Science.gov

Sample records for una planta procesadora

  1. Accidentes en plantas nucleares de electricidad y el riesgo de cáncer

    Cancer.gov

    Hoja informativa acerca de los riesgos del cáncer asociados con accidentes en plantas nucleares de electricidad. Incluye información para pacientes con cáncer que viven en una zona que puede haber sido afectada por un accidente en una planta nuclear.

  2. Plantas crucíferas y la prevención del cáncer

    Cancer.gov

    Hoja informativa que resume los resultados de estudios sobre las plantas crucíferas y el cáncer. Incluye una lista de plantas crucíferas y las guías alimentarias recomendadas para el consumo de verduras.

  3. Non-random sharing of Plantae genes.

    PubMed

    Chan, Cheong Xin; Bhattacharya, Debashish

    2011-05-01

    The power of eukaryote genomics relies strongly on taxon sampling. This point was underlined in a recent analysis of red algal genome evolution in which we tested the Plantae hypothesis that posits the monophyly of red, green (including plants) and glaucophyte algae. The inclusion of novel genome data from two mesophilic red algae enabled us to robustly demonstrate the sisterhood of red and green algae in the tree of life. Perhaps more exciting was the finding that >1,800 putative genes in the unicellular red alga Porphyridium cruentum showed evidence of gene-sharing with diverse lineages of eukaryotes and prokaryotes. Here we assessed the correlation between the putative functions of these shared genes and their susceptibility to transfer. It turns out that genes involved in complex interactive networks such as biological regulation and transcription/translation are less susceptible to endosymbiotic or horizontal gene transfer, when compared to genes with metabolic and transporter functions. PMID:21980581

  4. Time Gating of Chloroplast Autofluorescence Allows Clearer Fluorescence Imaging In Planta

    PubMed Central

    Kodama, Yutaka

    2016-01-01

    Chloroplast, an organelle facilitating photosynthesis, exhibits strong autofluorescence, which is an undesired background signal that restricts imaging experiments with exogenous fluorophore in plants. In this study, the autofluorescence was characterized in planta under confocal laser microscopy, and it was found that the time-gated imaging technique completely eliminates the autofluorescence. As a demonstration of the technique, a clearer signal of fluorescent protein-tagged phototropin, a blue-light photoreceptor localized at the chloroplast periphery, was visualized in planta. PMID:27027881

  5. Time Gating of Chloroplast Autofluorescence Allows Clearer Fluorescence Imaging In Planta.

    PubMed

    Kodama, Yutaka

    2016-01-01

    Chloroplast, an organelle facilitating photosynthesis, exhibits strong autofluorescence, which is an undesired background signal that restricts imaging experiments with exogenous fluorophore in plants. In this study, the autofluorescence was characterized in planta under confocal laser microscopy, and it was found that the time-gated imaging technique completely eliminates the autofluorescence. As a demonstration of the technique, a clearer signal of fluorescent protein-tagged phototropin, a blue-light photoreceptor localized at the chloroplast periphery, was visualized in planta. PMID:27027881

  6. Physiology of ex planta nitrogenase activity in Rhizobium japonicum

    SciTech Connect

    Agarwal, A.K.; Keister, D.L.

    1983-05-01

    Thirty-nine wild-type strains of Rhizobium japonicum have been studied for their ability to synthesize nitrogenase ex planta in defined liquid media under microaerobic conditions. Twenty-one produced more than trace amounts of acetylene reduction activity, but only a few of these yielded high activity. The oxygen response curves were similar for most of the nitrogenase-positive strains. The strains derepressible for activity had several phenotypic characteristics different from non-derepressible strains. These included slower growth and lower oxygen consumption under microaerobic conditions and lower extracellular polysaccharide production. Extracellular polysaccharide production during growth on gluconate in every nitrogenase-positive strain assayed was lower under both aerobic and microaerobic conditions than the non-depressible strains. These phenotypic characteristics may be representative of a genotype of a subspecies of R. japonicum. These studies were done in part to enlarge the base number of strains available for studies on the physiology, biochemistry, and genetics of nitrogen fixation. (35 Refs.)

  7. Una visita en Sud America

    NASA Astrophysics Data System (ADS)

    1982-09-01

    Oisfrute de una estadfa en el Hotel La Silla, el mejor hotel de Sud America con su tan unica atmosfera extraterrestre! Los espera su calificado personal de experimentados hoteleros, jefes de cocina, etc., ansiosos todos de satisfacer sus deseos hasta el mas mfnimo detalle. Naturalmente nuestro espacioso restaurant de tres estrellas ofrece un completo surtido de exquisitas comidas y deliciosos tragos (conocedores usualmente eligen "Oelicia Orion" 0 "Centauro Especial"). EI servicio cempleto durante 24 horas incluye nuestra ya mundialmente famosa "Cena de medianoche para los miradores de estrellas", por eso - no olvide: No pierda la oportunidad de una estadfa en EL HOTEL LA SILLA - una experiencia maravillosa!

  8. In vitro and in planta compatibility of insecticides and the endophytic entomopathogen, Lecanicillium lecanii.

    PubMed

    Gurulingappa, Pampapathy; Mc Gee, Peter; Sword, Gregory A

    2011-08-01

    In an attempt to clarify the potential role of endophytic fungi in integrated pest management, the compatibility of an endophytic isolate of Lecanicillium lecanii (Zimmermann) Gams & Zare (Hyphomycetes) with nine insecticides used against Aphis gossypii Glover (Homoptera : Aphididae) was examined both in vitro over 14 days and in planta. In the laboratory, most insecticides partially or completely inhibited the germination of conidia and growth of hyphae in nutrient-rich conditions. Endosulfan completely inhibited the germination of conidia and hyphal growth. In contrast, all insecticides were compatible with L. lecanii in planta, and the fungus was readily recovered from inoculated, colonized leaves. These data support the hypothesis that endophytic L. lecanii will be unaffected by insecticides and could be integrated in the management of pests in cotton. PMID:21424605

  9. In planta mutagenesis determines the functional regions of the wheat puroindoline proteins.

    PubMed

    Feiz, L; Beecher, B S; Martin, J M; Giroux, M J

    2009-11-01

    In planta analysis of protein function in a crop plant could lead to improvements in understanding protein structure/function relationships as well as selective agronomic or end product quality improvements. The requirements for successful in planta analysis are a high mutation rate, an efficient screening method, and a trait with high heritability. Two ideal targets for functional analysis are the Puroindoline a and Puroindoline b (Pina and Pinb, respectively) genes, which together compose the wheat (Triticum aestivum L.) Ha locus that controls grain texture and many wheat end-use properties. Puroindolines (PINs) together impart soft texture, and mutations in either PIN result in hard seed texture. Studies of the PINs' mode of action are limited by low allelic variation. To create new Pin alleles and identify critical function-determining regions, Pin point mutations were created in planta via EMS treatment of a soft wheat. Grain hardness of 46 unique PIN missense alleles was then measured using segregating F(2):F(3) populations. The impact of individual missense alleles upon PIN function, as measured by grain hardness, ranged from neutral (74%) to intermediate to function abolishing. The percentage of function-abolishing mutations among mutations occurring in both PINA and PINB was higher for PINB, indicating that PINB is more critical to overall Ha function. This is contrary to expectations in that PINB is not as well conserved as PINA. All function-abolishing mutations resulted from structure-disrupting mutations or from missense mutations occurring near the Tryptophan-rich region. This study demonstrates the feasibility of in planta functional analysis of wheat proteins and that the Tryptophan-rich region is the most important region of both PINA and PINB. PMID:19752217

  10. Time-weighted average SPME analysis for in planta determination of cVOCs.

    PubMed

    Sheehan, Emily M; Limmer, Matt A; Mayer, Philipp; Karlson, Ulrich Gosewinkel; Burken, Joel G

    2012-03-20

    The potential of phytoscreening for plume delineation at contaminated sites has promoted interest in innovative, sensitive contaminant sampling techniques. Solid-phase microextraction (SPME) methods have been developed, offering quick, undemanding, noninvasive sampling without the use of solvents. In this study, time-weighted average SPME (TWA-SPME) sampling was evaluated for in planta quantification of chlorinated solvents. TWA-SPME was found to have increased sensitivity over headspace and equilibrium SPME sampling. Using a variety of chlorinated solvents and a polydimethylsiloxane/carboxen (PDMS/CAR) SPME fiber, most compounds exhibited near linear or linear uptake over the sampling period. Smaller, less hydrophobic compounds exhibited more nonlinearity than larger, more hydrophobic molecules. Using a specifically designed in planta sampler, field sampling was conducted at a site contaminated with chlorinated solvents. Sampling with TWA-SPME produced instrument responses ranging from 5 to over 200 times higher than headspace tree core sampling. This work demonstrates that TWA-SPME can be used for in planta detection of a broad range of chlorinated solvents and methods can likely be applied to other volatile and semivolatile organic compounds. PMID:22332592

  11. Screening for in planta protein-protein interactions combining bimolecular fluorescence complementation with flow cytometry

    PubMed Central

    2012-01-01

    Understanding protein and gene function requires identifying interaction partners using biochemical, molecular or genetic tools. In plants, searching for novel protein-protein interactions is limited to protein purification assays, heterologous in vivo systems such as the yeast-two-hybrid or mutant screens. Ideally one would be able to search for novel protein partners in living plant cells. We demonstrate that it is possible to screen for novel protein-protein interactions from a random library in protoplasted Arabidopsis plant cells and recover some of the interacting partners. Our screen is based on capturing the bi-molecular complementation of mYFP between an YN-bait fusion partner and a completely random prey YC-cDNA library with FACS. The candidate interactions were confirmed using in planta BiFC assays and in planta FRET-FLIM assays. From this work, we show that the well characterized protein Calcium Dependent Protein Kinase 3 (CPK3) interacts with APX3, HMGB5, ORP2A and a ricin B-related lectin domain containing protein At2g39050. This is one of the first randomin planta screens to be successfully employed. PMID:22789293

  12. In planta anthocyanin degradation by a vacuolar class III peroxidase in Brunfelsia calycina flowers.

    PubMed

    Zipor, Gadi; Duarte, Patrícia; Carqueijeiro, Inês; Shahar, Liat; Ovadia, Rinat; Teper-Bamnolker, Paula; Eshel, Dani; Levin, Yishai; Doron-Faigenboim, Adi; Sottomayor, Mariana; Oren-Shamir, Michal

    2015-01-01

    In contrast to detailed knowledge regarding the biosynthesis of anthocyanins, the largest group of plant pigments, little is known about their in planta degradation. It has been suggested that anthocyanin degradation is enzymatically controlled and induced when beneficial to the plant. Here we investigated the enzymatic process in Brunfelsia calycina flowers, as they changed color from purple to white. We characterized the enzymatic process by which B. calycina protein extracts degrade anthocyanins. A candidate peroxidase was partially purified and characterized and its intracellular localization was determined. The transcript sequence of this peroxidase was fully identified. A basic peroxidase, BcPrx01, is responsible for the in planta degradation of anthocyanins in B. calycina flowers. BcPrx01 has the ability to degrade complex anthocyanins, it co-localizes with these pigments in the vacuoles of petals, and both the mRNA and protein levels of BcPrx01 are greatly induced parallel to the degradation of anthocyanins. Both isoelectric focusing (IEF) gel analysis and 3D structure prediction indicated that BcPrx01 is cationic. Identification of BcPrx01 is a significant breakthrough both in the understanding of anthocyanin catabolism in plants and in the field of peroxidases, where such a consistent relationship between expression levels, in planta subcellular localization and activity has seldom been demonstrated. PMID:25256351

  13. i Planta

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This application was designed for farmers, agriculture researchers, and gardeners. The application includes a calculator, a data logger, and a GPS mapping function. It was designed for vegetable farming but can be used for open field applications. The calculator solves for the amount of pesticide,...

  14. Comparative analysis of the in vitro and in planta secretomes from Mycosphaerella fijiensis isolates.

    PubMed

    Escobar-Tovar, Lina; Guzmán-Quesada, Mauricio; Sandoval-Fernández, Jorge A; Gómez-Lim, Miguel A

    2015-06-01

    Black Sigatoka, a devastating disease of bananas and plantains worldwide, is caused by the fungus Mycosphaerella fijiensis. Several banana cultivars such as 'Yangambi Km 5' and Calcutta IV, have been known to be resistant to the fungus, but the resistance has been broken in 'Yangambi Km 5' in Costa Rica. Since the resistance of this variety still persists in Mexico, the aim of this study was to compare the in vitro and in planta secretomes from two avirulent and virulent M. fijiensis isolates using proteomics and bioinformatics approaches. We aimed to identify differentially expressed proteins in fungal isolates that differ in pathogenicity and that might be responsible for breaking the resistance in 'Yangambi Km 5'. We were able to identify 90 protein spots in the secretomes of fungal isolates encoding 42 unique proteins and 35 differential spots between them. Proteins involved in carbohydrate transport and metabolism were more prevalent. Several proteases, pathogenicity-related, ROS detoxification and unknown proteins were also highly or specifically expressed by the virulent isolate in vitro or during in planta infection. An unknown protein representing a virulence factor candidate was also identified. These results demonstrated that the secretome reflects major differences between both M. fijiensis isolates. PMID:25986542

  15. Simultaneous specific in planta visualization of root-colonizing fungi using fluorescence in situ hybridization (FISH).

    PubMed

    Vági, Pál; Knapp, Dániel G; Kósa, Annamária; Seress, Diána; Horváth, Áron N; Kovács, Gábor M

    2014-05-01

    In planta detection of mutualistic, endophytic, and pathogenic fungi commonly colonizing roots and other plant organs is not a routine task. We aimed to use fluorescence in situ hybridization (FISH) for simultaneous specific detection of different fungi colonizing the same tissue. We have adapted ribosomal RNA (rRNA) FISH for visualization of common mycorrhizal (arbuscular- and ectomycorrhiza) and endophytic fungi within roots of different plant species. Beside general probes, we designed and used specific ones hybridizing to the large subunit of rRNA with fluorescent dyes chosen to avoid or reduce the interference with the autofluorescence of plant tissues. We report here an optimized efficient protocol of rRNA FISH and the use of both epifluorescence and confocal laser scanning microscopy for simultaneous specific differential detection of those fungi colonizing the same root. The method could be applied for the characterization of other plant-fungal interactions, too. In planta FISH with specific probes labeled with appropriate fluorescent dyes could be used not only in basic research but to detect plant colonizing pathogenic fungi in their latent life-period. PMID:24221902

  16. Dynamic root exudation of sorgoleone and its in planta mechanism of action

    PubMed Central

    Dayan, Franck E.; Howell, J'Lynn; Weidenhamer, Jeffrey D.

    2009-01-01

    The oily droplets exuded from the root hairs of sorghum are composed of a 1:1 ratio of sorgoleone and its lipid resorcinol analogue. The production of these droplets appears to be suppressed when c. 20 μg of exudate mg−1 root dry weight accumulates at the tip of the root hairs. However, more exudate is produced following gentle washing of the roots with water, suggesting that the biosynthesis of lipid benzoquinones and resorcinols is a dynamic process. Sorgoleone interferes with several molecular target sites, including photosynthetic electron transport, in in vitro assays. However, the in planta mechanism of action of sorgoleone remains controversial because it is not clear whether this lipid benzoquinone exuding from the roots of sorghum is taken up by roots of the receiving plants and translocated to their foliage where it must enter the chloroplast and inhibit PSII in the thylakoid membrane. Experiments designed to test the in planta mode of action of sorgoleone demonstrated that it has no effect on the photosynthesis of older plants, but inhibits photosynthesis in germinating seedlings. Sorgoleone is not translocated acropetally in older plants, but can be absorbed through the hypocotyl and cotyledonary tissues. Therefore, the mode of action of sorgoleone may be the result of inhibition of photosynthesis in young seedlings in concert with inhibition of its other molecular target sites in older plants. PMID:19357432

  17. Bcmimp1, a Botrytis cinerea Gene Transiently Expressed in planta, Encodes a Mitochondrial Protein

    PubMed Central

    Benito-Pescador, David; Santander, Daniela; Arranz, M.; Díaz-Mínguez, José M.; Eslava, Arturo P.; van Kan, Jan A. L.; Benito, Ernesto P.

    2016-01-01

    Botrytis cinerea is a widespread necrotrophic fungus which infects more than 200 plant species. In an attempt to characterize the physiological status of the fungus in planta and to identify genetic factors contributing to its ability to infect the host cells, a differential gene expression analysis during the interaction B. cinerea-tomato was carried out. Gene Bcmimp1 codes for a mRNA detected by differential display in the course of this analysis. During the interaction with the host, it shows a transient expression pattern with maximal expression levels during the colonization and maceration of the infected tissues. Bioinformatic analysis suggested that BCMIMP1 is an integral membrane protein located in the mitochondrial inner membrane. Co-localization experiments with a BCMIMP1-GFP fusion protein confirmed that the protein is targeted to the mitochondria. ΔBcmimp1 mutants do not show obvious phenotypic differences during saprophytic growth and their infection ability was unaltered as compared to the wild-type. Interestingly, the mutants produced increased levels of reactive oxygen species, likely as a consequence of disturbed mitochondrial function. Although Bcmimp1 expression is enhanced in planta it cannot be considered a pathogenicity factor. PMID:26952144

  18. In-planta Sporulation Capacity Enhances Infectivity and Rhizospheric Competitiveness of Frankia Strains.

    PubMed

    Cotin-Galvan, Laetitia; Pozzi, Adrien C; Schwob, Guillaume; Fournier, Pascale; Fernandez, Maria P; Herrera-Belaroussi, Aude

    2016-01-01

    Frankia Sp+ strains maintain their ability to sporulate in symbiosis with actinorhizal plants, producing abundant sporangia inside host plant cells, in contrast to Sp- strains, which are unable to perform in-planta sporulation. We herein examined the role of in-planta sporulation in Frankia infectivity and competitiveness for root infection. Fifteen strains belonging to different Sp+ and Sp- phylogenetic lineages were inoculated on seedlings of Alnus glutinosa (Ag) and A. incana (Ai). Strain competitiveness was investigated by performing Sp-/Sp+ co-inoculations. Plant inoculations were standardized using crushed nodules obtained under laboratory-controlled conditions (same plant species, age, and environmental factors). Specific oligonucleotide primers were developed to identify Frankia Sp+ and/or Sp- strains in the resulting nodules. Single inoculation experiments showed that (i) infectivity by Sp+ strains was significantly greater than that by Sp- strains, (ii) genetically divergent Sp+ strains exhibited different infective abilities, and (iii) Sp+ and Sp- strains showed different host preferences according to the origin (host species) of the inocula. Co-inoculations of Sp+ and Sp- strains revealed the greater competitiveness of Sp+ strains (98.3 to 100% of Sp+ nodules, with up to 15.6% nodules containing both Sp+ and Sp- strains). The results of the present study highlight differences in Sp+/Sp- strain ecological behaviors and provide new insights to strengthen the obligate symbiont hypothesis for Sp+ strains. PMID:26726131

  19. In-planta Sporulation Capacity Enhances Infectivity and Rhizospheric Competitiveness of Frankia Strains

    PubMed Central

    Cotin-Galvan, Laetitia; Pozzi, Adrien C.; Schwob, Guillaume; Fournier, Pascale; Fernandez, Maria P.; Herrera-Belaroussi, Aude

    2016-01-01

    Frankia Sp+ strains maintain their ability to sporulate in symbiosis with actinorhizal plants, producing abundant sporangia inside host plant cells, in contrast to Sp− strains, which are unable to perform in-planta sporulation. We herein examined the role of in-planta sporulation in Frankia infectivity and competitiveness for root infection. Fifteen strains belonging to different Sp+ and Sp− phylogenetic lineages were inoculated on seedlings of Alnus glutinosa (Ag) and A. incana (Ai). Strain competitiveness was investigated by performing Sp−/Sp+ co-inoculations. Plant inoculations were standardized using crushed nodules obtained under laboratory-controlled conditions (same plant species, age, and environmental factors). Specific oligonucleotide primers were developed to identify Frankia Sp+ and/or Sp− strains in the resulting nodules. Single inoculation experiments showed that (i) infectivity by Sp+ strains was significantly greater than that by Sp− strains, (ii) genetically divergent Sp+ strains exhibited different infective abilities, and (iii) Sp+ and Sp− strains showed different host preferences according to the origin (host species) of the inocula. Co-inoculations of Sp+ and Sp− strains revealed the greater competitiveness of Sp+ strains (98.3 to 100% of Sp+ nodules, with up to 15.6% nodules containing both Sp+ and Sp− strains). The results of the present study highlight differences in Sp+/Sp− strain ecological behaviors and provide new insights to strengthen the obligate symbiont hypothesis for Sp+ strains. PMID:26726131

  20. A method for obtaining RNA from Hemileia vastatrix appressoria produced in planta, suitable for transcriptomic analyses.

    PubMed

    Loureiro, Andreia; Gil Azinheira, Helena; do Céu Silva, Maria; Talhinhas, Pedro

    2015-11-01

    Appressoria are the first infection structures developed by rust fungi and require specific topographic signals from the host for their differentiation. The ease in obtaining appressoria in vitro for these biotrophic fungi led to studies concerning gene expression and gene discovery at appressorial level, avoiding the need to distinguish plant and fungal transcripts. However, in some pathosystems, it was observed that gene expression in appressoria seems to be influenced by host-derived signals, suggesting that transcriptomic analyses performed from in planta differentiated appressoria would be potentially more informative than those from in vitro differentiated appressoria. Nevertheless analysing appressorial RNA obtained from in planta samples is often hampered by an excessive dilution of fungal RNA within plant RNA, besides uncertainty regarding the fungal or plant origin of RNA from highly conserved genes. To circumvent these difficulties, we have recovered Hemileia vastatrix appressoria from Arabica coffee leaf surface using a film of nitrocellulose dissolved in butyl and ethyl acetates (nail polish), and extracted fungal RNA from the polish peel. RNA thus obtained is of good quality and usable for cDNA synthesis and transcriptomic (quantitative PCR) studies. This method could provide the means to investigate specific host-induced appressoria-related fungal pathogenicity factors. PMID:26466882

  1. Mexican papita viroid and tomato planta macho viroid belong to a single species in the genus Pospiviroid

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Tomato planta macho viroid (TPMVd) and Mexican papita viroid (MPVd) are two closely-related (>90% sequence identity) members of the genus Pospiviroid. Their current status as separate species is based upon the reported ability of TPMVd to replicate in Gomphrena globosa and the inability of this viro...

  2. An efficient method for visualization and growth of fluorescent Xanthomonas oryzae pv. oryzae in planta

    PubMed Central

    Han, Sang-Wook; Park, Chang-Jin; Lee, Sang-Won; Ronald, Pamela C

    2008-01-01

    Background Xanthomonas oryzae pv. oryzae, the causal agent of bacterial blight disease, is a serious pathogen of rice. Here we describe a fluorescent marker system to study virulence and pathogenicity of X. oryzae pv. oryzae. Results A fluorescent X. oryzae pv. oryzae Philippine race 6 strain expressing green fluorescent protein (GFP) (PXO99GFP) was generated using the gfp gene under the control of the neomycin promoter in the vector, pPneo-gfp. The PXO99GFPstrain displayed identical virulence and avirulence properties as the wild type control strain, PXO99. Using fluorescent microscopy, bacterial multiplication and colonization were directly observed in rice xylem vessels. Accurate and rapid determination of bacterial growth was assessed using fluoremetry and an Enzyme-Linked ImmunoSorbant Assay (ELISA). Conclusion Our results indicate that the fluorescent marker system is useful for assessing bacterial infection and monitoring bacterial multiplication in planta. PMID:18826644

  3. In Planta Response of Arabidopsis to Photothermal Impact Mediated by Gold Nanoparticles.

    PubMed

    Koo, Yeonjong; Lukianova-Hleb, Ekaterina Y; Pan, Joann; Thompson, Sean M; Lapotko, Dmitri O; Braam, Janet

    2016-02-01

    Biological responses to photothermal effects of gold nanoparticles (GNPs) have been demonstrated and employed for various applications in diverse systems except for one important class - plants. Here, the uptake of GNPs through Arabidopsis thaliana roots and translocation to leaves are reported. Successful plasmonic nanobubble generation and acoustic signal detection in planta is demonstrated. Furthermore, Arabidopsis leaves harboring GNPs and exposed to continuous laser or noncoherent light show elevated temperatures across the leaf surface and induced expression of heat-shock regulated genes. Overall, these results demonstrate that Arabidopsis can readily take up GNPs through the roots and translocate the particles to leaf tissues. Once within leaves, GNPs can act as photothermal agents for on-demand remote activation of localized biological processes in plants. PMID:26662357

  4. In Planta Microsphere-Based Lateral Flow Leaf Biosensor in Maize.

    PubMed

    Wen, Jessica T; Castro, Carlos; Tsutsui, Hideaki

    2015-08-01

    Low-cost and quick detection of biotic stresses is critically important for protection of staple food crops such as maize in smallholder farms in developing countries, where access to improved seed varieties, fertilizers, and pesticides is limited due to financial and geographical reasons. Here, we report a new lateral flow detection technology directly integrated in a maize leaf, in which microspheres conjugated with analyte-specific capture antibodies are non-invasively injected. The antibody-conjugated microspheres capture and detect an analyte in a concentration-specific manner. In this study, we optimized microsphere size for effective infiltration and immobilization in the leaf, and further demonstrated detection of a fluorescent mock biomarker, fluorescein, in a live maize plant. This in planta lateral flow biosensor is the first of its kind and is expected to provide a low-cost and user-friendly detection method for biotic stresses in the field. PMID:25271045

  5. In planta transformation method for T-DNA transfer in orchids

    NASA Astrophysics Data System (ADS)

    Semiarti, Endang; Purwantoro, Aziz; Mercuriani, Ixora S.; Anggriasari, Anida M.; Jang, Seonghoe; Suhandono, Sony; Machida, Yasunori; Machida, Chiyoko

    2014-03-01

    Transgenic plant technology is an efficient tool to study the function of gene(s) in plant. The most popular and widely used technique is Agrobacterium-mediated transformation in which cocultivation was done by immersing the plant tissues/organ in overnight bacterial cultured for about 30 minutes to one hour under in vitro condition. In this experiment, we developed more easier technique that omitted the in vitro step during cocultivation with Agrobacterium, namely in planta transformation method. Pollinaria (compact pollen mass of orchid) of Phalaenopsis amabilis and Spathoglottis plicata orchids were used as target explants that were immersed into bacterial culture for 30 minutes, then dried up the pollinaria, the transformed pollinaria was used to pollinate orchid flowers. The T-DNA used for this experiments were Ubipro∷PaFT/A. tumefaciens GV3101 for P. amabilis and MeEF1α2 pro∷GUS/ A. tumefaciens LBA 4404 for S.plicata. Seeds that were produced from pollinated flowers were grown onto 10 mg/l hygromicin containing NP (New Phalaenopsis) medium. The existance of transgene in putative transformant protocorm (developing orchid embryo) genome was confirmed using PCR with specific primers of either PaFT or GUS genes. Histochemical GUS assay was also performed to the putative transformants. The result showed that transformation frequencies were 2.1 % in P. amabilis, and 0,53% in S. plicata. These results indicates that in planta transformation method could be used for Agrobacterium-mediated genetic transformation, with advantage easier and more secure work from contaminants than that of the in vitro method.

  6. Analysis of in planta Expressed Orphan Genes in the Rice Blast Fungus Magnaporthe oryzae

    PubMed Central

    Sadat, Abu; Jeon, Junhyun; Mir, Albely Afifa; Kim, Seongbeom; Choi, Jaeyoung; Lee, Yong-Hwan

    2014-01-01

    Genomes contain a large number of unique genes which have not been found in other species. Although the origin of such “orphan” genes remains unclear, they are thought to be involved in species-specific adaptive processes. Here, we analyzed seven orphan genes (MoSPC1 to MoSPC7) prioritized based on in planta expressed sequence tag data in the rice blast fungus, Magnaporthe oryzae. Expression analysis using qRT-PCR confirmed the expression of four genes (MoSPC1, MoSPC2, MoSPC3 and MoSPC7) during plant infection. However, individual deletion mutants of these four genes did not differ from the wild-type strain for all phenotypes examined, including pathogenicity. The length, GC contents, codon adaptation index and expression during mycelial growth of the four genes suggest that these genes formed during the evolutionary history of M. oryzae. Synteny analyses using closely related fungal species corroborated the notion that these genes evolved de novo in the M. oryzae genome. In this report, we discuss our inability to detect phenotypic changes in the four deletion mutants. Based on these results, the four orphan genes may be products of de novo gene birth processes, and their adaptive potential is in the course of being tested for retention or extinction through natural selection. PMID:25506301

  7. Cytorhabdovirus P protein suppresses RISC-mediated cleavage and RNA silencing amplification in planta.

    PubMed

    Mann, Krin S; Johnson, Karyn N; Carroll, Bernard J; Dietzgen, Ralf G

    2016-03-01

    Plant viruses have evolved to undermine the RNA silencing pathway by expressing suppressor protein(s) that interfere with one or more key components of this antiviral defense. Here we show that the recently identified RNA silencing suppressor (RSS) of lettuce necrotic yellows virus (LNYV), phosphoprotein P, binds to RNA silencing machinery proteins AGO1, AGO2, AGO4, RDR6 and SGS3 in protein-protein interaction assays when transiently expressed. In planta, we demonstrate that LNYV P inhibits miRNA-guided AGO1 cleavage and translational repression, and RDR6/SGS3-dependent amplification of silencing. Analysis of LNYV P deletion mutants identified a C-terminal protein domain essential for both local RNA silencing suppression and interaction with AGO1, AGO2, AGO4, RDR6 and SGS3. In contrast to other viral RSS known to disrupt AGO activity, LNYV P sequence does not contain any recognizable GW/WG or F-box motifs. This suggests that LNYV P may represent a new class of AGO binding proteins. PMID:26808923

  8. Detection and visualization of an exopolysaccharide produced by Xylella fastidiosa in vitro and in planta.

    PubMed

    Roper, M Caroline; Greve, L Carl; Labavitch, John M; Kirkpatrick, Bruce C

    2007-11-01

    Many phytopathogenic bacteria, such as Ralstonia solanacearum, Pantoea stewartii, and Xanthomonas campestris, produce exopolysaccharides (EPSs) that aid in virulence, colonization, and survival. EPS can also contribute to host xylem vessel blockage. The genome of Xylella fastidiosa, the causal agent of Pierce's disease (PD) of grapevine, contains an operon that is strikingly similar to the X. campestris gum operon, which is responsible for the production of xanthan gum. Based on this information, it has been hypothesized that X. fastidiosa is capable of producing an EPS similar in structure and composition to xanthan gum but lacking the terminal mannose residue. In this study, we raised polyclonal antibodies against a modified xanthan gum polymer similar to the predicted X. fastidiosa EPS polymer. We used enzyme-linked immunosorbent assay to quantify production of EPS from X. fastidiosa cells grown in vitro and immunolocalization microscopy to examine the distribution of X. fastidiosa EPS in biofilms formed in vitro and in planta and assessed the contribution of X. fastidiosa EPS to the vascular occlusions seen in PD-infected grapevines. PMID:17827325

  9. Cloning of immunodominant membrane protein genes of phytoplasmas and their in planta expression.

    PubMed

    Kakizawa, Shigeyuki; Oshima, Kenro; Ishii, Yoshiko; Hoshi, Ayaka; Maejima, Kensaku; Jung, Hee-Young; Yamaji, Yasuyuki; Namba, Shigetou

    2009-04-01

    Phytoplasmas are plant pathogenic bacteria that cause devastating yield losses in diverse crops worldwide. Although the understanding of the pathogen biology is important in agriculture, the inability to culture phytoplasmas has hindered their full characterization. Previous studies demonstrated that immunodominant membrane proteins could be classified into three types, immunodominant membrane protein (Imp), immunodominant membrane protein A (IdpA), and antigenic membrane protein (Amp), and they are nonhomologous to each other. Here, cloning and sequencing of imp-containing genomic fragments were performed for several groups of phytoplasma including the aster yellows and rice yellow dwarf groups, for which an imp sequence has not previously been reported. Sequence comparison analysis revealed that Imps are highly variable among phytoplasmas, and clear positive selection was observed in several Imps, suggesting that Imp has important roles in host-phytoplasma interactions. As onion yellows (OY) phytoplasma was known to have Amp as the immunodominant membrane protein, the protein accumulation level of Imp in planta was measured compared with that of Amp. The resulting accumulation of Imp was calculated as approximately one-tenth that of Amp, being consistent with the immunodominant property of Amp in OY. It is suggested that an ancestral type of immunodominant membrane protein could be Imp, and subsequently the expression level of Amp or IdpA is increased in several phytoplasma groups. PMID:19222574

  10. Development of 4-methoxy-7-nitroindolinyl (MNI)-caged auxins which are extremely stable in planta.

    PubMed

    Hayashi, Ken-Ichiro; Kusaka, Naoyuki; Yamasaki, Soma; Zhao, Yunde; Nozaki, Hiroshi

    2015-10-15

    Phytohormone auxin is a master regulator in plant growth and development. Regulation of cellular auxin level plays a central role in plant development. Auxin polar transport system modulates an auxin gradient that determines plant developmental process in response to environmental conditions and developmental programs. Photolabile caged auxins allow optical control of artificial auxin gradients at cellular resolution. Especially, two-photon uncaging system achieves high spatiotemporal control of photolysis reaction at two-photon cross-section. However, the development of caged versions of auxin has been limited by the instability of the caged auxins to higher plant metabolic activities. Here, we describe the synthesis and application of highly stable caged auxins, 4-methoxy-7-nitroindolinyl (MNI)-caged auxins. Natural auxin, indole 3-acetic acid, and two synthetic auxins, 1-NAA and 2,4-D were caged by MNI caging group. MNI-caged auxins showed a high stability in planta and a rapid release the original auxin when photolyzed. We demonstrated that optical control of auxin-responsive gene expression and auxin-related physiological responses by using MNI-caged auxins. We anticipate that MNI-caged auxins will be an effective tool for high-resolution control of endogenous auxin level. PMID:26364943

  11. Development of 4-methoxy-7-nitroindolinyl (MNI)-caged auxins which are extremely stable in planta

    PubMed Central

    Hayashi, Ken-ichiro; Kusaka, Naoyuki; Yamasaki, Soma; Zhao, Yunde; Nozaki, Hiroshi

    2015-01-01

    Phytohormone auxin is a master regulator in plant growth and development. Regulation of cellular auxin level plays a central role in plant development. Auxin polar transport system modulates an auxin gradient that determines plant developmental process in response to environmental conditions and developmental programs. Photolabile caged auxins allow optical control of artificial auxin gradients at cellular resolution. Especially, two-photon uncaging system achieves high spatiotemporal control of photolysis reaction at two-photon cross-section. However, the development of caged versions of auxin has been limited by the instability of the caged auxins to higher plant metabolic activities. Here, we describe the synthesis and application of highly stable caged auxins, 4-methoxy-7-nitroindolinyl (MNI)-caged auxins. Natural auxin, indole 3-acetic acid, and two synthetic auxins, 1-NAA and 2,4-D were caged by MNI caging group. MNI-caged auxins showed a high stability in planta and a rapid release the original auxin when photolyzed. We demonstrated that optical control of auxin-responsive gene expression and auxin-related physiological responses by using MNI-caged auxins. We anticipate that MNI-caged auxins will be an effective tool for high-resolution control of endogenous auxin level. PMID:26364943

  12. TYLCV-Is movement in planta does not require V2 protein

    SciTech Connect

    Hak, Hagit; Levy, Yael; Chandran, Sam A.; Belausov, Eduard; Loyter, Abraham; Lapidot, Moshe; Gafni, Yedidya

    2015-03-15

    Tomato yellow leaf curl virus (TYLCV), a major tomato pathogen causing extensive crop losses, is a whitefly-transmitted geminivirus. V2 mutants of TYLCV-Is and related viruses tend to induce symptomless infection with attenuated viral DNA levels, while accumulating close to wild-type DNA levels in protoplasts, suggesting V2 as a movement protein. The discovery of plant-silencing mechanisms and viral silencing suppressors, V2 included, led us to reconsider V2's involvement in viral movement. We studied two mutant versions of the virus, one impaired in V2 silencing-suppression activity, and another carrying a non-translatable V2. While both mutant viruses spread in the infected plant to newly emerged leaves at the same rate as the wild-type virus, their DNA-accumulation levels were tenfold lower than in the wild-type virus. Thus, we suggest that the setback in virus proliferation, previously ascribed to a movement impediment, is due to lack of silencing-suppression activity. - Highlights: • TYLCV-Is V2 protein is localized in distinct microbodies throughout the cell cytoplasm, around the nucleus and in association with cytoplasmic strands but is not associated with the plasmodesmata. • Disruption of RNA-silencing suppression activity of TYLCV-Is V2 protein causes low titer of the virus in the infected plants. • The movement of TYLCV-Is in planta does not require a functional V2 protein.

  13. Advances in alfalfa mosaic virus-mediated expression of anthrax antigen in planta

    SciTech Connect

    Brodzik, R.; Bandurska, K.; Deka, D.; Golovkin, M.; Koprowski, H. . E-mail: h_koprowski@jefferson.edu

    2005-12-16

    Plant viruses show great potential for production of pharmaceuticals in plants. Such viruses can harbor a small antigenic peptide(s) as a part of their coat proteins (CP) and elicit an antigen-specific immune response. Here, we report the high yield and consistency in production of recombinant alfalfa mosaic virus (AlMV) particles for specific presentation of the small loop 15 amino acid epitope from domain-4 of the Bacillus anthracis protective antigen (PA-D4s). The epitope was inserted immediately after the first 25 N-terminal amino acids of AlMV CP to retain genome activation and binding of CP to viral RNAs. Recombinant AlMV particles were efficiently produced in tobacco, easily purified for immunological analysis, and exhibited extended stability and systemic proliferation in planta. Intraperitional injections of mice with recombinant plant virus particles harboring the PA-D4s epitope elicited a distinct immune response. Western blotting and ELISA analysis showed that sera from immunized mice recognized both native PA antigen and the AlMV CP.

  14. Advances in alfalfa mosaic virus-mediated expression of anthrax antigen in planta.

    PubMed

    Brodzik, R; Bandurska, K; Deka, D; Golovkin, M; Koprowski, H

    2005-12-16

    Plant viruses show great potential for production of pharmaceuticals in plants. Such viruses can harbor a small antigenic peptide(s) as a part of their coat proteins (CP) and elicit an antigen-specific immune response. Here, we report the high yield and consistency in production of recombinant alfalfa mosaic virus (AlMV) particles for specific presentation of the small loop 15 amino acid epitope from domain-4 of the Bacillus anthracis protective antigen (PA-D4s). The epitope was inserted immediately after the first 25 N-terminal amino acids of AlMV CP to retain genome activation and binding of CP to viral RNAs. Recombinant AlMV particles were efficiently produced in tobacco, easily purified for immunological analysis, and exhibited extended stability and systemic proliferation in planta. Intraperitional injections of mice with recombinant plant virus particles harboring the PA-D4s epitope elicited a distinct immune response. Western blotting and ELISA analysis showed that sera from immunized mice recognized both native PA antigen and the AlMV CP. PMID:16236249

  15. Bioinspired engineering study of Plantae vascules for self-healing composite structures

    PubMed Central

    Trask, R. S.; Bond, I. P.

    2010-01-01

    This paper presents the first conceptual study into creating a Plantae-inspired vascular network within a fibre-reinforced polymer composite laminate, which provides an ongoing self-healing functionality without incurring a mass penalty. Through the application of a ‘lost-wax’ technique, orthogonal hollow vascules, inspired by the ‘ray cell’ structures found in ring porous hardwoods, were successfully introduced within a carbon fibre-reinforced epoxy polymer composite laminate. The influence on fibre architecture and mechanical behaviour of single vascules (located on the laminate centreline) when aligned parallel and transverse to the local host ply was characterized experimentally using a compression-after-impact test methodology. Ultrasonic C-scanning and high-resolution micro-CT X-ray was undertaken to identify the influence of and interaction between the internal vasculature and impact damage. The results clearly show that damage morphology is influenced by vascule orientation and that a 10 J low-velocity impact damage event is sufficient to breach the vasculature; a prerequisite for any subsequent self-healing function. The residual compressive strength after a 10 J impact was found to be dependent upon vascule orientation. In general, residual compressive strength decreased to 70 per cent of undamaged strength when vasculature was aligned parallel to the local host ply and a value of 63 per cent when aligned transverse. This bioinspired engineering study has illustrated the potential that a vasculature concept has to offer in terms of providing a self-healing function with minimum mass penalty, without initiating premature failure within a composite structure. PMID:19955122

  16. In planta comparative transcriptomics of host-adapted strains of Ralstonia solanacearum.

    PubMed

    Ailloud, Florent; Lowe, Tiffany M; Robène, Isabelle; Cruveiller, Stéphane; Allen, Caitilyn; Prior, Philippe

    2016-01-01

    Background. Ralstonia solanacearum is an economically important plant pathogen with an unusually large host range. The Moko (banana) and NPB (not pathogenic to banana) strain groups are closely related but are adapted to distinct hosts. Previous comparative genomics studies uncovered very few differences that could account for the host range difference between these pathotypes. To better understand the basis of this host specificity, we used RNAseq to profile the transcriptomes of an R. solanacearum Moko strain and an NPB strain under in vitro and in planta conditions. Results. RNAs were sequenced from bacteria grown in rich and minimal media, and from bacteria extracted from mid-stage infected tomato, banana and melon plants. We computed differential expression between each pair of conditions to identify constitutive and host-specific gene expression differences between Moko and NPB. We found that type III secreted effectors were globally up-regulated upon plant cell contact in the NPB strain compared with the Moko strain. Genes encoding siderophore biosynthesis and nitrogen assimilation genes were highly up-regulated in the NPB strain during melon pathogenesis, while denitrification genes were up-regulated in the Moko strain during banana pathogenesis. The relatively lower expression of oxidases and the denitrification pathway during banana pathogenesis suggests that R. solanacearum experiences higher oxygen levels in banana pseudostems than in tomato or melon xylem. Conclusions. This study provides the first report of differential gene expression associated with host range variation. Despite minimal genomic divergence, the pathogenesis of Moko and NPB strains is characterized by striking differences in expression of virulence- and metabolism-related genes. PMID:26788428

  17. 3D domain swapping causes extensive multimerisation of human interleukin-10 when expressed in planta.

    PubMed

    Westerhof, Lotte B; Wilbers, Ruud H P; Roosien, Jan; van de Velde, Jan; Goverse, Aska; Bakker, Jaap; Schots, Arjen

    2012-01-01

    Heterologous expression platforms of biopharmaceutical proteins have been significantly improved over the last decade. Further improvement can be established by examining the intrinsic properties of proteins. Interleukin-10 (IL-10) is an anti-inflammatory cytokine with a short half-life that plays an important role in re-establishing immune homeostasis. This homodimeric protein of 36 kDa has significant therapeutic potential to treat inflammatory and autoimmune diseases. In this study we show that the major production bottleneck of human IL-10 is not protein instability as previously suggested, but extensive multimerisation due to its intrinsic 3D domain swapping characteristic. Extensive multimerisation of human IL-10 could be visualised as granules in planta. On the other hand, mouse IL-10 hardly multimerised, which could be largely attributed to its glycosylation. By introducing a short glycine-serine-linker between the fourth and fifth alpha helix of human IL-10 a stable monomeric form of IL-10 (hIL-10(mono)) was created that no longer multimerised and increased yield up to 20-fold. However, hIL-10(mono) no longer had the ability to reduce pro-inflammatory cytokine secretion from lipopolysaccharide-stimulated macrophages. Forcing dimerisation restored biological activity. This was achieved by fusing human IL-10(mono) to the C-terminal end of constant domains 2 and 3 of human immunoglobulin A (Fcα), a natural dimer. Stable dimeric forms of IL-10, like Fcα-IL-10, may not only be a better format for improved production, but also a more suitable format for medical applications. PMID:23049703

  18. In planta comparative transcriptomics of host-adapted strains of Ralstonia solanacearum

    PubMed Central

    Ailloud, Florent; Lowe, Tiffany M.; Robène, Isabelle; Cruveiller, Stéphane; Allen, Caitilyn

    2016-01-01

    Background. Ralstonia solanacearum is an economically important plant pathogen with an unusually large host range. The Moko (banana) and NPB (not pathogenic to banana) strain groups are closely related but are adapted to distinct hosts. Previous comparative genomics studies uncovered very few differences that could account for the host range difference between these pathotypes. To better understand the basis of this host specificity, we used RNAseq to profile the transcriptomes of an R. solanacearum Moko strain and an NPB strain under in vitro and in planta conditions. Results. RNAs were sequenced from bacteria grown in rich and minimal media, and from bacteria extracted from mid-stage infected tomato, banana and melon plants. We computed differential expression between each pair of conditions to identify constitutive and host-specific gene expression differences between Moko and NPB. We found that type III secreted effectors were globally up-regulated upon plant cell contact in the NPB strain compared with the Moko strain. Genes encoding siderophore biosynthesis and nitrogen assimilation genes were highly up-regulated in the NPB strain during melon pathogenesis, while denitrification genes were up-regulated in the Moko strain during banana pathogenesis. The relatively lower expression of oxidases and the denitrification pathway during banana pathogenesis suggests that R. solanacearum experiences higher oxygen levels in banana pseudostems than in tomato or melon xylem. Conclusions. This study provides the first report of differential gene expression associated with host range variation. Despite minimal genomic divergence, the pathogenesis of Moko and NPB strains is characterized by striking differences in expression of virulence- and metabolism-related genes. PMID:26788428

  19. In planta reduction of maize seedling stalk lesions by the bacterial endophyte Bacillus mojavensis.

    PubMed

    Bacon, Charles W; Hinton, Dorothy M

    2011-06-01

    Maize (Zea mays L.) is susceptible to infection by Fusarium verticillioides through autoinfection and alloinfection, resulting in diseases and contamination of maize kernels with the fumonisin mycotoxins. Attempts at controlling this fungus are currently being done with biocontrol agents such as bacteria, and this includes bacterial endophytes, such as Bacillus mojavensis . In addition to producing fumonisins, which are phytotoxic and mycotoxic, F. verticillioides also produces fusaric acid, which acts both as a phytotoxin and as an antibiotic. The question now is Can B. mojavensis reduce lesion development in maize during the alloinfection process, simulated by internode injection of the fungus? Mutant strains of B. mojavensis that tolerate fusaric acid were used in a growth room study to determine the development of stalk lesions, indicative of maize seedling blight, by co-inoculations with a wild-type strain of F. verticillioides and with non-fusaric acid producing mutants of F. verticillioides. Lesions were measured on 14-day-old maize stalks consisting of treatment groups inoculated with and without mutants and wild-type strains of bacteria and fungi. The results indicate that the fusaric-acid-tolerant B. mojavensis mutant reduced stalk lesions, suggesting an in planta role for this substance as an antibiotic. Further, lesion development occurred in maize infected with F. verticillioides mutants that do not produce fusaric acid, indicating a role for other phytotoxins, such as the fumonisins. Thus, additional pathological components should be examined before strains of B. mojavensis can be identified as being effective as a biocontrol agent, particularly for the control of seedling disease of maize. PMID:21635192

  20. Bioinspired engineering study of Plantae vascules for self-healing composite structures.

    PubMed

    Trask, R S; Bond, I P

    2010-06-01

    This paper presents the first conceptual study into creating a Plantae-inspired vascular network within a fibre-reinforced polymer composite laminate, which provides an ongoing self-healing functionality without incurring a mass penalty. Through the application of a 'lost-wax' technique, orthogonal hollow vascules, inspired by the 'ray cell' structures found in ring porous hardwoods, were successfully introduced within a carbon fibre-reinforced epoxy polymer composite laminate. The influence on fibre architecture and mechanical behaviour of single vascules (located on the laminate centreline) when aligned parallel and transverse to the local host ply was characterized experimentally using a compression-after-impact test methodology. Ultrasonic C-scanning and high-resolution micro-CT X-ray was undertaken to identify the influence of and interaction between the internal vasculature and impact damage. The results clearly show that damage morphology is influenced by vascule orientation and that a 10 J low-velocity impact damage event is sufficient to breach the vasculature; a prerequisite for any subsequent self-healing function. The residual compressive strength after a 10 J impact was found to be dependent upon vascule orientation. In general, residual compressive strength decreased to 70 per cent of undamaged strength when vasculature was aligned parallel to the local host ply and a value of 63 per cent when aligned transverse. This bioinspired engineering study has illustrated the potential that a vasculature concept has to offer in terms of providing a self-healing function with minimum mass penalty, without initiating premature failure within a composite structure. PMID:19955122

  1. Significant reduction of BiFC non-specific assembly facilitates in planta assessment of heterotrimeric G-protein interactors

    PubMed Central

    Gookin, Timothy E; Assmann, Sarah M

    2014-01-01

    Protein networks and signaling cascades are key mechanisms for intra- and intercellular signal transduction. Identifying the interacting partners of a protein can provide vital clues regarding its physiological role. The bimolecular fluorescence complementation (BiFC) assay has become a routine tool for in vivo analysis of protein–protein interactions and their subcellular location. Although the BiFC system has improved since its inception, the available options for in planta analysis are still subject to very low signal-to-noise ratios, and a systematic comparison of BiFC confounding background signals has been lacking. Background signals can obscure weak interactions, provide false positives, and decrease confidence in true positives. To overcome these problems, we performed an extensive in planta analysis of published BiFC fragments used in metazoa and plants, and then developed an optimized single vector BiFC system which utilizes monomeric Venus (mVenus) split at residue 210, and contains an integrated mTurquoise2 marker to precisely identify transformed cells in order to distinguish true negatives. Here we provide our streamlined double ORF expression (pDOE) BiFC system, and show that our advance in BiFC methodology functions even with an internally fused mVenus210 fragment. We illustrate the efficacy of the system by providing direct visualization of Arabidopsis MLO1 interacting with a calmodulin-like (CML) protein, and by showing that heterotrimeric G-protein subunits Gα (GPA1) and Gβ (AGB1) interact in plant cells. We further demonstrate that GPA1 and AGB1 each physically interact with PLDα1 in planta, and that mutation of the so-called PLDα1 ‘DRY’ motif abolishes both of these interactions. PMID:25187041

  2. Significant reduction of BiFC non-specific assembly facilitates in planta assessment of heterotrimeric G-protein interactors.

    PubMed

    Gookin, Timothy E; Assmann, Sarah M

    2014-11-01

    Protein networks and signaling cascades are key mechanisms for intra- and intercellular signal transduction. Identifying the interacting partners of a protein can provide vital clues regarding its physiological role. The bimolecular fluorescence complementation (BiFC) assay has become a routine tool for in vivo analysis of protein-protein interactions and their subcellular location. Although the BiFC system has improved since its inception, the available options for in planta analysis are still subject to very low signal-to-noise ratios, and a systematic comparison of BiFC confounding background signals has been lacking. Background signals can obscure weak interactions, provide false positives, and decrease confidence in true positives. To overcome these problems, we performed an extensive in planta analysis of published BiFC fragments used in metazoa and plants, and then developed an optimized single vector BiFC system which utilizes monomeric Venus (mVenus) split at residue 210, and contains an integrated mTurquoise2 marker to precisely identify transformed cells in order to distinguish true negatives. Here we provide our streamlined double ORF expression (pDOE) BiFC system, and show that our advance in BiFC methodology functions even with an internally fused mVenus210 fragment. We illustrate the efficacy of the system by providing direct visualization of Arabidopsis MLO1 interacting with a calmodulin-like (CML) protein, and by showing that heterotrimeric G-protein subunits Gα (GPA1) and Gβ (AGB1) interact in plant cells. We further demonstrate that GPA1 and AGB1 each physically interact with PLDα1 in planta, and that mutation of the so-called PLDα1 'DRY' motif abolishes both of these interactions. PMID:25187041

  3. Nodularin, a cyanobacterial toxin, is synthesized in planta by symbiotic Nostoc sp.

    PubMed

    Gehringer, Michelle M; Adler, Lewis; Roberts, Alexandra A; Moffitt, Michelle C; Mihali, Troco K; Mills, Toby J T; Fieker, Claus; Neilan, Brett A

    2012-10-01

    The nitrogen-fixing bacterium, Nostoc, is a commonly occurring cyanobacterium often found in symbiotic associations. We investigated the potential of cycad cyanobacterial endosymbionts to synthesize microcystin/nodularin. Endosymbiont DNA was screened for the aminotransferase domain of the toxin biosynthesis gene clusters. Five endosymbionts carrying the gene were screened for bioactivity. Extracts of two isolates inhibited protein phosphatase 2A and were further analyzed using electrospray ionization mass spectrometry (ESI-MS)/MS. Nostoc sp. 'Macrozamia riedlei 65.1' and Nostoc sp. 'Macrozamia serpentina 73.1' both contained nodularin. High performance liquid chromatography (HPLC) HESI-MS/MS analysis confirmed the presence of nodularin at 9.55±2.4 ng μg-1 chlorophyll a in Nostoc sp. 'Macrozamia riedlei 65.1' and 12.5±8.4 ng μg-1 Chl a in Nostoc sp. 'Macrozamia serpentina 73.1' extracts. Further scans indicated the presence of the rare isoform [L-Har(2)] nodularin, which contains L-homoarginine instead of L-arginine. Nodularin was also present at 1.34±0.74 ng ml(-1) (approximately 3 pmol per g plant ww) in the methanol root extracts of M. riedlei MZ65, while the presence of [L-Har(2)] nodularin in the roots of M. serpentina MZ73 was suggested by HPLC HESI-MS/MS analysis. The ndaA-B and ndaF genomic regions were sequenced to confirm the presence of the hybrid polyketide/non-ribosomal gene cluster. A seven amino-acid insertion into the NdaA-C1 domain of N. spumigena NSOR10 protein was observed in all endosymbiont-derived sequences, suggesting the transfer of the nda cluster from N. spumigena to terrestrial Nostoc species. This study demonstrates the synthesis of nodularin and [L-Har(2)] nodularin in a non-Nodularia species and the production of cyanobacterial hepatotoxin by a symbiont in planta. PMID:22456448

  4. In planta gene expression analysis of Xanthomonas oryzae pathovar oryzae, African strain MAI1

    PubMed Central

    2010-01-01

    Background Bacterial leaf blight causes significant yield losses in rice crops throughout Asia and Africa. Although both the Asian and African strains of the pathogen, Xanthomonas oryzae pv. oryzae (Xoo), induce similar symptoms, they are nevertheless genetically different, with the African strains being more closely related to the Asian X. oryzae pv. oryzicola (Xoc). Results Changes in gene expression of the African Xoo strain MAI1 in the susceptible rice cultivar Nipponbare were profiled, using an SSH Xoo DNA microarray. Microarray hybridization was performed comparing bacteria recovered from plant tissues at 1, 3, and 6 days after inoculation (dai) with bacteria grown in vitro. A total of 710 bacterial genes were found to be differentially expressed, with 407 up-regulated and 303 down-regulated. Expression profiling indicated that less than 20% of the 710 bacterial transcripts were induced in the first 24 h after inoculation, whereas 63% were differentially expressed at 6 dai. The 710 differentially expressed genes were one-end sequenced. 535 sequences were obtained from which 147 non-redundant sequences were identified. Differentially expressed genes were related to metabolism, secretion and transport, pathogen adherence to plant tissues, plant cell-wall degradation, IS elements, and virulence. In addition, various other genes encoding proteins with unknown function or showing no similarity to other proteins were also induced. The Xoo MAI1 non-redundant set of sequences was compared against several X. oryzae genomes, revealing a specific group of genes that was present only in MAI1. Numerous IS elements were also found to be differentially expressed. Quantitative real-time PCR confirmed 86% of the identified profile on a set of 14 genes selected according to the microarray analysis. Conclusions This is the first report to compare the expression of Xoo genes in planta across different time points during infection. This work shows that as-yet-unidentified and

  5. In-planta sporulation phenotype: a major life history trait to understand the evolution of Alnus-infective Frankia strains.

    PubMed

    Pozzi, Adrien C; Bautista-Guerrero, Hector H; Nouioui, Imen; Cotin-Galvan, Laëtitia; Pepin, Régis; Fournier, Pascale; Menu, Frédéric; Fernandez, Maria P; Herrera-Belaroussi, Aude

    2015-09-01

    Two major types of Frankia strains are usually recognized, based on the ability to sporulate in-planta: spore-positive (Sp+) and spore-negative (Sp-). We carried out a study of Sp+ and Sp- Frankia strains based on nodules collected on Alnus glutinosa, Alnus incana and Alnus viridis. The nodules were phenotyped using improved histology methods, and endophytic Frankia strain genotype was determined using a multilocus sequence analysis approach. An additional sampling was done to assess the relation between Sp+ phenotype frequency and genetic diversity of Frankia strains at the alder stand scale. Our results revealed that (i) Sp+ and Sp- Alnus-infective Frankia strains are genetically different even when sampled from the same alder stand and the same host-plant species; (ii) there are at least two distinct phylogenetic lineages of Sp+ Frankia that cluster according to the host-plant species and without regard of geographic distance and (iii) genetic diversity of Sp+ strains is very low at the alder stand scale compared with Sp- strains. Difference in evolutionary history and genetic diversity between Sp+ and Sp- Frankia allows us to discuss the possible ecological role of in-planta sporulation. PMID:25335453

  6. Plastid-localized amino acid biosynthetic pathways of Plantae are predominantly composed of non-cyanobacterial enzymes

    PubMed Central

    Reyes-Prieto, Adrian; Moustafa, Ahmed

    2012-01-01

    Studies of photosynthetic eukaryotes have revealed that the evolution of plastids from cyanobacteria involved the recruitment of non-cyanobacterial proteins. Our phylogenetic survey of >100 Arabidopsis nuclear-encoded plastid enzymes involved in amino acid biosynthesis identified only 21 unambiguous cyanobacterial-derived proteins. Some of the several non-cyanobacterial plastid enzymes have a shared phylogenetic origin in the three Plantae lineages. We hypothesize that during the evolution of plastids some enzymes encoded in the host nuclear genome were mistargeted into the plastid. Then, the activity of those foreign enzymes was sustained by both the plastid metabolites and interactions with the native cyanobacterial enzymes. Some of the novel enzymatic activities were favored by selective compartmentation of additional complementary enzymes. The mosaic phylogenetic composition of the plastid amino acid biosynthetic pathways and the reduced number of plastid-encoded proteins of non-cyanobacterial origin suggest that enzyme recruitment underlies the recompartmentation of metabolic routes during the evolution of plastids. PMID:23233874

  7. In planta expression of A. cellulolyticus Cel5A endocellulase reduces cell wall recalcitrance in tobacco and maize

    PubMed Central

    2011-01-01

    The glycoside hydrolase family 5 endocellulase, E1 (Cel5A), from Acidothermus cellulolyticus was transformed into both Nicotiana tabacum and Zea mays with expression targeted to the cell wall under a constitutive promoter. Here we explore the possibility that in planta expression of endocellulases will allow these enzymes to access their substrates during cell wall construction, rendering cellulose more amenable to pretreatment and enzyme digestion. Tobacco and maize plants were healthy and developed normally compared with the wild type (WT). After thermochemical pretreatment and enzyme digestion, transformed plants were clearly more digestible than WT, requiring lower pretreatment severity to achieve comparable conversion levels. Furthermore, the decreased recalcitrance was not due to post-pretreatment residual E1 activity and could not be reproduced by the addition of exogenous E1 to the biomass prior to pretreatment, indicating that the expression of E1 during cell wall construction altered the inherent recalcitrance of the cell wall. PMID:21269444

  8. Development of fiber optic spectroscopy for in-vitro and in-planta detection of fluorescent proteins

    NASA Astrophysics Data System (ADS)

    Liew, Oi Wah; Chen, Jun-Wei; Asundi, Anand K.

    2001-10-01

    The objective of this project is to apply photonics technology to bio-safety management of genetically modified (GM) plants. The conventional method for screening GM plants is through selection using antibiotic resistance markers. There is public concern with such approaches and these are associated with food safety issues, escape of antibiotic resistance genes to pathogenic microorganisms and interference with antibiotic therapy. Thus, the strategy taken in this project is to replace antibiotic resistance markers with fluorescent protein markers that allow for rapid and non-invasive optical screening of genetically modified plants. In this paper, fibre optic spectroscopy was developed to detect and quantify recombinant green (EGFP) and red (DsRED) fluorescent proteins in vitro and in planta. In vitro detection was first carried out to optimize the sensitivity of the optical system. The bacterial expression vectors carrying the coding regions of EGFP and DsRED were introduced into Escherichia coli host cells and fluorescent proteins were produced following induction with IPTG. Soluble EGFP and DsRED proteins were isolated from lysed bacterial cells and serially diluted for quantitative analysis by fibre optic spectroscopy using different light sources, namely, blue LED (475 nm), tungsten halogen (350 - 1000 nm) and double frequency Nd:YAG green laser (532 nm). Fluorescence near the expected emission wavelengths could be detected up to 320X dilution for EGFP and DsRED with blue LED and 532 nm green laser, respectively, as the excitation source. Tungsten halogen was found to be unsuitable for excitation of both EGFP and DsRED. EGFP was successfully purified by size separation under non-denaturing electrophoretic conditions and quantified. The minimum concentration of EGFP detectable with blue LED excitation was 5 mg/ml. To determine the capability of spectroscopy detection in planta, transgenic potato hairy roots and whole modified plant lines expressing the

  9. The application of laser microdissection to in planta gene expression profiling of the maize anthracnose stalk rot fungus Colletotrichum graminicola.

    PubMed

    Tang, Weihua; Coughlan, Sean; Crane, Edmund; Beatty, Mary; Duvick, Jon

    2006-11-01

    Laser microdissection (LM) offers a potential means for deep sampling of a fungal plant-pathogen transcriptome during the infection process using whole-genome DNA microarrays. The use of a fluorescent protein-expressing fungus can greatly facilitate the identification of fungal structures for LM sampling. However, fixation methods that preserve both tissue histology and protein fluorescence, and that also yield RNA of suitable quality for microarray applications, have not been reported. We developed a microwave-accelerated acetone fixation, paraffin-embedding method that fulfills these requirements and used it to prepare mature maize stalk tissues infected with an Anemonia majano cyan fluorescent protein-expressing isolate of the anthracnose stalk rot fungus Colletotrichum graminicola. We successfully used LM to isolate individual maize cells associated with C. graminicola hyphae at an early stage of infection. The LM-derived RNA, after two-round linear amplification, was of sufficient quality and quantity for global expression profiling using a fungal microarray. Comparing replicated LM samples representing an early stage of stalk cell infection with samples from in vitro-germinated conidia, we identified 437 and 370 C. graminicola genes showing significant up- or downregulation, respectively. We confirmed the differential expression of several representative transcripts by quantitative reverse-transcriptase polymerase chain reaction (RT-PCR) and documented extensive overlap of this dataset with a PCR-subtraction library enriched for C. graminicola transcripts in planta. Our results demonstrate that LM is feasible for in planta pathogen expression profiling and can reveal clues about fungal genes involved in pathogenesis. The method in this report may be advantageous for visualizing a variety of cellular features that depend on a high degree of histochemical preservation and RNA integrity prior to LM. PMID:17073306

  10. In planta Transformed Cumin (Cuminum cyminum L.) Plants, Overexpressing the SbNHX1 Gene Showed Enhanced Salt Endurance.

    PubMed

    Pandey, Sonika; Patel, Manish Kumar; Mishra, Avinash; Jha, Bhavanath

    2016-01-01

    Cumin is an annual, herbaceous, medicinal, aromatic, spice glycophyte that contains diverse applications as a food and flavoring additive, and therapeutic agents. An efficient, less time consuming, Agrobacterium-mediated, a tissue culture-independent in planta genetic transformation method was established for the first time using cumin seeds. The SbNHX1 gene, cloned from an extreme halophyte Salicornia brachiata was transformed in cumin using optimized in planta transformation method. The SbNHX1 gene encodes a vacuolar Na+/H+ antiporter and is involved in the compartmentalization of excess Na+ ions into the vacuole and maintenance of ion homeostasis Transgenic cumin plants were confirmed by PCR using gene (SbNHX1, uidA and hptII) specific primers. The single gene integration event and overexpression of the gene were confirmed by Southern hybridization and competitive RT-PCR, respectively. Transgenic lines L3 and L13 showed high expression of the SbNHX1 gene compared to L6 whereas moderate expression was detected in L5 and L10 transgenic lines. Transgenic lines (L3, L5, L10 and L13), overexpressing the SbNHX1 gene, showed higher photosynthetic pigments (chlorophyll a, b and carotenoid), and lower electrolytic leakage, lipid peroxidation (MDA content) and proline content as compared to wild type plants under salinity stress. Though transgenic lines were also affected by salinity stress but performed better compared to WT plants. The ectopic expression of the SbNHX1 gene confirmed enhanced salinity stress tolerance in cumin as compared to wild type plants under stress condition. The present study is the first report of engineering salt tolerance in cumin, so far and the plant may be utilized for the cultivation in saline areas. PMID:27411057

  11. Phytohormonal networks promote differentiation of fiber initials on pre-anthesis cotton ovules grown in vitro and in planta.

    PubMed

    Kim, Hee Jin; Hinchliffe, Doug J; Triplett, Barbara A; Chen, Z Jeffrey; Stelly, David M; Yeater, Kathleen M; Moon, Hong S; Gilbert, Matthew K; Thyssen, Gregory N; Turley, Rickie B; Fang, David D

    2015-01-01

    The number of cotton (Gossypium sp.) ovule epidermal cells differentiating into fiber initials is an important factor affecting cotton yield and fiber quality. Despite extensive efforts in determining the molecular mechanisms regulating fiber initial differentiation, only a few genes responsible for fiber initial differentiation have been discovered. To identify putative genes directly involved in the fiber initiation process, we used a cotton ovule culture technique that controls the timing of fiber initial differentiation by exogenous phytohormone application in combination with comparative expression analyses between wild type and three fiberless mutants. The addition of exogenous auxin and gibberellins to pre-anthesis wild type ovules that did not have visible fiber initials increased the expression of genes affecting auxin, ethylene, ABA and jasmonic acid signaling pathways within 1 h after treatment. Most transcripts expressed differentially by the phytohormone treatment in vitro were also differentially expressed in the ovules of wild type and fiberless mutants that were grown in planta. In addition to MYB25-like, a gene that was previously shown to be associated with the differentiation of fiber initials, several other differentially expressed genes, including auxin/indole-3-acetic acid (AUX/IAA) involved in auxin signaling, ACC oxidase involved in ethylene biosynthesis, and abscisic acid (ABA) 8'-hydroxylase an enzyme that controls the rate of ABA catabolism, were co-regulated in the pre-anthesis ovules of both wild type and fiberless mutants. These results support the hypothesis that phytohormonal signaling networks regulate the temporal expression of genes responsible for differentiation of cotton fiber initials in vitro and in planta. PMID:25927364

  12. Phytohormonal Networks Promote Differentiation of Fiber Initials on Pre-Anthesis Cotton Ovules Grown In Vitro and In Planta

    PubMed Central

    Kim, Hee Jin; Hinchliffe, Doug J.; Triplett, Barbara A.; Chen, Z. Jeffrey; Stelly, David M.; Yeater, Kathleen M.; Moon, Hong S.; Gilbert, Matthew K.; Thyssen, Gregory N.; Turley, Rickie B.; Fang, David D.

    2015-01-01

    The number of cotton (Gossypium sp.) ovule epidermal cells differentiating into fiber initials is an important factor affecting cotton yield and fiber quality. Despite extensive efforts in determining the molecular mechanisms regulating fiber initial differentiation, only a few genes responsible for fiber initial differentiation have been discovered. To identify putative genes directly involved in the fiber initiation process, we used a cotton ovule culture technique that controls the timing of fiber initial differentiation by exogenous phytohormone application in combination with comparative expression analyses between wild type and three fiberless mutants. The addition of exogenous auxin and gibberellins to pre-anthesis wild type ovules that did not have visible fiber initials increased the expression of genes affecting auxin, ethylene, ABA and jasmonic acid signaling pathways within 1 h after treatment. Most transcripts expressed differentially by the phytohormone treatment in vitro were also differentially expressed in the ovules of wild type and fiberless mutants that were grown in planta. In addition to MYB25-like, a gene that was previously shown to be associated with the differentiation of fiber initials, several other differentially expressed genes, including auxin/indole-3-acetic acid (AUX/IAA) involved in auxin signaling, ACC oxidase involved in ethylene biosynthesis, and abscisic acid (ABA) 8'-hydroxylase an enzyme that controls the rate of ABA catabolism, were co-regulated in the pre-anthesis ovules of both wild type and fiberless mutants. These results support the hypothesis that phytohormonal signaling networks regulate the temporal expression of genes responsible for differentiation of cotton fiber initials in vitro and in planta. PMID:25927364

  13. In planta Transformed Cumin (Cuminum cyminum L.) Plants, Overexpressing the SbNHX1 Gene Showed Enhanced Salt Endurance

    PubMed Central

    Pandey, Sonika; Patel, Manish Kumar; Jha, Bhavanath

    2016-01-01

    Cumin is an annual, herbaceous, medicinal, aromatic, spice glycophyte that contains diverse applications as a food and flavoring additive, and therapeutic agents. An efficient, less time consuming, Agrobacterium-mediated, a tissue culture-independent in planta genetic transformation method was established for the first time using cumin seeds. The SbNHX1 gene, cloned from an extreme halophyte Salicornia brachiata was transformed in cumin using optimized in planta transformation method. The SbNHX1 gene encodes a vacuolar Na+/H+ antiporter and is involved in the compartmentalization of excess Na+ ions into the vacuole and maintenance of ion homeostasis Transgenic cumin plants were confirmed by PCR using gene (SbNHX1, uidA and hptII) specific primers. The single gene integration event and overexpression of the gene were confirmed by Southern hybridization and competitive RT-PCR, respectively. Transgenic lines L3 and L13 showed high expression of the SbNHX1 gene compared to L6 whereas moderate expression was detected in L5 and L10 transgenic lines. Transgenic lines (L3, L5, L10 and L13), overexpressing the SbNHX1 gene, showed higher photosynthetic pigments (chlorophyll a, b and carotenoid), and lower electrolytic leakage, lipid peroxidation (MDA content) and proline content as compared to wild type plants under salinity stress. Though transgenic lines were also affected by salinity stress but performed better compared to WT plants. The ectopic expression of the SbNHX1 gene confirmed enhanced salinity stress tolerance in cumin as compared to wild type plants under stress condition. The present study is the first report of engineering salt tolerance in cumin, so far and the plant may be utilized for the cultivation in saline areas. PMID:27411057

  14. Detection of plum pox potyviral protein-protein interactions in planta using an optimized mRFP-based bimolecular fluorescence complementation system.

    PubMed

    Zilian, Eva; Maiss, Edgar

    2011-12-01

    In previous studies, protein interaction maps of different potyviruses have been generated using yeast two-hybrid (YTH) systems, and these maps have demonstrated a high diversity of interactions of potyviral proteins. Using an optimized bimolecular fluorescence complementation (BiFC) system, a complete interaction matrix for proteins of a potyvirus was developed for the first time under in planta conditions with ten proteins from plum pox virus (PPV). In total, 52 of 100 possible interactions were detected, including the self-interactions of CI, 6K2, VPg, NIa-Pro, NIb and CP, which is more interactions than have ever been detected for any other potyvirus in a YTH approach. Moreover, the BiFC system was shown to be able to localize the protein interactions, which was typified for the protein self-interactions indicated above. Additionally, experiments were carried out with the P3N-PIPO protein, revealing an interaction with CI but not with CP and supporting the involvement of P3N-PIPO in the cell-to-cell movement of potyviruses. No self-interaction of the PPV helper component-proteinase (HC-Pro) was detected using BiFC in planta. Therefore, additional experiments with turnip mosaic virus (TuMV) HC-Pro, PPV_HC-Pro and their mutants were conducted. The self-interaction of TuMV_HCpro, as recently demonstrated, and the self-interaction of the TuMV_ and PPV_HC-Pro mutants were shown by BiFC in planta, indicating that HC-Pro self-interactions may be species-specific. BiFC is a very useful and reliable method for the detection and localization of protein interactions in planta, thus enabling investigations under more natural conditions than studies in yeast cells. PMID:21880839

  15. Construction of EGFP-labeling system for visualizing the infection process of Xanthomonas axonopodis pv. citri in planta.

    PubMed

    Liu, Li-Ping; Deng, Zi-Niu; Qu, Jin-Wang; Yan, Jia-Wen; Catara, Vittoria; Li, Da-Zhi; Long, Gui-You; Li, Na

    2012-09-01

    Xanthomonas axonopodis pv. citri (Xac) is the causal agent of citrus bacterial canker, an economically important disease to world citrus industry. To monitor the infection process of Xac in different citrus plants, the enhanced green florescent protein (EGFP) visualizing system was constructed to visualize the propagation and localization in planta. First, the wild-type Xac was isolated from the diseased leaves of susceptible 'Bingtang' sweet orange, and then the isolated Xac was labeled with EGFP by triparental mating. After PCR identification, the growth kinetics and pathogenicity of the transformants were analyzed in comparison with the wild-type Xac. The EGFP-labeled bacteria were inoculated by spraying on the surface and infiltration in the mesophyll of 'Bingtang' sweet orange leaves. The bacterial cell multiplication and diffusion processes were observed directly under confocal laser scanning microscope at different intervals after inoculation. The results indicated that the EGFP-labeled Xac releasing clear green fluorescence light under fluorescent microscope showed the infection process and had the same pathogenicity as the wild type to citrus. Consequently, the labeled Xac demonstrated the ability as an efficient tool to monitor the pathogen infection. PMID:22674174

  16. In planta mechanism of action of leptospermone: impact of its physico-chemical properties on uptake, translocation, and metabolism.

    PubMed

    Owens, Daniel K; Nanayakkara, N P Dhammika; Dayan, Franck E

    2013-02-01

    Leptospermone is a natural β-triketone that specifically inhibits the enzyme p-hydrophyphenylpyruvate dioxygenase, the same molecular target site as that of the commercial herbicide mesotrione. The β-triketone-rich essential oil of Leptospermum scoparium has both preemergence and postemergence herbicidal activity, resulting in bleaching of treated plants and dramatic growth reduction. Radiolabeled leptospermone was synthesized to investigate the in planta mechanism of action of this natural herbicide. Approximately 50 % of the absorbed leptospermone was translocated to the foliage suggesting rapid acropetal movement of the molecule. On the other hand, very little leptospermone was translocated away from the point of application on the foliage, indicating poor phloem mobility. These observations are consistent with the physico-chemical properties of leptospermone, such as its experimentally measured logP and pK a values, and molecular mass, number of hydrogen donors and acceptors, and number of rotatable bonds. Consequently, leptospermone is taken up readily by roots and translocated to reach its molecular target site. This provides additional evidence that the anecdotal observation of allelopathic suppression of plant growth under β-triketone-producing species may be due to the release of these phytotoxins in soils. PMID:23314892

  17. In Planta Single-Molecule Pull-Down Reveals Tetrameric Stoichiometry of HD-ZIPIII:LITTLE ZIPPER Complexes[OPEN

    PubMed Central

    Husbands, Aman Y.; Aggarwal, Vasudha; Ha, Taekjip; Timmermans, Marja C.P.

    2016-01-01

    Deciphering complex biological processes markedly benefits from approaches that directly assess the underlying biomolecular interactions. Most commonly used approaches to monitor protein-protein interactions typically provide nonquantitative readouts that lack statistical power and do not yield information on the heterogeneity or stoichiometry of protein complexes. Single-molecule pull-down (SiMPull) uses single-molecule fluorescence detection to mitigate these disadvantages and can quantitatively interrogate interactions between proteins and other compounds, such as nucleic acids, small molecule ligands, and lipids. Here, we establish SiMPull in plants using the HOMEODOMAIN LEUCINE ZIPPER III (HD-ZIPIII) and LITTLE ZIPPER (ZPR) interaction as proof-of-principle. Colocalization analysis of fluorophore-tagged HD-ZIPIII and ZPR proteins provides strong statistical evidence of complex formation. In addition, we use SiMPull to directly quantify YFP and mCherry maturation probabilities, showing these differ substantially from values obtained in mammalian systems. Leveraging these probabilities, in conjunction with fluorophore photobleaching assays on over 2000 individual complexes, we determined HD-ZIPIII:ZPR stoichiometry. Intriguingly, these complexes appear as heterotetramers, comprising two HD-ZIPIII and two ZPR molecules, rather than heterodimers as described in the current model. This surprising result raises new questions about the regulation of these key developmental factors and is illustrative of the unique contribution SiMPull is poised to make to in planta protein interaction studies. PMID:27385814

  18. Overexpression of the rice carotenoid cleavage dioxygenase 1 gene in Golden Rice endosperm suggests apocarotenoids as substrates in planta.

    PubMed

    Ilg, Andrea; Yu, Qiuju; Schaub, Patrick; Beyer, Peter; Al-Babili, Salim

    2010-08-01

    Carotenoids are converted by carotenoid cleavage dioxygenases that catalyze oxidative cleavage reactions leading to apocarotenoids. However, apocarotenoids can also be further truncated by some members of this enzyme family. The plant carotenoid cleavage dioxygenase 1 (CCD1) subfamily is known to degrade both carotenoids and apocarotenoids in vitro, leading to different volatile compounds. In this study, we investigated the impact of the rice CCD1 (OsCCD1) on the pigmentation of Golden Rice 2 (GR2), a genetically modified rice variety accumulating carotenoids in the endosperm. For this purpose, the corresponding cDNA was introduced into the rice genome under the control of an endosperm-specific promoter in sense and anti-sense orientations. Despite high expression levels of OsCCD1 in sense plants, pigment analysis revealed carotenoid levels and patterns comparable to those of GR2, pleading against carotenoids as substrates in rice endosperm. In support, similar carotenoid contents were determined in anti-sense plants. To check whether OsCCD1 overexpressed in GR2 endosperm is active, in vitro assays were performed with apocarotenoid substrates. HPLC analysis confirmed the cleavage activity of introduced OsCCD1. Our data indicate that apocarotenoids rather than carotenoids are the substrates of OsCCD1 in planta. PMID:20549230

  19. In planta chemical cross-linking and mass spectrometry analysis of protein structure and interaction in Arabidopsis.

    PubMed

    Zhu, Xinliang; Yu, Fengchao; Yang, Zhu; Liu, Shichang; Dai, Chen; Lu, Xiaoyun; Liu, Chenyu; Yu, Weichuan; Li, Ning

    2016-07-01

    Site-specific chemical cross-linking in combination with mass spectrometry analysis has emerged as a powerful proteomic approach for studying the three-dimensional structure of protein complexes and in mapping protein-protein interactions (PPIs). Building on the success of MS analysis of in vitro cross-linked proteins, which has been widely used to investigate specific interactions of bait proteins and their targets in various organisms, we report a workflow for in vivo chemical cross-linking and MS analysis in a multicellular eukaryote. This approach optimizes the in vivo protein cross-linking conditions in Arabidopsis thaliana, establishes a MudPIT procedure for the enrichment of cross-linked peptides, and develops an integrated software program, exhaustive cross-linked peptides identification tool (ECL), to identify the MS spectra of in planta chemical cross-linked peptides. In total, two pairs of in vivo cross-linked peptides of high confidence have been identified from two independent biological replicates. This work demarks the beginning of an alternative proteomic approach in the study of in vivo protein tertiary structure and PPIs in multicellular eukaryotes. PMID:27198063

  20. Functional characterisation of metal(loid) processes in planta through the integration of synchrotron techniques and plant molecular biology

    PubMed Central

    Donner, Erica; Punshon, Tracy; Guerinot, Mary Lou; Lombi, Enzo

    2013-01-01

    Functional characterisation of the genes regulating metal(loid) homeostasis in plants is a major focus of crop biofortification, phytoremediation, and food security research. This paper focuses on the potential for advancing plant metal(loid) research by combining molecular biology and synchrotron-based techniques. Recent advances in x-ray focussing optics and fluorescence detection have greatly improved the potential of synchrotron techniques for plant science research, allowing metal(loids) to be imaged in vivo in hydrated plant tissues at sub-micron resolution. Laterally resolved metal(loid) speciation can also be determined. By using molecular techniques to probe the location of gene expression and protein localisation and combining it with this synchrotron-derived data, functional information can be effectively and efficiently assigned to specific genes. This paper provides a review of the state of the art in this field, and provides examples as to how synchrotron-based methods can be combined with molecular techniques to facilitate functional characterisation of genes in planta. PMID:22200921

  1. Sucrose phosphate synthase and sucrose phosphate phosphatase interact in planta and promote plant growth and biomass accumulation

    PubMed Central

    Maloney, Victoria J.; Park, Ji-Young; Unda, Faride; Mansfield, Shawn D.

    2015-01-01

    Bioinformatic analysis indicates that sucrose phosphate synthase (SPS) contains a putative C-terminal sucrose phosphate phosphatase (SPP)-like domain that may facilitates the binding of SPP. If an SPS–SPP enzyme complex exists, it may provide sucrose biosynthesis with an additional level of regulation, forming a direct metabolic channel for sucrose-6-phosphate between these two enzymes. Herein, the formation of an enzyme complex between SPS and SPP was examined, and the results from yeast two-hybrid experiments suggest that there is indeed an association between these proteins. In addition, in planta bioluminescence resonance energy transfer (BRET) was observed in Arabidopsis seedlings, providing physical evidence for a protein interaction in live cells and in real time. Finally, bimolecular fluorescence complementation (BiFC) was employed in an attempt to detect SPS–SPP interactions visually. The findings clearly demonstrated that SPS interacts with SPP and that this interaction impacts soluble carbohydrate pools and affects carbon partitioning to starch. Moreover, a fusion construct between the two genes promotes plant growth in both transgenic Arabidopsis and hybrid poplar. PMID:25873678

  2. In planta transformation of sorghum (Sorghum bicolor (L.) Moench) using TPS1 gene for enhancing tolerance to abiotic stresses.

    PubMed

    Yellisetty, Varalaxmi; Reddy, L A; Mandapaka, Maheswari

    2015-09-01

    An in planta transformation protocol for sorghum (Sorghum bicolor (L.) Moench) using shoot apical meristem of germinating seedlings is reported in this study. Agrobacterium tumefaciens strain, LBA4404 with pCAMBIA1303 vector and construct pCAMBIA1303TPS1 were individually used for transformation. Since, the transgene is integrated into the cells of already differentiated tissues, the T 0 plants were chimeric and stable integration was observed in T1 generation. β-Glucuronidase (GUS) expression in the seedlings and spikelets of emerging cob was the first indication of transformability in T0 generation which was further confirmed by PCR analysis using hpt and TPS1 gene-specific primers. Screening on 25 mg/L hygromycin combined with PCR analysis was used for selection of transformants in the T1 generation. Transformation efficiencies ranged between 34-38% and 26-34% using pCAMBIA1303 vector and construct pCAMBIA1303TPS1, respectively. Molecular characterization of the T2 transgenics using PCR, RT-PCR and Southern blot analyses further revealed the integration, expression and inheritance of the transgene. These results indicate the feasibility of the method to generate transgenics with pCAM-BIA1303 vector and construct pCAMBIA1303TPS1. The abiotic stress tolerance of TPS1 transgenics developed in the present study was evident by the ability of the transformants to tolerate 200 mM NaCl as well as higher root growth and biomass. PMID:26440081

  3. An optimized mRFP-based bimolecular fluorescence complementation system for the detection of protein-protein interactions in planta.

    PubMed

    Zilian, Eva; Maiss, Edgar

    2011-06-01

    An existing bimolecular fluorescence complementation (BiFC) system, based on a monomeric red fluorescent protein (mRFP), has been optimized for the investigation of protein-protein interactions in planta. The expression plasmids, encoding the N-terminal amino acids (aa) 1-168 and the C-terminal aa 169-225 of the mRFP, allow N- or C-terminal fusion of a split mRFP, with the genes of interest. Two major improvements over the original vectors have been made. Firstly, the coding sequence of a GGGSGGG-linker has been integrated between mRFP sequences and the genes of interest. Secondly, a modified mini binary vector (∼3.5 kb) was introduced as the backbone for the plant expression plasmids. Based on the results of yeast two-hybrid studies with plant viral proteins, interaction of viral proteins was tested in Nicotiana benthamiana plants and monitored by confocal laser scanning microscopy (CLSM). Plum pox virus coat protein and mutants thereof served as controls. The system was validated using the N-protein of Capsicum chlorosis virus for which a self-interaction was shown for the first time, the Tobacco mosaic virus coat protein and BC1 and BV1 of the Tomato yellow leaf curl Thailand virus. This optimized BiFC system provides a convenient alternative to other BiFC, as well as yeast two-hybrid assays, for detecting protein-protein interactions. PMID:21473882

  4. Transient expression in Nicotiana benthamiana fluorescent marker lines provides enhanced definition of protein localization, movement and interactions in planta.

    PubMed

    Martin, Kathleen; Kopperud, Kristin; Chakrabarty, Romit; Banerjee, Rituparna; Brooks, Robert; Goodin, Michael M

    2009-07-01

    Here, we report on the construction of a novel series of Gateway-compatible plant transformation vectors containing genes encoding autofluorescent proteins, including Cerulean, Dendra2, DRONPA, TagRFP and Venus, for the expression of protein fusions in plant cells. To assist users in the selection of vectors, we have determined the relative in planta photostability and brightness of nine autofluorescent proteins (AFPs), and have compared the use of DRONPA and Dendra2 in photoactivation and photoconversion experiments. Additionally, we have generated transgenic Nicotiana benthamiana lines that express fluorescent protein markers targeted to nuclei, endoplasmic reticulum or actin filaments. We show that conducting bimolecular fluorescence complementation assays in plants that constitutively express cyan fluorescent protein fused to histone 2B provides enhanced data quality and content over assays conducted without the benefit of a subcellular marker. In addition to testing protein interactions, we demonstrate that our transgenic lines that express red fluorescent protein markers offer exceptional support in experiments aimed at defining nuclear or endomembrane localization. Taken together, the new combination of pSITE-BiFC and pSITEII vectors for studying intracellular protein interaction, localization and movement, in conjunction with our transgenic marker lines, constitute powerful tools for the plant biology community. PMID:19309457

  5. A rapid, highly efficient and economical method of Agrobacterium-mediated in planta transient transformation in living onion epidermis.

    PubMed

    Xu, Kedong; Huang, Xiaohui; Wu, Manman; Wang, Yan; Chang, Yunxia; Liu, Kun; Zhang, Ju; Zhang, Yi; Zhang, Fuli; Yi, Liming; Li, Tingting; Wang, Ruiyue; Tan, Guangxuan; Li, Chengwei

    2014-01-01

    Transient transformation is simpler, more efficient and economical in analyzing protein subcellular localization than stable transformation. Fluorescent fusion proteins were often used in transient transformation to follow the in vivo behavior of proteins. Onion epidermis, which has large, living and transparent cells in a monolayer, is suitable to visualize fluorescent fusion proteins. The often used transient transformation methods included particle bombardment, protoplast transfection and Agrobacterium-mediated transformation. Particle bombardment in onion epidermis was successfully established, however, it was expensive, biolistic equipment dependent and with low transformation efficiency. We developed a highly efficient in planta transient transformation method in onion epidermis by using a special agroinfiltration method, which could be fulfilled within 5 days from the pretreatment of onion bulb to the best time-point for analyzing gene expression. The transformation conditions were optimized to achieve 43.87% transformation efficiency in living onion epidermis. The developed method has advantages in cost, time-consuming, equipment dependency and transformation efficiency in contrast with those methods of particle bombardment in onion epidermal cells, protoplast transfection and Agrobacterium-mediated transient transformation in leaf epidermal cells of other plants. It will facilitate the analysis of protein subcellular localization on a large scale. PMID:24416168

  6. Positive-selection and ligation-independent cloning vectors for large scale in planta expression for plant functional genomics.

    PubMed

    Oh, Sang-Keun; Kim, Saet-Byul; Yeom, Seon-In; Lee, Hyun-Ah; Choi, Doil

    2010-12-01

    Transient expression is an easy, rapid and powerful technique for producing proteins of interest in plants. Recombinational cloning is highly efficient but has disadvantages, including complicated, time consuming cloning procedures and expensive enzymes for large-scale gene cloning. To overcome these limitations, we developed new ligation-independent cloning (LIC) vectors derived from binary vectors including tobacco mosaic virus (pJL-TRBO), potato virus X (pGR106) and the pBI121 vector-based pMBP1. LIC vectors were modified to enable directional cloning of PCR products without restriction enzyme digestion or ligation reactions. In addition, the ccdB gene, which encodes a potent cell-killing protein, was introduced between the two LIC adapter sites in the pJL-LIC, pGR-LIC, and pMBP-LIC vectors for the efficient selection of recombinant clones. This new vector does not require restriction enzymes, alkaline phosphatase, or DNA ligase for cloning. To clone, the three LIC vectors are digested with SnaBI and treated with T4 DNA polymerase, which includes 3' to 5' exonuclease activity in the presence of only one dNTP (dGTP for the inserts and dCTP for the vector). To make recombinants, the vector plasmid and the insert PCR fragment were annealed at room temperature for 20 min prior to transformation into the host. Bacterial transformation was accomplished with 100% efficiency. To validate the new LIC vector systems, we were used to coexpressed the Phytophthora AVR and potato resistance (R) genes in N. benthamiana by infiltration of Agrobacterium. Coexpressed AVR and R genes in N. benthamiana induced the typical hypersensitive cell death resulting from in vivo interaction of the two proteins. These LIC vectors could be efficiently used for high-throughput cloning and laboratory-scale in planta expression. These vectors could provide a powerful tool for high-throughput transient expression assays for functional genomic studies in plants. PMID:21340673

  7. Barcoding the kingdom Plantae: new PCR primers for ITS regions of plants with improved universality and specificity.

    PubMed

    Cheng, Tao; Xu, Chao; Lei, Li; Li, Changhao; Zhang, Yu; Zhou, Shiliang

    2016-01-01

    The internal transcribed spacer (ITS) of nuclear ribosomal DNA is one of the most commonly used DNA markers in plant phylogenetic and DNA barcoding analyses, and it has been recommended as a core plant DNA barcode. Despite this popularity, the universality and specificity of PCR primers for the ITS region are not satisfactory, resulting in amplification and sequencing difficulties. By thoroughly surveying and analysing the 18S, 5.8S and 26S sequences of Plantae and Fungi from GenBank, we designed new universal and plant-specific PCR primers for amplifying the whole ITS region and a part of it (ITS1 or ITS2) of plants. In silico analyses of the new and the existing ITS primers based on these highly representative data sets indicated that (i) the newly designed universal primers are suitable for over 95% of plants in most groups; and (ii) the plant-specific primers are suitable for over 85% of plants in most groups without amplification of fungi. A total of 335 samples from 219 angiosperm families, 11 gymnosperm families, 24 fern and lycophyte families, 16 moss families and 17 fungus families were used to test the performances of these primers. In vitro PCR produced similar results to those from the in silico analyses. Our new primer pairs gave PCR improvements up to 30% compared with common-used ones. The new universal ITS primers will find wide application in both plant and fungal biology, and the new plant-specific ITS primers will, by eliminating PCR amplification of nonplant templates, significantly improve the quality of ITS sequence information collections in plant molecular systematics and DNA barcoding. PMID:26084789

  8. In planta analysis of a cis-regulatory cytokinin response motif in Arabidopsis and identification of a novel enhancer sequence.

    PubMed

    Ramireddy, Eswarayya; Brenner, Wolfram G; Pfeifer, Andreas; Heyl, Alexander; Schmülling, Thomas

    2013-07-01

    The phytohormone cytokinin plays a key role in regulating plant growth and development, and is involved in numerous physiological responses to environmental changes. The type-B response regulators, which regulate the transcription of cytokinin response genes, are a part of the cytokinin signaling system. Arabidopsis thaliana encodes 11 type-B response regulators (type-B ARRs), and some of them were shown to bind in vitro to the core cytokinin response motif (CRM) 5'-(A/G)GAT(T/C)-3' or, in the case of ARR1, to an extended motif (ECRM), 5'-AAGAT(T/C)TT-3'. Here we obtained in planta proof for the functionality of the latter motif. Promoter deletion analysis of the primary cytokinin response gene ARR6 showed that a combination of two extended motifs within the promoter is required to mediate the full transcriptional activation by ARR1 and other type-B ARRs. CRMs were found to be over-represented in the vicinity of ECRMs in the promoters of cytokinin-regulated genes, suggesting their functional relevance. Moreover, an evolutionarily conserved 27 bp long T-rich region between -220 and -193 bp was identified and shown to be required for the full activation by type-B ARRs and the response to cytokinin. This novel enhancer is not bound by the DNA-binding domain of ARR1, indicating that additional proteins might be involved in mediating the transcriptional cytokinin response. Furthermore, genome-wide expression profiling identified genes, among them ARR16, whose induction by cytokinin depends on both ARR1 and other specific type-B ARRs. This together with the ECRM/CRM sequence clustering indicates cooperative action of different type-B ARRs for the activation of particular target genes. PMID:23620480

  9. Efficient In Planta Detection and Dissection of De Novo Mutation Events in the Arabidopsis thaliana Disease Resistance Gene UNI.

    PubMed

    Ogawa, Tomohiko; Mori, Akiko; Igari, Kadunari; Morita, Miyo Terao; Tasaka, Masao; Uchida, Naoyuki

    2016-06-01

    Plants possess disease resistance (R) proteins encoded by R genes, and each R protein recognizes a specific pathogen factor(s) for immunity. Interestingly, a remarkably high degree of polymorphisms in R genes, which are traces of past mutation events during evolution, suggest the rapid diversification of R genes. However, little is known about molecular aspects that facilitate the rapid change of R genes because of the lack of tools that enable us to monitor de novo R gene mutations efficiently in an experimentally feasible time scale, especially in living plants. Here we introduce a model assay system that enables efficient in planta detection of de novo mutation events in the Arabidopsis thaliana R gene UNI in one generation. The uni-1D mutant harbors a gain-of-function allele of the UNI gene. uni-1D heterozygous individuals originally exhibit dwarfism with abnormally short stems. However, interestingly, morphologically normal stems sometimes emerge spontaneously from the uni-1D plants, and the morphologically reverted tissues carry additional de novo mutations in the UNI gene. Strikingly, under an extreme condition, almost half of the examined population shows the reversion phenomenon. By taking advantage of this phenomenon, we demonstrate that the reversion frequency is remarkably sensitive to a variety of fluctuations in DNA stability, underlying a mutable tendency of the UNI gene. We also reveal that activities of the salicylic acid pathway and DNA damage sensor pathway are involved in the reversion phenomenon. Thus, we provide an experimentally feasible model tool to explore factors and conditions that significantly affect the R gene mutation phenomenon. PMID:27016096

  10. Consentimiento informado: una praxis dialogica para la investigacion

    PubMed Central

    Mondragon-Barrios, Liliana

    2009-01-01

    El consentimiento informado es un proceso, en el que una persona acepta participar en una investigation, conociendo los riesgos, beneficios, consecuencias o problemas que se puedan presenter durante el desarrollo de la misma. El objetivo de este trabajo es reunir las caracteristicas del proceso de consentimiento informado (PCI) pare que su discernimiento y cumplimiento posibilite el ejercicio etc° dialogico, reflexivo y responsable del investigador. Se presentan los resultados de una extensa revision de les elementos del PCI, desde sus cornponentes hasta su fundamento etico y legal, incluyendo los mites y realidades que existen sobre el formato de consentimiento informado come recurso legal de protection. El consentimiento informado no es un formato establecido que los investigadores repliquen, se trata de que la praxis del PCI sea una tarea cotidiana dentro de la investigation con seres humanos, como comunicaciOn deliberative y critica, responsable y comprometida entre dos agentes morales, investigador-investigado. PMID:19507477

  11. Methodology for producing 100 tons of fuel peat for a cement plant test burn. Metodologia para producir 100 toneladas de turba combustible para una prueba de quema en uns planta de cemento

    SciTech Connect

    Allen, A.B.; Malavassi, L.; Ramirez, O.; Thayer, G. , Plymouth, NC; RECOPE, San Jose; Los Alamos National Lab., NM )

    1989-05-01

    As a part of the Agency for International Development-funded cooperative project between Los Alamos and Costa Rica, the burning characteristics of Costa Rican peat were to be tested in an application. The cement plant owned by Industria National de Cemento in Cartago has a capability to handle solid fuel and was chosen for the burn demonstration. The Jungle No. 1 Peat Deposit near El Cairo was chosen as the site of the peat excavation. This peat production methodology study covers project site selection, installation of an access road and clearing of the jungle vegetation, removal of an upper layer of organic peat, excavation of fuel-grade peat, transport of the peat to the drying site, and drying and stockpiling of the finished product. As of this date the peat removal for the demonstration project has been started, and a description of the operation is included as an appendix to this paper. 10 figs.

  12. Bikinin-like inhibitors targeting GSK3/Shaggy-like kinases: characterisation of novel compounds and elucidation of their catabolism in planta

    PubMed Central

    2014-01-01

    Background Plant GSK-3/Shaggy-like kinases are key players in brassinosteroid (BR) signalling which impact on plant development and participate in response to wounding, pathogens and salt stress. Bikinin was previously identified in a chemical genetics screen as an inhibitor targeting these kinases. To dissect the structural elements crucial for inhibition of GSK-3/Shaggy-like kinases by bikinin and to isolate more potent compounds we synthesised a number of related substances and tested their inhibitory activity in vitro and in vivo using Arabidopsis thaliana. Results A pyridine ring with an amido succinic acid residue in position 2 and a halogen in position 5 were crucial for inhibitory activity. The compound with an iodine substituent in position 5, denoted iodobikinin, was most active in inhibiting BIN2 activity in vitro and efficiently induced brassinosteroid-like responses in vivo. Its methyl ester, methyliodobikinin, showed improved cell permeability, making it highly potent in vivo although it had lower activity in vitro. HPLC analysis revealed that the methyl residue was rapidly cleaved off in planta liberating active iodobikinin. In addition, we provide evidence that iodobikinin and bikinin are inactivated in planta by conjugation with glutamic acid or malic acid and that the latter process is catalysed by the malate transferase SNG1. Conclusion Brassinosteroids participate in regulation of many aspects of plant development and in responses to environmental cues. Thus compounds modulating their action are valuable tools to study such processes and may be an interesting opportunity to modify plant growth and performance in horticulture and agronomy. Here we report the development of bikinin derivatives with increased potency that can activate BR signalling and mimic BR action. Methyliodobikinin was 3.4 times more active in vivo than bikinin. The main reason for the superior activity of methyliodobikinin, the most potent compound, is its enhanced plant

  13. Fruta Planta label

    Center for Drug Evaluation (CDER)

    ... Fii ììE~ J jJimît'¥ (2005 J j¡0125~ ( ~!!mlE J 1, l¡íi'Iß~JI, ¡;~Wl ß~JI, F€ß~JI, ~ )X ;¡;U~ ~ 9!lC/, ~ .& lit 1!. íll;;IHH'H'H;l i' *1!o 2, ii!f., 1I11L Jl-.1' f'l!ßi J.ll 0 r ...

  14. Una búsqueda de la identidad regional em Astrometria

    NASA Astrophysics Data System (ADS)

    Abad, C.

    2003-11-01

    La idea sobre una reunión latino-americana que congregara a las personas que desarrollan su trabajo en el área de la astrometría, se gestó a partir de una de las reuniones que el proyecto QUEST realizaba anualmente en Mérida (Venezuela). El proyecto QUEST (Quasars Equatorial Survey Team) celebraba su IV Reunión Anual, en la cual se hacía una revisón de la marcha del proyecto durante el último año. Además, se exponían los logros científicos y técnicos alcanzados hasta el momento y se planificaba el futuro del mismo para el período anual siguiente.

  15. Una técnica para filtrar patrones de fringing

    NASA Astrophysics Data System (ADS)

    Ostrov, P. G.

    Se presenta una nueva técnica para filtrar los patrones de fringing producidos en los CCDs tipo RCA. El método consiste en construir un mapa con los ángulos de inclinación de las franjas en cada punto de la imagen. Este mapa es ulteriormente utilizado para alinear con el patrón de interferencia una ventana estrecha, sobre la que se aplica un filtro de mediana. Este procedimiento permite eliminar la mayor parte del ruido del patrón de fringing sin destruirlo.

  16. Use of attenuated total reflection Fourier transform infrared (ATR FT-IR) spectroscopy in direct, non-destructive, and rapid assessment of developmental cotton fibers grown in planta and in culture

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Cotton fibers are routinely harvested from cotton plants (in planta), and their end-use qualities depend on their development stages. Cotton fibers are also cultured at controlled laboratory environments, so that cotton researchers can investigate many aspects of experimental protocols in cotton bre...

  17. Transient Expression of Secretory IgA In Planta is Optimal Using a Multi-Gene Vector and may be Further Enhanced by Improving Joining Chain Incorporation

    PubMed Central

    Westerhof, Lotte B.; Wilbers, Ruud H. P.; van Raaij, Debbie R.; van Wijk, Christina Z.; Goverse, Aska; Bakker, Jaap; Schots, Arjen

    2016-01-01

    Secretory IgA (sIgA) is a crucial antibody in host defense at mucosal surfaces. It is a promising antibody isotype in a variety of therapeutic settings such as passive vaccination and treatment of inflammatory disorders. However, heterologous production of this heteromultimeric protein complex is still suboptimal. The challenge is the coordinate expression of the four required polypeptides; the alpha heavy chain, the light chain, the joining chain, and part of the polymeric-Ig-receptor called the secretory component, in a 4:4:1:1 ratio. We evaluated the transient expression of three sIgAκ variants, harboring the heavy chain isotype α1, α2m1, or α2m2, of the clinical antibody Ustekinumab in planta. Ustekinumab is directed against the p40 subunit that is shared by the pro-inflammatory cytokines interleukin (IL)-12 and IL-23. A sIgA variant of this antibody may enable localized treatment of inflammatory bowel disease. Of the three different sIgA variants we obtained the highest yield with sIgA1κ reaching up to 373 μg sIgA/mg total soluble protein. The use of a multi-cassette vector containing all four expression cassettes was most efficient. However, not the expression strategy, but the incorporation of the joining chain turned out to be the limiting step for sIgA production. Our data demonstrate that transient expression in planta is suitable for the economic production of heteromultimeric protein complexes such as sIgA. PMID:26793201

  18. Entropía de la información: una herramienta útil

    NASA Astrophysics Data System (ADS)

    Cincotta, P.

    En este trabajo se presenta una muy breve introducción al concepto de Entropía de la Información y se muestran distintas aplicaciones de una misma técnica para estudiar problemas tan variados como la determinación de periodicidad en una serie de tiempo arbitraria o de estimar el grado de caoticidad de una trayectoria en un sistema dinámico de N grados de libertad.

  19. CONTAMINACIÓN AMBIENTAL, VARIABILIDAD CLIMÁTICA Y CAMBIO CLIMÁTICO: UNA REVISIÓN DEL IMPACTO EN LA SALUD DE LA POBLACIÓN PERUANA

    PubMed Central

    Gonzales, Gustavo F.; Zevallos, Alisson; Gonzales-Castañeda, Cynthia; Nuñez, Denisse; Gastañaga, Carmen; Cabezas, César; Naeher, Luke; Levy, Karen; Steenland, Kyle

    2015-01-01

    RESUMEN El presente artículo es una revisión sobre la contaminación del agua, el aire y el efecto del cambio climático en la salud de la población peruana. Uno de los principales contaminantes del aire es el material particulado menor de 2,5 μ (PM 2,5), en la ciudad de Lima, anualmente 2300 muertes prematuras son atribuibles a este contaminante. Otro problema es la contaminación del aire domiciliario por el uso de cocinas con combustible de biomasa, donde la exposición excesiva a PM 2,5 dentro de las casas es responsable de aproximadamente 3000 muertes prematuras anuales entre adultos, con otro número desconocido de muertes entre niños debido a infecciones respiratorias. La contaminación del agua tiene como principales causas los desagües vertidos directamente a los ríos, minerales (arsénico) de varias fuentes, y fallas de las plantas de tratamiento. En el Perú, el cambio climático puede impactar en la frecuencia y severidad del fenómeno de El Niño oscilación del sur (ENSO) que se ha asociado con un incremento en los casos de enfermedades como cólera, malaria y dengue. El cambio climático incrementa la temperatura y puede extender las áreas afectadas por enfermedades transmitidas por vectores, además de tener efecto en la disponibilidad del agua y en la contaminación del aire. En conclusión, el Perú, pasa por una transición de factores de riesgo ambientales, donde coexisten riesgos tradicionales y modernos, y persisten los problemas infecciosos y crónicos, algunos de los cuales se asocian con problemas de contaminación de agua y de aire. PMID:25418656

  20. In Planta Biocontrol of Pectobacterium atrosepticum by Rhodococcus erythropolis Involves Silencing of Pathogen Communication by the Rhodococcal Gamma-Lactone Catabolic Pathway

    PubMed Central

    Barbey, Corinne; Crépin, Alexandre; Bergeau, Dorian; Ouchiha, Asma; Mijouin, Lily; Taupin, Laure; Orange, Nicole; Feuilloley, Marc; Dufour, Alain; Burini, Jean-François; Latour, Xavier

    2013-01-01

    The virulence of numerous Gram-negative bacteria is under the control of a quorum sensing process based on synthesis and perception of N-acyl homoserine lactones. Rhodococcus erythropolis, a Gram-positive bacterium, has recently been proposed as a biocontrol agent for plant protection against soft-rot bacteria, including Pectobacterium. Here, we show that the γ-lactone catabolic pathway of R. erythropolis disrupts Pectobacterium communication and prevents plant soft-rot. We report the first characterization and demonstration of N-acyl homoserine lactone quenching in planta. In particular, we describe the transcription of the R. erythropolis lactonase gene, encoding the key enzyme of this pathway, and the subsequent lactone breakdown. The role of this catabolic pathway in biocontrol activity was confirmed by deletion of the lactonase gene from R. erythropolis and also its heterologous expression in Escherichia coli. The γ-lactone catabolic pathway is induced by pathogen communication rather than by pathogen invasion. This is thus a novel and unusual biocontrol pathway, differing from those previously described as protecting plants from phytopathogens. These findings also suggest the existence of an additional pathway contributing to plant protection. PMID:23805254

  1. In Planta Biocontrol of Pectobacterium atrosepticum by Rhodococcus erythropolis Involves Silencing of Pathogen Communication by the Rhodococcal Gamma-Lactone Catabolic Pathway.

    PubMed

    Barbey, Corinne; Crépin, Alexandre; Bergeau, Dorian; Ouchiha, Asma; Mijouin, Lily; Taupin, Laure; Orange, Nicole; Feuilloley, Marc; Dufour, Alain; Burini, Jean-François; Latour, Xavier

    2013-01-01

    The virulence of numerous Gram-negative bacteria is under the control of a quorum sensing process based on synthesis and perception of N-acyl homoserine lactones. Rhodococcus erythropolis, a Gram-positive bacterium, has recently been proposed as a biocontrol agent for plant protection against soft-rot bacteria, including Pectobacterium. Here, we show that the γ-lactone catabolic pathway of R. erythropolis disrupts Pectobacterium communication and prevents plant soft-rot. We report the first characterization and demonstration of N-acyl homoserine lactone quenching in planta. In particular, we describe the transcription of the R. erythropolis lactonase gene, encoding the key enzyme of this pathway, and the subsequent lactone breakdown. The role of this catabolic pathway in biocontrol activity was confirmed by deletion of the lactonase gene from R. erythropolis and also its heterologous expression in Escherichia coli. The γ-lactone catabolic pathway is induced by pathogen communication rather than by pathogen invasion. This is thus a novel and unusual biocontrol pathway, differing from those previously described as protecting plants from phytopathogens. These findings also suggest the existence of an additional pathway contributing to plant protection. PMID:23805254

  2. Colonization process of olive tissues by Verticillium dahliae and its in planta interaction with the biocontrol root endophyte Pseudomonas fluorescens PICF7

    PubMed Central

    Prieto, Pilar; Navarro‐Raya, Carmen; Valverde‐Corredor, Antonio; Amyotte, Stefan G.; Dobinson, Katherine F.; Mercado‐Blanco, Jesús

    2009-01-01

    Summary The colonization process of Olea europaea by the defoliating pathotype of Verticillium dahliae, and the in planta interaction with the endophytic, biocontrol strain Pseudomonas fluorescens PICF7 were determined. Differential fluorescent protein tagging was used for the simultaneous visualization of P. fluorescens PICF7 and V. dahliae in olive tissues. Olive plants were bacterized with PICF7 and then transferred to V. dahliae‐infested soil. Monitoring olive colonization events by V. dahliae and its interaction with PICF7 was conducted using a non‐gnotobiotic system, confocal laser scanner microscopy and tissue vibratoming sections. A yellow fluorescently tagged V. dahliae derivative (VDAT‐36I) was obtained by Agrobacterium tumefaciens‐mediated transformation. Isolate VDAT‐36I quickly colonized olive root surface, successfully invaded root cortex and vascular tissues via macro‐ and micro‐breakages, and progressed to the aerial parts of the plant through xylem vessel cells. Strain PICF7 used root hairs as preferred penetration site, and once established on/in root tissues, hindered pathogen colonization. For the first time using this approach, the entire colonization process of a woody plant by V. dahliae is reported. Early and localized root surface and root endophytic colonization by P. fluorescens PICF7 is needed to impair full progress of verticillium wilt epidemics in olive. PMID:21255281

  3. Specific in planta recognition of two GKLR proteins of the downy mildew Bremia lactucae revealed in a large effector screen in lettuce.

    PubMed

    Stassen, Joost H M; den Boer, Erik; Vergeer, Pim W J; Andel, Annemiek; Ellendorff, Ursula; Pelgrom, Koen; Pel, Mathieu; Schut, Johan; Zonneveld, Olaf; Jeuken, Marieke J W; Van den Ackerveken, Guido

    2013-11-01

    Breeding lettuce (Lactuca sativa) for resistance to the downy mildew pathogen Bremia lactucae is mainly achieved by introgression of dominant downy mildew resistance (Dm) genes. New Bremia races quickly render Dm genes ineffective, possibly by mutation of recognized host-translocated effectors or by suppression of effector-triggered immunity. We have previously identified 34 potential RXLR(-like) effector proteins of B. lactucae that were here tested for specific recognition within a collection of 129 B. lactucae-resistant Lactuca lines. Two effectors triggered a hypersensitive response: BLG01 in 52 lines, predominantly L. saligna, and BLG03 in two L. sativa lines containing Dm2 resistance. The N-terminal sequences of BLG01 and BLG03, containing the signal peptide and GKLR variant of the RXLR translocation motif, are not required for in planta recognition but function in effector delivery. The locus responsible for BLG01 recognition maps to the bottom of lettuce chromosome 9, whereas recognition of BLG03 maps in the RGC2 cluster on chromosome 2. Lactuca lines that recognize the BLG effectors are not resistant to Bremia isolate Bl:24 that expresses both BLG genes, suggesting that Bl:24 can suppress the triggered immune responses. In contrast, lettuce segregants displaying Dm2-mediated resistance to Bremia isolate Bl:5 are responsive to BLG03, suggesting that BLG03 is a candidate Avr2 protein. PMID:23883357

  4. In Planta Stage-Specific Fungal Gene Profiling Elucidates the Molecular Strategies of Fusarium graminearum Growing inside Wheat Coleoptiles[W][OA

    PubMed Central

    Zhang, Xiao-Wei; Jia, Lei-Jie; Zhang, Yan; Jiang, Gang; Li, Xuan; Zhang, Dong; Tang, Wei-Hua

    2012-01-01

    The ascomycete Fusarium graminearum is a destructive fungal pathogen of wheat (Triticum aestivum). To better understand how this pathogen proliferates within the host plant, we tracked pathogen growth inside wheat coleoptiles and then examined pathogen gene expression inside wheat coleoptiles at 16, 40, and 64 h after inoculation (HAI) using laser capture microdissection and microarray analysis. We identified 344 genes that were preferentially expressed during invasive growth in planta. Gene expression profiles for 134 putative plant cell wall–degrading enzyme genes suggest that there was limited cell wall degradation at 16 HAI and extensive degradation at 64 HAI. Expression profiles for genes encoding reactive oxygen species (ROS)–related enzymes suggest that F. graminearum primarily scavenges extracellular ROS before a later burst of extracellular ROS is produced by F. graminearum enzymes. Expression patterns of genes involved in primary metabolic pathways suggest that F. graminearum relies on the glyoxylate cycle at an early stage of plant infection. A secondary metabolite biosynthesis gene cluster was specifically induced at 64 HAI and was required for virulence. Our results indicate that F. graminearum initiates infection of coleoptiles using covert penetration strategies and switches to overt cellular destruction of tissues at an advanced stage of infection. PMID:23266949

  5. [Valutare le esperienze positive e negative: la validazione di una nuova misura del benessere in una popolazione italiana].

    PubMed

    Corno, Giulia; Molinari, Guadalupe; Baños, Rosa Maria

    2016-01-01

    RIASSUNTO. Lo scopo del presente studio è quello di esplorare le proprietà psicometriche di uno strumento di misurazione dell'affetto, la Scala di Esperienze Positive e Negative (SPANE), all'interno di una popolazione italiana. Gli esiti dell'analisi fattoriale confermativa comprovano l'attesa struttura a due fattori, sentimenti positivi e negativi. Le correlazioni con altre dimensioni (per es., ansia, depressione, affetto, aspettative future) confermano i risultati ottenuti con le precedenti versioni della scala: affetto negativo, ansia, depressione e aspettative future negative correlano positivamente con la subscala di esperienze negative e negativamente con la subscala di esperienze positive dello SPANE. In conclusione, i risultati del nostro studio dimostrano che la versione italiana dello SPANE presenta caratteristiche psicometriche simili a quelle mostrate dalla versione orginale e da successive validazioni dello strumento in altre lingue. La scala presenta, inoltre, affidabilità e validità fattoriale. Lo SPANE è un indice utile dal punto di vista clinico che può fornire informazioni rilevanti circa l'esperienza emotiva e il benessere della persona. Nonostante ulteriori studi siano necessari per confermare le caratteristiche psicometriche della scala, la presente validazione della versione italiana dello SPANE può contribuire ad ampliare la ricerca nell'ambito del benessere in una popolazione Italiana. PMID:27362822

  6. Radioastronomía: Una Mirada Más Amplia

    NASA Astrophysics Data System (ADS)

    Bianchi, Viviana

    2004-12-01

    The existence of the celestial bodies is known due to the light they emit. But light is a small part of a bigger phenomenon known as electromagnetic radiation. The representation of all kinds of electromagnetic radiation as a function of its frequency is called the electromagnetic spectrum. If we only studied the visible part of the spectrum, we would ignore a great deal of information. The celestial bodies emit radiation in all the regions of the electromagnetic spectrum, with very different intensity. To capture these data, astronomers design new types of telescopes that capture radiation at different wavelengths, among them the radiotelescopes that, as the name indicates, detect radio waves. The radiotelescope can be compared to a regular radio set. A radio station sends information by means of radio waves. The radio set captures these waves by means of its antenna, processes it and reproduces the information (music, etc.) in audible form. In the field of radioastronomy, interesting observations can be carried out: radio emission from stars, nebulas, pulsars, distant galaxies and stars, radiogalaxies, the Milky Way, the Sun, Jupiter, etc. La presencia de los cuerpos celestes se conoce gracias a que emiten luz. Pero la luz constituye solo una pequeña parte de un fenómeno mucho mas amplio conocido como radiación electromagnética. La representación de todas las clases de radiación como función de su frecuencia se denomina espectro electromagnético. Si solo estudiáramos la parte visual del espectro ignoraríamos una gran cantidad de información. Los cuerpos celestes emiten radiación en todas las regiones del espectro electromagnético, aunque con muy distinta intensidad. Para captar estos datos, los astrónomos deben diseñar nuevos tipos de telescopios que puedan captar la radiación de distintas longitudes de ondas, entre ellos los radiotelescopios, qué como su nombre lo indica detectan ondas de radio. El radiotelescopio puede compararse a un aparato

  7. Endogenous silencing of Puccinia triticina pathogenicity genes through in planta-expressed sequences leads to the suppression of rust diseases on wheat.

    PubMed

    Panwar, Vinay; McCallum, Brent; Bakkeren, Guus

    2013-02-01

    Rust fungi are destructive plant pathogens. The draft genomes of several wheat-infecting species have been released and potential pathogenicity genes identified through comparative analyses to fungal pathogens that are amenable to genetic manipulation. Functional gene analysis tools are needed to understand the infection process of these obligate parasites and to confirm whether predicted pathogenicity genes could become targets for disease control. We have modified an Agrobacterium tumefaciens-mediated in planta-induced transient gene silencing (PITGS) assay for use in Triticum spp. (wheat), and used this assay to target predicted wheat leaf rust fungus, Puccinia triticina (Pt) pathogenicity genes, a MAP kinase (PtMAPK1), a cyclophilin (PtCYC1) and calcineurin B (PtCNB), to analyze their roles in disease. Agroinfiltration effectively delivered hairpin silencing constructs in wheat, leading to the generation of fungal gene-specific siRNA molecules in infiltrated leaves, and resulting in up to 70% reduction in transcription of the endogenous target genes in superinfected Pt. In vivo silencing caused severe disease suppression, compromising fungal growth and sporulation, as viewed by confocal microscopy and measured by reductions in fungal biomass and emergence of uredinia. Interestingly, using the same gene constructs, suppression of infection by Puccinia graminis and Puccinia striiformis was also achieved. Our results show that A. tumefaciens-mediated PITGS can be used as a reverse-genetics tool to discover gene function in rust fungi. This proof-of-concept study indicates that the targeted fungal transcripts might be important in pathogenesis, and could potentially be used as promising targets for developing RNA interference-based resistance against rust fungi. PMID:23110316

  8. Boosting In Planta Production of Antigens Derived from the Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) and Subsequent Evaluation of Their Immunogenicity

    PubMed Central

    Piron, Robin; De Koker, Stefaan; De Paepe, Annelies; Goossens, Julie; Grooten, Johan; Nauwynck, Hans; Depicker, Ann

    2014-01-01

    Porcine reproductive and respiratory syndrome (PRRS) is a disease of swine, caused by an arterivirus, the PRRS virus (PRRSV). This virus infects pigs worldwide and causes huge economic losses. Due to genetic drift, current vaccines are losing their power. Adaptable vaccines could provide a solution to this problem. This study aims at producing in planta a set of antigens derived from the PRRSV glycoproteins (GPs) to be included in a subunit vaccine. We selected the GP3, GP4 and GP5 and optimized these for production in an Arabidopsis seed platform by removing transmembrane domains (Tm) and/or adding stabilizing protein domains, such as the green fluorescent protein (GFP) and immunoglobulin (IgG) ‘Fragment crystallizable’ (Fc) chains. Accumulation of the GPs with and without Tm was low, reaching no more than 0.10% of total soluble protein (TSP) in homozygous seed. However, addition of stabilizing domains boosted accumulation up to a maximum of 2.74% of TSP when GFP was used, and albeit less effectively, also the Fc chains of the porcine IgG3 and murine IgG2a increased antigen accumulation, to 0.96% and 1.81% of TSP respectively, while the murine IgG3 Fc chain did not. Antigens with Tm were less susceptible to these manipulations to increase yield. All antigens were produced in the endoplasmic reticulum and accordingly, they carried high-mannose N-glycans. The immunogenicity of several of those antigens was assessed and we show that vaccination with purified antigens did elicit the production of antibodies with virus neutralizing activity in mice but not in pigs. PMID:24614617

  9. A ubiquitin carboxyl extension protein secreted from a plant-parasitic nematode Globodera rostochiensis is cleaved in planta to promote plant parasitism.

    PubMed

    Chronis, Demosthenis; Chen, Shiyan; Lu, Shunwen; Hewezi, Tarek; Carpenter, Sara C D; Loria, Rosemary; Baum, Thomas J; Wang, Xiaohong

    2013-04-01

    Nematode effector proteins originating from esophageal gland cells play central roles in suppressing plant defenses and in formation of the plant feeding cells that are required for growth and development of cyst nematodes. A gene (GrUBCEP12) encoding a unique ubiquitin carboxyl extension protein (UBCEP) that consists of a signal peptide for secretion, a mono-ubiquitin domain, and a 12 amino acid carboxyl extension protein (CEP12) domain was cloned from the potato cyst nematode Globodera rostochiensis. This GrUBCEP12 gene was expressed exclusively within the nematode's dorsal esophageal gland cell, and was up-regulated in the parasitic second-stage juvenile, correlating with the time when feeding cell formation is initiated. We showed that specific GrUBCEP12 knockdown via RNA interference reduced nematode parasitic success, and that over-expression of the secreted Gr(Δ) (SP) UBCEP12 protein in potato resulted in increased nematode susceptibility, providing direct evidence that this secreted effector is involved in plant parasitism. Using transient expression assays in Nicotiana benthamiana, we found that Gr(Δ) (SP) UBCEP12 is processed into free ubiquitin and a CEP12 peptide (GrCEP12) in planta, and that GrCEP12 suppresses resistance gene-mediated cell death. A target search showed that expression of RPN2a, a gene encoding a subunit of the 26S proteasome, was dramatically suppressed in Gr(Δ) (SP) UBCEP12 but not GrCEP12 over-expression plants when compared with control plants. Together, these results suggest that, when delivered into host plant cells, Gr(Δ) (SP) UBCEP12 becomes two functional units, one acting to suppress plant immunity and the other potentially affecting the host 26S proteasome, to promote feeding cell formation. PMID:23346875

  10. Virulence and in planta movement of Xanthomonas hortorum pv. pelargonii are affected by the diffusible signal factor (DSF)-dependent quorum sensing system.

    PubMed

    Barel, Victoria; Chalupowicz, Laura; Barash, Isaac; Sharabani, Galit; Reuven, Michal; Dror, Orit; Burdman, Saul; Manulis-Sasson, Shulamit

    2015-09-01

    Xanthomonas hortorum pv. pelargonii (Xhp), the causal agent of bacterial blight in pelargonium, is the most threatening bacterial disease of this ornamental worldwide. To gain an insight into the regulation of virulence in Xhp, we have disrupted the quorum sensing (QS) genes, which mediate the biosynthesis and sensing of the diffusible signal factor (DSF). Mutations in rpfF (encoding the DSF synthase) and rpfC (encoding the histidine sensor kinase of the two-component system RfpC/RpfG) and overexpression of rpfF showed a significant reduction in incidence and severity of the disease on pelargonium. Confocal laser scanning microscopy images of inoculated plants with a green fluorescent protein (GFP)-labelled wild-type strain showed that the pathogen is homogeneously dispersed in the lumen of xylem vessels, reaching the apex and invading the intercellular spaces of the leaf mesophyll tissue within 21 days. In contrast, the rpfF and rpfC knockout mutants, as well as the rpfF-overexpressing strain, remained confined to the vicinity of the inoculation site. The rpfF and rpfC mutants formed large incoherent aggregates in the xylem vessels that might interfere with upward movement of the bacterium within the plant. Both mutants also formed extended aggregates under in vitro conditions, whereas the wild-type strain formed microcolonies. Expression levels of putative virulence genes in planta were substantially reduced within 48 h after inoculation with the QS mutants when compared with the wild-type. The results presented indicate that an optimal DSF concentration is crucial for successful colonization and virulence of Xhp in pelargonium. PMID:25530086

  11. RiPEIP1, a gene from the arbuscular mycorrhizal fungus Rhizophagus irregularis, is preferentially expressed in planta and may be involved in root colonization.

    PubMed

    Fiorilli, Valentina; Belmondo, Simone; Khouja, Hassine Radhouane; Abbà, Simona; Faccio, Antonella; Daghino, Stefania; Lanfranco, Luisa

    2016-08-01

    Transcriptomics and genomics data recently obtained from the arbuscular mycorrhizal (AM) fungus Rhizophagus irregularis have offered new opportunities to decipher the contribution of the fungal partner to the establishment of the symbiotic association. The large number of genes which do not show similarity to known proteins witnesses the uniqueness of this group of plant-associated fungi. In this work, we characterize a gene that was called RiPEIP1 (Preferentially Expressed In Planta). Its expression is strongly induced in the intraradical phase, including arbuscules, and follows the expression profile of the Medicago truncatula phosphate transporter MtPT4, a molecular marker of a functional symbiosis. Indeed, mtpt4 mutant plants, which exhibit low mycorrhizal colonization and an accelerated arbuscule turnover, also show a reduced RiPEIP1 mRNA abundance. To further characterize RiPEIP1, in the absence of genetic transformation protocols for AM fungi, we took advantage of two different fungal heterologous systems. When expressed as a GFP fusion in yeast cells, RiPEIP1 localizes in the endomembrane system, in particular to the endoplasmic reticulum, which is consistent with the in silico prediction of four transmembrane domains. We then generated RiPEIP1-expressing strains of the fungus Oidiodendron maius, ericoid endomycorrhizal fungus for which transformation protocols are available. Roots of Vaccinium myrtillus colonized by RiPEIP1-expressing transgenic strains showed a higher mycorrhization level compared to roots colonized by the O. maius wild-type strain, suggesting that RiPEIP1 may regulate the root colonization process. PMID:27075897

  12. In planta assessment of the role of thioredoxin h proteins in the regulation of S-locus receptor kinase signaling in transgenic Arabidopsis.

    PubMed

    Yamamoto, Masaya; Nasrallah, June B

    2013-11-01

    The self-incompatibility (SI) response of the Brassicaceae is mediated by allele-specific interaction between the stigma-localized S-locus receptor kinase (SRK) and its ligand, the pollen coat-localized S-locus cysteine-rich protein (SCR). Based on work in Brassica spp., the thioredoxin h-like proteins THL1 and THL2, which interact with SRK, have been proposed to function as oxidoreductases that negatively regulate SRK catalytic activity. By preventing the spontaneous activation of SRK in the absence of SCR ligand, these thioredoxins are thought to be essential for the success of cross pollinations in self-incompatible plants. However, the in planta role of thioredoxins in the regulation of SI signaling has not been conclusively demonstrated. Here, we addressed this issue using Arabidopsis thaliana plants transformed with the SRKb-SCRb gene pair isolated from self-incompatible Arabidopsis lyrata. These plants express an intense SI response, allowing us to exploit the extensive tools and resources available in A. thaliana for analysis of SI signaling. To test the hypothesis that SRK is redox regulated by thioredoxin h, we expressed a mutant form of SRKb lacking a transmembrane-localized cysteine residue thought to be essential for the SRK-thioredoxin h interaction. We also analyzed transfer DNA insertion mutants in the A. thaliana orthologs of THL1 and THL2. In neither case did we observe an effect on the pollination responses of SRKb-expressing stigmas toward incompatible or compatible pollen. Our results are consistent with the conclusion that, contrary to their proposed role, thioredoxin h proteins are not required to prevent the spontaneous activation of SRK in the A. thaliana stigma. PMID:24077073

  13. Boosting in planta production of antigens derived from the porcine reproductive and respiratory syndrome virus (PRRSV) and subsequent evaluation of their immunogenicity.

    PubMed

    Piron, Robin; De Koker, Stefaan; De Paepe, Annelies; Goossens, Julie; Grooten, Johan; Nauwynck, Hans; Depicker, Ann

    2014-01-01

    Porcine reproductive and respiratory syndrome (PRRS) is a disease of swine, caused by an arterivirus, the PRRS virus (PRRSV). This virus infects pigs worldwide and causes huge economic losses. Due to genetic drift, current vaccines are losing their power. Adaptable vaccines could provide a solution to this problem. This study aims at producing in planta a set of antigens derived from the PRRSV glycoproteins (GPs) to be included in a subunit vaccine. We selected the GP3, GP4 and GP5 and optimized these for production in an Arabidopsis seed platform by removing transmembrane domains (Tm) and/or adding stabilizing protein domains, such as the green fluorescent protein (GFP) and immunoglobulin (IgG) 'Fragment crystallizable' (Fc) chains. Accumulation of the GPs with and without Tm was low, reaching no more than 0.10% of total soluble protein (TSP) in homozygous seed. However, addition of stabilizing domains boosted accumulation up to a maximum of 2.74% of TSP when GFP was used, and albeit less effectively, also the Fc chains of the porcine IgG3 and murine IgG2a increased antigen accumulation, to 0.96% and 1.81% of TSP respectively, while the murine IgG3 Fc chain did not. Antigens with Tm were less susceptible to these manipulations to increase yield. All antigens were produced in the endoplasmic reticulum and accordingly, they carried high-mannose N-glycans. The immunogenicity of several of those antigens was assessed and we show that vaccination with purified antigens did elicit the production of antibodies with virus neutralizing activity in mice but not in pigs. PMID:24614617

  14. Tdp studies and tests for C. A. Energia Electrica de Venezuela (enelven) at planta ramon laguna, units RL-17 and RL-10. Volume 2. Unit RL-10 boiler condition assessment report. Export trade information

    SciTech Connect

    Not Available

    1991-03-28

    The study, conducted by Babcock and Wilcox, was funded by the U.S. Trade and Development agency on behalf of Enelven. In order to maximize generated power output and minimize operating costs at Planta Ramon Laguna, tests were done to evaluate the condition of equipment at the plant. In order to identify any damage and determine the operating output of each unit, assessments were done of the furnaces, boilers, generators and boiler feed pumps being used in the plant. The report presents the results of these tests. This is the second of three volumes and it includes the following section: (1) Condition Assessment of Unit RL-10 Boiler.

  15. Differential subcellular targeting of recombinant human α₁-proteinase inhibitor influences yield, biological activity and in planta stability of the protein in transgenic tomato plants.

    PubMed

    Jha, Shweta; Agarwal, Saurabh; Sanyal, Indraneel; Jain, G K; Amla, D V

    2012-11-01

    The response of protein accumulation site on yield, biological activity and in planta stability of therapeutic recombinant human proteinase inhibitor (α₁-PI) was analyzed via targeting to different subcellular locations, like endoplasmic reticulum (ER), apoplast, vacuole and cytosol in leaves of transgenic tomato plants. In situ localization of the recombinant α₁-PI protein in transgenic plant cells was monitored by immunohistochemical staining. Maximum accumulation of recombinant α₁-PI in T₀ and T₁ transgenic tomato plants was achieved from 1.5 to 3.2% of total soluble protein (TSP) by retention in ER lumen, followed by vacuole and apoplast, whereas cytosolic targeting resulted into degradation of the protein. The plant-derived recombinant α₁-PI showed biological activity for elastase inhibition, as monitored by residual porcine pancreatic elastase (PPE) activity assay and band-shift assay. Recombinant α₁-PI was purified from transgenic tomato plants with high yield, homogeneity and biological activity. Purified protein appeared as a single band of ∼48-50 kDa on SDS-PAGE with pI value ranging between 5.1 and 5.3. Results of mass spectrometry and optical spectroscopy of purified recombinant α₁-PI revealed the structural integrity of the recombinant protein comparable to native serum α₁-PI. Enzymatic deglycosylation and lectin-binding assays with the purified recombinant α₁-PI showed compartment-specific N-glycosylation of the protein targeted to ER, apoplast and vacuole. Conformational studies based on urea-induced denaturation and circular dichroism (CD) spectroscopy revealed relatively lower stability of the recombinant α₁-PI protein, compared to its serum counterpart. Pharmacokinetic evaluation of plant derived recombinant and human plasma-purified α₁-PI in rat, by intravenous route, revealed significantly faster plasma clearance and lower area under curve (AUC) of recombinant protein. Our data suggested significance of

  16. Genomic homologous recombination in planta.

    PubMed Central

    Gal, S; Pisan, B; Hohn, T; Grimsley, N; Hohn, B

    1991-01-01

    A system for monitoring intrachromosomal homologous recombination in whole plants is described. A multimer of cauliflower mosaic virus (CaMV) sequences, arranged such that CaMV could only be produced by recombination, was integrated into Brassica napus nuclear DNA. This set-up allowed scoring of recombination events by the appearance of viral symptoms. The repeated homologous regions were derived from two different strains of CaMV so that different recombinant viruses (i.e. different recombination events) could be distinguished. In most of the transgenic plants, a single major virus species was detected. About half of the transgenic plants contained viruses of the same type, suggesting a hotspot for recombination. The remainder of the plants contained viruses with cross-over sites distributed throughout the rest of the homologous sequence. Sequence analysis of two recombinant molecules suggest that mismatch repair is linked to the recombination process. Images PMID:2026150

  17. Tdp studies and tests for C. A. Energia Electrica de Venezuela (enelven) at planta ramon laguna, units RL-17 and RL-10. Volume 1. Executive summary, RL-17 test report, and gas conversion proposals. Export trade information

    SciTech Connect

    Not Available

    1991-03-28

    The study, conducted by Babcock and Wilcox, was funded by the U.S. Trade and Development agency on behalf of Enelven. In order to maximize generated power output and minimize operating costs at Planta Ramon Laguna, tests were done to evaluate the condition of equipment at the plant. In order to identify any damage and determine the operating output of each unit, assessments were done of the furnaces, boilers, generators and boiler feed pumps being used in the plant. The report presents the results of these tests. This is the first of three volumes and it is divided into the following sections: (1) Executive Summary; (2) Hydrogen Damage Assessment; (3) RL-17 Gas Conversion Proposal; (4) RL-10 and RL-11 Gas Conversion Proposals.

  18. Una propuesta para el desarrollo de un arreglo de síntesis de apertura

    NASA Astrophysics Data System (ADS)

    Arnal, E. M.

    Los estudios llevados a cabo en la transición del hidrógeno neutro a λ~21-cm han contribuído a incrementar nuestro conocimiento acerca de las propiedades globales del medio interestelar, sea este galáctico o extragaláctico. Avances en este campo han sido provocados, a menudo, por la puesta en servicio de radiotelescopios que poseen una mayor resolución angular. Aquí se presenta una propuesta para desarrollar un nuevo instrumento, un interferómetro, que permitirá abrir nuevas líneas de investigación. Este instrumento combinará la técnica de síntesis de apertura con la de espectroscopía de correlación digital, para alcanzar una resolución angular de 1' y un campo de visión de ~1o.7.

  19. Cultivating "Una Persona Educada: A Sentipensante" (Sensing/Thinking) Vision of Education

    ERIC Educational Resources Information Center

    Rendon, Laura I.

    2011-01-01

    This essay focuses on the need to educate the new "persona educada", a dignified, honorable person with a good measure of social and personal responsibility who also possesses the habits of the mind and heart. To cultivate "una persona educada" requires a newly formed vision of education and pedagogy. Examples of three entrenched agreements that…

  20. Batkoa apiculata (Thaxter) Humber affecting Anopheles (Diptera: Culicidae) in the municipality of Una, Southern Bahia, Brazil

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Surveys for fungal pathogens affecting adult mosquitoes from the genus Anopheles were conducted in flooded and swamp-like natural breeding sites near residences in the center and suburbs of the city of Una as well as the nearby village of Outeiro in southern Bahia. Surveys of 54 mosquito breeding si...

  1. Landscape Terms and Place Names in the Trobriand Islands--The Kaile'una Subset

    ERIC Educational Resources Information Center

    Senft, Gunter

    2008-01-01

    After a brief introduction to the topic the paper first gives an overview of Kilivila landscape terms and then presents the inventory of names for villages, wells, island points, reef-channels and gardens on Kaile'una Island, one of the Trobriand Islands in the Milne Bay Province of Papua New Guinea. The data on the meaning of the place names…

  2. An updated model for nitrate uptake modelling in plants. I. Functional component: cross-combination of flow–force interpretation of nitrate uptake isotherms, and environmental and in planta regulation of nitrate influx

    PubMed Central

    Le Deunff, Erwan; Malagoli, Philippe

    2014-01-01

    Background and Aims In spite of major breakthroughs in the last three decades in the identification of root nitrate uptake transporters in plants and the associated regulation of nitrate transport activities, a simplified and operational modelling approach for nitrate uptake is still lacking. This is due mainly to the difficulty in linking the various regulations of nitrate transport that act at different levels of time and on different spatial scales. Methods A cross-combination of a Flow–Force approach applied to nitrate influx isotherms and experimentally determined environmental and in planta regulation is used to model nitrate in oilseed rape, Brassica napus. In contrast to ‘Enzyme–Substrate’ interpretations, a Flow–Force modelling approach considers the root as a single catalytic structure and does not infer hypothetical cellular processes among nitrate transporter activities across cellular layers in the mature roots. In addition, this approach accounts for the driving force on ion transport based on the gradient of electrochemical potential, which is more appropriate from a thermodynamic viewpoint. Key Results and Conclusions Use of a Flow–Force formalism on nitrate influx isotherms leads to the development of a new conceptual mechanistic basis to model more accurately N uptake by a winter oilseed rape crop under field conditions during the whole growth cycle. This forms the functional component of a proposed new structure–function mechanistic model of N uptake. PMID:24638820

  3. In Planta Mutagenesis of Src Homology 3 Domain-like Fold of NdhS, a Ferredoxin-binding Subunit of the Chloroplast NADH Dehydrogenase-like Complex in Arabidopsis

    PubMed Central

    Yamamoto, Hiroshi; Shikanai, Toshiharu

    2013-01-01

    Chloroplast NADH dehydrogenase-like (NDH) complex mediates cyclic electron transport around photosystem I and chlororespiration in angiosperms. The Src homology 3 domain (SH3)-like fold protein NdhS/CRR31 is an NDH subunit that is necessary for high affinity binding of ferredoxin, indicating that chloroplast NDH functions as a ferredoxin:plastoquinone oxidoreductase. However, the mechanism of the interaction between NdhS and ferredoxin is unclear. In this study, we analyzed their interaction in planta by using site-directed mutagenesis of NdhS. In general, binding of ferredoxin to its target proteins depends on electrostatic interaction. In silico analysis predicted the presence of a positively charged pocket in the SH3-like domain of NdhS, where nine charged residues are highly conserved among plants. Systematic alteration of these sites with neutral glutamine revealed that only arginine 193 was required for high NDH activity in vivo. Further replacement of arginine 193 with negatively charged aspartate or glutamate or hydrophobic alanine significantly decreased the efficiency of ferredoxin-dependent plastoquinone reduction by NDH in ruptured chloroplasts. Similar results were obtained in in vivo analyses of NDH activity and electron transport. From these results, we propose that the positive charge of arginine 193 in the SH3-like domain of NdhS is critical for electrostatic interaction with ferredoxin in vivo. PMID:24225949

  4. Assessing channel migration on decadal scales at Una inlet (Peruíbe, Brazil)

    NASA Astrophysics Data System (ADS)

    Chagas, F. M.; Ribeiro, J.; Italiani, D. M.; Siegle, E.; Coastal Dynamics Laboratory

    2013-05-01

    Inlets are often cited as ecologically important areas, been frequent subject of study in marine sciences and engineering. Controlling forces of these complex systems are the tidal variation, waves, sediment budget, river outflow and local morphology. Over years, migration can occur alongshore or crosshore as dynamic equilibrium occurs or one of dominant forces varies. The goal of this study is to analyze decadal migration of Una inlet over 1962 to 2006, by means of aerial photographs comparison and computational measurements of coastline variation, indicating physical controlling patterns that act over southern coastal area of Sao Paulo state. Una River (also locally named as Una do Prelado) is part of the main drainage basin of Juréia Ecological Reserve at Peruíbe, southern coast of Sao Paulo state, Brazil. It is 57 km long surrounding almost completely the Juréia Massif, reaching the sea through the Barra do Una inlet. Morphologically, Una beach at Barra do Una can be classified as intermediate or dissipative, composed mainly by very fine sand, well selected due to waves and microtidal currents. Available aerial photographs range from 1962 to 2006. These images have been analyzed and three mosaics were selected to represent the decadal evolution. Images were georeferenced through the ENVI software by RST method using nearest neighbor interpolation. Both inlet margins were digitalized considering the vegetation line as limit border of water-land. For migration calculations, a hand-picked longitudinal transect was established over the inlet, in which 10 parallel segments automatic calculate the horizontal variations over digitalized data at ArgGIS 9.3 system. Through this method, the main direction of channel migration (lateral movement southwest-northeast) was obtained. Upstream transects showed small variations of line margins, typically under 40 m, indicating a relative lateral stability area on the inlet. From transect #4 downstream direction, margin

  5. La binaria LSS 3074 y su entorno: ?`una nueva asociación OB?

    NASA Astrophysics Data System (ADS)

    Niemela, V.; Morrell, N.; Corti, M.

    En este trabajo presentamos un nuevo análisis orbital de LSS~3074, junto con tipos espectrales y velocidades radiales de estrellas que podrían constituir con ella una nueva asociación OB. La estrella O4f LSS3074 fue descubierta como binaria espectroscópica de corto período y líneas dobles por Morrell & Niemela (1990, ASP Conf. Ser. 7, 57). Posteriormente, Haefner et~al.(1994, IBVS 3969) encontraron variaciones fotométricas, estimando una inclinación orbital entre 50o y 55o. Teniendo en cuenta la importancia de obtener valores empíricos para las masas de estrellas O tempranas, y considerando la gran dispersión existente entre los valores observados y su discrepancia con los predichos por los modelos teóricos, hemos obtenido nuevas observaciones espectroscópicas de este sistema, con el propósito de mejorar los elementos orbitales derivados en la solución preliminar. Además, como las estrellas O tempranas suelen formar parte de cúmulos y asociaciones OB, hemos llevado a cabo una investigación espectroscópica de varias estrellas tempranas que podrían estar físicamente relacionadas con LSS~3074.

  6. A gene in the Pseudomonas syringae pv. tomato Hrp pathogenicity island conserved effector locus, hopPtoA1, contributes to efficient formation of bacterial colonies in planta and is duplicated elsewhere in the genome.

    PubMed

    Badel, J L; Charkowski, A O; Deng, W L; Collmer, A

    2002-10-01

    The ability of Pseudomonas syringae to grow in planta is thought to be dependent upon the Hrp (type III secretion) system and multiple effector proteins that this system injects into plant cells. ORF5 in the conserved effector locus of the P. syringae pv. tomato DC3000 Hrp pathogenicity island was shown to encode a Hrp-secreted protein and to have a similarly secreted homolog encoded in an effector-rich pathogenicity island located elsewhere in the genome. These putative effector genes were designated hopPtoA1 and hopPtoA2, respectively. DNA gel blot analysis revealed that sequences hybridizing with hopPtoA1 were widespread among P. syringae pathovars, and some strains, like DC3000, appear to have two copies of the gene. uidA transcriptional fusions revealed that expression of hopPtoA1 and hopPtoA2 can be activated by the HrpL alternative sigma factor. hopPtoA1 and hopPtoA1/hopPtoA2 double mutants were not obviously different from wild-type P. syringae pv. tomato DC3000 in their ability to produce symptoms or to increase their total population size in host tomato and Arabidopsis leaves. However, confocal laser-scanning microscopy of GFP (green fluorescent protein)-labeled bacteria in Arabidopsis leaves 2 days after inoculation revealed that the frequency of undeveloped individual colonies was higher in the hopPtoA1 mutant and even higher in the hopPtoA1/hopPtoA2 double mutant. These results suggest that hopPtoA1 and hopPtoA2 contribute redundantly to the formation of P. syringae pv. tomato DC3000 colonies in Arabidopsis leaves. PMID:12437299

  7. In planta processing and glycosylation of a nematode CLAVATA3/ENDOSPERM SURROUNDING REGION-like effector and its interaction with a host CLAVATA2-like receptor to promote parasitism.

    PubMed

    Chen, Shiyan; Lang, Ping; Chronis, Demosthenis; Zhang, Sheng; De Jong, Walter S; Mitchum, Melissa G; Wang, Xiaohong

    2015-01-01

    Like other biotrophic plant pathogens, plant-parasitic nematodes secrete effector proteins into host cells to facilitate infection. Effector proteins that mimic plant CLAVATA3/ENDOSPERM SURROUNDING REGION-related (CLE) proteins have been identified in several cyst nematodes, including the potato cyst nematode (PCN); however, the mechanistic details of this cross-kingdom mimicry are poorly understood. Plant CLEs are posttranslationally modified and proteolytically processed to function as bioactive ligands critical to various aspects of plant development. Using ectopic expression coupled with nanoliquid chromatography-tandem mass spectrometry analysis, we show that the in planta mature form of proGrCLE1, a multidomain CLE effector secreted by PCN during infection, is a 12-amino acid arabinosylated glycopeptide (named GrCLE1-1Hyp4,7g) with striking structural similarity to mature plant CLE peptides. This glycopeptide is more resistant to hydrolytic degradation and binds with higher affinity to a CLAVATA2-like receptor (StCLV2) from potato (Solanum tuberosum) than its nonglycosylated forms. We further show that StCLV2 is highly up-regulated at nematode infection sites and that transgenic potatoes with reduced StCLV2 expression are less susceptible to PCN infection, indicating that interference of the CLV2-mediated signaling pathway confers nematode resistance in crop plants. These results strongly suggest that phytonematodes have evolved to utilize host cellular posttranslational modification and processing machinery for the activation of CLE effectors following secretion into plant cells and highlight the significance of arabinosylation in regulating nematode CLE effector activity. Our finding also provides evidence that multidomain CLEs are modified and processed similarly to single-domain CLEs, adding new insight into CLE maturation in plants. PMID:25416475

  8. Pasado, presente y futuro de la epidemiología. Una perspective latinoamericana

    PubMed Central

    Morabia, Alfredo

    2015-01-01

    Este artículo intenta contestar tres preguntas. Sobre el pasado: ¿Por qué no existió una epidemiología precolombina? Sobre el presente: ¿Cuáles son los orígenes de la epidemiología moderna, incluyendo sus raíces sudamericanas? Y sobre el futuro, escogí un título surrealista para enfatizar el hecho que estoy consciente de que es siempre delicado hacer predicciones: ¿Por qué fenómenos complejos son los “objetos oscuros del deseo” epidemiológico? PMID:25124247

  9. Heterochromatin heterogeneity in Hypostomus prope unae (Steindachner, 1878) (Siluriformes, Loricariidae)from Northeastern Brazil

    PubMed Central

    Bitencourt, J.A.; Affonso, P.R.A.M.; Giuliano-Caetano, L.; Dias, A.L.

    2011-01-01

    Abstract Cytogenetic analyses using C-banding and chromosomal digestion by several restriction enzymes were carried out in four populations (named A, B, C and D) of Hypostomus prope unae (Loricariidae, Hypostominae) from Contas river basin, northeastern Brazil. These populations share 2n=76 and single NORs on the second metacentric pair but exclusive karyotype forms for each locality. Populations A and B presented conspicuous terminal and interstitial heterochromatic blocks on most of acrocentric chromosomes and equivalent to NORs with differences in both position and bearing pair. Population D showed evident marks at interstitial regions and interspersed with nucleolar region while population C presented interstitial and terminal heterochromatin segments, non-coincident with NORs. The banding pattern after digestion with the endonucleases Alu I, Bam HI, Hae III and Dde I revealed a remarkable heterogeneity within heterochromatin, allowing the identification of distinctive clusters of repeated DNA in the studied populations, besides specific patterns along euchromatic regions. The analysis using restriction enzymes has proved to be highly informative, characterizing population differences and peculiarities in the genome organization of Hypostomus prope unae. PMID:24260639

  10. Observaciones del disco solar y de una protuberancia quiescente en radiación ultravioleta

    NASA Astrophysics Data System (ADS)

    Cirigliano, D.; Vial, J.-C.; Rovira, M.

    Observaciones del disco solar y de una protuberancia quiescente en el rango de longitudes de onda ultravioleta fueron obtenidas con el instrumento CDS (Coronal Diagnostic Spectrograph) y SUMER (Solar Ultraviolet Measurements of emitted radiation) a bordo de la sonda SOHO. El propósito es investigar las velocidades macroscópicas de varias especies metálicas que se observan tanto en el disco solar como en el plasma de las protuberancias. Para calcular las velocidades del disco solar aplicamos una técnica mixta para modelar la distribución de estructuras en UV en el Sol quieto. Las velocidades macroscópicas en las protuberancias se calcularon a partir de los corrimientos Doppler en cada línea espectral y luego se tomaron las del disco solar como referencia. Obtuvimos valores absolutos para las velocidades macroscópicas entre 5 y 40 km/seg. También detectamos comportamientos diferentes en las velocidades de las protuberancias en el centro con respecto a los bordes.

  11. Desarrollo curricular, conciencia ambiental y tecnologia para estudiantes de intermedia: Una investigacion en accion

    NASA Astrophysics Data System (ADS)

    Rodriguez Ramos, Teresita

    Se llevó a cabo una investigación en acción con los propósitos de 1) documentar las relaciones de las tecnologías de la información y la comunicación en las clases de ciencias de escuela intermedia como elemento de apoyo cuando se aborda el tema ambiental y sus conceptos pertinentes, a partir de las observaciones de la investigadora, así como las entrevistas y diarios reflexivos de los estudiantes de una escuela intermedia en la zona metropolitana, y luego 2) diseñar una unidad instruccional sobre el tema ambiental que integre actividades tecnologías para el curso de ciencias de la escuela intermedia según el modelo PROCIC y las observaciones que hayan iniciado los estudiantes participantes. Finalmente, se plantearon las implicaciones educativas para el currículo del Programa de Ciencias al instrumentar este modelo de unidad mediante PROCIC, e integrado la tecnología y el tema ambiental. Los hallazgos se analizaron y se categorizaron de acuerdo con las preguntas de investigación. El hallazgo principal de la investigación aborda las cuatro relaciones centrales en las que se articula la utilización de las tecnologías y sus aplicaciones en la clase de ciencias. Estas cuatro relaciones que recogen la posición de los estudiantes son: 1) Perspectiva de los estudiantes hacia la tecnología. 2) Participación de los estudiantes en los aspectos docentes. 3) Aprendizaje estudiantil sobre el ambiente, y 4) Conciencia ambiental en relación con la vida diaria. Estas relaciones ponen de manifiesto,cómo se plantea en las implicaciones, la necesidad de más investigación en acción en la sala de clases, la importancia—como tema transversal—de la conciencia ambiental mediante la tecnología al construir conocimientos significativos dentro y fuera de la escuela, asó como, valorar la investigación y la dialogicidad en la sala de clases como actividades que obligan al reexamen de la práctica didáctica en su formas curriculares de objetivos, recursos

  12. Simple quantification of in planta fungal biomass.

    PubMed

    Ayliffe, Michael; Periyannan, Sambasivam K; Feechan, Angela; Dry, Ian; Schumann, Ulrike; Lagudah, Evans; Pryor, Anthony

    2014-01-01

    An accurate assessment of the disease resistance status of plants to fungal pathogens is an essential requirement for the development of resistant crop plants. Many disease resistance phenotypes are partial rather than obvious immunity and are frequently scored using subjective qualitative estimates of pathogen development or plant disease symptoms. Here we report a method for the accurate comparison of total fungal biomass in plant tissues. This method, called the WAC assay, is based upon the specific binding of the plant lectin wheat germ agglutinin to fungal chitin. The assay is simple, high-throughput, and sensitive enough to discriminate between single Puccinia graminis f.sp tritici infection sites on a wheat leaf segment. It greatly lends itself to replication as large volumes of tissue can be pooled from independent experiments and assayed to provide truly representative quantification, or, alternatively, fungal growth on a single, small leaf segment can be quantified. In addition, as the assay is based upon a microscopic technique, pathogen infection sites can also be examined at high magnification prior to quantification if desired and average infection site areas are determined. Previously, we have demonstrated the application of the WAC assay for quantifying the growth of several different pathogen species in both glasshouse grown material and large-scale field plots. Details of this method are provided within. PMID:24643560

  13. In para totale...una cosa da panico...sulla lingua dei giovani in Italia (In para totale...una cosa da panico...The Language of Young People in Italy).

    ERIC Educational Resources Information Center

    Marcato, Carla

    1997-01-01

    Describes and analyzes the language of young people in Italy today. Particular focus is on the expressions using "para" (e.g., "in para totale" = to be very bored or worried) and the phrase "una cosa da panico" (something terrible or its opposite something wonderful). (CFM)

  14. [Influence of antioxidant phenovin and immunomodulator Una de gato on free radical oxidation at parodontitis].

    PubMed

    Gogebashvili, N N; Dzhashi, L M; Datunashvili, I V; Intskirveli, N A; Kiparoidze, L I

    2009-04-01

    The aim of the research: detection of changes in free radical oxidation at treatment of parodontitis with combination of preparations - antioxidant Phenovin and immunomodulator Una de gato. Reactive compounds of nitrogen, oxygen and lipids in saliva, blood and gingival tissue of patients suffering from moderate form of parodontitis has been studied by means of the electronic paramagnetic resonance (EPR) method and spin-traps (DETC, DMPO, PBN - Sigma). In patients with parodontitis content of free NO in saliva and blood increases, while in gingival tissue - decreases. In saliva, blood and gingival tissue of patients intense EPR signals of superoxidradicals (O2(-)) and lipoperoxides (LOO(-)) has been revealed indicating intensification of processes of lipid peroxidation in oral cavity, as well as in whole organism of patient. Exaggerated synthesis of NO in saliva and blood of patient is determined by high- expression of inducible NO-synthase triggered by oxidative stress, and increased activity of neuronal NO-synthase in saliva as a result of high concentrations of metacholine and P substance intensely secreted at parodontitis. Decreased content of free NO in gingival tissue of patients with parodontitis compared to control is the result of biological degradation of nitric oxide (conversion of NO into peroxinitrite on the background of intense oxidative stress in oral cavity) and nitrosylation of mitochondrial electron transport proteins of gingival tissue (characteristic for parodontitis) with further decrease in content of free oxide, suppression of intensity of mitochondrial respiration, energogenesis, development of ischemia in oral tissue leading to further initiation of destructive processes and progression of parodontitis. Treatment with combination of preparations - Phenovin and Una de gato decreased intensity of oxidative stress in organism of patients and reduced destructive processes of tissues in oral cavity. Relative normalization of oxidative metabolism

  15. La reconstrucción del seno después de una mastectomía

    Cancer.gov

    Hoja informativa que describe la reconstrucción del seno (mama) después de una mastectomía. Contiene información sobre las opciones quirúrgicas, los cuidados de seguimiento y los exámenes de detección del cáncer de seno después de la reconstrucción.

  16. Tdp studies and tests for C. A. Energia Electrica de Venezuela (enelven) at planta ramon laguna, units RL-17 and RL-10. Volume 3. Unit RL-10 turbine generator condition assessment report and units RL-10 and RL-11 boiler feed pump conditon assessment report. Export trade information

    SciTech Connect

    Not Available

    1991-03-28

    The study, conducted by Babcock and Wilcox, was funded by the U.S. Trade and Development agency on behalf of Enelven. In order to maximize generated power output and minimize operating costs at Planta Ramon Laguna, tests were done to evaluate the condition of equipment at the plant. In order to identify any damage and determine the operating output of each unit, assessments were done of the furnaces, boilers, generators and boiler feed pumps being used in the plant. The report presents the results of these tests. This is the last of three volumes and it is divided into the following sections: (1) Condition Assessment of Unit RL-10 Turbine-Generator; (2) Condition Assessment of Unit RL-10 and RL-11 Boiler Feed Pumps.

  17. In planta mutagenesis of Src homology 3 domain-like fold of NdhS, a ferredoxin-binding subunit of the chloroplast NADH dehydrogenase-like complex in Arabidopsis: a conserved Arg-193 plays a critical role in ferredoxin binding.

    PubMed

    Yamamoto, Hiroshi; Shikanai, Toshiharu

    2013-12-20

    Chloroplast NADH dehydrogenase-like (NDH) complex mediates cyclic electron transport around photosystem I and chlororespiration in angiosperms. The Src homology 3 domain (SH3)-like fold protein NdhS/CRR31 is an NDH subunit that is necessary for high affinity binding of ferredoxin, indicating that chloroplast NDH functions as a ferredoxin:plastoquinone oxidoreductase. However, the mechanism of the interaction between NdhS and ferredoxin is unclear. In this study, we analyzed their interaction in planta by using site-directed mutagenesis of NdhS. In general, binding of ferredoxin to its target proteins depends on electrostatic interaction. In silico analysis predicted the presence of a positively charged pocket in the SH3-like domain of NdhS, where nine charged residues are highly conserved among plants. Systematic alteration of these sites with neutral glutamine revealed that only arginine 193 was required for high NDH activity in vivo. Further replacement of arginine 193 with negatively charged aspartate or glutamate or hydrophobic alanine significantly decreased the efficiency of ferredoxin-dependent plastoquinone reduction by NDH in ruptured chloroplasts. Similar results were obtained in in vivo analyses of NDH activity and electron transport. From these results, we propose that the positive charge of arginine 193 in the SH3-like domain of NdhS is critical for electrostatic interaction with ferredoxin in vivo. PMID:24225949

  18. Regiones de formación de la línea de He II λ 1640 en una atmósfera en expansión

    NASA Astrophysics Data System (ADS)

    Venero, R.; Cidale, L.; Ringuelet, A.

    Se calculan perfiles de la línea del He II λ1640 empleando un modelo de atmósfera en expansión con una estructura cromosférica para determinar la región de formación de la línea. El problema se trata en geometría esférica, fuera de equilibrio termodinámico. La ecuación de transporte radiativo se resuelve rigurosamente en el marco de referencia del fluído para distintas leyes de velocidad, monótonamente crecientes, y diferentes leyes de temperatura. Se buscan relaciones globales entre las características del perfil y los parámetros físicos atribuídos a la atmósfera estelar, con la finalidad de definir una herramienta de diagnóstico de las propiedades del medio.

  19. Informe Anual a la Nación sobre el Estado del Cáncer con una sección especial sobre la prevalencia d

    Cancer.gov

    El Informe Anual a la Nación sobre el Estado del Cáncer (1975 a 2010), mostró un descenso más acelerado que en años anteriores de los índices de mortalidad por cáncer de pulmón. También contiene una sección especial que destaca los efectos significativos

  20. El Instituto Nacional del Cáncer de EE. UU. y Perú firman una Declaración de Intención

    Cancer.gov

    El Instituto Nacional del Cáncer y la República del Perú firmaron una declaración de intención para compartir el interés en fomentar la investigación biomédica de oncología, basándose en la colaboración mutua de ambas entidades, así como el objetivo común

  1. Determinación de la orientación global SAO-Hipparcos mediante una expansión en armónicos vectoriales

    NASA Astrophysics Data System (ADS)

    Cionco, R. G.; Vucetich, H.; Orellana, R.; Arias, E. F.

    En base a las diferencias de posición y movimientos propios de 101352 estrellas con posición SAO observadas por HIPPARCOS y utilizando la naturaleza vectorial de esas diferencias, determinamos 6 parámetros de orientación global (3 de rotación y 3 de desplazamiento axial), para los sistemas de referencia asociados a los marcos mencionados, mediante una descomposición en serie de armónicos vectoriales ortogonales.

  2. Tratamiento Quirúrgico de los Meningiomas del Foramen Óptico, Técnicay Resultados de una Serie de 18 Pacientes

    PubMed Central

    Goldschmidt, Ezequiel; Ajler, Pablo; Campero, Álvaro; Landriel, Federico; Sposito, Maximiliano; Carrizo, Antonio

    2014-01-01

    Introducción: los meningiomas del foramen óptico producen un rápido deterioro de la función visual aún cuando su tamaño es pequeño, por eso su diagnóstico y manejo difiere del resto de los meningiomas clinoideos. El propósito de este estudio es presentar la técnica y los resultados de nuestro manejo quirúrgico de meningiomas foraminales (MF). Pacientes y Métodos: se llevó a cabo una revisión de las historias clínicas de 47 pacientes con meningiomas primarios intraorbitarios. Se realizaron 52 cirugías en los pacientes con MF. Se empleó una craneotomía fronto-orbitaria, seguida de una descompresión extradural del canal óptico, resección del componente intraorbitario y exploración intradural del nervio óptico. Resultados: de los 12 pacientes con MF que presentaban la visión conservada, la agudeza visual fue preservada en 7 casos, mejoró en 2, y empeoró en 3. En 18 pacientes, el principal síntoma fue exoftalmos y en 35 pacientes ceguera unilateral. Ocurrieron 6 recurrencias, 2 a 10 años después de la resección quirúrgica. Cinco de ellos fueron reoperados. Se indicó radioterapia después de la recurrencia en 3 pacientes. Conclusión: el manejo de los MF continúa siendo controvertido y frecuentemente se propone un tratamiento conservador. Basados en nuestros hallazgos de frecuente extensión intracraneal, proponemos realizar una resección total o subtotal del tumor, preservando el nervio óptico en pacientes con visión prequirúrgica conservada. PMID:25165616

  3. Anthropization on the Cerrado biome in the Brazilian Uruçuí-Una Ecological Station estimated from orbital images.

    PubMed

    Pereira, A C; Gama, V F

    2010-11-01

    In this study we analysed the dynamics of deforestation and burnings during the dry seasons from 2003 to 2008 in the Uruçuí-Una Ecological Station (UUES) and its buffer zone, located in the Cerrado biome of the southwest of Piauí, a Brazil's State, based on images from the orbital sensors CCD/CBERS-2 and TM/Landsat-5. Two dates from each of the years were interpreted and analysed: one in the middle of the dry season and one at the end. The deforested areas were expanded through the period analysed and were larger in the buffer zone, suggesting a relative protection of the UUES. New cut-offs were predictable because of the early opening of roads that would become their limits. The burning scars were larger at the end of the dry season as a consequence of the management and implementation of agricultures and pastures. In 2004 and 2007 these scars were larger probably because of the increase of dry phytomass that every three years is big enough to spread the fire originated in the anthropogenic burnings through the native vegetation. This scenario reaffirms the need for: stronger enforcement in order to stop anthropisation in the UUES and a management plan, absent for this unit so far. These proceedings are urgent also because the UUES is located in one of the most preserved regions of the Cerrado and controversially where intense anthropisation in ongoing, which stresses the lack, need and urgency of biological conservation proceedings for the Piauí's southeastern Cerrado. PMID:21180901

  4. Geomorphological significance of the palaeodrainage network on a karst plateau: The Una-Korana plateau, Dinaric karst, Croatia

    NASA Astrophysics Data System (ADS)

    Bočić, Neven; Pahernik, Mladen; Mihevc, Andrej

    2015-10-01

    Karst plateaus often have a complex geological and geomorphological history. It is widely accepted that their development requires a long period of karst denudation. This study reconstructs the palaeodrainage network of a karst plateau, analyses its properties and establishes its geomorphological significance. The main purpose of this research was to deepen our understanding of a key stage in the evolution of karst plateaus - the transition from a fluvial land surface to one dominated by karst surface processes. The study was conducted on a large part of the Una-Korana plateau, the largest plateau in the Dinaric karst. The majority of the plateau is made of carbonate rocks of Triassic, Jurassic and Cretaceous age that set the conditions for the development of the karst. We have reconstructed the palaeodrainage network based on 1:25,000 topographic maps with 10 m contours. The transition of the surface drainage network to the underground karst drainage network is still in progress, so, depending on the degree of karstification, the drainage network was divided into three categories: active, dry and relict. It was found that 90.5% of the pre-existing drainage network has undergone some degree of karstification. The active surface drainage network gradually shifted to a dry network, then to a relict network. The surface drainage network is gradually replaced by a dense network of dolines. Today, the flat and karstified inter-fluvial area is drained underground towards the main watercourses and these drain the entire region over the surface towards the Pannonian basin. This is the largest known karst palaeodrainage network in the Dinaric karst that has been reconstructed in this way.

  5. Seismicity in 2010 and major earthquakes recorded and located in Costa Rica from 1983 until 2012, by the local OVSICORI-UNA seismic network

    NASA Astrophysics Data System (ADS)

    Ronnie, Q.; Segura, J.; Burgoa, B.; Jimenez, W.; McNally, K. C.

    2013-05-01

    This work is the result of the analysis of existing information in the earthquake database of the Observatorio Sismológico y Vulcanológico de Costa Rica, Universidad Nacional (OVSICORI-UNA), and seeks disclosure of basic seismological information recorded and processed in 2010. In this year there was a transition between the software used to record, store and locate earthquakes. During the first three months of 2010, we used Earthworm (http://folkworm.ceri.memphis.edu/ew-doc), SEISAN (Haskov y Ottemoller, 1999) and Hypocenter (Lienert y Haskov, 1995) to capture, store and locate the earthquakes, respectively; in April 2010, ANTELOPE (http://www.brtt.com/software.html) start to be used for recording and storing and GENLOC (Fan at al, 2006) and LOCSAT (Bratt and Bache 1988), to locate earthquakes. GENLOC was used for local events and LOCSAT for regional and distant earthquakes. The local earthquakes were located using the 1D velocity model of Quintero and Kissling (2001) and for regional and distant earthquakes IASPEI91 (Kennett and Engdahl, 1991) was used. All the events for 2010 and shown in this work were rechecked by the authors. We located 3903 earthquakes in and around Costa Rica and 746 regional and distant seismic events were recorded (see Figure 1). In this work we also give a summary of major earthquakes recorded and located by OVSICORI-UNA network between 1983 and 2012. Seismicity recorded by OVSICORI-UNA network in 2010

  6. Diseno de una Actividad de Aprendizaje Basada en la Argumentacion Dialogica en un curso Virtual de Biotecnologia y su Incidencia en el Desarrollo de Competencias Cientificas

    NASA Astrophysics Data System (ADS)

    Ortiz Benavides, Fedra Lorena

    El proposito de la investigacion fue evaluar la efectividad de una actividad de aprendizaje basado en la argumentacion dialogica en linea y su incidencia en el desarrollo de competencias cientificas. Se fundamenta en la teoria del aprendizaje socio cultural de Vigotsky (1984), los principios del diseno instruccional de la cognicion situada por Hung y Der-Thang (2001) y como estrategia se aplico la argumentacion dialogica utilizando el Modelo Argumentativo de Toulmin MAT (1984). El diseno experimental comparo dos grupos de estudiantes A y B en el curso virtual de Biotecnologia. El grupo A (experimental) desarrollo la discusion a partir de la estrategia disenada para este estudio y el grupo B (control) realizo la discusion desde las actividades tradicionales. El desarrollo de la competencia argumentativa se valoro con el instrumento de evaluacion para argumentacion dialogica en linea propuesta por Clark y Sampson (2008). La evaluacion de las competencias cientificas se realizo a partir de una postprueba. Los datos fueron analizados con pruebas estadisticas no parametricas. Los resultados de la investigacion, indicaron diferencias significativas en el nivel de la competencia argumental en el grupo experimental en comparacion al grupo control. Igualmente se demostro que existe una relacion positiva entre el nivel de desarrollo de la competencia argumentativa y el nivel de desarrollo de las competencias cientificas.

  7. "Una Luna Brilla."

    ERIC Educational Resources Information Center

    Teaching Music, 2002

    2002-01-01

    Focuses on the "World's Largest Concerts" listing various songs to be sung by students. Includes teaching tips in the form of quotes from music teachers on aspects of music education, such as teaching rhythm. Includes information on how to access the songs. (CMK)

  8. Identification of Suitable Water Harvesting Zones Based on Geomorphic Resources for Drought Areas: A Case Study of Una District, Himachal Pradesh, India.

    NASA Astrophysics Data System (ADS)

    Prakasam, D. C., Jr.; Zaman, B.

    2014-12-01

    Water is one of the most vital natural resource and its availability and quality determine ecosystem productivity, both for agricultural and natural systems. Una district is one of the major potential agricultural districts in Himachal Pradesh, India. More than 70% of the population of this district is engaged in agriculture and allied sectors and major crops grown are maize, wheat, rice, sugarcane, pulses and vegetables. The region faces drought every year and about 90 per cent of the area is water stressed. This has resulted in crop loss and shortage of food and fodder. The sources of drinking water, small ponds and bowlies dry-up during summer season resulting in scarcity of drinking water. Una district receives rainfall during monsoons from June to September and also during non-monsoon period (winter). The annual average rainfall in the area is about 1040 mm with 55 average rainy days. But due to heavy surface run-off the farmers not able to cultivate the crops more than once in a year. Past research indicate that the geomorphology of the Una district might be responsible for such droughts as it controls the surface as well as ground water resources. The research proposes to develop a water stress model for Una district using the geomorphic parameters, water resource and land use land cover data of the study area. Using Survey of India topographical maps (1:50000), the geomorphic parameters are extracted. The spatial layers of these parameters i.e. drainage density, slope, relative relief, ruggedness index, surface water body's frequency are created in GIS. A time series of normalized remotely sensed data of the study area is used for land use land cover classification and analyses. Based on the results from the water stress model, the drought/water stress areas and water harvesting zones are identified and documented. The results of this research will help the general population in resolving the drinking water problem to a certain extent and also the

  9. In Planta mutagenesis determines the functional regions of puroindoline proteins.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    PINA and PINB functionally comprise the wheat (Triticum aestivum L.) Ha locus which controls grain texture and many wheat end-use properties. PINs in their functional forms impart soft texture and a mutation in either PIN is found in all hard wheats. Studies of the PINs mode of action is limited ...

  10. Selective Effect of Pesticides on Plant--A Review.

    PubMed

    Parween, Talat; Jan, Sumira; Mahmooduzzafar, Sumira; Fatma, Tasneem; Siddiqui, Zahid Hameed

    2016-01-01

    This review represents systematic and integrated picture of pesticide exposure to plant and its effect on growth and metabolism. Decades ago, agrochemicals were introduced aiming at enhancing crop yields and protecting crops from pests. Due to adaptation and resistance developed by pests to chemicals, every year higher amounts and new chemical compounds are used to protect crops, causing undesired side effects and raising the costs of food production. Biological chemical free agriculture is gaining also more and more support but it is still not able to respond to the need for producing massive amounts of food. The use of agrochemicals, including pesticides, remains a common practice especially in tropical regions and South countries. Cheap compounds, such as DDT, HCH, and Lindane, that are environmentally persistent, are today banned from agriculture use in developed countries, but remain popular in developing countries. As a consequence, persistent residues of these chemicals contaminate food and disperse in the environment. Therefore, the thrust of this paper was to review the application of pesticides effect early from germination to growth of the plant, leading to alteration in biochemical, physiological and different enzymatic and non-enzymatic antioxidants which ultimately affect the yield and resulted in residues in plant, vegetables, and fruits. PMID:25216296

  11. Reconexión magnética en una región activa en decaimiento

    NASA Astrophysics Data System (ADS)

    Mandrini, C. H.; Démoulin, P.; van Driel-Gesztelyi, L.; Schmieder, B.; Bagalá, L. G.; Rovira, M. G.

    Se han obtenido observaciones desde Tierra (Hα y magnetogramas) coordinadas con el Soft X-ray Telescope (SXT), a bordo del satélite japonés Yohkoh, de una región activa bipolar en decaimiento. Estos datos constituyen la base para el estudio de un punto brillante en rayos X (PBX) y de la actividad relacionada con el mismo en distintas capas de la atmósfera solar. Las observaciones muestran que el PBX está relacionado con la aparición de un bipolo menor (~ 1020 Mx) y que su abrillantamiento continuo, así como sus aumentos de brillo esporádicos (``fulguraciones"), son el resultado de la reconexión entre el pequeño arco emergente (visto en Hα como un sistema de filamentos arqueados, SFA) y arcos mayores asociados al campo facular preexistente. Se ha extrapolado el campo magnético observado en la aproximación libre de fuerzas lineal y se ha seguido su evolución a lo largo de la vida del PBX. Se ha calculado la posición de las cuasiseparatrices (CSs) a partir del campo modelado. Las líneas de campo extrapoladas, cuyas bases fotosféricas se encuentran a ambos lados de las CSs, están de acuerdo con las estructuras cromosféricas y coronales observadas. Se ha calculado el espesor de la CD ubicada a lo largo de la polaridad negativa emergente, encontrándose que su variación está de acuerdo con la evolución de la intensidad del PBX; la CS es muy delgada durante la vida del PBX (~ 100 m), mientras que su espesor aumenta considerablemente (>= 10^4 m) cuando el PBX desaparece de las imágenes del SXT. Estos resultados señalan que el abrillantamiento y las ``fulguraciones" de este PBX se deben al proceso de reconexión en 3D que tiene lugar en las CSs.

  12. Cigarrillos de mentol | Smokefree Español

    Cancer.gov

    ¿Qué es el mentol? El mentol es una sustancia encontrada naturalmente en las plantas de menta, tales como la menta fresca y la hierbabuena. Deja una sensación refrescante y se usa frecuentemente para aliviar el dolor y la irritación leve, y para prevenir infecciones [1].

  13. EL PROGRAMA CITI: UNA ALTERNATIVA PARA LA CAPACITACIÓN EN ÉTICA DE LA INVESTIGACIÓN EN AMÉRICA LATINA

    PubMed Central

    Litewka, Sergio; Goodman, Kenneth; Braunschweiger, y Paul

    2011-01-01

    El Programa CITI (Collaborative Institutional Training Initiative) es un curso basado en Internet y desarrollado por voluntarios, que brinda material educacional sobre protección de sujetos humanos en investigaciones biomédicas y de conducta, buenas prácticas clínicas, conducta responsable de la investigación y trato humanitario a animales de laboratorio. Es un esfuerzo conjunto del Fred Hutchinson Cancer Research Center y la Universidad de Miami. Con la colaboración de expertos locales, gran parte del contenido se ha adaptado para América Latina, se ha traducido al español y portugués y se han establecido acuerdos de colaboración con instituciones especializadas en bioética en Chile, Brasil, Costa Rica y Perú. Los autores presentan una metodología accesible para la capacitación en ética de la investigación y una justificación de su uso y difusión en América Latina PMID:21687817

  14. Síndrome del Outlet Torácico: ¿Una Patología Siempre Quirúrgica? Análisis de una Serie de 31 Cirugías Realizadas por Vía Supraclavicular Serie clínica

    PubMed Central

    Socolovsky, Mariano; Di Masi, Gilda; Binaghi, Daniela; Campero, Álvaro; Páez, Miguel Domínguez; Dubrovsky, Alberto

    2014-01-01

    Introducción: El síndrome de outlet torácico es una compresión del plexo braquial que suscita polémica. Se clasifica en Outlet Torácico Verdadero o neurogénico (OTV) y Outlet Torácico Disputado o no neurogénico (OTD). El primero presenta síntomas motores en la mano, mientras que el segundo sólo síntomas sensitivos en el miembro superior. El objetivo de este trabajo es analizar los resultados obtenidos en una serie de 31 cirugías. Métodos: Se analizaron las cirugías de nervios efectuadas entre 2003-2012, tomando los diagnósticos de outlet torácico cuyo período de seguimiento post-operatorio mínimo fuera de 6 meses. Se buscaron los siguientes datos: edad, sexo, presencia de síntomas sensitivos y/o motores, clasificación, resultado de los estudios neurofisiológicos y de imágenes, resultado de la cirugía, complicaciones post-operatorias y recidivas. Resultados: Se incluyeron 31 cirugías realizadas en 30 pacientes, 9 OTV (8 mujeres) de 24.3 años, y 21 con OTD (18 mujeres) de 37.4 años de edad en promedio. Un 90% presentaron alteraciones neurofisiológicas preoperatorias, y 66,6% imagenológicas. En el intraoperatorio, el 100% de los OTV presentó una alteración anatómica relacionada con la sintomatología, hecho observado sólo en el 36.7% de los OTD operados. El 87,5% de los OTV mejoraron sensitivamente, mientras que 77,7% mejoraron la atrofia. Por el contrario, 45.4% de los OTD mejoraron permanentemente, 36.3% no tuvieron cambios, 13.6% mejoraron transitoriamente y 4.5% (un caso) empeoró. Las complicaciones post-operatorias fueron más frecuentes aunque transitorias en el grupo de OTV (3 casos sobre 9 operados, 33.3%) que en los OTD (3 casos sobre 22, un 13.6%). Conclusión: El OTV suele mayormente mejorar luego de la cirugía, igual que el OTD aunque en una proporción mucho menor. Estos hallazgos coinciden con otros reportes recientes de esta patología. PMID:25165614

  15. La preparacion en ciencia de los candidatos a maestros del nivel elemental primario segun la reforma de la educacion cientifica en Puerto Rico: Una propuesta de secuencia curricular

    NASA Astrophysics Data System (ADS)

    Rodriguez Plaza, Evelyn

    El proposito de esta investigacion fue identificar los componentes de la preparacion en ciencia que deben recibir los estudiantes del Bachillerato en Artes en Educacion Elemental, Nivel Primario, de acuerdo a los documentos que dirigen la reforma de la educacion cientifica en Puerto Rico. Tambien, se identificaron los componentes de los cursos que forman parte de la preparacion en ciencia de estos estudiantes. Se compararon los componentes de la preparacion en ciencia y los componentes de los cursos para determinar congruencias y discrepancias. Con los datos recopilados se identificaron los componentes de los cursos de una secuencia curricular para la preparacion en ciencia de los candidatos a maestros del nivel elemental primario. La secuencia curricular que se propone en esta investigacion incluye cursos de contenido cientifico y de metodologia en la ensenanza de la ciencia disenados para satisfacer las necesidades de los candidatos. Se recomienda que en los procesos para el diseno, la implantacion y la evaluacion de estos cursos participen profesores de ciencia, profesores de educacion y maestros del nivel elemental primario. Todos los cursos de la secuencia curricular deben tener un enfoque constructivista. Las experiencias educativas que se incluyan en los cursos deben aspirar a desarrollar en los candidatos los atributos de la cultura cientifica y actitudes positivas hacia la ciencia y hacia la ensenanza de esta disciplina. El modelaje por parte de los profesores que ensenen los cursos de la secuencia curricular es fundamental en el desarrollo profesional de los candidatos. Se recomienda que en los cursos de contenido cientifico se estudien los conceptos y los conocimientos cientificos que forman parte del curriculo de Kindergarten a tercer grado de forma integrada y con una profundidad universitaria. Estos cursos deben tener un enfoque interdisciplinario e incluir el estudio de la naturaleza de la ciencia y un componente de laboratorio para desarrollar los

  16. Cobertura de los sistemas de pensiones y factores asociados al acceso a una pensión de jubilación en México

    PubMed Central

    Murillo-López, Sandra; Venegas-Martínez, Francisco

    2016-01-01

    Resumen Objetivos: obtener estimaciones de indicadores de cobertura de las pensiones por jubilación o retiro para la población mexicana de 65 y más años, y evaluar el impacto que tienen los sistemas de pensiones en las transiciones al retiro de los adultos en edades medias y avanzadas en México. Para ello se utilizan datos microeconómicos provenientes de la Encuesta Nacional de Salud y Envejecimiento. Mediante análisis econométrico se identifican los factores sociodemográficos, económicos, laborales e institucionales que están asociados al acceso a una pensión de jubilación, o bien, a la dependencia de otras fuentes de ingresos. Se encontró que, en México, las transiciones al retiro del mercado de trabajo en las etapas avanzadas del ciclo de vida son limitadas debido a las características eminentemente contributivas de los esquemas de pensiones, los cuales favorecen a la población con trayectorias laborales formales y más estables asociadas a: características de género, oportunidades educativas y posibilidades de inserción en el mercado laboral. PMID:27524936

  17. A screen of chemical modifications identifies position-specific modification by UNA to most potently reduce siRNA off-target effects

    PubMed Central

    Bramsen, Jesper B.; Pakula, Malgorzata M.; Hansen, Thomas B.; Bus, Claus; Langkjær, Niels; Odadzic, Dalibor; Smicius, Romualdas; Wengel, Suzy L.; Chattopadhyaya, Jyoti; Engels, Joachim W.; Herdewijn, Piet; Wengel, Jesper; Kjems, Jørgen

    2010-01-01

    Small interfering RNAs (siRNAs) are now established as the preferred tool to inhibit gene function in mammalian cells yet trigger unintended gene silencing due to their inherent miRNA-like behavior. Such off-target effects are primarily mediated by the sequence-specific interaction between the siRNA seed regions (position 2–8 of either siRNA strand counting from the 5′-end) and complementary sequences in the 3′UTR of (off-) targets. It was previously shown that chemical modification of siRNAs can reduce off-targeting but only very few modifications have been tested leaving more to be identified. Here we developed a luciferase reporter-based assay suitable to monitor siRNA off-targeting in a high throughput manner using stable cell lines. We investigated the impact of chemically modifying single nucleotide positions within the siRNA seed on siRNA function and off-targeting using 10 different types of chemical modifications, three different target sequences and three siRNA concentrations. We found several differently modified siRNAs to exercise reduced off-targeting yet incorporation of the strongly destabilizing unlocked nucleic acid (UNA) modification into position 7 of the siRNA most potently reduced off-targeting for all tested sequences. Notably, such position-specific destabilization of siRNA–target interactions did not significantly reduce siRNA potency and is therefore well suited for future siRNA designs especially for applications in vivo where siRNA concentrations, expectedly, will be low. PMID:20453030

  18. A'Una ("Let's Go!")

    ERIC Educational Resources Information Center

    Brescia, Bill

    The guide, intended to aid students (grades four and above) in making their bodies a good home for their spirit, provides a variety of athletic activities and traditional Indian recipes for use in the classroom or in cultural programs. Apart from running (the major American Indian means of maintaining physical fitness), instructions are given for…

  19. TEMAS PARA UNA BIOETICA LATINOAMERICANA

    PubMed Central

    Yunta, Eduardo Rodriguez

    2009-01-01

    El presente trabajo reflexiona sobre el fundamento filosófico de la bioética como disciplina, método y movimiento desde su contexto histórico y en relación con el contexto social y cultural de la vivencia latinoamericana. PMID:20209030

  20. Learning Astronomy by Playing in a Park. (Spanish Title: Aprender AstronoMía Jugando en Una Plaza.) Aprender Astronomia Brincando em Uma Praça

    NASA Astrophysics Data System (ADS)

    Camino, Néstor

    2012-12-01

    Some public-square games are presented in this paper, considered as didactic modules to help children imagine astronomical processes, based on the concept that learning in Astronomy should be developed to strengthen the relationship of our body with three-dimensional space and time, much in the same way we experience when observing the actual sky, holding a permanent "dialogue" between the actual world and what is to be learned. The games presented (merry-go-rounds and slides) were designed to work on the astronomical concepts related to the translation of the Earth around the Sun, the phases of the Moon and gravity, and on what is perceived by an observer about those phenomena. The description of each game, their physical and astronomical foundations, and a critical comment about their didactical importance are the key parts of the paper. Finally, a recommendation is given about the role teachers should play to be essential partners in the process of learning Astronomy by means of the interaction with these games. Se presentan en este trabajo algunos juegos de plaza, considerados como módulos didácticos para imaginar procesos astronómicos, a partir de la concepción de que el aprendizaje en Astronomía debe desarrollarse fortaleciendo la relación del propio cuerpo con el espacio tridimensional y con el tiempo, tal como se vive al observar el cielo, construyendo un "diálogo" entre el mundo real y los aprendizajes a construir. Los juegos presentados (calesitas y toboganes) fueron diseñados para trabajar sobre la traslación de la Tierra en torno al Sol, las fases de la Luna y la gravedad, y sobre lo que un observador percibe de los mismos. Se da la descripción de cada juego, se discuten sus fundamentos físicos y astronómicos, y se desarrolla una crítica didáctica de los mismos. Finalmente, se comenta el rol que deberían tener los docentes en el acompañamiento a los aprendices en el proceso de interacción con los juegos presentados. Apresentam-se neste

  1. Revision curricular a partir de un analisis comparativo de las discrepancias en los curriculos de una escuela de optometria en Puerto Rico con las competencias requeridas para las agencias de revalida y acreditacion 2004

    NASA Astrophysics Data System (ADS)

    Rivera Pacheco, Andres

    El proposito de esta investigacion, un estudio cualitativo de caso, fue comparar y contrastar el curriculo vigente de la Escuela de Optometria de la UIAPR con las competencias y estandares requeridos por las agencias de acreditacion y de revalida. Con este proposito, decidimos realizar una revision y un analisis de documentos: el prontuario de cada uno de los cursos de los curriculos implantados en el 1993 y en el 2001; las competencias y estandares establecidos por las agencias de revalida y de acreditacion; y las estadisticas en las que se analiza el porcentaje de estudiantes que aprueban cada una de las partes de los examenes de revalida entre el 1998 al 2003. Se realizaron entrevistas dirigidas para dar apoyo y complementar la revision y el analisis de estos documentos. Los participantes de las entrevistas fueron tres estudiantes de la clase de optometria del 2004 (ultima clase del curriculo del 1993); tres estudiantes de la clase de optometria del 2005 (primera clase graduanda del curriculo vigente) y tres profesores y/o directores de los Departamentos de Ciencias Basicas, Ciencias Clinicas y Cuidado al Paciente. Esta investigacion se enmarco en el modelo de evaluacion curricular de discrepancia de Malcolm Provus y en el modelo de desarrollo basado en competencias. Uno de los hallazgos mas importantes del estudio es que los cambios que se implantaron al curriculo del 2001 no han logrado que los estudiantes mejoren su ejecucion en los examenes de revalida. Por otro lado, se encontro que el curriculo vigente atiende completamente los estandares de la practica de Optometria, pero no las competencias. Esta informacion fue validada mediante el uso de una tabla de cotejo para el analisis de los cursos y de la informacion obtenida de las entrevistas. El estudio determina y concluye que existen discrepancias entre los prontuarios de los cursos del curriculo y las competencias requeridas por la agencia de revalida. Segundo, que el Departamento de Ciencias Basicas es el

  2. Adapting a Common Photographic Camera to Take Pictures of the Sky. (Spanish Title: Adaptando Una Camara Fotografica Comun Para Obtener Fotografias del Cielo.) Adaptando Uma Câmera Fotográfica Manual Simples Para Fotografar o Céu

    NASA Astrophysics Data System (ADS)

    Danhoni Neves, Marcos Cesar; Pereira, Ricardo Francisco

    2007-12-01

    In this paper will be introduced a method of astrophotography using a non-reflex photographic camera (a low-cost method). It will be revised some photographic processes commonly used nowadays for comparison with the aims of this paper. En este trabajo será introducido un método de astrofotografia que utiliza una cámara fotográfica non-reflex (un método de bajo costo). Serán revisados algunos procesos fotográficos comúnmente utilizados actualmente para comparación con los objetivos de este trabajo. O presente artigo procura introduzir um método de astrofotografia utilizando uma câmera fotográfica não reflex, de baixo custo. É feita uma revisão do processo fotográfico comumente empregado para fins de comparação com os objetivos pretendidos no presente trabalho.

  3. In planta distribution and quantification of Asiatic strain of citrus Huanglongbing pathogen

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Huanglongbing is one of the most devastating diseases caused by an uncultured phloem limited bacterium, Candidatus Liberibacter spp. In this study, a combination of traditional and real-time PCR targeting the putative DNA polymerase and 16S rDNA sequence, respectively, were used to examine the distr...

  4. In planta reduction of maize seedling stalk lesions by the bacterial endophyte Bacillus mojavensis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Due to autoinfection and alloinfection, maize is susceptible to infection by Fusarium verticillioides resulting in diseases and contamination of maize kernels with the fumonisin mycotoxins. Attempts at controlling this fungus are currently being done with biocontrol agents such as bacteria and this...

  5. "In planta" regulation of the "Arabidopsis" Ca(2+)/H(+) antiporter CAX1

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Vacuolar localized Ca(2+)/H(+) exchangers such as "Arabidopsis thaliana" cation exchanger 1 (CAX1) play important roles in Ca(2+) homeostasis. When expressed in yeast, CAX1 is regulated via an N-terminal autoinhibitory domain. In yeast expression assays, a 36 amino acid N-terminal truncation of CAX1...

  6. Plant Tissue Preparation for the Detection of an Endophytic Fungus In Planta.

    PubMed

    McKinnon, Aimee C

    2016-01-01

    The identification of fungal endophytes often relies on culturing isolates from surface-sterilized plant tissue. However, molecular techniques have enabled the rapid detection and identification of targeted endophyte species, and next-generation sequencing technology provides an opportunity to obtain comprehensive information on endophytic communities, directly from plant tissue. In order to achieve accurate detection from internal tissues, surface microbes and associated deoxyribonucleic acid (DNA) must be eliminated, with particular consideration for the type of plant tissue and the efficacy of the surface sterilization procedure used. The methodology described later was developed specifically for detection of DNA from the entomopathogenic fungal endophyte Beauveria bassiana (Vuillemin) (Ascomycota: Hypocreales) in various tissues of Zea mays (L.). However, the protocol may be easily applied to other fungi and bacterial endophytes. Included is a stringent sodium hypochlorite-based surface sterilization protocol for plant material in preparation for polymerase chain reaction (PCR) to detect target DNA within plant tissue. Included are a treatment for dealing with surface DNA contamination and a novel procedure for assessing the efficacy of surface sterilization using PCR. PMID:27565499

  7. Floral Benzenoid Carboxyl Methyltransferases: From in Vitro to in Planta Function

    SciTech Connect

    Effmert,U.; Saschenbrecker, S.; Ross, J.; Negre, F.; Fraser, C.; Noel, J.; Dudareva, N.; Piechulla, B.

    2005-01-01

    Benzenoid carboxyl methyltransferases synthesize methyl esters (e.g., methyl benzoate and methyl salicylate), which are constituents of aromas and scents of many plant species and play important roles in plant communication with the surrounding environment. Within the past five years, eleven such carboxyl methyltransferases were isolated and most of them were comprehensively investigated at the biochemical, molecular and structural level. Two types of enzymes can be distinguished according to their substrate preferences: the SAMT-type enzymes isolated from Clarkia breweri, Stephanotis floribunda, Antirrhinum majus, Hoya carnosa, and Petunia hybrida, which have a higher catalytic efficiency and preference for salicylic acid, while BAMT-type enzymes from A. majus, Arabidopsis thaliana, Arabidopsis lyrata, and Nicotiana suaveolens prefer benzoic acid. The elucidation of C. breweri SAMT's three-dimensional structure allowed a detailed modelling of the active sites of the carboxyl methyltransferases and revealed that the SAM binding pocket is highly conserved among these enzymes while the methyl acceptor binding site exhibits some variability, allowing a classification into SAMT-type and BAMT-type enzymes. The analysis of expression patterns coupled with biochemical characterization showed that these carboxyl methyltransferases are involved either in floral scent biosynthesis or in plant defense responses. While the latter can be induced by biotic or abiotic stress, the genes responsible for floral scent synthesis exhibit developmental and rhythmic expression pattern. The nature of the product and efficiency of its formation in plants depend on the availability of substrates, the catalytic efficiency of the enzyme toward benzoic acid and/or salicylic acid, and the transcriptional, translational, and post-translational regulation at the enzyme level. The biochemical properties of benzenoid carboxyl methyltransferases suggest that the genes involved in plant defenses might represent the ancestor for the presently existing floral genes which during evolution gained different expression profiles and encoded enzymes with the ability to accept structurally similar substrates.

  8. Binary 2in1 Vectors Improve in Planta (Co)localization and Dynamic Protein Interaction Studies.

    PubMed

    Hecker, Andreas; Wallmeroth, Niklas; Peter, Sébastien; Blatt, Michael R; Harter, Klaus; Grefen, Christopher

    2015-07-01

    Fluorescence-based protein-protein interaction techniques are vital tools for understanding in vivo cellular functions on a mechanistic level. However, only under the condition of highly efficient (co)transformation and accumulation can techniques such as Förster resonance energy transfer (FRET) realize their potential for providing highly accurate and quantitative interaction data. FRET as a fluorescence-based method unifies several advantages, such as measuring in an in vivo environment, real-time context, and the ability to include transient interactions as well as detecting the mere proximity of proteins. Here, we introduce a novel vector set that incorporates the benefit of the recombination-based 2in1 cloning system with the latest state-of-the-art fluorescent proteins for optimal coaccumulation and FRET output studies. We demonstrate its utility across a range of methods. Merging the 2in1 cloning system with new-generation FRET fluorophore pairs allows for enhanced detection, speeds up the preparation of clones, and enables colocalization studies and the identification of meaningful protein-protein interactions in vivo. PMID:25971551

  9. Chrysanthemyl diphosphate synthase operates in planta as a bifunctional enzyme with chrysanthemol synthase activity.

    PubMed

    Yang, Ting; Gao, Liping; Hu, Hao; Stoopen, Geert; Wang, Caiyun; Jongsma, Maarten A

    2014-12-26

    Chrysanthemyl diphosphate synthase (CDS) is the first pathway-specific enzyme in the biosynthesis of pyrethrins, the most widely used plant-derived pesticide. CDS catalyzes c1'-2-3 cyclopropanation reactions of two molecules of dimethylallyl diphosphate (DMAPP) to yield chrysanthemyl diphosphate (CPP). Three proteins are known to catalyze this cyclopropanation reaction of terpene precursors. Two of them, phytoene and squalene synthase, are bifunctional enzymes with both prenyltransferase and terpene synthase activity. CDS, the other member, has been reported to perform only the prenyltransferase step. Here we show that the NDXXD catalytic motif of CDS, under the lower substrate conditions prevalent in plants, also catalyzes the next step, converting CPP into chrysanthemol by hydrolyzing the diphosphate moiety. The enzymatic hydrolysis reaction followed conventional Michaelis-Menten kinetics, with a Km value for CPP of 196 μm. For the chrysanthemol synthase activity, DMAPP competed with CPP as substrate. The DMAPP concentration required for half-maximal activity to produce chrysanthemol was ∼100 μm, and significant substrate inhibition was observed at elevated DMAPP concentrations. The N-terminal peptide of CDS was identified as a plastid-targeting peptide. Transgenic tobacco plants overexpressing CDS emitted chrysanthemol at a rate of 0.12-0.16 μg h(-1) g(-1) fresh weight. We propose that CDS should be renamed a chrysanthemol synthase utilizing DMAPP as substrate. PMID:25378387

  10. Dynamic Root Exudation of Sorgoleone and Its in Planta Mechanism of Action

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The oily droplets exuding from the root hairs of sorghum are composed of a 1:1 ratio of sorgoleone and its lipid resorcinol analog. The production and release appear to be suppressed when 15 to 20 µg/mg dry weight accumulates at the tip of the root hairs. However, more exudate is produced followin...

  11. Stable plasmid vectors for complementation of Xylella fastidiosa mutants in planta

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Current understanding of the mechanisms of Pierce’s Disease development has been significantly advanced by molecular genetic studies of the causal agent, Xylella fastidiosa (Xf). Plasmid vectors are an essential tool for studies of bacterial genetics and pathogenesis. However, most commonly used pla...

  12. Quantification of Sclerotinia homoeocarpa overwintering in planta and detection in commercial seed

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Dollar spot is the most economically important disease of amenity turf grasses in the United States, yet little is known about the source of primary inoculum for this disease. With the exception of a few isolates from the United Kingdom, Sclerotinia homoeocarpa, the causal agent of dollar spot, does...

  13. Origin and evolution of metal P-type ATPases in Plantae (Archaeplastida)

    PubMed Central

    Hanikenne, Marc; Baurain, Denis

    2013-01-01

    Metal ATPases are a subfamily of P-type ATPases involved in the transport of metal cations across biological membranes. They all share an architecture featuring eight transmembrane domains in pairs of two and are found in prokaryotes as well as in a variety of Eukaryotes. In Arabidopsis thaliana, eight metal P-type ATPases have been described, four being specific to copper transport and four displaying a broader metal specificity, including zinc, cadmium, and possibly copper and calcium. So far, few efforts have been devoted to elucidating the origin and evolution of these proteins in Eukaryotes. In this work, we use large-scale phylogenetics to show that metal P-type ATPases form a homogenous group among P-type ATPases and that their specialization into either monovalent (Cu) or divalent (Zn, Cd…) metal transport stems from a gene duplication that took place early in the evolution of Life. Then, we demonstrate that the four subgroups of plant metal ATPases all have a different evolutionary origin and a specific taxonomic distribution, only one tracing back to the cyanobacterial progenitor of the chloroplast. Finally, we examine the subsequent evolution of these proteins in green plants and conclude that the genes thoroughly characterized in model organisms are often the result of lineage-specific gene duplications, which calls for caution when attempting to infer function from sequence similarity alone in non-model organisms. PMID:24575101

  14. In Planta Proteomics and Proteogenomics of the Biotrophic Barley Fungal Pathogen Blumeria graminis f. sp. hordei*

    PubMed Central

    Bindschedler, Laurence V.; Burgis, Timothy A.; Mills, Davinia J. S.; Ho, Jenny T. C.; Cramer, Rainer; Spanu, Pietro D.

    2009-01-01

    To further our understanding of powdery mildew biology during infection, we undertook a systematic shotgun proteomics analysis of the obligate biotroph Blumeria graminis f. sp. hordei at different stages of development in the host. Moreover we used a proteogenomics approach to feed information into the annotation of the newly sequenced genome. We analyzed and compared the proteomes from three stages of development representing different functions during the plant-dependent vegetative life cycle of this fungus. We identified 441 proteins in ungerminated spores, 775 proteins in epiphytic sporulating hyphae, and 47 proteins from haustoria inside barley leaf epidermal cells and used the data to aid annotation of the B. graminis f. sp. hordei genome. We also compared the differences in the protein complement of these key stages. Although confirming some of the previously reported findings and models derived from the analysis of transcriptome dynamics, our results also suggest that the intracellular haustoria are subject to stress possibly as a result of the plant defense strategy, including the production of reactive oxygen species. In addition, a number of small haustorial proteins with a predicted N-terminal signal peptide for secretion were identified in infected tissues: these represent candidate effector proteins that may play a role in controlling host metabolism and immunity. PMID:19602707

  15. Wide Screening of Phage-Displayed Libraries Identifies Immune Targets in Planta

    PubMed Central

    Rioja, Cristina; Van Wees, Saskia C.; Charlton, Keith A.; Pieterse, Corné M. J.; Lorenzo, Oscar; García-Sánchez, Susana

    2013-01-01

    Microbe-Associated Molecular Patterns and virulence effectors are recognized by plants as a first step to mount a defence response against potential pathogens. This recognition involves a large family of extracellular membrane receptors and other immune proteins located in different sub-cellular compartments. We have used phage-display technology to express and select for Arabidopsis proteins able to bind bacterial pathogens. To rapidly identify microbe-bound phage, we developed a monitoring method based on microarrays. This combined strategy allowed for a genome-wide screening of plant proteins involved in pathogen perception. Two phage libraries for high-throughput selection were constructed from cDNA of plants infected with Pseudomonas aeruginosa PA14, or from combined samples of the virulent isolate DC3000 of Pseudomonas syringae pv. tomato and its avirulent variant avrRpt2. These three pathosystems represent different degrees in the specificity of plant-microbe interactions. Libraries cover up to 2×107 different plant transcripts that can be displayed as functional proteins on the surface of T7 bacteriophage. A number of these were selected in a bio-panning assay for binding to Pseudomonas cells. Among the selected clones we isolated the ethylene response factor ATERF-1, which was able to bind the three bacterial strains in competition assays. ATERF-1 was rapidly exported from the nucleus upon infiltration of either alive or heat-killed Pseudomonas. Moreover, aterf-1 mutants exhibited enhanced susceptibility to infection. These findings suggest that ATERF-1 contains a microbe-recognition domain with a role in plant defence. To identify other putative pathogen-binding proteins on a genome-wide scale, the copy number of selected-vs.-total clones was compared by hybridizing phage cDNAs with Arabidopsis microarrays. Microarray analysis revealed a set of 472 candidates with significant fold change. Within this set defence-related genes, including well-known targets of bacterial effectors, are over-represented. Other genes non-previously related to defence can be associated through this study with general or strain-specific recognition of Pseudomonas. PMID:23372747

  16. Regioselectivity of glucosylation of caffeic acid by a UDP-glucose:glucosyltransferase is maintained in planta.

    PubMed Central

    Lim, Eng-Kiat; Higgins, Gillian S; Li, Yi; Bowles, Dianna J

    2003-01-01

    Caffeic acid is a phenylpropanoid playing an important role in the pathways leading to lignin synthesis and the production of a wide variety of secondary metabolites. The compound is also an antioxidant and has potential utility as a general protectant against free radicals. Three glucosylated forms of caffeic acid are known to exist: the 3- O - and 4- O -glucosides and the glucose ester. This study describes for the first time a glucosyltransferase [UDP-glucose:glucosyltransferase (UGT)] that is specific for the 3-hydroxyl, and not the 4-hydroxyl, position of caffeic acid. The UGT sequence of Arabidopsis, UGT71C1, has been expressed as a recombinant fusion protein in Escherichia coli, purified and assayed against a range of substrates in vitro. The assay confirmed that caffeic acid as the preferred substrate when compared with other hydroxycinnamates, although UGT71C1 also exhibited substantial activity towards flavonoid substrates, known to have structural features that can be recognized by many different UGTs. The expression of UGT71C1 in transgenic Arabidopsis was driven by the constitutive cauliflower mosaic virus 35 S (CaMV35S) promoter. Nine independent transgenic lines were taken to homozygosity and characterized by Northern-blot analysis, assay of enzyme activity in leaf extracts and HPLC analysis of the glucosides. The level of expression of UGT71C1 was enhanced considerably in several lines, leading to a higher level of the corresponding enzyme activity and a higher level of caffeoyl-3- O -glucoside. The data are discussed in the context of the utility of UGTs for natural product biotransformations. PMID:12741958

  17. Angiogenic activity of latex from Euphorbia tirucalli Linnaeus 1753 (Plantae, Euphorbiaceae).

    PubMed

    Bessa, G; Melo-Reis, P R; Araújo, L A; Mrué, F; Freitas, G B; Brandão, M L; Silva Júnior, N J

    2015-08-01

    To assess the pro-angiogenic activity of Euphorbia tirucalli, commonly known as "avelós" plant, we performed a series of tests by applying an aqueous E. tirucalli latex solution (10 mg/mL) to the chorioallantoic membranes (CAMs) of 80 fertilized chicken eggs incubated in a temperature- and humidity-controlled automatic incubator. The results indicated that the aqueous latex solution increased vascular network formation compared to that with the negative control (p < 0.05) and the inhibitor control (p < 0.05). This suggests that under the experimental conditions tested, the aqueous latex solution induced an inflammatory response leading to neoangiogenesis. PMID:26465734

  18. Chrysanthemyl Diphosphate Synthase Operates in Planta as a Bifunctional Enzyme with Chrysanthemol Synthase Activity*

    PubMed Central

    Yang, Ting; Gao, Liping; Hu, Hao; Stoopen, Geert; Wang, Caiyun; Jongsma, Maarten A.

    2014-01-01

    Chrysanthemyl diphosphate synthase (CDS) is the first pathway-specific enzyme in the biosynthesis of pyrethrins, the most widely used plant-derived pesticide. CDS catalyzes c1′-2-3 cyclopropanation reactions of two molecules of dimethylallyl diphosphate (DMAPP) to yield chrysanthemyl diphosphate (CPP). Three proteins are known to catalyze this cyclopropanation reaction of terpene precursors. Two of them, phytoene and squalene synthase, are bifunctional enzymes with both prenyltransferase and terpene synthase activity. CDS, the other member, has been reported to perform only the prenyltransferase step. Here we show that the NDXXD catalytic motif of CDS, under the lower substrate conditions prevalent in plants, also catalyzes the next step, converting CPP into chrysanthemol by hydrolyzing the diphosphate moiety. The enzymatic hydrolysis reaction followed conventional Michaelis-Menten kinetics, with a Km value for CPP of 196 μm. For the chrysanthemol synthase activity, DMAPP competed with CPP as substrate. The DMAPP concentration required for half-maximal activity to produce chrysanthemol was ∼100 μm, and significant substrate inhibition was observed at elevated DMAPP concentrations. The N-terminal peptide of CDS was identified as a plastid-targeting peptide. Transgenic tobacco plants overexpressing CDS emitted chrysanthemol at a rate of 0.12–0.16 μg h−1 g−1 fresh weight. We propose that CDS should be renamed a chrysanthemol synthase utilizing DMAPP as substrate. PMID:25378387

  19. In planta changes in protein phosphorylation induced by the plant hormone abscisic acid

    PubMed Central

    Kline, Kelli G.; Barrett-Wilt, Gregory A.; Sussman, Michael R.

    2010-01-01

    Abscisic acid (ABA) is a hormone that controls seed dormancy and germination as well as the overall plant response to important environmental stresses such as drought. Recent studies have demonstrated that the ABA-bound receptor binds to and inhibits a class of protein phosphatases. To identify more broadly the phosphoproteins affected by this hormone in vivo, we used 14N/15N metabolic labeling to perform a quantitative untargeted mass spectrometric analysis of the Arabidopsis thaliana phosphoproteome following ABA treatment. We found that 50 different phosphopeptides had their phosphorylation state significantly altered by ABA over a treatment period lasting 5–30 min. Among these changes were increases in phosphorylation of subfamily 2 SNF1-related kinases and ABA-responsive basic leucine zipper transcription factors implicated in ABA signaling by previous in vitro studies. Furthermore, four members of the aquaporin family showed decreased phosphorylation at a carboxy-terminal serine which is predicted to cause closure of the water-transporting aquaporin gate, consistent with ABA's role in ameliorating the effect of drought. Finally, more than 20 proteins not previously known to be involved with ABA were found to have significantly altered phosphorylation levels. Many of these changes are phosphorylation decreases, indicating that an expanded model of ABA signaling, beyond simple phosphatase inhibition, may be necessary. This quantitative proteomics dataset provides a more comprehensive, albeit incomplete, view both of the protein targets whose biochemical activities are likely to be controlled by ABA and of the nature of the emerging phosphorylation and dephosphorylation cascades triggered by this hormone. PMID:20733066

  20. In planta localisation patterns of MADS domain proteins during floral development in Arabidopsis thaliana

    PubMed Central

    Urbanus, Susan L; de Folter, Stefan; Shchennikova, Anna V; Kaufmann, Kerstin; Immink, Richard GH; Angenent, Gerco C

    2009-01-01

    Background MADS domain transcription factors play important roles in various developmental processes in flowering plants. Members of this family play a prominent role in the transition to flowering and the specification of floral organ identity. Several studies reported mRNA expression patterns of the genes encoding these MADS domain proteins, however, these studies do not provide the necessary information on the temporal and spatial localisation of the proteins. We have made GREEN FLUORESCENT PROTEIN (GFP) translational fusions with the four MADS domain proteins SEPALLATA3, AGAMOUS, FRUITFULL and APETALA1 from the model plant Arabidopsis thaliana and analysed the protein localisation patterns in living plant tissues by confocal laser scanning microscopy (CLSM). Results We unravelled the protein localisation patterns of the four MADS domain proteins at a cellular and subcellular level in inflorescence and floral meristems, during development of the early flower bud stages, and during further differentiation of the floral organs. The protein localisation patterns revealed a few deviations from known mRNA expression patterns, suggesting a non-cell autonomous action of these factors or alternative control mechanisms. In addition, we observed a change in the subcellular localisation of SEPALLATA3 from a predominantly nuclear localisation to a more cytoplasmic localisation, occurring specifically during petal and stamen development. Furthermore, we show that the down-regulation of the homeodomain transcription factor WUSCHEL in ovular tissues is preceded by the occurrence of both AGAMOUS and SEPALLATA3 proteins, supporting the hypothesis that both proteins together suppress WUSCHEL expression in the ovule. Conclusion This approach provides a highly detailed in situ map of MADS domain protein presence during early and later stages of floral development. The subcellular localisation of the transcription factors in the cytoplasm, as observed at certain stages during development, points to mechanisms other than transcriptional control. Together this information is essential to understand the role of these proteins in the regulatory processes that drive floral development and leads to new hypotheses. PMID:19138429

  1. Staphylococcal abscess caused by trauma with a rosebush aculeus (Plantae, Rosaceae): apropos of a case.

    PubMed

    Bernardes Filho, Fred; Alves, Andreia Oliveira; Martins, Gustavo; Sasso, Letícia Soares; Gama, Carolina Mendonça; Cenci, Gardênia Borges

    2013-01-01

    Cutaneous abscess is a localized collection of pus in the dermis and subcutaneous tissue usually caused by trauma. The authors report the case of a 30-year-old male patient, gardener, that presents an ulcerated plaque in the third right finger, caused by an aculeus plant wound. The examination of the lesion's exudate ruled out the existence of fungi and showed the presence of Staphylococcus aureus. The authors emphasize the sporotrichosis as an important differential diagnosis, especially in endemic areas, and the indication of the exudate culture is discussed. PMID:24474127

  2. Staphylococcal abscess caused by trauma with a rosebush aculeus (Plantae, Rosaceae): apropos of a case*

    PubMed Central

    Bernardes Filho, Fred; Alves, Andreia Oliveira; Martins, Gustavo; Sasso, Letícia Soares; Gama, Carolina Mendonça; Cenci, Gardênia Borges

    2013-01-01

    Cutaneous abscess is a localized collection of pus in the dermis and subcutaneous tissue usually caused by trauma. The authors report the case of a 30-year-old male patient, gardener, that presents an ulcerated plaque in the third right finger, caused by an aculeus plant wound. The examination of the lesion's exudate ruled out the existence of fungi and showed the presence of Staphylococcus aureus. The authors emphasize the sporotrichosis as an important differential diagnosis, especially in endemic areas, and the indication of the exudate culture is discussed. PMID:24474127

  3. Regulation and mechanism of potassium release from barley roots: an in planta 42K+ analysis.

    PubMed

    Coskun, Devrim; Britto, Dev T; Kronzucker, Herbert J

    2010-12-01

    Potassium (K(+) ) flux into plant cells is a well-characterized ion transport phenomenon. By contrast, little is known about the mechanisms and regulation of K(+) flux from the cell. Here, we present a radioisotopic analysis of K(+) fluxes from roots of intact barley (Hordeum vulgare), in the context of recent discoveries in the molecular biology and electrophysiology of this process. Plants were labelled with (42)K(+), and kinetics of its release from roots were monitored at low (0.1 mM) or high (1.0 mM) external K concentration, [K(+)](ext), and with the application of channel modulators and nutrient shifts. At 0.1 (but not 1.0) mM [K(+)], where K(+) efflux is thought to be mediated by K(+)-outward-rectifying channels, (42)K(+) efflux was inhibited by the channel blockers barium (Ba(2+)), caesium (Cs(+)), tetraethylammonium (TEA(+)), and lanthanum (La(3+)). Ammonium and nitrate (10 mM) stimulated and inhibited (42)K(+) efflux, respectively, while 10 mM [K(+)](ext) or [Rb(+) ](ext) decreased it. No evidence for the involvement of ATP-binding cassettes, nonselective cation channels, or active K(+)-efflux pumps was found. Our study provides new evidence for the thermodynamic transition between high- and low-affinity transport, from the efflux perspective, identifying the operation of channels at low [K(+)], and the cessation of transmembrane efflux at high [K(+)]. PMID:20731780

  4. Chemical compounds effective against the citrus huanglongbing bacterium 'Candidatus Liberibacter asiaticus' in planta

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus huanglongbing (HLB) is one of the most destructive diseases of citrus worldwide, and is threatening the survival of the Floridian citrus industry. Currently, there is no established cure for this century-old and emerging disease. The new antibiotic combination of penicillin and streptomycin (...

  5. The Papaver rhoeas S determinants confer self-incompatibility to Arabidopsis thaliana in planta.

    PubMed

    Lin, Zongcheng; Eaves, Deborah J; Sanchez-Moran, Eugenio; Franklin, F Christopher H; Franklin-Tong, Vernonica E

    2015-11-01

    Self-incompatibility (SI) is a major genetically controlled system used to prevent inbreeding in higher plants. S determinants regulate allele-specific rejection of "self" pollen by the pistil. SI is an important model system for cell-to-cell recognition and signaling and could be potentially useful for first-generation (F1) hybrid breeding. To date, the transfer of S determinants has used the complementation of orthologs to "restore" SI in close relatives. We expressed the Papaver rhoeas S determinants PrsS and PrpS in Arabidopsis thaliana. This enabled pistils to reject pollen expressing cognate PrpS. Moreover, plants coexpressing cognate PrpS and PrsS exhibit robust SI. This demonstrates that PrsS and PrpS are sufficient for a functional synthetic S locus in vivo. This transfer of novel S determinants into a highly divergent species (>140 million years apart) with no orthologs suggests their potential utility in crop production. PMID:26542572

  6. The Moon Phases in a Paper Box. (Spanish Title: Las Fases de la Luna en Una Caja de Cartón.) As Fases da Lua Numa Caixa de Papelão

    NASA Astrophysics Data System (ADS)

    de Fátima O. Saraiva, Maria; Amador, Cláudio B.; Kemper, Érico; Goulart, Paulo; Muller, Angela

    2007-12-01

    We present a very simple concrete model to demonstrate the concept of phases of an illuminated body. The main objective of our model is to help the understanding of the Moon phases as viewed from the perspective of an observer on Earth. The material allows the visualization of two important effects: (1st) even though all the time half Moon is illuminated by the Sun, we see different fractions of the illuminated Moon surface, depending on our angle of sight; (2nd) the orientation of the convex part of the Moon in the crescent and waning phases on the sky also depends on our perspective from Earth. The use of a closed box allows one to see the contrast among the different phases with no need of a dark room. We also present a text on the Moon phases, emphasizing the dependence of the aspect of the bright part on the angle of sight. En este trabajo proponemos la construcción de material didáctico de bajo costo para demostración del concepto de fases de un cuerpo iluminado. El principal objetivo de nuestro material es facilitar la comprensión de las fases de la Luna desde la perspectiva de un observador en la Tierra. El material ayuda la visualización de dos efectos importantes: (1º) a pesar de tener siempre la mitad de la Luna (representada por una bolita de espuma plástica o de ping-pong), iluminada por el Sol ( representado por una fuente de luz natural o artificial), vemos diferentes fracciones de su superficie iluminada, dependiendo del ángulo por el cual la vemos; (2º) la orientación del borde convexo de la Luna en las fases Creciente y Menguante también depende de la perspectiva por la cual la miramos desde la Tierra. El uso de una caja cerrada permite observar el contraste entre las diferentes fases sin necesidad de estar en un recinto oscuro. Presentamos también un texto explicativo sobre las fases de la Luna, enfatizando la dependencia de la apariencia de la parte iluminada con el ángulo de visión. Neste trabalho propomos a construção de

  7. Autoreporte de exposición a publicidad y promoción de tabaco en una cohorte de fumadores mexicanos: Antes y durante la publicación de la Ley General para el Control del tabaco en 2008

    PubMed Central

    Hernández, Rosaura Pérez; Thrasher, James F.; Bolaños, Rosibel Rodríguez; Gutiérrez, Inti Barrientos; Hernández, Norma A Ibañez

    2015-01-01

    Resumen Objetivo Determinar en población fumadora el nivel de exposición a la mercadotecnia por parte de la Industria Tabacalera (IT), a través de diferentes métodos de promocionar sus productos de tabaco, antes y durante la publicación de la Ley General para el Control del Tabaco (LGCT) en 2008. Material y métodos Estudio de cohorte en fumadores adultos (n=941 pre-LGCT y n=1051 post-LGCT) de cuatro ciudades mexicanas. Se realizaron análisis multivariados mediante modelos de ecuaciones de estimación generalizada (GEE). Resultados Se incremento el autoreporte de recepción de muestras gratis de cigarros (3.7% a 8.1%), ropa o artículos con marcas o logos (3.6% a 6.4%), haber visto información sobre eventos especiales (1.9% a 4.7%), y bares, antros y discos para mayores de edad (21.4% a 28%). Se observaron decrementos de publicidad en exteriores (54.7% a 47.2%). Conclusión Es necesaria una política integral con prohibiciones totales de la publicidad y promoción de los productos de tabaco que integre mayor vigilancia y sanciones para lograr la disminución y prevención del consumo de tabaco. PMID:22689158

  8. Basic Concepts of Astronomy: a Methodological Proposal. (Spanish Title: Conceptos Básicos de Astronomía: Una Propuesta Metodológica.) Conceitos Básicos de Astronomia: Uma Proposta Metodológica

    NASA Astrophysics Data System (ADS)

    Darroz, Luiz Marcelo; Heineck, Renato; Samudio Pérez, Carlos Ariel

    2011-12-01

    In this report, the development of a methodological proposal which approaches basic concepts of astronomy-grounded pedagogically on Meaningful Learning is described. The proposal, which consists of four meetings, was developed by teachers and academics of the course of Professor in Physics of the University of Passo Fundo (UPF), through an extension course to a group of highschool students of a public school of the town of Passo Fundo, RS. The work was focused into basic concepts of astronomy. The signs of Meaningful Learning have been obtained by means of research and evaluation tools that were applied at the end of each meeting. The evaluation of the proposal has been conducted by means of a final questionnaire which was answered by the participants at the end ofthe development of activities. By means of the results obtained from the different instruments, and the comments made by the participants during the activities and by means of the high rates of approval obtained in the final questionnaire, we think that the proposal reached the established goals and it may be repeated with the certainty of success. En este relato se describe una propuesta de desarrollo metodológico que aborda conceptos básicos de astronomía fundamentada pedagógicamente en el Aprendizaje Significativo. La propuesta que comprende cuatro encuentros, fue desarrollada por profesores y académicos del curso de Licenciatura en Física de la Universidad de Passo Fundo (UPF), a través de un curso de extensión para un grupo de Liceo del 6º año de una Escuela Pública de la ciudad de Passo Fundo/RS. El trabajo tuvo como eje principal los "conceptos básicos de astronomía". Los indicios de Aprendizaje Significativo fueron obtenidos por instrumentos de pesquisa y evaluación, siempre aplicados después de cada encuentro. La evaluación de la propuesta fue hecha a través de un cuestionario final y contestado por los participantes al finalizar el desarrollo de actividades. Por los resultados

  9. Estudio epidemiológico de sucesos traumáticos, trastorno de estrés post-traumático y otros trastornos psiquiátricos en una muestra representativa de Chile

    PubMed Central

    Pérez Benítez, Carlos I.; Vicente, Benjamin; Zlotnick, Caron; Kohn, Robert; Johnson, Jennifer; Valdivia, Sandra; Rioseco, Pedro

    2010-01-01

    RESUMEN Durante la década de 1990 en los Estados Unidos (EU), el conocimiento sobre el trastorno de estrés post-traumático (TEPT) evolucionó de estudios específicos en un principio, sobre veteranos de guerra y sobre víctimas de desastres, a estudios epidemiológicos más tarde, sin embargo, la epidemiología del TEPT en países en desarrollo ha sido un área poco estudiada hasta ahora. Los expertos en el área de trauma han propuesto que los sucesos traumáticos que ocurren en la niñez son más perjudiciales para la salud mental que aquellos que ocurren más tarde en la vida. Este trabajo revisa los resultados de un estudio epidemiológico llevado a cabo en Chile. Específicamente, se revisan los resultados sobre las tasas de prevalencia del TEPT, traumas asociados más frecuentemente con él, así como la comorbilidad de este trastorno con otros trastornos psiquiátricos a lo largo de la vida. Igualmente se analizaron las diferencias del TEPT en cada sexo, así como la exposición a traumas en una muestra representativa de chilenos. Además se comparó la prevalencia de trastornos psiquiátricos en personas que sufrieron su primer trauma durante la niñez, durante la edad adulta, o que no reportaron traumas durante su vida. En estos estudios epidemiológicos se usaron módulos del TEPT y trastorno de personalidad antisocial (TPA) de la entrevista diagnóstica siguiendo los criterios del DSM-III-R (DIS–III-R). Para evaluar el resto de los trastornos psiquiátricos se usó la Entrevista Diagnóstica Internacional Compuesta (CIDI). Estos instrumentos fueron administrados en tres ciudades chilenas a 2390 personas mayores de 15 años. Para estimar los errores estándares (EE) debido al diseño de la muestra y a la necesidad de ajuste se usó el método Taylor de linearización seriada. También se usó un análisis de regresión logística para examinar la relación entre el TEPT, los factores demográficos de riesgo y el tipo de trauma. Además se utilizó la

  10. A Proposed Activity for a Meaningful Learning about the Moon Phases. (Breton Title: Uma Proposta de Atividade Para a Aprendizagem Significativa sobre as Fases da Lua.) Una Actividad Propuesta Para EL Aprendizaje Significativo Acerca de Las Fases de la Luna

    NASA Astrophysics Data System (ADS)

    Martins, Bruno Andrade; Langhi, Rodolfo

    2012-12-01

    ínua avaliação durante o processo, culmina com a elaboração de uma história em quadrinhos envolvendo as fases da Lua. Portanto, o objetivo deste trabalho é apresentar uma proposta de atividade didática diferenciada sustentada pelos princípios teóricos da aprendizagem significativa aplicada ao ensino das fases da Lua durante as aulas de Física no ensino médio. En este trabajo se presenta uno de los contenidos de la Astronomía y el evidente fracaso en la enseñanza del tema en la escuela secundaria, a pesar del hecho que los documentos oficiales apuntan para la necesidad de trabajar contenidos de Astronomía en este nivel. Entre los conceptos alternativos en Astronomía que los alumnos secundarios llevan consigo, aún después de terminados los estudios, destacamos aquí el fenómeno de las fases de la Luna. El desarrollo de diferentes estrategias en relación con los métodos tradicionales, dirigidas al proceso de enseñanza-aprendizaje en este tema fue considerado en este trabajo como uno de los resultados obtenidos a la luz de los referenciales del aprendizaje significativo, tal como fueron fundamentados por Ausubel. Según la propuesta que aquí se presenta, la participación activa de los estudiantes en la ejecución de una actividad experimental y otras actividades educativas destinadas a la evaluación continua durante el proceso culminó en la elaboración de una historieta respecto de las fases de la Luna. Por lo tanto, el objetivo de este trabajo es presentar una propuesta de actividad de enseñanza diferenciada con el apoyo de los principios teóricos del aprendizaje significativo aplicado a la enseñanza de las fases de la luna durante las clases de física del ciclo secundario.

  11. Suva Una? (What Is It Doing?)

    ERIC Educational Resources Information Center

    MacLean, Edna A.

    This primary level reader is part of a series of materials developed for beginning reading instruction in Barrow Inupiaq. Each page consists of a drawing and brief text. It is a revision of an earlier version of the same title. (AMH)

  12. Ciencia, Sociedad, Soluciones: Una Introduccion al USGS

    USGS Publications Warehouse

    U.S. Geological Survey

    2001-01-01

    El USGS sirve a la nacion de los Estados Unidos proveyendo informacion fidedigna para ? Describir y comprender la Tierra; ? Minimizar la perdida de vidas y propiedades por desastres naturales; ? Manejar los recursos hidrologicos, biologicos, energeticos y minerales; y ? Mejorar y proteger nuestra calidad de vida.

  13. In planta detection of Xanthomonas axonopodis pv. commiphorae using fyuA and rpoD genes.

    PubMed

    Samanta, Jatindra Nath; Mandal, Kunal

    2013-06-01

    Guggal is tapped for extraction of medicinally important oleo-gum-resin (guggul) by inoculating the stem bark with natural gum suspension containing pathogenic bacterium Xanthomonas axonopodis pv. commiphorae (Xac). The tree dies in the process. In absence of any specific medium for isolation of Xac, it is difficult to assess spread of the pathogen within the plant. A PCR based molecular detection technique usingfyuA and rpoD gene specific primers is described here. The primers amplified products only from Xac and not from host tissues or common saprophytes. The method was sensitive enough to produce positive signals for up to 4.4 bacterial cells or 2 pg target DNA per reaction mixture. However, PCR inhibitors present in plant tissues drastically reduced the limit of detection. A simple overnight incubation of surface sterilised plant tissues in nutrient medium was introduced to increase pathogen titre and to overcome this problem. This technique was successfully used to measure spread of Xac in plant tissues away from the site of inoculation. The pathogen showed preference for acropetal movement and did not spread to 7-8 cm below the site of inoculation till 15 days after inoculation. This suggests possibility to manage the disease through plant surgery. PMID:23926696

  14. Inhibitory effect of Xenorhabdus nematophila TB on plant pathogens Phytophthora capsici and Botrytis cinerea in vitro and in planta

    PubMed Central

    Fang, Xiangling; Zhang, Manrang; Tang, Qian; Wang, Yonghong; Zhang, Xing

    2014-01-01

    Entomopathogenic bacteria Xenorhabdus spp. produce secondary metabolites with potential antimicrobial activity for use in agricultural productions. This study evaluated the inhibitory effect of X. nematophila TB culture on plant pathogens Botrytis cinerea and Phytophthora capsici. The cell-free filtrate of TB culture showed strong inhibitory effects (>90%) on mycelial growth of both pathogens. The methanol-extracted bioactive compounds (methanol extract) of TB culture also had strong inhibitory effects on mycelial growth and spore germinations of both pathogens. The methanol extract (1000 μg/mL) and cell-free filtrate both showed strong therapeutic and protective effects (>70%) on grey mold both in detached tomato fruits and plants, and leaf scorch in pepper plants. This study demonstrates X. nematophila TB produces antimicrobial metabolites of strong activity on plant pathogens, with great potential for controlling tomato grey mold and pepper leaf scorch and being used in integrated disease control to reduce chemical application. PMID:24599183

  15. In planta production of the highly potent resveratrol analogue pterostilbene via stilbene synthase and O-methyltransferase co-expression

    SciTech Connect

    Rimando A. M.; Liu C.; Pan, Z.; Polashock, J. J.; Dayan, F. E., Mizuno, C. S.; Snook, M. E.; Baerson, S. R.

    2012-04-01

    Resveratrol and related stilbenes are thought to play important roles in defence responses in several plant species and have also generated considerable interest as nutraceuticals owing to their diverse health-promoting properties. Pterostilbene, a 3,5-dimethylether derivative of resveratrol, possesses properties similar to its parent compound and, additionally, exhibits significantly higher fungicidal activity in vitro and superior pharmacokinetic properties in vivo. Recombinant enzyme studies carried out using a previously characterized O-methyltransferase sequence from Sorghum bicolor (SbOMT3) demonstrated its ability to catalyse the A ring-specific 3,5-bis-O-methylation of resveratrol, yielding pterostilbene. A binary vector was constructed for the constitutive co-expression of SbOMT3 with a stilbene synthase sequence from peanut (AhSTS3) and used for the generation of stably transformed tobacco and Arabidopsis plants, resulting in the accumulation of pterostilbene in both species. A reduced floral pigmentation phenotype observed in multiple tobacco transformants was further investigated by reversed-phase HPLC analysis, revealing substantial decreases in both dihydroquercetin-derived flavonoids and phenylpropanoid-conjugated polyamines in pterostilbene-producing SbOMT3/AhSTS3 events. These results demonstrate the potential utility of this strategy for the generation of pterostilbene-producing crops and also underscore the need for the development of additional approaches for minimizing concomitant reductions in key phenylpropanoid-derived metabolites.

  16. Production of recombinant single chain antibodies (scFv) in vegetatively reproductive Kalanchoe pinnata by in planta transformation.

    PubMed

    Jung, Yuchul; Rhee, Yong; Auh, Chung-Kyoon; Shim, Hyekyung; Choi, Jung-Jin; Kwon, Suk-Tae; Yang, Joo-Sung; Kim, Donggiun; Kwon, Myung-Hee; Kim, Yong-Sung; Lee, Sukchan

    2009-10-01

    We developed an asexual reproductive plant, Kalanchoe pinnata, as a new bioreactor for plant-based molecular farming using a newly developed transformation method. Leaf crenate margins were pin-pricked to infect the plant with the Agrobacterium strain LBA4404 and vacuum infiltration was also applied to introduce the target gene into the plants. Subsequently, the young mother leaf produced new clones at the leaf crenate margins without the need for time- and labor-consuming tissue culture procedures. The average transformation rates were approximately 77 and 84% for pin-prickling and vacuum-infiltration methods, respectively. To functionally characterize an introduced target protein, a nucleic acid hydrolyzing recombinant 3D8 scFv was selected and the plant based 3D8 scFv proteins were purified and analyzed. Based on abzyme analysis, the purified protein expressed with this system had catalytic activity and exhibited all of properties of the protein produced in an E. coli system. This result suggested that vegetatively reproductive K. pinnata can be a novel and potent bioreactor for bio-pharmaceutical proteins. PMID:19688214

  17. Establishing and fine-tuning an in planta system for potato zebra chip disease research in California

    Technology Transfer Automated Retrieval System (TEKTRAN)

    We have acquired an APHIS-PPQ permit to work on “Candidatus Liberibacter solanacearum” at the USDA-ARS San Joaquin Valley Agricultural Sciences Center in Parlier, California. “Ca. L. solanacearum”, associated with Zebra chip disease (ZCD) of potato. This bacterium is currently unculturable in vitro....

  18. Development of a Mild Viral Expression System for Gain-Of-Function Study of Phytoplasma Effector In Planta.

    PubMed

    Hu, Sin-Fen; Huang, Yu-Hsin; Lin, Chan-Pin; Liu, Li-Yu Daisy; Hong, Syuan-Fei; Yang, Chiao-Yin; Lo, Hsiao-Feng; Tseng, Ting-Yu; Chen, Wei-Yao; Lin, Shih-Shun

    2015-01-01

    PHYL1 and SAP54 are orthologs of pathogenic effectors of Aster yellow witches'-broom (AYWB) phytoplasma and Peanut witches'-broom (PnWB) phytoplasma, respectively. These effectors cause virescence and phyllody symptoms (hereafter leafy flower) in phytoplasma-infected plants. T0 lines of transgenic Arabidopsis expressing the PHYL1 or SAP54 genes (PHYL1 or SAP54 plants) show a leafy flower phenotype and result in seedless, suggesting that PHYL1 and SAP54 interfere with reproduction stage that restrict gain-of-function studies in the next generation of transgenic plants. Turnip mosaic virus (TuMV) mild strain (TuGK) has an Arg182Lys mutation in the helper-component proteinase (HC-ProR182K) that blocks suppression of the miRNA pathway and prevents symptom development in TuGK-infected plants. We exploited TuGK as a viral vector for gain-of-function studies of PHYL1 and SAP54 in Arabidopsis plants. TuGK-PHYL1- and TuGK-SAP54-infected Arabidopsis plants produced identical leafy flower phenotypes and similar gene expression profiles as PHYL1 and SAP54 plants. In addition, the leafy flower formation rate was enhanced in TuGK-PHYL1- or TuGK-SAP54-infected Arabidopsis plants that compared with the T0 lines of PHYL1 plants. These results provide more evidence and novel directions for further studying the mechanism of PHYL1/SAP54-mediated leafy flower development. In addition, the TuGK vector is a good alternative in transgenic plant approaches for rapid gene expression in gain-of-function studies. PMID:26076458

  19. A RAPID INFECTION ASSAY FOR ARMILLARIA AND REAL-TIME PCR QUANTITATION OF THE FUNGAL BIOMASS IN PLANTA

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Slow and unreliable infection in the greenhouse has been a barrier to research on Armillaria root disease. The existing infection assay takes 7-18 months for detectable infection, during which time the inoculum often dies, resulting in unequal challenge among plants. As symptom expression and mort...

  20. Remote sensing of gene expression in Planta: transgenic plants as monitors of exogenous stress perception in extraterrestrial environments

    NASA Technical Reports Server (NTRS)

    Manak, Michael S.; Paul, Anna-Lisa; Sehnke, Paul C.; Ferl, Robert J.

    2002-01-01

    Transgenic arabidopsis plants containing the alcohol dehydrogenase (Adh) gene promoter fused to the green fluorescent protein (GFP) reporter gene were developed as biological sensors for monitoring physiological responses to unique environments. Plants were monitored in vivo during exposure to hypoxia, high salt, cold, and abcissic acid in experiments designed to characterize the utility and responses of the Adh/GFP biosensors. Plants in the presence of environmental stimuli that induced the Adh promoter responded by expressing GFP, which in turn generated a detectable fluorescent signal. The GFP signal degraded when the inducing stimulus was removed. Digital imaging of the Adh/GFP plants exposed to each of the exogenous stresses demonstrated that the stress-induced gene expression could be followed in real time. The experimental results established the feasibility of using a digital monitoring system for collecting gene expression data in real time from Transgenic Arabidopsis Gene Expression System (TAGES) biosensor plants during space exploration experiments.

  1. In planta sequential hydroxylation and glycosylation of a fungal phytotoxin: Avoiding cell death and overcoming the fungal invader

    PubMed Central

    Pedras, M. S. C.; Zaharia, I. L.; Gai, Y.; Zhou, Y.; Ward, D. E.

    2001-01-01

    To facilitate plant colonization, some pathogenic fungi produce phytotoxic metabolites that damage tissues; plants may be resistant to a particular pathogen if they produce an enzyme(s) that catalyzes detoxification of this metabolite(s). Alternaria blackspot is one of the most damaging and significant fungal diseases of brassica crops, with no source of resistance known within the Brassica species. Destruxin B is the major phytotoxin produced by the blackspot-causing fungus, Alternaria brassicae (Berkley) Saccardo. We have established that a blackspot-resistant species (Sinapis alba) metabolized 14C-labeled destruxin B to a less toxic product substantially faster than any of the susceptible species. The first metabolite, hydroxydestruxin B (14C-labeled), was further biotransformed to the β-d-glucosyl derivative at a slower rate. The structures of hydroxydestruxin B and β-d-glucosyl hydroxydestruxin B were deduced from their spectroscopic data [NMR, high resolution (HR)-MS, Fourier transform infrared (FTIR)] and confirmed by total chemical synthesis. Although these hydroxylation and glucosylation reactions occurred in both resistant (S. alba) and susceptible (Brassica napus, Brassica juncea, and Brassica rapa) species, hydroxylation was the rate limiting step in the susceptible species, whereas glucosylation was the rate limiting step in the resistant species. Remarkably, it was observed that the hydroxydestruxin B induced the biosynthesis of phytoalexins in blackspot-resistant species but not in susceptible species. This appears to be a unique example of phytotoxin detoxification and simultaneous phytoalexin elicitation by the detoxification product. Our studies suggest that S. alba can overcome the fungal invader through detoxification of destruxin B coupled with production of phytoalexins. PMID:11149945

  2. A visual identification key utilizing both gestalt and analytic approaches to identification of Carices present in North America (Plantae, Cyperaceae)

    PubMed Central

    2013-01-01

    Abstract Images are a critical part of the identification process because they enable direct, immediate and relatively unmediated comparisons between a specimen being identified and one or more reference specimens. The Carices Interactive Visual Identification Key (CIVIK) is a novel tool for identification of North American Carex species, the largest vascular plant genus in North America, and two less numerous closely-related genera, Cymophyllus and Kobresia. CIVIK incorporates 1288 high-resolution tiled image sets that allow users to zoom in to view minute structures that are crucial at times for identification in these genera. Morphological data are derived from the earlier Carex Interactive Identification Key (CIIK) which in turn used data from the Flora of North America treatments. In this new iteration, images can be viewed in a grid or histogram format, allowing multiple representations of data. In both formats the images are fully zoomable. PMID:24723777

  3. Binary 2in1 Vectors Improve in Planta (Co)localization and Dynamic Protein Interaction Studies1[OPEN

    PubMed Central

    Hecker, Andreas; Wallmeroth, Niklas; Peter, Sébastien; Blatt, Michael R.

    2015-01-01

    Fluorescence-based protein-protein interaction techniques are vital tools for understanding in vivo cellular functions on a mechanistic level. However, only under the condition of highly efficient (co)transformation and accumulation can techniques such as Förster resonance energy transfer (FRET) realize their potential for providing highly accurate and quantitative interaction data. FRET as a fluorescence-based method unifies several advantages, such as measuring in an in vivo environment, real-time context, and the ability to include transient interactions as well as detecting the mere proximity of proteins. Here, we introduce a novel vector set that incorporates the benefit of the recombination-based 2in1 cloning system with the latest state-of-the-art fluorescent proteins for optimal coaccumulation and FRET output studies. We demonstrate its utility across a range of methods. Merging the 2in1 cloning system with new-generation FRET fluorophore pairs allows for enhanced detection, speeds up the preparation of clones, and enables colocalization studies and the identification of meaningful protein-protein interactions in vivo. PMID:25971551

  4. Health risk assessment of air emissions from a municipal solid waste incineration plant--a case study.

    PubMed

    Cangialosi, Federico; Intini, Gianluca; Liberti, Lorenzo; Notarnicola, Michele; Stellacci, Paolo

    2008-01-01

    A health risk assessment of long-term emissions of carcinogenic and non-carcinogenic air pollutants has been carried out for the municipal solid waste incinerator (MSWI) of the city of Taranto, Italy. Ground level air concentrations and soil deposition of carcinogenic (Polychlorinated Dibenzo-p-Dioxins/Furans and Cd) and non-carcinogenic (Pb and Hg) pollutants have been estimated using a well documented atmospheric dispersion model. Health risk values for air inhalation, dermal contact, soil and food ingestion have been calculated based on a combination of these concentrations and a matrix of environmental exposure factors. Exposure of the surrounding population has been addressed for different release scenarios based on four pollutants, four exposure pathways and two receptor groups (children and adults). Spatial risk distribution and cancer excess cases projected from plant emissions have been compared with background mortality records. Estimated results based on the MSWI emissions show: (1) individual risks well below maximum acceptable levels, (2) very small incremental cancer risk compared with background level. PMID:17611096

  5. Phytohormonal networks promote differentiation of fiber initials on pre-anthesis cotton ovules grown in vitro and in planta

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The number of cotton (Gossypium sp.) ovule epidermal cells differentiating into fiber initials is an important factor affecting cotton yield and fiber quality. Despite extensive efforts in determining the molecular mechanisms regulating fiber initial differentiation, only a few genes responsible for...

  6. In planta mechanism of action of leptospermone: impact of its physico-chemical properties on uptake, translocation, and metabolism

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Leptospermone is a natural ß-triketone that specifically inhibits the enzyme p-hydrophyphenylpyruvate dioxygenase, the same molecular target site as that of the commercial herbicide mesotrione. The ß-triketone-rich essential oil of Leptospermum scoparium has both preemergence or postemergence herbi...

  7. Production and characterization of in planta transiently produced polygalacturanase from Aspergillus niger and its fusions with hydrophobin or ELP tags

    PubMed Central

    2014-01-01

    Background Pectinases play an important role in plant cell wall deconstruction and have potential in diverse industries such as food, wine, animal feed, textile, paper, fuel, and others. The demand for such enzymes is increasing exponentially, as are the efforts to improve their production and to implement their use in several industrial processes. The goal of this study was to examine the potential of producing polygalacturonase I from Aspergillus niger in plants and to investigate the effects of subcellular compartmentalization and protein fusions on its accumulation and activity. Results Polygalacturonase I from Aspergillus niger (AnPGI) was transiently produced in Nicotiana benthamiana by targeting it to five different cellular compartments: apoplast, endoplasmic reticulum (ER), vacuole, chloroplast and cytosol. Accumulation levels of 2.5%, 3.0%, and 1.9% of total soluble protein (TSP) were observed in the apoplast, ER, and vacuole, respectively, and specific activity was significantly higher in vacuole-targeted AnPGI compared to the same enzyme targeted to the ER or apoplast. No accumulation was found for AnPGI when targeted to the chloroplast or cytosol. Analysis of AnPGI fused with elastin-like polypeptide (ELP) revealed a significant increase in the protein accumulation level, especially when targeted to the vacuole where the protein doubles its accumulation to 3.6% of TSP, while the hydrophobin (HFBI) fusion impaired AnPGI accumulation and both tags impaired activity, albeit to different extents. The recombinant protein showed activity against polygalacturonic acid with optimum conditions at pH 5.0 and temperature from 30 to 50°C, depending on its fusion. In vivo analysis of reducing sugar content revealed a higher release of reducing sugars in plant tissue expressing recombinant AnPGI compared to wild type N. benthamiana leaves. Conclusion Our results demonstrate that subcellular compartmentalization of enzymes has an impact on both the target protein accumulation and its activity, especially in the case of proteins that undergo post-translational modifications, and should be taken into consideration when protein production strategies are designed. Using plants to produce heterologous enzymes for the degradation of a key component of the plant cell wall could reduce the cost of biomass pretreatment for the production of cellulosic biofuels. PMID:24970673

  8. In planta production of the highly potent resveratrol analogue pterostilbene via stilbene synthase and O-methyltransferase co-expression

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Stilbenoid phytoalexins such as resveratrol, play an important defense role in several plant species against pathogens such as Botrytis cinerea. In addition to their antimicrobial properties, resveratrol and related stilbenes have also generated considerable interest as nutraceuticals, due to their...

  9. An in planta-expressed polyketide synthase produces (R)-mellein in the wheat pathogen Parastagonospora nodorum.

    PubMed

    Chooi, Yit-Heng; Krill, Christian; Barrow, Russell A; Chen, Shasha; Trengove, Robert; Oliver, Richard P; Solomon, Peter S

    2015-01-01

    Parastagonospora nodorum is a pathogen of wheat that affects yields globally. Previous transcriptional analysis identified a partially reducing polyketide synthase (PR-PKS) gene, SNOG_00477 (SN477), in P. nodorum that is highly upregulated during infection of wheat leaves. Disruption of the corresponding SN477 gene resulted in the loss of production of two compounds, which we identified as (R)-mellein and (R)-O-methylmellein. Using a Saccharomyces cerevisiae yeast heterologous expression system, we successfully demonstrated that SN477 is the only enzyme required for the production of (R)-mellein. This is the first identification of a fungal PKS that is responsible for the synthesis of (R)-mellein. The P. nodorum ΔSN477 mutant did not show any significant difference from the wild-type strain in its virulence against wheat. However, (R)-mellein at 200 μg/ml inhibited the germination of wheat (Triticum aestivum) and barrel medic (Medicago truncatula) seeds. Comparative sequence analysis identified the presence of mellein synthase (MLNS) homologues in several Dothideomycetes and two sodariomycete genera. Phylogenetic analysis suggests that the MLNSs in fungi and bacteria evolved convergently from fungal and bacterial 6-methylsalicylic acid synthases. PMID:25326302

  10. An In Planta-Expressed Polyketide Synthase Produces (R)-Mellein in the Wheat Pathogen Parastagonospora nodorum

    PubMed Central

    Krill, Christian; Barrow, Russell A.; Chen, Shasha; Trengove, Robert; Oliver, Richard P.; Solomon, Peter S.

    2014-01-01

    Parastagonospora nodorum is a pathogen of wheat that affects yields globally. Previous transcriptional analysis identified a partially reducing polyketide synthase (PR-PKS) gene, SNOG_00477 (SN477), in P. nodorum that is highly upregulated during infection of wheat leaves. Disruption of the corresponding SN477 gene resulted in the loss of production of two compounds, which we identified as (R)-mellein and (R)-O-methylmellein. Using a Saccharomyces cerevisiae yeast heterologous expression system, we successfully demonstrated that SN477 is the only enzyme required for the production of (R)-mellein. This is the first identification of a fungal PKS that is responsible for the synthesis of (R)-mellein. The P. nodorum ΔSN477 mutant did not show any significant difference from the wild-type strain in its virulence against wheat. However, (R)-mellein at 200 μg/ml inhibited the germination of wheat (Triticum aestivum) and barrel medic (Medicago truncatula) seeds. Comparative sequence analysis identified the presence of mellein synthase (MLNS) homologues in several Dothideomycetes and two sodariomycete genera. Phylogenetic analysis suggests that the MLNSs in fungi and bacteria evolved convergently from fungal and bacterial 6-methylsalicylic acid synthases. PMID:25326302

  11. Development of a Mild Viral Expression System for Gain-Of-Function Study of Phytoplasma Effector In Planta

    PubMed Central

    Liu, Li-Yu Daisy; Hong, Syuan-Fei; Yang, Chiao-Yin; Lo, Hsiao-Feng; Tseng, Ting-Yu; Chen, Wei-Yao; Lin, Shih-Shun

    2015-01-01

    PHYL1 and SAP54 are orthologs of pathogenic effectors of Aster yellow witches’-broom (AYWB) phytoplasma and Peanut witches’-broom (PnWB) phytoplasma, respectively. These effectors cause virescence and phyllody symptoms (hereafter leafy flower) in phytoplasma-infected plants. T0 lines of transgenic Arabidopsis expressing the PHYL1 or SAP54 genes (PHYL1 or SAP54 plants) show a leafy flower phenotype and result in seedless, suggesting that PHYL1 and SAP54 interfere with reproduction stage that restrict gain-of-function studies in the next generation of transgenic plants. Turnip mosaic virus (TuMV) mild strain (TuGK) has an Arg182Lys mutation in the helper-component proteinase (HC-ProR182K) that blocks suppression of the miRNA pathway and prevents symptom development in TuGK-infected plants. We exploited TuGK as a viral vector for gain-of-function studies of PHYL1 and SAP54 in Arabidopsis plants. TuGK-PHYL1- and TuGK-SAP54-infected Arabidopsis plants produced identical leafy flower phenotypes and similar gene expression profiles as PHYL1 and SAP54 plants. In addition, the leafy flower formation rate was enhanced in TuGK-PHYL1- or TuGK-SAP54-infected Arabidopsis plants that compared with the T0 lines of PHYL1 plants. These results provide more evidence and novel directions for further studying the mechanism of PHYL1/SAP54-mediated leafy flower development. In addition, the TuGK vector is a good alternative in transgenic plant approaches for rapid gene expression in gain-of-function studies. PMID:26076458

  12. Inhibitory effect of Xenorhabdus nematophila TB on plant pathogens Phytophthora capsici and Botrytis cinerea in vitro and in planta.

    PubMed

    Fang, Xiangling; Zhang, Manrang; Tang, Qian; Wang, Yonghong; Zhang, Xing

    2014-01-01

    Entomopathogenic bacteria Xenorhabdus spp. produce secondary metabolites with potential antimicrobial activity for use in agricultural productions. This study evaluated the inhibitory effect of X. nematophila TB culture on plant pathogens Botrytis cinerea and Phytophthora capsici. The cell-free filtrate of TB culture showed strong inhibitory effects (>90%) on mycelial growth of both pathogens. The methanol-extracted bioactive compounds (methanol extract) of TB culture also had strong inhibitory effects on mycelial growth and spore germinations of both pathogens. The methanol extract (1000 μg/mL) and cell-free filtrate both showed strong therapeutic and protective effects (>70%) on grey mold both in detached tomato fruits and plants, and leaf scorch in pepper plants. This study demonstrates X. nematophila TB produces antimicrobial metabolites of strong activity on plant pathogens, with great potential for controlling tomato grey mold and pepper leaf scorch and being used in integrated disease control to reduce chemical application. PMID:24599183

  13. The Universe in a Box: Introduction to the Study of Astronomy in the Initial Formation of Physics Teachers. (Spanish Title: El Universo Representado en Una Caja: Introducción al Estudio de la Astronomía en la Formación Inicial de Profesores de Física.) O Universo Representado em Uma Caixa: Introdução ao Estudo da Astronomia NA Formação Inicial de Professores de Física

    NASA Astrophysics Data System (ADS)

    Longhini, Marcos Daniel

    2009-07-01

    This is a report of an activity of introduction to the study of Astronomy developed with a group of future Physics teachers at a Brazilian public university. Such activity had the goal of giving privileged emphasis to notions of spatiality, alternative conceptions of the participants and the process of interaction among peers, and consisted of the representation, in a three dimensional space, of the models of the universe that the participants had. The results, which were categorized as miscellaneous, geocentric, heliocentric and acentric models of the universe, were qualitatively analyzed. Analyses of the activity in the perspective of the participants are indicated and additional considerations are made regarding its use as a resource for teaching Astronomy and for teacher training. Este es el informe de una actividad para presentar un estudio introductorio de la Astronomía, desarrollado con una clase de futuros profesores de física en una universidad pública brasileña. Esta actividad tuvo como objetivo centrar las nociones de espacialidad, las concepciones alternativas de los participantes y el proceso de interacción entre pares, y consistió en la representación en un espacio tridimensional, de los modelos del universo que los participantes habían. Los resultados, que se clasificaron en universo miscelania, geocéntrico, heliocéntrico y acentrico, se analizaron cualitativamente. Son identificadas análisis de la actividad por los participantes, e hizo observaciones sobre su uso como recurso para la enseñanza de la astronomía y la formación de docentes. Trata-se do relato de uma atividade de introdução ao estudo da Astronomia, desenvolvida com uma turma de futuros professores de Física, em uma universidade pública brasileira. Tal atividade teve como meta privilegiar noções de espacialidade, as concepções alternativas dos participantes e o processo de interação entre pares e constou da representação, em um espaço tridimensional, dos

  14. The Understanding of Astronomy Concepts by Students from Basic Education of a Public School. (Spanish Title: El Entendimiento de Conceptos de Aastronmía Por Los Alumnos de Educación Básica en Una Escuela Pública.) O Entendimento de Conceitos de Astronomia Por Alunos da Educação Básica: O Caso de Uma Escola Pública Brasileira

    NASA Astrophysics Data System (ADS)

    Iria Machado, Daniel; dos Santos, Carlos

    2011-07-01

    We present the results obtained in a research on the comprehension of basic astronomical concepts, in which 561 students from fifth grade middle school to third grade high school of a public school of the city of Foz do Iguaçu (Brazil) took part. A test with 20 multiple-choice questions was applied to indentify the most common conceptions expressed by the students. This test was elaborated based on the literature about misconceptions and covered the following topics: the day-night cycle; the time zones; the seasons of the year; the phases of the Moon; the movement of the Moon; the apparent movement of the Sun in the celestial sphere; the eclipses; the dimensions and distances in the Universe; the brightness of the stars and its observation from Earth. Though a small progress was verified in the proportion of scientifically acceptable answers when comparing the eighth grade of middle school to the fifth, and the third grade of high school to the first, there was an overall predominance of alternative conceptions regarding most of the explored subjects, which persisted up to the last year of secondary school. The comparison to data found in this research made in other socio-cultural contexts revealed, in many aspects, similar notions and difficulties revealed by the students. Se presentan los resultados de una investigación sobre la comprensión de conceptos astronómicos básicos, en la cual participaron 561 estudiantes que cursaban entre el quinto grado de la enseñanza primaria y el tercer año de la enseñanza secundaria de una escuela pública de la ciudad de Foz do Iguaçu (Brasil). Se utilizó un test de 20 preguntas de opción múltiple para identificar las concepciones más comunes expresadas por los estudiantes. Este instrumento de recolección de datos se desarrolló en base a la literatura sobre las concepciones alternativas y trató los siguientes temas: el ciclo día-noche, los husos horarios, las estaciones del año, las fases de la Luna, el

  15. The Understanding of Astronomy Concepts by Students from Basic Education of a Public School. (Spanish Title: El Entendimiento de Conceptos de Aastronmía Por Los Alumnos de Educación Básica en Una Escuela Pública.) O Entendimento de Conceitos de Astronomia Por Alunos da Educação Básica: O Caso de Uma Escola Pública Brasileira

    NASA Astrophysics Data System (ADS)

    Iria Machado, Daniel; dos Santos, Carlos

    2011-07-01

    We present the results obtained in a research on the comprehension of basic astronomical concepts, in which 561 students from fifth grade middle school to third grade high school of a public school of the city of Foz do Iguaçu (Brazil) took part. A test with 20 multiple-choice questions was applied to indentify the most common conceptions expressed by the students. This test was elaborated based on the literature about misconceptions and covered the following topics: the day-night cycle; the time zones; the seasons of the year; the phases of the Moon; the movement of the Moon; the apparent movement of the Sun in the celestial sphere; the eclipses; the dimensions and distances in the Universe; the brightness of the stars and its observation from Earth. Though a small progress was verified in the proportion of scientifically acceptable answers when comparing the eighth grade of middle school to the fifth, and the third grade of high school to the first, there was an overall predominance of alternative conceptions regarding most of the explored subjects, which persisted up to the last year of secondary school. The comparison to data found in this research made in other socio-cultural contexts revealed, in many aspects, similar notions and difficulties revealed by the students. Se presentan los resultados de una investigación sobre la comprensión de conceptos astronómicos básicos, en la cual participaron 561 estudiantes que cursaban entre el quinto grado de la enseñanza primaria y el tercer año de la enseñanza secundaria de una escuela pública de la ciudad de Foz do Iguaçu (Brasil). Se utilizó un test de 20 preguntas de opción múltiple para identificar las concepciones más comunes expresadas por los estudiantes. Este instrumento de recolección de datos se desarrolló en base a la literatura sobre las concepciones alternativas y trató los siguientes temas: el ciclo día-noche, los husos horarios, las estaciones del año, las fases de la Luna, el

  16. La Ciencia de los Antiguos Mexicanos: Una Bibliografia Selecta

    ERIC Educational Resources Information Center

    Ortiz-Franco, Luis; Magana, Maria

    1973-01-01

    Fifty-five citations pertaining to the scientific and mathematic development of ancient Mexicans, particularly the Mayas, are given in this select bibliography. The introduction and descriptions of resource libraries in 8 States are in Spanish. (NQ)

  17. Una Perspectiva de Mexico: Its People, Places and History.

    ERIC Educational Resources Information Center

    Bianchi, Gary; Bianchi, Maria Eugenia Matute

    Evolving from a general commitment to the goals of cultural pluralism and bicultural education, this portfolio of 24 full color and halftone photographs aims to reinforce those curricular objectives which emphasize a respect for the value and individuality of different cultures and groups, affirm the right of an individual to maintain a bicultural…

  18. C/1999 S4 (LINEAR): la morte di una cometa

    NASA Astrophysics Data System (ADS)

    Foglia, Sergio; Ligustri, Rolando; Milani, Giannantonio

    2002-01-01

    The comet C/1999 S4 was expected to be a moderately bright object during summer 2000. It was observed by visual, photographic and CCD techniques. The derived light curve shows that the comet reached a brightness maximum (mag. 6 around July 26) corresponding to the total dissolution of its nucleus. From the analysis of the light curve a double maximum was detected, the dissolution event appearing to start around July 20. Several morphological changes were also observed during the apparition in relation to partial fragmentation events. The comet aspect completely changed during the final disruption. Relevant changes in the plasma tail were observed and they are probably related both to the activity of the nucleus and to the solar wind. A perspective anti-tail was detected in early July.

  19. Recuerdos de una revolucion cultural (Memories of a Cultural Revolution)

    ERIC Educational Resources Information Center

    Carvalho-Neto, Paulo de

    1973-01-01

    Describes elements of the modernism movement in Brazil; paper read at a symposium commemorating the fiftieth anniversay of the Week of Modern Art of Sao Paulo'' held February 11-12, 1972, at the University of California, Los Angeles, California. (DS)

  20. Esercitazioni Matematiche: Una Rivista Ad uso Degli Studenti Universitari.

    ERIC Educational Resources Information Center

    Micale, Biagio

    1992-01-01

    The history of the journal "Esercitazioni Matematiche" (1921-43) edited by the "Circolo Matematico di Catania" for university students is described. A selection of proposed problems with some of their solutions is reported. (Author)

  1. Una mirada hacia el pasado -- El Telescopio Espacial James Webb

    NASA Video Gallery

    La NASA planea usar el nuevo Telescopio Espacial James Webb para mirar el pasado. Al observar la luz de las estrellas que se formaron al principio del universo, la NASA está a punto de arrojar nuev...

  2. A Diagnostic Assessment for the Teaching of Astronomy. (Spanish Title: Una Evaluación Diagnóstica Para la Enseñanza de la Astronomia.) Uma Avaliação Diagnóstica Para o Ensino da Astronomia

    NASA Astrophysics Data System (ADS)

    Pacifico Ribeiro de Assis Silveira, Felipa; Soares Gomes de Sousa, Célia Maria; Moreira, Antonio

    2011-07-01

    This article aims to present the results of a diagnostic evaluation, used as a tool for collecting data on prior knowledge of scientific concepts needed to understand the topic "Earth and Universe", from a group comprised of 47 students of 6th grade. The educational method of diagnosing the student's prior knowledge before teaching them is founded on the Meaningful Learning theory, proposed by David Ausubel. The diagnostic evaluation consisted of 25 questions; the answers were analyzed and categorized, making possible their interpretation, which allows us to understand the meanings assigned by the student to these concepts. It follows from the diagnostic evaluation, that the majority of students surveyed had difficulties in exposing scientific concepts on the topic "Earth and Universe" when starting the 6th grade. However, we identified relevant ideas and representations that contributed to the re-signification of scientific concepts proposed for the teaching of the subject in this grade. The results of diagnostic evaluation served as reference to the organization of the syllabus, making possible the learning process by matching the sequence of teaching to the students" characteristics and context of the classroom. It is expected that other researchers in this topic can also use the instrument presented in this paper. Este artículo tiene como objetivo presentar los resultados de una evaluación diagnóstica, utilizada como una herramienta para recoger datos sobre el conocimiento previo de los conceptos científicos necesarios para comprender el tema "Tierra y Universo", de un grupo compuesto por 47 alumnos de 6º grado de la escuela primaria. La acción pedagógica del diagnóstico de los conocimientos previos del estudiante antes de enseñarles está justificada por la teoría del Aprendizaje Significativo, propuesta por David Ausubel. La evaluación de diagnóstico consistió en 25 preguntas, las respuestas fueron analizadas y clasificadas objetivando-se su

  3. A Diagnostic Assessment for the Teaching of Astronomy. (Spanish Title: Una Evaluación Diagnóstica Para la Enseñanza de la Astronomia.) Uma Avaliação Diagnóstica Para o Ensino da Astronomia

    NASA Astrophysics Data System (ADS)

    Pacifico Ribeiro de Assis Silveira, Felipa; Soares Gomes de Sousa, Célia Maria; Moreira, Antonio

    2011-07-01

    This article aims to present the results of a diagnostic evaluation, used as a tool for collecting data on prior knowledge of scientific concepts needed to understand the topic "Earth and Universe", from a group comprised of 47 students of 6th grade. The educational method of diagnosing the student's prior knowledge before teaching them is founded on the Meaningful Learning theory, proposed by David Ausubel. The diagnostic evaluation consisted of 25 questions; the answers were analyzed and categorized, making possible their interpretation, which allows us to understand the meanings assigned by the student to these concepts. It follows from the diagnostic evaluation, that the majority of students surveyed had difficulties in exposing scientific concepts on the topic "Earth and Universe" when starting the 6th grade. However, we identified relevant ideas and representations that contributed to the re-signification of scientific concepts proposed for the teaching of the subject in this grade. The results of diagnostic evaluation served as reference to the organization of the syllabus, making possible the learning process by matching the sequence of teaching to the students" characteristics and context of the classroom. It is expected that other researchers in this topic can also use the instrument presented in this paper. Este artículo tiene como objetivo presentar los resultados de una evaluación diagnóstica, utilizada como una herramienta para recoger datos sobre el conocimiento previo de los conceptos científicos necesarios para comprender el tema "Tierra y Universo", de un grupo compuesto por 47 alumnos de 6º grado de la escuela primaria. La acción pedagógica del diagnóstico de los conocimientos previos del estudiante antes de enseñarles está justificada por la teoría del Aprendizaje Significativo, propuesta por David Ausubel. La evaluación de diagnóstico consistió en 25 preguntas, las respuestas fueron analizadas y clasificadas objetivando-se su

  4. Phylogenetic Analysis of Downy Mildew Pathogens of Opium Poppy and PCR-Based In Planta and Seed Detection of Peronospora arborescens.

    PubMed

    Landa, Blanca B; Montes-Borrego, Miguel; Muñoz-Ledesma, Francisco J; Jiménez-Díaz, Rafael M

    2007-11-01

    ABSTRACT Severe downy mildew diseases of opium poppy (Papaver somniferum) can be caused by Peronospora arborescens and P. cristata, but differentiating between the two pathogens is difficult because they share morphological features and a similar host range. In Spain, where severe epidemics of downy mildew of opium poppy have occurred recently, the pathogen was identified as P. arborescens on the basis of morphological traits. In this current study, sequence homology and phylogenetic analyses of the internal transcribed spacer regions (ITS) of the ribosomal DNA (rDNA) were carried out with DNA from P. arborescens and P. cristata from diverse geographic origins, which suggested that only P. arborescens occurs in cultivated Papaver somniferum in Spain. Moreover, analyses of the rDNA ITS region from 27 samples of downy-mildew-affected tissues from all opium-poppy-growing regions in Spain showed that genetic diversity exists within P. arborescens populations in Spain and that these are phylogenetically distinct from P. cristata. P. cristata instead shares a more recent, common ancestor with a range of Peronospora species that includes those found on host plants that are not members of the Papaveraceae. Species-specific primers and a PCR assay protocol were developed that differentiated P. arborescens and P. cristata and proved useful for the detection of P. arborescens in symptomatic and asymptomatic opium poppy plant parts. Use of these primers demonstrated that P. arborescens can be transmitted in seeds and that commercial seed stocks collected from crops with high incidence of the disease were frequently infected. Field experiments conducted in microplots free from P. arborescens using seed stocks harvested from infected capsules further demonstrated that transmission from seedborne P. arborescens to opium poppy plants can occur. Therefore, the specific-PCR detection protocol developed in this study can be of use for epidemiological studies and diagnosing the pathogen in commercial seed stocks; thus facilitating the sanitary control of the disease and avoidance of the pathogen distribution in seeds. PMID:18943506

  5. In planta biocatalysis screen of P450s identifies 8-methoxypsoralen as a substrate for the CYP82C subfamily, yielding original chemical structures.

    PubMed

    Kruse, Tanya; Ho, Kwongling; Yoo, Hye-Dong; Johnson, Thomas; Hippely, Matt; Park, Joon-Hyun; Flavell, Richard; Bobzin, Steve

    2008-02-01

    An in vivo plant screen that allows for the analysis of exogenously applied substrates against transgenic Arabidopsis lines overexpressing individual cytochrome P450s has been developed. By deploying this screen with a subset of 91 P450s, we have identified an original substrate for members of the CYP82C subfamily. The therapeutic compound 8-methoxypsoralen was hydroxylated by plants overexpressing CYP82C2 or CYP82C4, forming 5-hydroxy-8-methoxypsoralen. Additionally, plants further modified this product to create a glycosylated compound, likely the compound 5-O-beta-D-glucopyranosyl-8-methoxypsoralen. The discovery of adducts of therapeutic compounds demonstrates the potential of this biocatalysis screening approach to create compounds that may be of pharmacological value. Additionally, this platform provides a means to expand the general knowledge base of P450 enzyme/substrate combinations and may provide valuable tools for a vast array of biocatalytic and bioremediation processes. PMID:18291319

  6. Beetle and plant arrow poisons of the Ju|'hoan and Hai||om San peoples of Namibia (Insecta, Coleoptera, Chrysomelidae; Plantae, Anacardiaceae, Apocynaceae, Burseraceae).

    PubMed

    Chaboo, Caroline S; Biesele, Megan; Hitchcock, Robert K; Weeks, Andrea

    2016-01-01

    The use of archery to hunt appears relatively late in human history. It is poorly understood but the application of poisons to arrows to increase lethality must have occurred shortly after developing bow hunting methods; these early multi-stage transitions represent cognitive shifts in human evolution. This paper is a synthesis of widely-scattered literature in anthropology, entomology, and chemistry, dealing with San ("Bushmen") arrow poisons. The term San (or Khoisan) covers many indigenous groups using so-called 'click languages' in southern Africa. Beetles are used for arrow poison by at least eight San groups and one non-San group. Fieldwork and interviews with Ju|'hoan and Hai||om hunters in Namibia revealed major differences in the nature and preparation of arrow poisons, bow and arrow construction, and poison antidote. Ju|'hoan hunters use leaf-beetle larvae of Diamphidia Gerstaecker and Polyclada Chevrolat (Chrysomelidae: Galerucinae: Alticini) collected from soil around the host plants Commiphora africana (A. Rich.) Engl. and Commiphora angolensis Engl. (Burseracaeae). In the Nyae Nyae area of Namibia, Ju|'hoan hunters use larvae of Diamphidia nigroornata Ståhl. Larvae and adults live above-ground on the plants and eat leaves, but the San collect the underground cocoons to extract the mature larvae. Larval hemolymph is mixed with saliva and applied to arrows. Hai||om hunters boil the milky plant sap of Adenium bohemianum Schinz (Apocynaceae) to reduce it to a thick paste that is applied to their arrows. The socio-cultural, historical, and ecological contexts of the various San groups may determine differences in the sources and preparation of poisons, bow and arrow technology, hunting behaviors, poison potency, and perhaps antidotes. PMID:27006594

  7. In planta processing and glycosylation of a nematode CLE effector and its interaction with a CLV2-like receptor to promote parasitism

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Like other biotrophic plant pathogens, plant-parasitic nematodes secrete effector proteins into host cells to facilitate infection. Effector proteins that mimic plant CLAVATA3/ESR (CLE)-like proteins have been identified in several cyst nematodes including the potato cyst nematode (PCN); however, th...

  8. The influence of antibiotic production and pre-emptive colonization on the population dynamics of Pantoea agglomerans (Erwinia herbicola) Eh1087 and Erwinia amylovora in planta.

    PubMed

    Giddens, Stephen R; Houliston, Gary J; Mahanty, H Khris

    2003-10-01

    Stigma colonization by Erwinia amylovora is the crucial first step in the development of most fire blight infections in apple and pear trees. Suppression at this point of the disease process by antagonists of E. amylovora, such as Pantoea agglomerans (Erwinia herbicola) strain Eh1087, is a rational approach to control fire blight. We tested the hypothesis that the ability of E. amylovora to compete with Eh1087 for colonization of a stigma is reduced by the potential for Eh1087 to produce the phenazine antibiotic, d-alanylgriseoluteic acid (AGA). In competition experiments on the stigmas of apple flowers, E. amylovora was significantly less successful against Eh1087 (AGA+) than against EhDeltaAGA (AGA-). Further experiments to test the importance of pre-emptive colonization of the stigma by either the pathogen or the antagonist suggested that AGA production significantly enhanced the competitiveness of Eh1087 when it was applied at the same time or 24 h before the pathogen. We also found that pre-emptive stigma colonization by either the pathogen or the antagonist resulted in a population that was resilient to subsequent invasion by a second species suggesting that niche exclusion has a dominant influence on the dynamics of bacterial populations on stigmas. PMID:14510856

  9. In planta imaging of Δ9-tetrahydrocannabinolic acid in Cannabis sativa L. with hyperspectral coherent anti-Stokes Raman scattering microscopy

    NASA Astrophysics Data System (ADS)

    Garbacik, Erik T.; Korai, Roza P.; Frater, Eric H.; Korterik, Jeroen P.; Otto, Cees; Offerhaus, Herman L.

    2013-04-01

    Nature has developed many pathways to produce medicinal products of extraordinary potency and specificity with significantly higher efficiencies than current synthetic methods can achieve. Identification of these mechanisms and their precise locations within plants could substantially increase the yield of a number of natural pharmaceutics. We report label-free imaging of Δ9-tetrahydrocannabinolic acid (THCa) in Cannabis sativa L. using coherent anti-Stokes Raman scattering microscopy. In line with previous observations we find high concentrations of THCa in pistillate flowering bodies and relatively low amounts within flowering bracts. Surprisingly, we find differences in the local morphologies of the THCa-containing bodies: organelles within bracts are large, diffuse, and spheroidal, whereas in pistillate flowers they are generally compact, dense, and have heterogeneous structures. We have also identified two distinct vibrational signatures associated with THCa, both in pure crystalline form and within Cannabis plants; at present the exact natures of these spectra remain an open question.

  10. Seed development and maturation in early spring-flowering Galanthus nivalis and Narcissus pseudonarcissus continues post-shedding with little evidence of maturation in planta

    PubMed Central

    Newton, Rosemary J.; Hay, Fiona R.; Ellis, Richard H.

    2013-01-01

    Background and Aims Seeds of the moist temperate woodland species Galanthus nivalis and Narcissus pseudonarcissus, dispersed during spring or early summer, germinated poorly in laboratory tests. Seed development and maturation were studied to better understand the progression from developmental to germinable mode in order to improve seed collection and germination practices in these and similar species. Methods Phenology, seed mass, moisture content and ability to germinate and tolerate desiccation were monitored during seed development until shedding. Embryo elongation within seeds was investigated during seed development and under several temperature regimes after shedding. Key Results Seeds were shed at high moisture content (>59 %) with little evidence that dry mass accumulation or embryo elongation were complete. Ability to germinate developed prior to the ability of some seeds to tolerate enforced desiccation. Germination was sporadic and slow. Embryo elongation occurred post-shedding in moist environments, most rapidly at 20 °C in G. nivalis and 15 °C in N. pseudonarcissus. The greatest germination also occurred in these regimes, 78 and 48 %, respectively, after 700 d. Conclusions Seeds of G. nivalis and N. pseudonarcissus were comparatively immature at shedding and substantial embryo elongation occurred post-shedding. Seeds showed limited desiccation tolerance at dispersal. PMID:23478943

  11. Vascular Streak Dieback of cacao in Southeast Asia detection and Melanesia: in planta detection of the pathogen and a new taxonomy

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Vascular Streak Dieback (VSD) disease of cacao (Theobroma cacao) in Southeast Asia and Melanesia is caused by a basidiomycete (Ceratobasidiales) fungus described in a monotypic genus as Oncobasidium theobromae (syn. =Thanatephorus theobromae). The symptoms of the disease include green-spotted chloro...

  12. Proteins with High Turnover Rate in Barley Leaves Estimated by Proteome Analysis Combined with in Planta Isotope Labeling1[W][OPEN

    PubMed Central

    Nelson, Clark J.; Alexova, Ralitza; Jacoby, Richard P.; Millar, A. Harvey

    2014-01-01

    Protein turnover is a key component in cellular homeostasis; however, there is little quantitative information on degradation kinetics for individual plant proteins. We have used 15N labeling of barley (Hordeum vulgare) plants and gas chromatography-mass spectrometry analysis of free amino acids and liquid chromatography-mass spectrometry analysis of proteins to track the enrichment of 15N into the amino acid pools in barley leaves and then into tryptic peptides derived from newly synthesized proteins. Using information on the rate of growth of barley leaves combined with the rate of degradation of 14N-labeled proteins, we calculate the turnover rates of 508 different proteins in barley and show that they vary by more than 100-fold. There was approximately a 9-h lag from label application until 15N incorporation could be reliably quantified in extracted peptides. Using this information and assuming constant translation rates for proteins during the time course, we were able to quantify degradation rates for several proteins that exhibit half-lives on the order of hours. Our workflow, involving a stringent series of mass spectrometry filtering steps, demonstrates that 15N labeling can be used for large-scale liquid chromatography-mass spectrometry studies of protein turnover in plants. We identify a series of abundant proteins in photosynthesis, photorespiration, and specific subunits of chlorophyll biosynthesis that turn over significantly more rapidly than the average protein involved in these processes. We also highlight a series of proteins that turn over as rapidly as the well-known D1 subunit of photosystem II. While these proteins need further verification for rapid degradation in vivo, they cluster in chlorophyll and thiamine biosynthesis. PMID:25082890

  13. Regulon Studies and In Planta Role of the BraI/R Quorum-Sensing System in the Plant-Beneficial Burkholderia Cluster

    PubMed Central

    Coutinho, Bruna G.; Mitter, Birgit; Talbi, Chouhra; Sessitsch, Angela; Bedmar, Eulogio J.; Halliday, Nigel; James, Euan K.; Cámara, Miguel

    2013-01-01

    The genus Burkholderia is composed of functionally diverse species, and it can be divided into several clusters. One of these, designated the plant-beneficial-environmental (PBE) Burkholderia cluster, is formed by nonpathogenic species, which in most cases have been found to be associated with plants. It was previously established that members of the PBE group share an N-acyl-homoserine lactone (AHL) quorum-sensing (QS) system, designated BraI/R, that produces and responds to 3-oxo-C14-HSL (OC14-HSL). Moreover, some of them also possess a second AHL QS system, designated XenI2/R2, producing and responding to 3-hydroxy-C8-HSL (OHC8-HSL). In the present study, we performed liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) analysis to determine which AHL molecules are produced by each QS system of this group of bacteria. The results showed that XenI2/R2 is mainly responsible for the production of OHC8-HSL and that the BraI/R system is involved in the production of several different AHLs. This analysis also revealed that Burkholderia phymatum STM815 produces greater amounts of AHLs than the other species tested. Further studies showed that the BraR protein of B. phymatum is more promiscuous than other BraR proteins, responding equally well to several different AHL molecules, even at low concentrations. Transcriptome studies with Burkholderia xenovorans LB400 and B. phymatum STM815 revealed that the BraI/R regulon is species specific, with exopolysaccharide production being the only common phenotype regulated by this system in the PBE cluster. In addition, BraI/R was shown not to be important for plant nodulation by B. phymatum strains or for endophytic colonization and growth promotion of maize by B. phytofirmans PsJN. PMID:23686262

  14. Taxonomic Treatment of Solanum Section Petota (Wild Potatoes) in Catálogo de Plantas Vasculares del Cono Sur (Argentina, Chile, Paraguay, Uruguay, y sur del Brasil)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Solanum section Petota (Solanaceae), which includes the cultivated potato (Solanum tuberosum) and its wild relatives, contains over 150 wild species distributed from the southwestern U.S.A. (38°N) to central Argentina and adjacent Chile (41°S). This catalog includes all species from the Southern Con...

  15. Development and Use of a Reverse Transcription-PCR Assay To Study Expression of Tri5 by Fusarium Species In Vitro and In Planta

    PubMed Central

    Doohan, F. M.; Weston, G.; Rezanoor, H. N.; Parry, D. W.; Nicholson, P.

    1999-01-01

    The Tri5 gene encodes trichodiene synthase, which catalyzes the first reaction in the trichothecene biosynthetic pathway. In vitro, a direct relationship was observed between Tri5 expression and the increase in deoxynivalenol production over time. We developed a reverse transcription (RT)-PCR assay to quantify Tri5 gene expression in trichothecene-producing strains of Fusarium species. We observed an increase in Tri5 expression following treatment of Fusarium culmorum with fungicides, and we also observed an inverse relationship between Tri5 expression and biomass, as measured by β-d-glucuronidase activity, during colonization of wheat (cv. Avalon) seedlings by F. culmorum. RT-PCR analysis also showed that for ears of wheat cv. Avalon inoculated with F. culmorum, there were different levels of Tri5 expression in grain and chaff at later growth stages. We used the Tri5-specific primers to develop a PCR assay to detect trichothecene-producing Fusarium species in infected plant material. PMID:10473385

  16. Species-specific ITS primers for the identification of Picoa juniperi and Picoa lefebvrei and using nested-PCR for detection of P. juniperi in planta.

    PubMed

    Jamali, Samad; Banihashemi, Zia

    2013-10-01

    Desert truffles, hypogeous Pezizales (Ascomycota), are difficult to identify due to evolutionary convergence of morphological characters among taxa that share a similar habitat and mode of spore dispersal. Also, during their symbiotic phase, these are barely distinguishable morphologically, and molecular probes are needed for their identification. We have developed a PCR-based method for the identification of Picoa juniperi and Picoa lefebvrei based on internal transcribed spacers of rDNA. Two PCR primers specific for P. lefebvrei (FLE/RLE) and two specific for P. juniperi (FJU/RJU) were designed. A collection of samples from different geographical areas representing diversity of these species were examined for unique regions of internal transcribed spacers 1, 2 and 5.8S gene of rDNA (ITS) compared to other closely related species. Annealing temperatures and extension times were optimized for each set of primers for maximum specificity and efficiency. They proved to be efficient to specifically detect the presence of P. juniperi and P. lefebvrei by PCR and neither set amplified purified DNA from other truffle species as well as some ascomycetous fungi. The partial small subunit of ribosomal DNA genes of P. juniperi were amplified with the genomic DNA extracted from Helianthemum ledifolium var. ledifolium roots by nested polymerase chain reaction (PCR) using the universal fungal primer pair ITS1/ITS4 and specific primer pair FTC/RTC, which was designed based on internal transcribed spacer 1, 2 and 5.8S gene of rDNA sequences of P juniperi. The nested-PCR was sensitive enough to re-amplify the direct-PCR product, resulting in a DNA fragment of 426 bp. The efficacy of nested-PCR showed that it could re-amplify the direct-PCR product and detect 200 fg genomic DNA. PMID:24065525

  17. The Pratylenchus penetrans Transcriptome as a Source for the Development of Alternative Control Strategies: Mining for Putative Genes Involved in Parasitism and Evaluation of in planta RNAi

    PubMed Central

    Vieira, Paulo; Eves-van den Akker, Sebastian; Verma, Ruchi; Wantoch, Sarah; Eisenback, Jonathan D.; Kamo, Kathryn

    2015-01-01

    The root lesion nematode Pratylenchus penetrans is considered one of the most economically important species within the genus. Host range studies have shown that nearly 400 plant species can be parasitized by this species. To obtain insight into the transcriptome of this migratory plant-parasitic nematode, we used Illumina mRNA sequencing analysis of a mixed population, as well as nematode reads detected in infected soybean roots 3 and 7 days after nematode infection. Over 140 million paired end reads were obtained for this species, and de novo assembly resulted in a total of 23,715 transcripts. Homology searches showed significant hit matches to 58% of the total number of transcripts using different protein and EST databases. In general, the transcriptome of P. penetrans follows common features reported for other root lesion nematode species. We also explored the efficacy of RNAi, delivered from the host, as a strategy to control P. penetrans, by targeted knock-down of selected nematode genes. Different comparisons were performed to identify putative nematode genes with a role in parasitism, resulting in the identification of transcripts with similarities to other nematode parasitism genes. Focusing on the predicted nematode secreted proteins found in this transcriptome, we observed specific members to be up-regulated at the early time points of infection. In the present study, we observed an enrichment of predicted secreted proteins along the early time points of parasitism by this species, with a significant number being pioneer candidate genes. A representative set of genes examined using RT-PCR confirms their expression during the host infection. The expression patterns of the different candidate genes raise the possibility that they might be involved in critical steps of P. penetrans parasitism. This analysis sheds light on the transcriptional changes that accompany plant infection by P. penetrans, and will aid in identifying potential gene targets for selection and use to design effective control strategies against root lesion nematodes. PMID:26658731

  18. Investigations of the in Planta Expression of Active Cellobiohydrolase I: Cooperative Research and Development Final Report, CRADA Number CRD-07-219

    SciTech Connect

    Himmel, M.

    2011-02-01

    It was the purpose of this project to determine if cellulases produced in transgenic plants could effectively be utilized in the production of ethanol and other feedstock chemicals from lignocellulosic substrates.

  19. In Planta Variation of Volatile Biosynthesis: An Alternative Biosynthetic Route to the Formation of the Pathogen-Induced Volatile Homoterpene DMNT via Triterpene Degradation in Arabidopsis Roots

    PubMed Central

    Sohrabi, Reza; Huh, Jung-Hyun; Badieyan, Somayesadat; Rakotondraibe, Liva Harinantenaina; Kliebenstein, Daniel J.; Sobrado, Pablo; Tholl, Dorothea

    2015-01-01

    Plant-derived volatile compounds such as terpenes exhibit substantial structural variation and serve multiple ecological functions. Despite their structural diversity, volatile terpenes are generally produced from a small number of core 5- to 20-carbon intermediates. Here, we present unexpected plasticity in volatile terpene biosynthesis by showing that irregular homo/norterpenes can arise from different biosynthetic routes in a tissue specific manner. While Arabidopsis thaliana and other angiosperms are known to produce the homoterpene (E)-4,8-dimethyl-1,3,7-nonatriene (DMNT) or its C16-analog (E,E)-4,8,12-trimethyl-1,3,7,11-tridecatetraene by the breakdown of sesquiterpene and diterpene tertiary alcohols in aboveground tissues, we demonstrate that Arabidopsis roots biosynthesize DMNT by the degradation of the C30 triterpene diol, arabidiol. The reaction is catalyzed by the Brassicaceae-specific cytochrome P450 monooxygenase CYP705A1 and is transiently induced in a jasmonate-dependent manner by infection with the root-rot pathogen Pythium irregulare. CYP705A1 clusters with the arabidiol synthase gene ABDS, and both genes are coexpressed constitutively in the root stele and meristematic tissue. We further provide in vitro and in vivo evidence for the role of the DMNT biosynthetic pathway in resistance against P. irregulare. Our results show biosynthetic plasticity in DMNT biosynthesis in land plants via the assembly of triterpene gene clusters and present biochemical and genetic evidence for volatile compound formation via triterpene degradation in plants. PMID:25724638

  20. Beetle and plant arrow poisons of the Ju|’hoan and Hai||om San peoples of Namibia (Insecta, Coleoptera, Chrysomelidae; Plantae, Anacardiaceae, Apocynaceae, Burseraceae)

    PubMed Central

    Chaboo, Caroline S.; Biesele, Megan; Hitchcock, Robert K.; Weeks, Andrea

    2016-01-01

    Abstract The use of archery to hunt appears relatively late in human history. It is poorly understood but the application of poisons to arrows to increase lethality must have occurred shortly after developing bow hunting methods; these early multi-stage transitions represent cognitive shifts in human evolution. This paper is a synthesis of widely-scattered literature in anthropology, entomology, and chemistry, dealing with San (“Bushmen”) arrow poisons. The term San (or Khoisan) covers many indigenous groups using so-called ‘click languages’ in southern Africa. Beetles are used for arrow poison by at least eight San groups and one non-San group. Fieldwork and interviews with Ju|’hoan and Hai||om hunters in Namibia revealed major differences in the nature and preparation of arrow poisons, bow and arrow construction, and poison antidote. Ju|’hoan hunters use leaf-beetle larvae of Diamphidia Gerstaecker and Polyclada Chevrolat (Chrysomelidae: Galerucinae: Alticini) collected from soil around the host plants Commiphora africana (A. Rich.) Engl. and Commiphora angolensis Engl. (Burseracaeae). In the Nyae Nyae area of Namibia, Ju|’hoan hunters use larvae of Diamphidia nigroornata Ståhl. Larvae and adults live above-ground on the plants and eat leaves, but the San collect the underground cocoons to extract the mature larvae. Larval hemolymph is mixed with saliva and applied to arrows. Hai||om hunters boil the milky plant sap of Adenium bohemianum Schinz (Apocynaceae) to reduce it to a thick paste that is applied to their arrows. The socio-cultural, historical, and ecological contexts of the various San groups may determine differences in the sources and preparation of poisons, bow and arrow technology, hunting behaviors, poison potency, and perhaps antidotes. PMID:27006594

  1. SECONDARY METABOLITE PRODUCTION BY THE FUNGAL PATHOGEN EUTYPA LATA: ANALYSIS OF EXTRACTS FROM GRAPEVINE CULTURES AND DETECTION OF THOSE METABOLITES IN PLANTA

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Eutypa dieback of grapevines is caused by the fungal pathogen Eutypa lata and reduces vineyard longevity worldwide. Early detection could reduce losses due to this disease, so our aim was to identify acetylenic phenol metabolites of E. lata that could prove suitable as chemical markers in an early d...

  2. Transient Co-Expression of Post-Transcriptional Gene Silencing Suppressors for Increased in Planta Expression of a Recombinant Anthrax Receptor Fusion Protein

    PubMed Central

    Arzola, Lucas; Chen, Junxing; Rattanaporn, Kittipong; Maclean, James M.; McDonald, Karen A.

    2011-01-01

    Potential epidemics of infectious diseases and the constant threat of bioterrorism demand rapid, scalable, and cost-efficient manufacturing of therapeutic proteins. Molecular farming of tobacco plants provides an alternative for the recombinant production of therapeutics. We have developed a transient production platform that uses Agrobacterium infiltration of Nicotiana benthamiana plants to express a novel anthrax receptor decoy protein (immunoadhesin), CMG2-Fc. This chimeric fusion protein, designed to protect against the deadly anthrax toxins, is composed of the von Willebrand factor A (VWA) domain of human capillary morphogenesis 2 (CMG2), an effective anthrax toxin receptor, and the Fc region of human immunoglobulin G (IgG). We evaluated, in N. benthamiana intact plants and detached leaves, the expression of CMG2-Fc under the control of the constitutive CaMV 35S promoter, and the co-expression of CMG2-Fc with nine different viral suppressors of post-transcriptional gene silencing (PTGS): p1, p10, p19, p21, p24, p25, p38, 2b, and HCPro. Overall, transient CMG2-Fc expression was higher on intact plants than detached leaves. Maximum expression was observed with p1 co-expression at 3.5 days post-infiltration (DPI), with a level of 0.56 g CMG2-Fc per kg of leaf fresh weight and 1.5% of the total soluble protein, a ten-fold increase in expression when compared to absence of suppression. Co-expression with the p25 PTGS suppressor also significantly increased the CMG2-Fc expression level after just 3.5 DPI. PMID:21954339

  3. Learning about the Sky from the Environment: An Experience Working Along One Year with Students of Elementary Education. (Spanish Title: Aprendiendo sobre EL Cielo Desde el Entorno: Una Experiencia Trabajando Durante un Año Junto a Estudiantes del Primario.) Aprendendo sobre o Céu a Partir do Entorno: Uma Experiência de Trabalho ao Longo de um Ano com Alunos de Ensino Fundamental

    NASA Astrophysics Data System (ADS)

    Longhini, Marcos Daniel; Gomide, Hanny Angeles

    2014-12-01

    Research developed with 95 students of the 6th year of elementary education in a public school of Uberlândia, Minas Gerais. It was a continuous work from February to December 2013, which led the students to participate in activities of observation of the environment, specifically the sky, analyzing the changes occurred. We focused on the study of variations in temperature, rainfall, day length, variations in the size of the shadows and changes in the aspect of the Moon. Our focus of analysis targeted the discussion of the knowledge that these students had about the topics indicated and as they entered the stage during the implementation of the proposal. The results showed a limited perception that students have of their environment, however, lately expanded due to the undertaken activities, especially in relation to the Moon. Working with systematic measure procedures reveals the careful handling of data so that they become understandable to students, and working with the shadows points towards the students first understand how shadows are formed, and then apply this knowledge to Astronomy. Finally, we conclude that the lived process consisted of an initial step of a work that should be encouraged for the subsequent years of training of these students. Proyecto de investigación desarrollado con 95 alumnos del sexto año de primaria en una escuela pública de Uberlândia, Minas Gerais. Fue un trabajo continuo, de febrero a diciembre de 2013, que llevó a los estudiantes a participar en actividades de observación de su entorno, entre ellas, el cielo, analizando los cambios ocurridos. Nos centramos en el estudio de las variaciones en la temperatura, las precipitaciones, la duración del día, las variaciones en el tamaño de las sombras y los cambios en los aspectos de la Luna. Nuestro foco de análisis se centró en discutir el conocimiento que estos estudiantes tenían sobre los temas indicados al inicio y término de la propuesta. Los resultados mostraron una

  4. Open-field host specificity test of Gratiana boliviana (Coleoptera: Chrysomelidae), a biological control agent of tropical soda apple (Solanaceae) in the United States

    SciTech Connect

    Gandolfo, D.; McKay, F.; Medal, J.C.; Cuda, J.P.

    2007-03-15

    An open-field experiment was conducted to assess the suitability of the South American leaf feeding beetle Gratiana boliviana Spaeth for biological control of Solanum viarum Dunal in the USA. An open-field test with eggplant, Solanum melongena L., was conducted on the campus of the University of Buenos Aires, Argentina, and a S. viarum control plot was established 40 km from the campus. One hundred adult beetles were released in each plot at the beginning of the experiment during the vegetative stage of the plants, and forty additional beetles were released in the S. melongena plot at the flowering stage. All the plants in each plot were checked twice a week and the number of adults, immatures, and eggs recorded. Results showed almost a complete rejection of eggplant by G. boliviana. No noticeable feeding damage was ever recorded on eggplant. The experiment was ended when the eggplants started to senesce or were severely damaged by whiteflies and spider mites. The results of this open-field experiment corroborate previous quarantine/laboratory host-specificity tests indicating that a host range expansion of G. boliviana to include eggplant is highly unlikely. Gratiana boliviana was approved for field release in May 2003 in the USA. To date, no non-target effects have been observed either on eggplant or native species of Solanum. (author) [Spanish] Una prueba de campo fue conducida para evaluar la especificidad del escarabajo suramericano defoliador Gratiana boliviana Spaeth para control biologico de Solanum viarum Dunal en los Estados Unidos. La prueba con berenjena se realizo en el campo experimental de la Universidad de Buenos Aires, Argentina, y una parcela control con S. viarum fue establecida a 40 km. Cien escarabajos adultos fueron liberados en cada parcela al inicio del experimento durante la fase vegetativa, y cuarenta escarabajos adicionales fueron liberados en la parcela de berenjena durante la floracion. Todas las plantas en cada parcela fueron

  5. Invertebrate fauna associated with Torpedograss, Panicum repens (Cyperales: Poaceae), in Lake Okeechobee, Florida, and prospects for biological control

    SciTech Connect

    Cuda, J.P.; Dunford, J.C.; Leavengood, J.M. Jr.

    2007-03-15

    Torpedograss, Panicum repens L., is an adventive, rhizomatous grass species that has become an invasive weed of terrestrial, wetland, and aquatic environments in tropical and subtropical regions worldwide. Until recently, strategies for controlling torpedograss in the USA have focused almost exclusively on mechanical and chemical methods, either alone or in combination, with varied results. A survey of the arthropods and nematodes currently associated with the plant in Lake Okeechobee, Florida, was conducted as part of a feasibility study to determine whether torpedograss is an appropriate target for a classical biological control program. Overall, approximately 4,000 arthropods and 400 nematode specimens were collected. Sweep, clipped vegetation, and soil core samples were dominated by representatives of the arthropod orders Hemiptera, Hymenoptera, Diptera, and Acari. Lesion nematodes of the genus Pratylenchus were commonly associated with the roots of torpedograss. None of the organisms collected were torpedograss specialists. Although classical biological control of torpedograss is feasible based on the extent of the infestation, economic losses, resistance to conventional controls, and the report of a potentially host specific natural enemy in India, the botanical position of this grass weed will require a formal risk assessment before proceeding with a classical biological control program. (author) [Spanish] La conota, Panicum repens L., es una especie foranea de pasto que produce rizomas que ha convertido en ser una maleza invasora de ambientes terrestres, pantanosos y acuaticos en regiones tropicales y subtropicales en todo el mundo. Hasta hace un tiempo reciente, las estrategias para controlar conota en los EEUU eran enfocadas casi exclusivamente en los metodos mecanicos y quimicos, solos o en combinacion, con resultados variables. Un muestreo de los artropodos y nematodos asociados corrientemente con esta planta en el Lago de Okeechobee, Florida, fue

  6. The Combine Project: An Experience in a Dual-Language Classroom = El proyecto de la cosechadora: una experiencia en una clase bilingue.

    ERIC Educational Resources Information Center

    Wilson, Rebecca

    This article, also available in Spanish, describes what happened when a bilingual kindergarten class in West Liberty, Iowa, investigated a combine. The dual-language Kindergarten program supports content area instruction in both Spanish and English. The first part of the article tells the story of the Combine Project, this class's first project…

  7. A quantitative synthesis of the medicinal ethnobotany of the Malinké of Mali and the Asháninka of Peru, with a new theoretical framework

    PubMed Central

    Bletter, Nathaniel

    2007-01-01

    from around the world to pinpoint the species with the highest potential efficacy to take into the laboratory and analyze further, ultimately saving much field and laboratory time and resources. Spanish abstract Las búsquedas que utilizan la etnomedicina y la taxonomía para descubrir nuevas plantas medicinales, pueden aumentar la probabilidad de éxito de encontrar compuestos químicos activos en plantas, en comparación con las búsquedas aleatorias. A pesar de lo anterior, en las últimas décadas, la etnobotánica no ha cumplido con las expectativas de proveer numerosas plantas medicinales y químicos nuevos una vez examinados en el laboratorio. Cada año se describen una plétora de plantas medicinales y sus usos, sin embargo las limitaciones de tiempo y recursos en los laboratorios, unidos al alto coste de los ensayos clínicos de las drogas potenciales, hacen muy difícil probar, aislar, y elucidar la estructura y el mecanismo de los compuestos de estas plantas. Se propone un nuevo marco teórico cuantitativo cuyo fin es focalizar la búsqueda de nueva plantas medicinales. Este marco teórico está basado en la hipótesis que las plantas cercanamente relacionadas, usadas para tratar enfermedades cercanamente relacionadas en culturas distantemente relacionadas, tienen una eficacia potencial más alta, debido a que es más probable que estos hallazgos sean descubrimientos independientes de compuestos químicos similares. Parte de esta hipótesis, que las escogencias racionales se hacen para elegir plantas medicinales similares y que la flora medicinal de dos culturas distantes es más similar que su flora general, se probó usando métodos estadísticos de remuestreo con datos de campo de la comunidad Malinké de Malí y de la Asháninka de Perú, y las enfermedades de paludismo, enfermedad africana del sueño, enfermedad de Chagas, leishmania, diabetes, eczema, asma, y fibromas uterinos. Se encontró, en este caso, que

  8. Establishing the Empirical Relationship Between Non-Science Majoring Undergraduate Learners' Spatial Thinking Skills and Their Conceptual Astronomy Knowledge. (Spanish Title: Estableciendo Una Relación Empírica Entre el Razonamiento Espacial de los Estudiantes de Graduación de Carreras no Científicas y su Conocimento Conceptual de la Astronomía.) Estabelecendo Uma Relação Empírica Entre o RacioCínio Espacial dos Estudantes de Graduação EM Carreiras Não Científicas e Seu Conhecimento Conceitual da Astronomia

    NASA Astrophysics Data System (ADS)

    Heyer, Inge; Slater, Stephanie J.; Slater, Timothy F.

    2013-12-01

    The astronomy education community has tacitly assumed that learning astronomy is a conceptual domain resting upon spatial thinking skills. As a first step to formally identify an empirical relationship, undergraduate students in a non-major introductory astronomy survey class at a mediumsized, Ph.D. granting, mid-western US university were given pre- and post-astronomy conceptual diagnostics and spatial reasoning diagnostics, Instruments used were the "Test Of Astronomy Standards" and "What Do You Know?" Using only fully matched data for analysis, our sample consisted of 86 undergraduate non-science majors. Students' normalized gains for astronomy surveys were low at .26 and .13 respectively. Students' spatial thinking was measured using an instrument designed specifically for this study. Correlations between the astronomy instruments' pre- to post-course gain scores and the spatial assessment instrument show moderate to strong relationships suggesting the relationship between spatial reasoning and astronomy ability can explain about 25% of the variation in student achievement. La comunidad de educación en astronomía ha supuesto de forma tácita que el aprendizaje de la astronomía consiste en un dominio conceptual fundamentado en el razonamiento espacial. Como un primer paso para identificar formalmente una relación empírica entre estas dos cosas, utilizamos como muestra los estudantes de graduación de carreras no científicas de un curso experimental en una universidad norteamericana del medioeste de porte mediano con programa de Doctorado em curso, en el cual estos estudiantes se sometieron a un diagnóstico de razonamiento espacial y conceptos astronómicos antes e después del mismo. Las herramientas utilizadas fueron el Test Of Astronomy Standards (TOAST) y el cuestionario What do you know? Utilizando solo los datos completamente consistentes para este análisis, nuestra muestra consistió en 86 estudantes de graduación. Las mejoras, depués de

  9. Habitos Alimentarios e Higiene Dental en una Poblacion Preescolar de Madrid.

    ERIC Educational Resources Information Center

    Lafarga, R. Tarazona; And Others

    1991-01-01

    Nutriment habits and oral hygiene in a preschool population in Madrid were studied by stratification in public or private schools. Significant differences were found only with regard to consumption of fish. Vegetable and fruit consumption was low in both groups, but not statistically significant. (SH)

  10. Tengo una Bomba: The Paralinguistic and Linguistic Conventions of the Oral Practice Chismeando.

    ERIC Educational Resources Information Center

    Hall, Joan Kelly

    1993-01-01

    This article offers a linguistic and paralinguistic explication of the oral practice of chismeando (gossiping) as engaged in by a group of women from the Dominican Republic. A culture-specific study of the structuring resources by which the participants construct, maintain, and/or modify their in-group identities in everyday oral practice is…

  11. Protegiendo a nuestros ninos: una guia de accion (Safeguarding Our Children: An Action Guide).

    ERIC Educational Resources Information Center

    American Institutes for Research, Washington, DC. Center for Effective Collaboration and Practice.

    Although most schools are safe, they all can be safer. Improving school safety requires the strategic investment of time and dollars--scarce resources for which there are competing demands. Fortunately, schools that strategically coordinate schoolwide efforts are more likely to improve academic performance as well as reduce behavior problems. This…

  12. "...Una Etapa Siempre Dificil": Concepts of Adolescence and Programs of Secondary Education in Mexico.

    ERIC Educational Resources Information Center

    Levinson, Bradley A.

    An American visiting a Mexican secundaria (equivalent to the U.S. junior high school) noted the strong presence of discourses on "adolescence." This prompted an ideological and historical review of the secundaria since its development in the postrevolutionary ethos of the 1920s-30s. Inspired by a U.S. model of adolescent education in the liberal…

  13. Borders and Identity: A Resource Guide for Teachers = Identidad y Fronteras: Una Guia para Maestros.

    ERIC Educational Resources Information Center

    Belauus, Betty; Botein, Emily; Cadaval, Olivia

    The materials in this resource guide include a four-part video, a poster-size cultural map with additional exercises, and the five sections of this guide. The unit, presented in English and Spanish, intends to introduce students to the peoples and cultures of the U.S.-Mexico border, to explore the concept of borders in their own communities, to…

  14. The Complete Chloroplast and Mitochondrial Genomes of the Green Macroalga Ulva sp. UNA00071828 (Ulvophyceae, Chlorophyta)

    PubMed Central

    Melton, James T.; Leliaert, Frederik; Tronholm, Ana; Lopez-Bautista, Juan M.

    2015-01-01

    Sequencing mitochondrial and chloroplast genomes has become an integral part in understanding the genomic machinery and the phylogenetic histories of green algae. Previously, only three chloroplast genomes (Oltmannsiellopsis viridis, Pseudendoclonium akinetum, and Bryopsis hypnoides) and two mitochondrial genomes (O. viridis and P. akinetum) from the class Ulvophyceae have been published. Here, we present the first chloroplast and mitochondrial genomes from the ecologically and economically important marine, green algal genus Ulva. The chloroplast genome of Ulva sp. was 99,983 bp in a circular-mapping molecule that lacked inverted repeats, and thus far, was the smallest ulvophycean plastid genome. This cpDNA was a highly compact, AT-rich genome that contained a total of 102 identified genes (71 protein-coding genes, 28 tRNA genes, and three ribosomal RNA genes). Additionally, five introns were annotated in four genes: atpA (1), petB (1), psbB (2), and rrl (1). The circular-mapping mitochondrial genome of Ulva sp. was 73,493 bp and follows the expanded pattern also seen in other ulvophyceans and trebouxiophyceans. The Ulva sp. mtDNA contained 29 protein-coding genes, 25 tRNA genes, and two rRNA genes for a total of 56 identifiable genes. Ten introns were annotated in this mtDNA: cox1 (4), atp1 (1), nad3 (1), nad5 (1), and rrs (3). Double-cut-and-join (DCJ) values showed that organellar genomes across Chlorophyta are highly rearranged, in contrast to the highly conserved organellar genomes of the red algae (Rhodophyta). A phylogenomic investigation of 51 plastid protein-coding genes showed that Ulvophyceae is not monophyletic, and also placed Oltmannsiellopsis (Oltmannsiellopsidales) and Tetraselmis (Chlorodendrophyceae) closely to Ulva (Ulvales) and Pseudendoclonium (Ulothrichales). PMID:25849557

  15. Confieso que Divulgo. Reflexiones y Experiencias de una Astrofísica

    NASA Astrophysics Data System (ADS)

    Rodríguez Hidalgo, I.

    Este artículo presenta algunas reflexiones en torno a la popularización de la Ciencia, desarrolladas a lo largo de mi trayectoria profesional, un camino inacabado desde la intuición al oficio. Tras revisar las señas de identidad de la divulgación científica, se exponen ideas, experiencias y recursos, cribados por la práctica y su posterior análisis crítico. Se destacan las actividades relacionadas con la Astronomía, que se cuentan entre las más espectaculares y gratificantes. Confessions of a popularizer: This paper presents some author's thoughts about scientific outreach, developed along her professional path, an unfinished way from intuition to trade. First, identity signs of outreach are revised; then, ideas, experiences and resources, sifted by practice and further critical analysis, are reviewed. Activities related to Astronomy, being one of the most spectacular and rewarding, are remarked 1

  16. Sistemas Eolicos Pequenos para Generacion de Electricidad; Una guia para consumidores en los EE.UU.

    SciTech Connect

    Not Available

    2007-09-01

    This Spanish version of the popular Small Wind Electric Systems: A U.S. Consumer's Guide provides consumers with information to help them determine whether a small wind electric system can provide all or a portion of the energy they need for their home or business based on their wind resource, energy needs, and economics. Topics include how to make a home more energy efficient, how to choose the correct turbine size, the parts of a wind electric system, how to determine whether enough wind resource exists, how to choose the best site for a turbine, how to connect a system to the utility grid, and whether it's possible to become independent of the utility grid using wind energy. In addition, the cover of the guide contains a list of contacts for more information.

  17. Sistemas Eolicos Pequenos para Generacion de Electricidad: Una guia para consumidores en Nuevo Mexico

    SciTech Connect

    Not Available

    2007-09-01

    This Spanish version of the popular Small Wind Electric Systems: A New Mexico Consumer's Guide provides consumers with information to help them determine whether a small wind electric system can provide all or a portion of the energy they need for their home or business based on their wind resource, energy needs, and economics. Topics include how to make a home more energy efficient, how to choose the correct turbine size, the parts of a wind electric system, how to determine whether enough wind resource exists, how to choose the best site for a turbine, how to connect a system to the utility grid, and whether it's possible to become independent of the utility grid using wind energy. In addition, the cover of the guide contains a list of contacts for more information.

  18. Una Guia Para Padres Para El Cuidado de Ninos (A Parent's Guide to Child Care).

    ERIC Educational Resources Information Center

    Oregon State Dept. of Human Resources, Salem.

    This booklet was designed to help parents in Oregon in finding and financing child care for their children. The first section of the booklet provides information on the Oregon Department of Human Resources' Adult and Family Services (AFS) Division's Aid for Dependent Children (ADC) and Employment Related Day Care (ERDC) programs. This section…

  19. Una nina adoptada su conducta en el jardin de infantes (An Adopted Girl's Conduct in Preschool).

    ERIC Educational Resources Information Center

    Truscello de Mason, Marile

    1994-01-01

    Reports observations made by a preschool teacher in Argentina about a four-year-old adopted girl in her preschool. The teacher noticed problems between the girl and her adoptive mother, who tried to prevent the girl from remembering her past. The teacher particularly noticed changes in the girl's drawings. (AA)

  20. PREVENCIÓN DEL VIH/SIDA EN LOS CIRCUITOS DE LEVANTE HSH: UNA ASIGNATURA PENDIENTE.

    PubMed

    Barreda, Victoria; Carballo-Dieguez, Alex; Marone, Rubén; Balán, Iván; Pando, María Ángeles; Avila, María Mercedes

    2010-12-01

    A partir de un relevamiento de tipo etnográfico, se describen lugares de encuentro de HSH en la Ciudad de Buenos Aires y sus prácticas sexuales. El reconocimiento de tales espacios, así como las características que asumen en ellos los encuentros sexuales entre los HSH, plantean obstáculos específicos en la adopción de comportamientos preventivos y, asimismo, generan nuevos desafíos para las actividades de prevención. Se plantean las dificultades y debates conceptuales que la misma categoría presenta, y sus consecuencias en el abordaje preventivo y teórico-metodológico para las ciencias sociales. Además, se proponen nuevos interrogantes acerca de los alcances y las limitaciones del modelo preventivo del VIH/Sida para HSH. PMID:21874154

  1. Areas Nacionales de Estudio Ambiental: Una Guia. (National Environmental Study Area: A Guide).

    ERIC Educational Resources Information Center

    National Park Service (Dept. of Interior), Washington, DC.

    This booklet, the Spanish version of SE 014 817, is a guide for teachers and resource managers who are interested in establishing National Environmental Study Areas (NESA) or interested in receiving NESA recognition for their on-going environmental study area programs. It outlines the characteristics and procedures of the program; the nature,…

  2. Historia Verdadera de Una Gota de Miel. (The True Story of a Drop of Honey).

    ERIC Educational Resources Information Center

    Galarza, Ernesto

    Part of the series "Coleccion Mini-Libros" (Mini-Book Collection), the booklet is a factual account of the life of a bee. Designed to provide a tool for education of the Spanish-speaking child, as well as to stimulate an interest in Spanish among the youth of our country, the publication is a compilation of photographs with a description of bees…

  3. Historia de vidas de profesoras de ciencias: Hacia una ensenanza cientifica de calidad

    NASA Astrophysics Data System (ADS)

    Vega Osorio, Melba

    The purpose of this research was to capture the memories, practices, thoughts and philosophies of education that four experienced Science teachers have accumulated through the years. The life story qualitative research design was used in order to collect more meaningful information from the participants amenable to in depth analysis and interpretation. The life stories interviews were conducted using an interview protocol especially designed for the study. The interviews were recorded and transcribed. The questions that guided this research are: 1. What are the meanings that science teachers give to their educational practices? 2. How did the life experiences of the participants contribute to their success as science teachers? 3. How did the participants' educational practices change and evolve over time? 4. What does teaching science mean in the life of the participants? 5. Which reflections arise from the work of the participants that impact the teaching of science? 6. What are the participants' conceptions about teachers' professional development? 7. What are the participants' contributions to the field of scientific education? Wolcott's (1994) DAI analytical model was used for the analysis of the information and the narratives were written following the three dimensions of the model: description, analysis and interpretation. According to the life stories of the participants, being a teacher requires more than teaching the content. To be a successful and an exemplary science teacher, they have to live the teaching vocation, with dedication, passion, commitment and love for teaching, for students and science; teachers have to give their best for the students, even if it means making personal sacrifices; they should keep updated in knowledge (content) and in educational innovations (strategies); emphasizing both the theoretical and the practical aspects in the field of science, and being an inspirational guide for students.

  4. [Clip Sheets from BOCES. Opportunities. Health. Careers. = Oportunidades. Salud. Una Camera En...

    ERIC Educational Resources Information Center

    State Univ. of New York, Geneseo. Coll. at Geneseo. Migrant Center.

    This collection of 83 clip sheets, or classroom handouts, was created to help U.S. migrants learn more about health, careers, and general "opportunities" including education programs. They are written in both English and Spanish and are presented in an easily understandable format. Health clip-sheet topics include the following: Abuse; AIDS;…

  5. Guia Practica para una Evaluacion Alternativa (Practical Guide to Alternative Assessment).

    ERIC Educational Resources Information Center

    Herman, Joan L.; Aschbacher, Pamela R.; Winters, Lynn

    Guidance is offered in Spanish on the creation and use of alternative assessment, and a process model is presented that links assessment with curriculum and instruction, based on contemporary theories of learning and cognition. The introductory chapter provides background on the purposes of assessment and the need for new alternatives, with an…

  6. Una experiencia europea de innovacion pedagogica basada en la escritura de un ibro.

    ERIC Educational Resources Information Center

    Montane, Mireia; Roc, Merce

    2002-01-01

    Describes a European project based on writing a book from an educational perspective. Participants included 18 schools from 13 countries. The study originated in a Spanish Comenius 3.1 project, which determined that university teams would visit "book writing" schools, supporting teachers and analyzing and publishing their experiences. After…

  7. Mediacion en educacion especial: Una guia para los padres (Special Education Mediation: A Guide for Parents).

    ERIC Educational Resources Information Center

    Consortium for Appropriate Dispute Resolution in Special Education (CADRE), Eugene, OR.

    Designed for Spanish-speaking parents of students with disabilities living in Oregon, this brochure describes the general mediation process that parents can use to resolve special education services disputes with schools. It begins by discussing what mediation is and the characteristics of a trained mediator. It addresses the requirement for…

  8. Chile: Una Vision Politica, Economica y Social (Chile: A Political, Economic, and Social View).

    ERIC Educational Resources Information Center

    Cortes-Hwang, Adriana

    1972-01-01

    This address seeks to explain in brief the historical background and political, economic, and social conditions leading to the democratic election of a Marxist president in Chile. A historical sketch of Chilean government from independence in 1810 is provided with a description of the situation just before Salvador Allende's election in 1969. Some…

  9. Community Resources Guide for Central Florida = Una Guia de Recursos en la Comunidad de Florida Central.

    ERIC Educational Resources Information Center

    Medina, Linda, Comp.

    Designed to orient Hispanic refugees to the services that are available in Central Florida, this bilingual guide consists of a section of general information on living and working in the United States and a section devoted to various public and private agencies. Provided first are addresses and phone numbers of various government agencies:…

  10. The Struggle for Mexico's First Gay-Straight Alliance: Como una Novela Real

    ERIC Educational Resources Information Center

    Macgillivray, Ian K.

    2006-01-01

    In 2004, a group of high school students at a private American school in Mexico City started the first gay-straight alliance in Mexico. A small group of conservative parents and a Mormon principal organized in opposition. This paper details the students' struggle to keep their club and offers lessons learned about student activism, school change,…