01

    The stem cell data presented and discussed during the symposium raise the hope that important medical progress can be made in several fields: neuro-degenerative diseases, those linked to cellular deficit, some aspects of aging linked to cellular degeneration, and the treatment of cancers that may harm normal tissues at risk of being infiltrated by malignant cells. Three main types of stem cells are available. (i) Those present in normal adult tissue: contrary to what was believed, some data suggest that certain adult stem cells have a great plasticity (they can differentiate into cells different from those in tissues from which they were taken) and can proliferate in vitro without losing their properties. Nevertheless, their use faces several obstacles: in ill or elderly subjects, then these cells can be limited in number or not multiply well in vitro. In this case, auto-grafting of the cells cannot be used. They must be sought in another subject, and allo-grafting causes difficult and sometimes insoluble problems of immunological tolerance. (ii) Embryonic stem cells from surplus human embryos, obtained by in vitro fertilisation, which the parents decide not to use: these cells have a great potential for proliferation and differentiation, but can also encounter problems of immunological intolerance. (iii) Cells obtained from cell nuclear transfer in oocytes: these cells are well tolerated, since they are genetically and immunologically identical to those of the host. All types of stem cells can be obtained with them. However, they do present problems. For obtaining them, female oocytes are needed, which could lead to their commercialization. Moreover, the first steps for obtaining these cells are identical to those used in reproductive cloning. It therefore appears that each type of cell raises difficult scientific and practical problems. More research is needed to overcome these obstacles and to determine which type of stem cell constitutes the best solution for

  16. Common stemness regulators of embryonic and cancer stem cells

    PubMed Central

    Hadjimichael, Christiana; Chanoumidou, Konstantina; Papadopoulou, Natalia; Arampatzi, Panagiota; Papamatheakis, Joseph; Kretsovali, Androniki

    2015-01-01

    Pluripotency of embryonic stem cells (ESCs) and induced pluripotent stem cells is regulated by a well characterized gene transcription circuitry. The circuitry is assembled by ESC specific transcription factors, signal transducing molecules and epigenetic regulators. Growing understanding of stem-like cells, albeit of more complex phenotypes, present in tumors (cancer stem cells), provides a common conceptual and research framework for basic and applied stem cell biology. In this review, we highlight current results on biomarkers, gene signatures, signaling pathways and epigenetic regulators that are common in embryonic and cancer stem cells. We discuss their role in determining the cell phenotype and finally, their potential use to design next generation biological and pharmaceutical approaches for regenerative medicine and cancer therapies. PMID:26516408

  17. Global STEM Navigators

    ERIC Educational Resources Information Center

    Dalimonte, Cathy

    2013-01-01

    In the STEM classroom, students can work in collaborative teams to build those essential skills needed for the 21st-century world. In project-based learning (PBL), teams of four to six students are often randomly selected to describe a realistic situation that may occur in today's workplace; this may be done by counting off in fours, fives,…

  18. Isolation and characterization of rhizosphere bacteria for the biocontrol of the damping-off disease of tomatoes in Tunisia.

    PubMed

    Hammami, Inés; Ben Hsouna, Anis; Hamdi, Naceur; Gdoura, Radhouane; Triki, Mohamed Ali

    2013-01-01

    Fluorescent Pseudomonas spp., isolated from tomato and pepper plants rhizosphere soil, was evaluated in vitro as a potential antagonist of fungal pathogens. Pseudomonas strains were tested against the causal agents of tomatoes damping-off (Sclerotinia sclerotiorum), root rot (Fusarium solani), and causal agents of stem canker and leaf blight (Alternaria alternata). For this purpose, dual culture antagonism assays were carried out on 25% tryptic soy agar, King B medium and potato dextrose agar to determine the effect of the strains on mycelial growth of the pathogens. In addition, strains were screened for their ability to produce exoenzymes and siderophores. All the strains significantly inhibited Alternaria alternata, particularly in 25% TSA medium. Antagonistic effect on Sclerotinia sclerotiorum and Fusarium solani was greater on King B medium. Protease was produced by 30% of the strains, but no strain produced cellulase or chitinase. Finally, the selected Pseudomonas strain, Psf5, was evaluated on tomato seedling development and as a potential candidate for controlling tomato damping-off caused by Sclerotinia sclerotiorum, under growth chamber conditions. In vivo studies resulted in significant increases in plant stand as well as in root dry weight. Psf5 was able to establish and survive in tomato plants rhizosphere after 40days following the planting of bacterized seeds.

  19. Stem cells in dentistry--part I: stem cell sources.

    PubMed

    Egusa, Hiroshi; Sonoyama, Wataru; Nishimura, Masahiro; Atsuta, Ikiru; Akiyama, Kentaro

    2012-07-01

    Stem cells can self-renew and produce different cell types, thus providing new strategies to regenerate missing tissues and treat diseases. In the field of dentistry, adult mesenchymal stem/stromal cells (MSCs) have been identified in several oral and maxillofacial tissues, which suggests that the oral tissues are a rich source of stem cells, and oral stem and mucosal cells are expected to provide an ideal source for genetically reprogrammed cells such as induced pluripotent stem (iPS) cells. Furthermore, oral tissues are expected to be not only a source but also a therapeutic target for stem cells, as stem cell and tissue engineering therapies in dentistry continue to attract increasing clinical interest. Part I of this review outlines various types of intra- and extra-oral tissue-derived stem cells with regard to clinical availability and applications in dentistry. Additionally, appropriate sources of stem cells for regenerative dentistry are discussed with regard to differentiation capacity, accessibility and possible immunomodulatory properties.

  20. Measuring stem cell circadian rhythm.

    PubMed

    Hrushesky, William; Rich, Ivan N

    2015-01-01

    Circadian rhythms are biological rhythms that occur within a 24-h time cycle. Sleep is a prime example of a circadian rhythm and with it melatonin production. Stem cell systems also demonstrate circadian rhythms. This is particularly the case for the proliferating cells within the system. In fact, all proliferating cell populations exhibit their own circadian rhythm, which has important implications for disease and the treatment of disease. Stem cell chronobiology is particularly important because the treatment of cancer can be significantly affected by the time of day a drug is administered. This protocol provides a basis for measuring hematopoietic stem cell circadian rhythm for future stem cell chronotherapeutic applications.

  1. Contrasting views on STEM employment

    NASA Astrophysics Data System (ADS)

    Jones, Matthew; Huber, Michael

    2015-07-01

    In reply to “Unemployed and STEM” (Careers, May pp46-47), in which Penny Jackson described how her efforts to find a job in science, technology, engineering or mathematics (STEM) met with little success at first, even though she has a BSc in physics and a Master's in accelerator science. And in reply to “STEM paradox revisited” (Editorial, June p15), on why employers are expressing concerns about STEM shortages at a time when many recent STEM graduates are struggling to find jobs.

  2. STEm Minority Graduate Program

    SciTech Connect

    Nicholas, Kaen E

    2012-09-20

    ABSTRACT The state of science, technology, engineering and math (STEM) education in the United States has seen some unfavorable assessments over the past decade. In early February, 2010 the House of Representatives heard testimony on undergraduate and graduate education. The message from the panel, which included experts from academia, STEM-based industries, and the National Science Foundation (NSF) was dire and required an urgent response. The experts along with the committee's chairperson, U. S. Representative Daniel Lipinski (D-IL) cited that the complexity of Science, Technology, Engineering, and Mathematics applications and coursework and the methodology utilized to teach these subjects are forcing students out of these disciplines. As the National Academies described in its 2007 report Rising Above the Gathering Storm, successful STEM education is not just an academic pursuit it's a necessity for competing in the knowledge-based economy that the United States had a key role in creating. The potential for action is being made available again as the America COMPETES Act of 2007 is up for reauthorization. Its initial focus was on STEM education at the K-12 levels, but efforts at the undergraduate and graduate levels are needed to retain students to fill the jobs left vacant as baby boomers retire. The Educational Advancement Alliance, Inc. (EAA) has for two decades created programs that have not only addressed the issues of ensuring that students are aptly prepared for college but have focused its efforts over the past decade on increasing the number of students who pursue degrees in STEM disciplines. For the EAA, the introduction of the wonders of science begins at the elementary and middle school level via the Learning Lab, a state-of-the-art mobile science laboratory that visits students in grades 4-6 at the various schools throughout Philadelphia and The Math/Tech Academy which meets on Saturdays for students in grades 5-7. For the past two years the EAA has

  3. An Ecological Model of STEM Education: Operationalizing STEM FOR ALL

    ERIC Educational Resources Information Center

    Basham, James D.; Israel, Maya; Maynard, Kathie

    2010-01-01

    This article describes a framework for how to provide more accessible, relevant, and effective instruction in science, technology, engineering, and mathematics (STEM) education to all students. The STEM for All initiative asserts that all students, including those with disabilities and other diverse learning needs, should be included in meaningful…

  4. Learning for STEM Literacy: STEM Literacy for Learning

    ERIC Educational Resources Information Center

    Zollman, Alan

    2012-01-01

    We are in the STEM generation whose comprehensive purpose is to resolve (1) societal needs for new technological and scientific advances; (2) economic needs for national security; and (3) personal needs to become a fulfilled, productive, knowledgeable citizen. STEM specifically refers to science, technology, engineering, and mathematics, but now…

  5. Aging, metabolism and stem cells: Spotlight on muscle stem cells.

    PubMed

    García-Prat, Laura; Muñoz-Cánoves, Pura

    2017-04-15

    All tissues and organs undergo a progressive regenerative decline as they age. This decline has been mainly attributed to loss of stem cell number and/or function, and both stem cell-intrinsic changes and alterations in local niches and/or systemic environment over time are known to contribute to the stem cell aging phenotype. Advancing in the molecular understanding of the deterioration of stem cell cells with aging is key for targeting the specific causes of tissue regenerative dysfunction at advanced stages of life. Here, we revise exciting recent findings on why stem cells age and the consequences on tissue regeneration, with a special focus on regeneration of skeletal muscle. We also highlight newly identified common molecular pathways affecting diverse types of aging stem cells, such as altered proteostasis, metabolism, or senescence entry, and discuss the questions raised by these findings. Finally, we comment on emerging stem cell rejuvenation strategies, principally emanating from studies on muscle stem cells, which will surely burst tissue regeneration research for future benefit of the increasing human aging population.

  6. Stem Cells, Redox Signaling, and Stem Cell Aging

    PubMed Central

    Liang, Raymond

    2014-01-01

    Abstract Significance: Functional stem cell decline has been postulated to result in loss of maintenance of tissue homeostasis leading to organismal decline and diseases of aging. Recent Advances: Recent findings implicate redox metabolism in the control of stem cell pool and stem cell aging. Although reactive oxygen species (ROS) are better known for their damaging properties to DNA, proteins and lipids, recent findings suggest that ROS may also be an integral physiological mediator of cellular signaling in primary cells. Critical Issues: Here we review recent published work on major signaling pathways and transcription factors that are regulated by ROS and mediate ROS regulation of stem cell fate. We will specifically focus on how alterations in this regulation may be implicated in disease and particularly in diseases of stem cell aging. In general, based on the work described here we propose a model in which ROS function as stem cell rheostat. Future Directions: Future work in elucidating how ROS control stem cell cycling, apoptotic machinery, and lineage determination should shed light on mechanisms whereby ROS may control stem cell aging. Antioxid. Redox Signal. 20, 1902–1916. PMID:24383555

  7. STEM TIPS: Supporting the Beginning Secondary STEM Teacher

    ERIC Educational Resources Information Center

    Jones, Griff; Dana, Thomas; LaFramenta, Joanne; Adams, Thomasenia Lott; Arnold, Jason Dean

    2016-01-01

    The STEM TIPS mobile-ready support platform gives institutions or school districts the ability to provide immediate and customized mentoring to teachers through multiple tiers of web-based support and resources. Using the results of a needs assessment, STEM TIPS was created and launched in partnership with 18 Florida school districts. Further…

  8. Advancing the STEM Workforce through STEM-Centric Career Development

    ERIC Educational Resources Information Center

    Feller, Rich

    2011-01-01

    Preparing for the future is not what it used to be. Yet, advising students, preparing lessons, and promoting the value of STEM options remains constant. As a result, technical and engineering educators seek clarity about the future of careers, career development, and ways to promote STEM options. Recently, the ITEEA conference allowed the author…

  9. 6. VIEW SHOWING UPRIGHT OUTLET GATE WHEEL, STEM AND STEM ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    6. VIEW SHOWING UPRIGHT OUTLET GATE WHEEL, STEM AND STEM GUIDE, WITH LOG ACCESS STRUCTURE, LOOKING WEST - High Mountain Dams in Upalco Unit, Bluebell Lake Dam, Ashley National Forest, 11.2 miles Northwest of Swift Creek Campground, Mountain Home, Duchesne County, UT

  10. 5. VIEW SHOWING UPRIGHT OUTLET GATE WHEEL, STEM AND STEM ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    5. VIEW SHOWING UPRIGHT OUTLET GATE WHEEL, STEM AND STEM GUIDE, WITH LOG ACCESS STRUCTURE, LOOKING SOUTHEAST - High Mountain Dams in Upalco Unit, Bluebell Lake Dam, Ashley National Forest, 11.2 miles Northwest of Swift Creek Campground, Mountain Home, Duchesne County, UT

  11. Are STEM High School Students Entering the STEM Pipeline?

    ERIC Educational Resources Information Center

    Franco, M. Suzanne; Patel, Nimisha H.; Lindsey, Jill

    2012-01-01

    This study compared the career skills and interests for students in two STEM schools to national data. Students completed the KUDER skills assessment and career planning online tools. Results were compared across school, grade level, and sex. The results provided evidence that STEM high school students expressed career intents in predominately…

  12. The advantages of hair follicle pluripotent stem cells over embryonic stem cells and induced pluripotent stem cells for regenerative medicine.

    PubMed

    Amoh, Yasuyuki; Katsuoka, Kensei; Hoffman, Robert M

    2010-12-01

    Multipotent adult stem cells have many potential therapeutic applications. Our recent findings suggest that hair follicles are a promising source of easily accessible multipotent stem cells. Stem cells in the hair follicle area express the neural stem cell marker nestin, suggesting that hair-follicle stem cells and neural stem cells have common features. Nestin-expressing hair follicle stem cells can form neurons and other cell types, and thus adult hair follicle stem cells could have important therapeutic applications, particularly for neurologic diseases. Transplanted hair follicle stem cells promote the functional recovery of injured peripheral nerve and spinal cord. Recent findings suggest that direct transplantation of hair-follicle stem cells without culture can promote nerve repair, which makes them potentially clinically practical. Human hair follicle stem cells as well as mouse hair follicle stem cells promote nerve repair and can be applied to test the hypothesis that human hair follicle stem cells can provide a readily available source of neurologically therapeutic stem cells. The use of hair follicle stem cells for nerve regeneration overcomes critical problems of embryonic stem cells or induced pluripotent stem cells in that the hair follicle stem cells are multipotent, readily accessible, non-oncogenic, and are not associated with ethical issues.

  13. Stem Cell Research

    SciTech Connect

    Verfaillie, Catherine

    2009-01-23

    We have identified a population of primitive cells in normal human post-natal bone marrow that can, at the single cell level, differentiate in many ways and also proliferate extensively. These cells can differentiate in vitro into most mesodermal cell types (for example, bone cells, and others), as well as cells into cells of the nervous system. The finding that stem cells exist in post-natal tissues with previously unknown proliferation and differentiation potential opens up the possibility of using them to treat a host of degenerative, traumatic or congenital diseases.

  14. Gastrointestinal stem cell up-to-date.

    PubMed

    Pirvulet, V

    2015-01-01

    Cellular and tissue regeneration in the gastrointestinal tract depends on stem cells with properties of self-renewal, clonogenicity, and multipotency. Progress in stem cell research and the identification of potential gastric, intestinal, colonic stem cells new markers and the signaling pathways provide hope for the use of stem cells in regenerative medicine and treatments for disease. This review provides an overview of the different types of stem cells, focusing on tissue-restricted adult stem cells.

  15. Building STEM Opportunities for All

    ERIC Educational Resources Information Center

    Lynch, Sharon J.; Peters-Burton, Erin; Ford, Michael

    2015-01-01

    In response to a report from the President's Council of Advisors on Science and Technology, President Obama issued a challenge to the U.S. education system to create more than 1,000 new STEM-focused schools, including 200 high schools. Inclusive STEM-focused high schools--which focus their efforts on females, minorities, and students who are…

  16. Engaging Students in STEM Education

    ERIC Educational Resources Information Center

    Kennedy, T. J.; Odell, M. R. L.

    2014-01-01

    With the "flattening" of the global economy in the 21st century, the teaching of Science, Technology, Engineering, and Mathematics (STEM) has taken on new importance as economic competition has become truly global. STEM education has evolved into a meta-discipline, an integrated effort that removes the traditional barriers between these…

  17. Excising the Root from STEM

    ERIC Educational Resources Information Center

    Lock, Roger

    2009-01-01

    There are a number of well-intentioned STEM initiatives, some designed to improve the recruitment and retention of science teachers. Sometimes it appears that the initiators are remote from direct contact with the "grass roots" issues that feed the "stem" on which the blossoms of young enthusiastic recruits to the science teaching profession are…

  18. Deconstruction Geography: A STEM Approach

    ERIC Educational Resources Information Center

    Gehlhar, Adam M.; Duffield, Stacy K.

    2015-01-01

    This article will define the engineering design process used to create an integrated curriculum at STEM Center Middle School, and it features the planning, implementation, and revision of the Deconstruction Geography unit. The Science Technology Engineering and Math (STEM) Center opened in the fall of 2009 as a way to relieve overcrowding at the…

  19. Trouble at the stem end

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Potato tubers have an axial, stem to bud polarity that is apparent at all stages of growth and development. This polarity is most evident in elongate potatoes, but is also found in “round” varieties. These differences between the stem end and other regions of the tuber establish the context for rese...

  20. University Festival Promotes STEM Education

    ERIC Educational Resources Information Center

    Quagliata, Andrew B.

    2015-01-01

    STEM education is argued as an essential ingredient in preparing our children for careers of the future. This study describes a university festival that includes the promotion of STEM-related career interests in young people among its goals. A total of 203 participants between the age of 7 and 17 completed both pre-event and post-event surveys. In…