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Sample records for campestris pv vesicatoria

  1. Xanthomonas campestris pv. vesicatoria Secretes Proteases and Xylanases via the Xps Type II Secretion System and Outer Membrane Vesicles.

    PubMed

    Solé, Magali; Scheibner, Felix; Hoffmeister, Anne-Katrin; Hartmann, Nadine; Hause, Gerd; Rother, Annekatrin; Jordan, Michael; Lautier, Martine; Arlat, Matthieu; Büttner, Daniela

    2015-09-01

    Many plant-pathogenic bacteria utilize type II secretion (T2S) systems to secrete degradative enzymes into the extracellular milieu. T2S substrates presumably mediate the degradation of plant cell wall components during the host-pathogen interaction and thus promote bacterial virulence. Previously, the Xps-T2S system from Xanthomonas campestris pv. vesicatoria was shown to contribute to extracellular protease activity and the secretion of a virulence-associated xylanase. The identities and functions of additional T2S substrates from X. campestris pv. vesicatoria, however, are still unknown. In the present study, the analysis of 25 candidate proteins from X. campestris pv. vesicatoria led to the identification of two type II secreted predicted xylanases, a putative protease and a lipase which was previously identified as a virulence factor of X. campestris pv. vesicatoria. Studies with mutant strains revealed that the identified xylanases and the protease contribute to virulence and in planta growth of X. campestris pv. vesicatoria. When analyzed in the related pathogen X. campestris pv. campestris, several T2S substrates from X. campestris pv. vesicatoria were secreted independently of the T2S systems, presumably because of differences in the T2S substrate specificities of the two pathogens. Furthermore, in X. campestris pv. vesicatoria T2S mutants, secretion of T2S substrates was not completely absent, suggesting the contribution of additional transport systems to protein secretion. In line with this hypothesis, T2S substrates were detected in outer membrane vesicles, which were frequently observed for X. campestris pv. vesicatoria. We, therefore, propose that extracellular virulence-associated enzymes from X. campestris pv. vesicatoria are targeted to the Xps-T2S system and to outer membrane vesicles. The virulence of plant-pathogenic bacteria often depends on TS2 systems, which secrete degradative enzymes into the extracellular milieu. T2S substrates are being

  2. Xanthomonas campestris pv. vesicatoria Secretes Proteases and Xylanases via the Xps Type II Secretion System and Outer Membrane Vesicles

    PubMed Central

    Solé, Magali; Scheibner, Felix; Hoffmeister, Anne-Katrin; Hartmann, Nadine; Hause, Gerd; Rother, Annekatrin; Jordan, Michael; Lautier, Martine; Arlat, Matthieu

    2015-01-01

    ABSTRACT Many plant-pathogenic bacteria utilize type II secretion (T2S) systems to secrete degradative enzymes into the extracellular milieu. T2S substrates presumably mediate the degradation of plant cell wall components during the host-pathogen interaction and thus promote bacterial virulence. Previously, the Xps-T2S system from Xanthomonas campestris pv. vesicatoria was shown to contribute to extracellular protease activity and the secretion of a virulence-associated xylanase. The identities and functions of additional T2S substrates from X. campestris pv. vesicatoria, however, are still unknown. In the present study, the analysis of 25 candidate proteins from X. campestris pv. vesicatoria led to the identification of two type II secreted predicted xylanases, a putative protease and a lipase which was previously identified as a virulence factor of X. campestris pv. vesicatoria. Studies with mutant strains revealed that the identified xylanases and the protease contribute to virulence and in planta growth of X. campestris pv. vesicatoria. When analyzed in the related pathogen X. campestris pv. campestris, several T2S substrates from X. campestris pv. vesicatoria were secreted independently of the T2S systems, presumably because of differences in the T2S substrate specificities of the two pathogens. Furthermore, in X. campestris pv. vesicatoria T2S mutants, secretion of T2S substrates was not completely absent, suggesting the contribution of additional transport systems to protein secretion. In line with this hypothesis, T2S substrates were detected in outer membrane vesicles, which were frequently observed for X. campestris pv. vesicatoria. We, therefore, propose that extracellular virulence-associated enzymes from X. campestris pv. vesicatoria are targeted to the Xps-T2S system and to outer membrane vesicles. IMPORTANCE The virulence of plant-pathogenic bacteria often depends on TS2 systems, which secrete degradative enzymes into the extracellular milieu. T2S

  3. DNA probes for detection of copper resistance genes in Xanthomonas campestris pv. Vesicatoria

    SciTech Connect

    Garde, S.; Bender, C.L. )

    1991-08-01

    The copper resistance (Cu{sup r}) genes encoded on pXV10A, a 190-kb plasmid in Xanthomonas campestris pv. vesicatoria XV10, were isolated on a 44-kb cosmid clone designated pCuR1. Tn5 mutagenesis of pCuR1 indicated that a 4.0-kb region was required for copper resistance. Three restriction fragments located within the 4.0-kb region demonstrated high specificity for the Cu{sup r} genes present in X. campestris pv. vesicatoria and will be useful in monitoring the presence of these genes in the environment.

  4. Importance of opgHXcv of Xanthomonas campestris pv. vesicatoria in host-parasite interactions.

    PubMed

    Minsavage, G V; Mudgett, M B; Stall, R E; Jones, J B

    2004-02-01

    Tn5 insertion mutants of Xanthomonas campestris pv. vesicatoria were inoculated into tomato and screened for reduced virulence. One mutant exhibited reduced aggressiveness and attenuated growth in planta. Southern blot analyses indicated that the mutant carried a single Tn5 insertion not associated with previously cloned pathogenicity-related genes of X. campestris pv. vesicatoria. The wild-type phenotype of this mutant was restored by one recombinant plasmid (pOPG361) selected from a genomic library of X. campestris pv. vesicatoria 91-118. Tn3-gus insertion mutagenesis and sequence analyses of a subclone of pOPG361 identified a 1,929-bp open reading frame (ORF) essential for complementation of the mutants. The predicted protein encoded by this ORF was highly homologous to the previously reported pathogenicity-related HrpM protein of Pseudomonas syringae pv. syringae and OpgH of Erwinia chrysanthemi. Based on homology, the new locus was designated opgHXcv. Manipulation of the osmotic potential in the intercellular spaces of tomato leaves by addition of mannitol at low concentrations (25 to 50 mM) compensates for the opgHXcv mutation.

  5. 40 CFR 180.1261 - Xanthomonas campestris pv. vesicatoria and Pseudomonas syringae pv. tomato specific Bacteriophages.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... and Pseudomonas syringae pv. tomato specific Bacteriophages. 180.1261 Section 180.1261 Protection of.... vesicatoria and Pseudomonas syringae pv. tomato specific Bacteriophages. An exemption from the requirement of... syringae pv. tomato specific bacteriophages in or on pepper and tomato. ...

  6. 40 CFR 180.1261 - Xanthomonas campestris pv. vesicatoria and Pseudomonas syringae pv. tomato specific Bacteriophages.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... and Pseudomonas syringae pv. tomato specific Bacteriophages. 180.1261 Section 180.1261 Protection of.... vesicatoria and Pseudomonas syringae pv. tomato specific Bacteriophages. An exemption from the requirement of... syringae pv. tomato specific bacteriophages in or on pepper and tomato....

  7. 40 CFR 180.1261 - Xanthomonas campestris pv. vesicatoria and Pseudomonas syringae pv. tomato specific Bacteriophages.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... and Pseudomonas syringae pv. tomato specific Bacteriophages. 180.1261 Section 180.1261 Protection of.... vesicatoria and Pseudomonas syringae pv. tomato specific Bacteriophages. An exemption from the requirement of... syringae pv. tomato specific bacteriophages in or on pepper and tomato....

  8. 40 CFR 180.1261 - Xanthomonas campestris pv. vesicatoria and Pseudomonas syringae pv. tomato specific Bacteriophages.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... and Pseudomonas syringae pv. tomato specific Bacteriophages. 180.1261 Section 180.1261 Protection of.... vesicatoria and Pseudomonas syringae pv. tomato specific Bacteriophages. An exemption from the requirement of... syringae pv. tomato specific bacteriophages in or on pepper and tomato....

  9. 40 CFR 180.1261 - Xanthomonas campestris pv. vesicatoria and Pseudomonas syringae pv. tomato specific Bacteriophages.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... and Pseudomonas syringae pv. tomato specific Bacteriophages. 180.1261 Section 180.1261 Protection of.... vesicatoria and Pseudomonas syringae pv. tomato specific Bacteriophages. An exemption from the requirement of... syringae pv. tomato specific bacteriophages in or on pepper and tomato....

  10. Response to Xanthomonas campestris pv. vesicatoria in tomato involves regulation of ethylene receptor gene expression.

    PubMed

    Ciardi, J A; Tieman, D M; Lund, S T; Jones, J B; Stall, R E; Klee, H J

    2000-05-01

    Although ethylene regulates a wide range of defense-related genes, its role in plant defense varies greatly among different plant-microbe interactions. We compared ethylene's role in plant response to virulent and avirulent strains of Xanthomonas campestris pv. vesicatoria in tomato (Lycopersicon esculentum Mill.). The ethylene-insensitive Never ripe (Nr) mutant displays increased tolerance to the virulent strain, while maintaining resistance to the avirulent strain. Expression of the ethylene receptor genes NR and LeETR4 was induced by infection with both virulent and avirulent strains; however, the induction of LeETR4 expression by the avirulent strain was blocked in the Nr mutant. To determine whether ethylene receptor levels affect symptom development, transgenic plants overexpressing a wild-type NR cDNA were infected with virulent X. campestris pv. vesicatoria. Like the Nr mutant, the NR overexpressors displayed greatly reduced necrosis in response to this pathogen. NR overexpression also reduced ethylene sensitivity in seedlings and mature plants, indicating that, like LeETR4, this receptor is a negative regulator of ethylene response. Therefore, pathogen-induced increases in ethylene receptors may limit the spread of necrosis by reducing ethylene sensitivity.

  11. Response to Xanthomonas campestris pv. vesicatoria in Tomato Involves Regulation of Ethylene Receptor Gene Expression1

    PubMed Central

    Ciardi, Joseph A.; Tieman, Denise M.; Lund, Steven T.; Jones, Jeffrey B.; Stall, Robert E.; Klee, Harry J.

    2000-01-01

    Although ethylene regulates a wide range of defense-related genes, its role in plant defense varies greatly among different plant-microbe interactions. We compared ethylene's role in plant response to virulent and avirulent strains of Xanthomonas campestris pv. vesicatoria in tomato (Lycopersicon esculentum Mill.). The ethylene-insensitive Never ripe (Nr) mutant displays increased tolerance to the virulent strain, while maintaining resistance to the avirulent strain. Expression of the ethylene receptor genes NR and LeETR4 was induced by infection with both virulent and avirulent strains; however, the induction of LeETR4 expression by the avirulent strain was blocked in the Nr mutant. To determine whether ethylene receptor levels affect symptom development, transgenic plants overexpressing a wild-type NR cDNA were infected with virulent X. campestris pv. vesicatoria. Like the Nr mutant, the NR overexpressors displayed greatly reduced necrosis in response to this pathogen. NR overexpression also reduced ethylene sensitivity in seedlings and mature plants, indicating that, like LeETR4, this receptor is a negative regulator of ethylene response. Therefore, pathogen-induced increases in ethylene receptors may limit the spread of necrosis by reducing ethylene sensitivity. PMID:10806227

  12. Domain structure of HrpE, the Hrp pilus subunit of Xanthomonas campestris pv. vesicatoria.

    PubMed

    Weber, Ernst; Koebnik, Ralf

    2005-09-01

    The plant-pathogenic bacterium Xanthomonas campestris pv. vesicatoria possesses a type III secretion (TTS) system necessary for pathogenicity in susceptible hosts and induction of the hypersensitive response in resistant plants. This specialized protein transport system is encoded by a 23-kb hrp (hypersensitive response and pathogenicity) gene cluster. X. campestris pv. vesicatoria produces filamentous structures, Hrp pili, at the cell surface under hrp-inducing conditions. The Hrp pilus acts as a cell surface appendage of the TTS system and serves as a conduit for the transfer of bacterial effector proteins into the plant cell cytosol. The major pilus component, the HrpE pilin, is unique to xanthomonads and is encoded within the hrp gene cluster. In this study, functional domains of HrpE were mapped by linker-scanning mutagenesis and by reporter protein fusions to an N-terminally truncated avirulence protein (AvrBs3Delta2). Thirteen five-amino-acid peptide insertion mutants were obtained and could be grouped into six phenotypic classes. Three permissive mutations were mapped in the N-terminal half of HrpE, which is weakly conserved within the HrpE protein family. Four dominant-negative peptide insertions in the strongly conserved C-terminal region suggest that this domain is critical for oligomerization of the pilus subunits. Reporter protein fusions revealed that the N-terminal 17 amino acid residues act as an efficient TTS signal. From these results, we postulate a three-domain structure of HrpE with an N-terminal secretion signal, a surface-exposed variable region of the N-terminal half, and a C-terminal polymerization domain. Comparisons with a mutant study of HrpA, the Hrp pilin from Pseudomonas syringae pv. tomato DC3000, and hydrophobicity plot analyses of several nonhomologous Hrp pilins suggest a common architecture of Hrp pilins of different plant-pathogenic bacteria.

  13. Volatile organic compounds produced by the phytopathogenic bacterium Xanthomonas campestris pv. vesicatoria 85-10.

    PubMed

    Weise, Teresa; Kai, Marco; Gummesson, Anja; Troeger, Armin; von Reuß, Stephan; Piepenborn, Silvia; Kosterka, Francine; Sklorz, Martin; Zimmermann, Ralf; Francke, Wittko; Piechulla, Birgit

    2012-01-01

    Xanthomonas campestris is a phytopathogenic bacterium and causes many diseases of agricultural relevance. Volatiles were shown to be important in inter- and intraorganismic attraction and defense reactions. Recently it became apparent that also bacteria emit a plethora of volatiles, which influence other organisms such as invertebrates, plants and fungi. As a first step to study volatile-based bacterial-plant interactions, the emission profile of Xanthomonas c. pv. vesicatoria 85-10 was determined by using GC/MS and PTR-MS techniques. More than 50 compounds were emitted by this species, the majority comprising ketones and methylketones. The structure of the dominant compound, 10-methylundecan-2-one, was assigned on the basis of its analytical data, obtained by GC/MS and verified by comparison of these data with those of a synthetic reference sample. Application of commercially available decan-2-one, undecan-2-one, dodecan-2-one, and the newly synthesized 10-methylundecan-2-one in bi-partite Petri dish bioassays revealed growth promotions in low quantities (0.01 to 10 μmol), whereas decan-2-one at 100 μmol caused growth inhibitions of the fungus Rhizoctonia solani. Volatile emission profiles of the bacteria were different for growth on media (nutrient broth) with or without glucose.

  14. Aconitase B is required for optimal growth of Xanthomonas campestris pv. vesicatoria in pepper plants.

    PubMed

    Kirchberg, Janine; Büttner, Daniela; Thiemer, Barbara; Sawers, R Gary

    2012-01-01

    The aerobic plant pathogenic bacterium Xanthomonas campestris pv. vesicatoria (Xcv) colonizes the intercellular spaces of pepper and tomato. One enzyme that might contribute to the successful proliferation of Xcv in the host is the iron-sulfur protein aconitase, which catalyzes the conversion of citrate to isocitrate in the tricarboxylic acid (TCA) cycle and might also sense reactive oxygen species (ROS) and changes in cellular iron levels. Xcv contains three putative aconitases, two of which, acnA and acnB, are encoded by a single chromosomal locus. The focus of this study is aconitase B (AcnB). acnB is co-transcribed with two genes, XCV1925 and XCV1926, encoding putative nucleic acid-binding proteins. In vitro growth of acnB mutants was like wild type, whereas in planta growth and symptom formation in pepper plants were impaired. While acnA, XCV1925 or XCV1926 mutants showed a wild-type phenotype with respect to bacterial growth and in planta symptom formation, proliferation of the acnB mutant in susceptible pepper plants was significantly impaired. Furthermore, the deletion of acnB led to reduced HR induction in resistant pepper plants and an increased susceptibility to the superoxide-generating compound menadione. As AcnB complemented the growth deficiency of an Escherichia coli aconitase mutant, it is likely to be an active aconitase. We therefore propose that optimal growth and survival of Xcv in pepper plants depends on AcnB, which might be required for the utilization of citrate as carbon source and could also help protect the bacterium against oxidative stress.

  15. Aconitase B Is Required for Optimal Growth of Xanthomonas campestris pv. vesicatoria in Pepper Plants

    PubMed Central

    Kirchberg, Janine; Büttner, Daniela; Thiemer, Barbara; Sawers, R. Gary

    2012-01-01

    The aerobic plant pathogenic bacterium Xanthomonas campestris pv. vesicatoria (Xcv) colonizes the intercellular spaces of pepper and tomato. One enzyme that might contribute to the successful proliferation of Xcv in the host is the iron-sulfur protein aconitase, which catalyzes the conversion of citrate to isocitrate in the tricarboxylic acid (TCA) cycle and might also sense reactive oxygen species (ROS) and changes in cellular iron levels. Xcv contains three putative aconitases, two of which, acnA and acnB, are encoded by a single chromosomal locus. The focus of this study is aconitase B (AcnB). acnB is co-transcribed with two genes, XCV1925 and XCV1926, encoding putative nucleic acid-binding proteins. In vitro growth of acnB mutants was like wild type, whereas in planta growth and symptom formation in pepper plants were impaired. While acnA, XCV1925 or XCV1926 mutants showed a wild-type phenotype with respect to bacterial growth and in planta symptom formation, proliferation of the acnB mutant in susceptible pepper plants was significantly impaired. Furthermore, the deletion of acnB led to reduced HR induction in resistant pepper plants and an increased susceptibility to the superoxide-generating compound menadione. As AcnB complemented the growth deficiency of an Escherichia coli aconitase mutant, it is likely to be an active aconitase. We therefore propose that optimal growth and survival of Xcv in pepper plants depends on AcnB, which might be required for the utilization of citrate as carbon source and could also help protect the bacterium against oxidative stress. PMID:22493725

  16. Inducible Expression of the De-Novo Designed Antimicrobial Peptide SP1-1 in Tomato Confers Resistance to Xanthomonas campestris pv. vesicatoria.

    PubMed

    Herrera Diaz, Areli; Kovacs, Izabella; Lindermayr, Christian

    2016-01-01

    Antimicrobial peptides (AMPs) are small peptides with less than 50 amino acids and are part of the innate immune response in almost all organisms, including bacteria, vertebrates, invertebrates and plants. AMPs are active against a broad-spectrum of pathogens. The inducible expression of AMPs in plants is a promising approach to combat plant pathogens with minimal negative side effects, such as phytotoxicity or infertility. In this study, inducible expression of the de-novo designed AMP SP1-1 in Micro Tom tomato protected tomato fruits against bacterial spot disease caused by Xanthomonas campestris pv. vesicatoria. The peptide SP1-1 was targeted to the apoplast which is the primary infection site for plant pathogens, by fusing SP1-1 peptide to the signal peptide RsAFP1 of radish (Raphanus sativus). The pathogen inducibility of the expression was enabled by using an optimized inducible 4XW2/4XS promoter. As a result, the tomato fruits of independently generated SP1-1 transgenic lines were significantly more resistant to X. campestris pv. vesicatoria than WT tomato fruits. In transgenic lines, bacterial infection was reduced up to 65% in comparison to the infection of WT plants. Our study demonstrates that the combination of the 4XW2/4XS cis-element from parsley with the synthetic antimicrobial peptide SP1-1 is a good alternative to protect tomato fruits against infections with X. campestris pv. vesicatoria.

  17. Inducible Expression of the De-Novo Designed Antimicrobial Peptide SP1-1 in Tomato Confers Resistance to Xanthomonas campestris pv. vesicatoria

    PubMed Central

    Herrera Diaz, Areli; Kovacs, Izabella; Lindermayr, Christian

    2016-01-01

    Antimicrobial peptides (AMPs) are small peptides with less than 50 amino acids and are part of the innate immune response in almost all organisms, including bacteria, vertebrates, invertebrates and plants. AMPs are active against a broad-spectrum of pathogens. The inducible expression of AMPs in plants is a promising approach to combat plant pathogens with minimal negative side effects, such as phytotoxicity or infertility. In this study, inducible expression of the de-novo designed AMP SP1-1 in Micro Tom tomato protected tomato fruits against bacterial spot disease caused by Xanthomonas campestris pv. vesicatoria. The peptide SP1-1 was targeted to the apoplast which is the primary infection site for plant pathogens, by fusing SP1-1 peptide to the signal peptide RsAFP1 of radish (Raphanus sativus). The pathogen inducibility of the expression was enabled by using an optimized inducible 4XW2/4XS promoter. As a result, the tomato fruits of independently generated SP1-1 transgenic lines were significantly more resistant to X. campestris pv. vesicatoria than WT tomato fruits. In transgenic lines, bacterial infection was reduced up to 65% in comparison to the infection of WT plants. Our study demonstrates that the combination of the 4XW2/4XS cis-element from parsley with the synthetic antimicrobial peptide SP1-1 is a good alternative to protect tomato fruits against infections with X. campestris pv. vesicatoria. PMID:27706237

  18. Functional characterization of the Xcs and Xps type II secretion systems from the plant pathogenic bacterium Xanthomonas campestris pv vesicatoria.

    PubMed

    Szczesny, Robert; Jordan, Matthias; Schramm, Claudia; Schulz, Steve; Cogez, Virginie; Bonas, Ulla; Büttner, Daniela

    2010-09-01

    *Type II secretion (T2S) systems of many plant-pathogenic bacteria often secrete cell wall-degrading enzymes into the plant apoplast. *Here, we show that the Xps-T2S system from the plant pathogen Xanthomonas campestris pv vesicatoria (Xcv) promotes disease and contributes to the translocation of effector proteins that are delivered into the plant cell by the type III secretion (T3S) system. *The Xcs-T2S system instead lacks an obvious virulence function. However, individual xcs genes can partially complement mutants in homologous xps genes, indicating that they encode functional components of T2S systems. Enzyme activity assays showed that the Xps system contributes to secretion of proteases and xylanases. We identified the virulence-associated xylanase XynC as a substrate of the Xps system. However, homologs of known T2S substrates from other Xanthomonas spp. are not secreted by the T2S systems from Xcv. Thus, T2S systems from Xanthomonas spp. appear to differ significantly in their substrate specificities. *Transcript analyses revealed that expression of xps genes in Xcv is activated by HrpG and HrpX, key regulators of the T3S system. By contrast, expression of xynC and extracellular protease and xylanase activities are repressed by HrpG and HrpX, suggesting that components and substrates of the Xps system are differentially regulated.

  19. Characterization of Citrus sinensis transcription factors closely associated with the non-host response to Xanthomonas campestris pv. vesicatoria.

    PubMed

    Daurelio, Lucas D; Romero, María S; Petrocelli, Silvana; Merelo, Paz; Cortadi, Adriana A; Talón, Manuel; Tadeo, Francisco R; Orellano, Elena G

    2013-07-01

    Plants, when exposed to certain pathogens, may display a form of genotype-independent resistance, known as non-host response. In this study, the response of Citrus sinensis (sweet orange) leaves to Xanthomonas campestris pv. vesicatoria (Xcv), a pepper and tomato pathogenic bacterium, was analyzed through biochemical assays and cDNA microarray hybridization and compared with Asiatic citrus canker infection caused by Xanthomonas citri subsp. citri. Citrus leaves exposed to the non-host bacterium Xcv showed hypersensitive response (HR) symptoms (cell death), a defense mechanism common in plants but poorly understood in citrus. The HR response was accompanied by differentially expressed genes that are associated with biotic stress and cell death. Moreover, 58 transcription factors (TFs) were differentially regulated by Xcv in citrus leaves, including 26 TFs from the stress-associated families AP2-EREBP, bZip, Myb and WRKY. Remarkably, in silico analysis of the distribution of expressed sequence tags revealed that 10 of the 58 TFs, belonging to C2C2-GATA, C2H2, CCAAT, HSF, NAC and WRKY gene families, were specifically over-represented in citrus stress cDNA libraries. This study identified candidate TF genes for the regulation of key steps during the citrus non-host HR. Furthermore, these TFs might be useful in future strategies of molecular breeding for citrus disease resistance. Copyright © 2013 Elsevier GmbH. All rights reserved.

  20. The Xanthomonas campestris pv. vesicatoria Type-3 Effector XopB Inhibits Plant Defence Responses by Interfering with ROS Production.

    PubMed

    Priller, Johannes Peter Roman; Reid, Stephen; Konein, Patrick; Dietrich, Petra; Sonnewald, Sophia

    2016-01-01

    The bacterial pathogen Xanthomonas campestris pv. vesicatoria 85-10 (Xcv) translocates about 30 type-3 effector proteins (T3Es) into pepper plants (Capsicum annuum) to suppress plant immune responses. Among them is XopB which interferes with PTI, ETI and sugar-mediated defence responses, but the underlying molecular mechanisms and direct targets are unknown so far. Here, we examined the XopB-mediated suppression of plant defence responses in more detail. Infection of susceptible pepper plants with Xcv lacking xopB resulted in delayed symptom development compared to Xcv wild type infection concomitant with an increased formation of salicylic acid (SA) and expression of pathogenesis-related (PR) genes. Expression of xopB in Arabidopsis thaliana promoted the growth of the virulent Pseudomonas syringae pv. tomato (Pst) DC3000 strain. This was paralleled by a decreased SA-pool and a lower induction of SA-dependent PR gene expression. The expression pattern of early flg22-responsive marker genes indicated that MAPK signalling was not altered in the presence of XopB. However, XopB inhibited the flg22-triggered burst of reactive oxygen species (ROS). Consequently, the transcript accumulation of AtOXI1, a ROS-dependent marker gene, was reduced in xopB-expressing Arabidopsis plants as well as callose deposition. The lower ROS production correlated with a low level of basal and flg22-triggered expression of apoplastic peroxidases and the NADPH oxidase RBOHD. Conversely, deletion of xopB in Xcv caused a higher production of ROS in leaves of susceptible pepper plants. Together our results demonstrate that XopB modulates ROS responses and might thereby compromise plant defence.

  1. The Xanthomonas campestris pv. vesicatoria Type-3 Effector XopB Inhibits Plant Defence Responses by Interfering with ROS Production

    PubMed Central

    Priller, Johannes Peter Roman; Reid, Stephen; Konein, Patrick; Dietrich, Petra

    2016-01-01

    The bacterial pathogen Xanthomonas campestris pv. vesicatoria 85–10 (Xcv) translocates about 30 type-3 effector proteins (T3Es) into pepper plants (Capsicum annuum) to suppress plant immune responses. Among them is XopB which interferes with PTI, ETI and sugar-mediated defence responses, but the underlying molecular mechanisms and direct targets are unknown so far. Here, we examined the XopB-mediated suppression of plant defence responses in more detail. Infection of susceptible pepper plants with Xcv lacking xopB resulted in delayed symptom development compared to Xcv wild type infection concomitant with an increased formation of salicylic acid (SA) and expression of pathogenesis-related (PR) genes. Expression of xopB in Arabidopsis thaliana promoted the growth of the virulent Pseudomonas syringae pv. tomato (Pst) DC3000 strain. This was paralleled by a decreased SA-pool and a lower induction of SA-dependent PR gene expression. The expression pattern of early flg22-responsive marker genes indicated that MAPK signalling was not altered in the presence of XopB. However, XopB inhibited the flg22-triggered burst of reactive oxygen species (ROS). Consequently, the transcript accumulation of AtOXI1, a ROS-dependent marker gene, was reduced in xopB-expressing Arabidopsis plants as well as callose deposition. The lower ROS production correlated with a low level of basal and flg22-triggered expression of apoplastic peroxidases and the NADPH oxidase RBOHD. Conversely, deletion of xopB in Xcv caused a higher production of ROS in leaves of susceptible pepper plants. Together our results demonstrate that XopB modulates ROS responses and might thereby compromise plant defence. PMID:27398933

  2. The type III-dependent Hrp pilus is required for productive interaction of Xanthomonas campestris pv. vesicatoria with pepper host plants.

    PubMed

    Weber, Ernst; Ojanen-Reuhs, Tuula; Huguet, Elisabeth; Hause, Gerd; Romantschuk, Martin; Korhonen, Timo K; Bonas, Ulla; Koebnik, Ralf

    2005-04-01

    The plant pathogenic bacterium Xanthomonas campestris pv. vesicatoria expresses a type III secretion system that is necessary for both pathogenicity in susceptible hosts and the induction of the hypersensitive response in resistant plants. This specialized protein transport system is encoded by a 23-kb hrp (hypersensitive response and pathogenicity) gene cluster. Here we show that X. campestris pv. vesicatoria produces filamentous structures, the Hrp pili, at the cell surface under hrp-inducing conditions. Analysis of purified Hrp pili and immunoelectron microscopy revealed that the major component of the Hrp pilus is the HrpE protein which is encoded in the hrp gene cluster. Sequence homologues of hrpE are only found in other xanthomonads. However, hrpE is syntenic to the hrpY gene from another plant pathogen, Ralstonia solanacearum. Bioinformatic analyses suggest that all major Hrp pilus subunits from gram-negative plant pathogens may share the same structural organization, i.e., a predominant alpha-helical structure. Analysis of nonpolar mutants in hrpE demonstrated that the Hrp pilus is essential for the productive interaction of X. campestris pv. vesicatoria with pepper host plants. Furthermore, a functional Hrp pilus is required for type III-dependent protein secretion. Immunoelectron microscopy revealed that type III-secreted proteins, such as HrpF and AvrBs3, are in close contact with the Hrp pilus during and/or after their secretion. By systematic analysis of nonpolar hrp/hrc (hrp conserved) and hpa (hrp associated) mutants, we found that Hpa proteins as well as the translocon protein HrpF are dispensable for pilus assembly, while all other Hrp and Hrc proteins are required. Hence, there are no other conserved Hrp or Hrc proteins that act downstream of HrpE during type III-dependent protein translocation.

  3. Development of a Model to Predict the Primary Infection Date of Bacterial Spot (Xanthomonas campestris pv. vesicatoria) on Hot Pepper

    PubMed Central

    Kim, Ji-Hoon; Kang, Wee-Soo; Yun, Sung-Chul

    2014-01-01

    A population model of bacterial spot caused by Xanthomonas campestris pv. vesicatoria on hot pepper was developed to predict the primary disease infection date. The model estimated the pathogen population on the surface and within the leaf of the host based on the wetness period and temperature. For successful infection, at least 5,000 cells/ml of the bacterial population were required. Also, wind and rain were necessary according to regression analyses of the monitored data. Bacterial spot on the model is initiated when the pathogen population exceeds 1015 cells/g within the leaf. The developed model was validated using 94 assessed samples from 2000 to 2007 obtained from monitored fields. Based on the validation study, the predicted initial infection dates varied based on the year rather than the location. Differences in initial infection dates between the model predictions and the monitored data in the field were minimal. For example, predicted infection dates for 7 locations were within the same month as the actual infection dates, 11 locations were within 1 month of the actual infection, and only 3 locations were more than 2 months apart from the actual infection. The predicted infection dates were mapped from 2009 to 2012; 2011 was the most severe year. Although the model was not sensitive enough to predict disease severity of less than 0.1% in the field, our model predicted bacterial spot severity of 1% or more. Therefore, this model can be applied in the field to determine when bacterial spot control is required. PMID:25288995

  4. Development of a Model to Predict the Primary Infection Date of Bacterial Spot (Xanthomonas campestris pv. vesicatoria) on Hot Pepper.

    PubMed

    Kim, Ji-Hoon; Kang, Wee-Soo; Yun, Sung-Chul

    2014-06-01

    A population model of bacterial spot caused by Xanthomonas campestris pv. vesicatoria on hot pepper was developed to predict the primary disease infection date. The model estimated the pathogen population on the surface and within the leaf of the host based on the wetness period and temperature. For successful infection, at least 5,000 cells/ml of the bacterial population were required. Also, wind and rain were necessary according to regression analyses of the monitored data. Bacterial spot on the model is initiated when the pathogen population exceeds 10(15) cells/g within the leaf. The developed model was validated using 94 assessed samples from 2000 to 2007 obtained from monitored fields. Based on the validation study, the predicted initial infection dates varied based on the year rather than the location. Differences in initial infection dates between the model predictions and the monitored data in the field were minimal. For example, predicted infection dates for 7 locations were within the same month as the actual infection dates, 11 locations were within 1 month of the actual infection, and only 3 locations were more than 2 months apart from the actual infection. The predicted infection dates were mapped from 2009 to 2012; 2011 was the most severe year. Although the model was not sensitive enough to predict disease severity of less than 0.1% in the field, our model predicted bacterial spot severity of 1% or more. Therefore, this model can be applied in the field to determine when bacterial spot control is required.

  5. Reduced expression of the tomato ethylene receptor gene LeETR4 enhances the hypersensitive response to Xanthomonas campestris pv. vesicatoria.

    PubMed

    Ciardi, J A; Tieman, D M; Jones, J B; Klee, H J

    2001-04-01

    The hypersensitive response (HR) involves rapid death of cells at the site of pathogen infection and is thought to limit pathogen growth through the plant. Ethylene regulates senescence and developmental programmed cell death, but its role in hypersensitive cell death is less clear. Expression of two ethylene receptor genes, NR and LeETR4, is induced in tomato (Lycopersicon esculentum cv. Mill) leaves during an HR to Xanthomonas campestris pv. vesicatoria, with the greatest increase observed in LeETR4. LeETR4 antisense plants previously were shown to exhibit increased sensitivity to ethylene. These plants also exhibit greatly reduced induction of LeETR4 expression during infection and an accelerated HR at inoculum concentrations ranging from 10(5) to 10(7) CFU/ml. Increases in ethylene synthesis and pathogenesis-related gene expression are greater and more rapid in infected LeETR4 antisense plants, indicating an enhanced defense response. Populations of avirulent X. campestris pv. vesicatoria decrease more quickly and to a lower level in the transgenic plants, indicating a greater resistance to this pathogen. Because the ethylene action inhibitor 1-methylcyclopropene alleviates the enhanced HR phenotype in LeETR4 antisense plants, these changes in pathogen response are a result of increased ethylene sensitivity.

  6. Regulation of Cell Wall-Bound Invertase in Pepper Leaves by Xanthomonas campestris pv. vesicatoria Type Three Effectors

    PubMed Central

    Sonnewald, Sophia; Priller, Johannes P. R.; Schuster, Julia; Glickmann, Eric; Hajirezaei, Mohammed-Reza; Siebig, Stefan; Mudgett, Mary Beth; Sonnewald, Uwe

    2012-01-01

    Xanthomonas campestris pv. vesicatoria (Xcv) possess a type 3 secretion system (T3SS) to deliver effector proteins into its Solanaceous host plants. These proteins are involved in suppression of plant defense and in reprogramming of plant metabolism to favour bacterial propagation. There is increasing evidence that hexoses contribute to defense responses. They act as substrates for metabolic processes and as metabolic semaphores to regulate gene expression. Especially an increase in the apoplastic hexose-to-sucrose ratio has been suggested to strengthen plant defense. This shift is brought about by the activity of cell wall-bound invertase (cw-Inv). We examined the possibility that Xcv may employ type 3 effector (T3E) proteins to suppress cw-Inv activity during infection. Indeed, pepper leaves infected with a T3SS-deficient Xcv strain showed a higher level of cw-Inv mRNA and enzyme activity relative to Xcv wild type infected leaves. Higher cw-Inv activity was paralleled by an increase in hexoses and mRNA abundance for the pathogenesis-related gene PRQ. These results suggest that Xcv suppresses cw-Inv activity in a T3SS-dependent manner, most likely to prevent sugar-mediated defense signals. To identify Xcv T3Es that regulate cw-Inv activity, a screen was performed with eighteen Xcv strains, each deficient in an individual T3E. Seven Xcv T3E deletion strains caused a significant change in cw-Inv activity compared to Xcv wild type. Among them, Xcv lacking the xopB gene (Xcv ΔxopB) caused the most prominent increase in cw-Inv activity. Deletion of xopB increased the mRNA abundance of PRQ in Xcv ΔxopB-infected pepper leaves, but not of Pti5 and Acre31, two PAMP-triggered immunity markers. Inducible expression of XopB in transgenic tobacco inhibited Xcv-mediated induction of cw-Inv activity observed in wild type plants and resulted in severe developmental phenotypes. Together, these data suggest that XopB interferes with cw-Inv activity in planta to suppress sugar

  7. Regulation of cell wall-bound invertase in pepper leaves by Xanthomonas campestris pv. vesicatoria type three effectors.

    PubMed

    Sonnewald, Sophia; Priller, Johannes P R; Schuster, Julia; Glickmann, Eric; Hajirezaei, Mohammed-Reza; Siebig, Stefan; Mudgett, Mary Beth; Sonnewald, Uwe

    2012-01-01

    Xanthomonas campestris pv. vesicatoria (Xcv) possess a type 3 secretion system (T3SS) to deliver effector proteins into its Solanaceous host plants. These proteins are involved in suppression of plant defense and in reprogramming of plant metabolism to favour bacterial propagation. There is increasing evidence that hexoses contribute to defense responses. They act as substrates for metabolic processes and as metabolic semaphores to regulate gene expression. Especially an increase in the apoplastic hexose-to-sucrose ratio has been suggested to strengthen plant defense. This shift is brought about by the activity of cell wall-bound invertase (cw-Inv). We examined the possibility that Xcv may employ type 3 effector (T3E) proteins to suppress cw-Inv activity during infection. Indeed, pepper leaves infected with a T3SS-deficient Xcv strain showed a higher level of cw-Inv mRNA and enzyme activity relative to Xcv wild type infected leaves. Higher cw-Inv activity was paralleled by an increase in hexoses and mRNA abundance for the pathogenesis-related gene PRQ. These results suggest that Xcv suppresses cw-Inv activity in a T3SS-dependent manner, most likely to prevent sugar-mediated defense signals. To identify Xcv T3Es that regulate cw-Inv activity, a screen was performed with eighteen Xcv strains, each deficient in an individual T3E. Seven Xcv T3E deletion strains caused a significant change in cw-Inv activity compared to Xcv wild type. Among them, Xcv lacking the xopB gene (Xcv ΔxopB) caused the most prominent increase in cw-Inv activity. Deletion of xopB increased the mRNA abundance of PRQ in Xcv ΔxopB-infected pepper leaves, but not of Pti5 and Acre31, two PAMP-triggered immunity markers. Inducible expression of XopB in transgenic tobacco inhibited Xcv-mediated induction of cw-Inv activity observed in wild type plants and resulted in severe developmental phenotypes. Together, these data suggest that XopB interferes with cw-Inv activity in planta to suppress sugar

  8. Spontaneous and induced mutations in a single open reading frame alter both virulence and avirulence in Xanthomonas campestris pv. vesicatoria avrBs2.

    PubMed

    Swords, K M; Dahlbeck, D; Kearney, B; Roy, M; Staskawicz, B J

    1996-08-01

    Molecular characterization of the avrBs2 locus from Xanthomonas campestris pv. vesicatoria has revealed that expression of this gene triggers disease resistance in Bs2 pepper (Capsicum annuum) plants and contributes to virulence of the pathogen. Deletion analysis and site-directed mutagenesis established the avrBs2 gene as a 2,190-bp open reading frame encoding a putative 80.1-kDa protein. Two classes of Xanthomonas pathogens evading Bs2 host resistance and displaying reduced fitness were found to be specifically mutated in avrBs2. Members of one class contained a 5-bp insertion, while the second class was distinguished by a divergent 3' region of avrBs2; both mutant classes were complemented in trans by a plasmid-borne copy of avrBs2. A divergent avrBs2 homolog was cloned from the Brassica pathogen X. campestris pv. campestris. The predicted AvrBs2 proteins from the two Xanthomonas pathovars were strongly conserved and had predicted sequence similarity with both Agrobacterium tumefaciens agrocinopine synthase and Escherichia coli UgpQ, two enzymes involved in the synthesis or hydrolysis of phosphodiester linkages. On the basis of homology with agrocinopine synthase and UgpQ and the dual phenotype of avirulence and virulence, several models for the function of AvrBs2 are proposed.

  9. hpaA mutants of Xanthomonas campestris pv. vesicatoria are affected in pathogenicity but retain the ability to induce host-specific hypersensitive reaction.

    PubMed

    Huguet, E; Hahn, K; Wengelnik, K; Bonas, U

    1998-09-01

    Xanthomonas campestris pv. vesicatoria is the causal agent of bacterial spot disease on pepper and tomato plants. We reported previously that the main hrp (hypersensitive reaction and pathogenicity) gene cluster in X. c. pv. vesicatoria contains six transcription units, designated hrpA to hrpF. We present here the sequence of the hrpD operon and an analysis of non-polar mutants in each of the six genes. Three genes, hrcQ, hrcR and hrcS, are predicted to encode conserved components of type III protein secretion systems in plant and mammalian pathogenic bacteria. For hrpD5 and hrpD6, homologues have only been found in Ralstonia solanacearum. Interestingly, the hrpD operon contains one gene, hpaA (for hrp-associated), which is specifically required for disease development. hpaA mutants are affected in pathogenicity, but retain in part the ability to induce avirulence gene-mediated, host-specific hypersensitive reaction (HR). In addition, HpaA was found to contain two functional nuclear localization signals, which are important for the interaction with the plant. We propose that HpaA is an effector protein that may be translocated into the host cell via the Hrp secretion pathway.

  10. Specific Binding of the Xanthomonas campestris pv. vesicatoria AraC-Type Transcriptional Activator HrpX to Plant-Inducible Promoter Boxes▿

    PubMed Central

    Koebnik, Ralf; Krüger, Antje; Thieme, Frank; Urban, Alexander; Bonas, Ulla

    2006-01-01

    The pathogenicity of the plant-pathogenic bacterium Xanthomonas campestris pv. vesicatoria depends on a type III secretion system which is encoded by the 23-kb hrp (hypersensitive response and pathogenicity) gene cluster. Expression of the hrp operons is strongly induced in planta and in a special minimal medium and depends on two regulatory proteins, HrpG and HrpX. In this study, DNA affinity enrichment was used to demonstrate that the AraC-type transcriptional activator HrpX binds to a conserved cis-regulatory element, the plant-inducible promoter (PIP) box (TTCGC-N15-TTCGC), present in the promoter regions of four hrp operons. No binding of HrpX was observed when DNA fragments lacking a PIP box were used. HrpX also bound to a DNA fragment containing an imperfect PIP box (TTCGC-N8-TTCGT). Dinucleotide replacements in each half-site of the PIP box strongly decreased binding of HrpX, while simultaneous dinucleotide replacements in both half-sites completely abolished binding. Based on the complete genome sequence of Xanthomonas campestris pv. vesicatoria, putative plant-inducible promoters consisting of a PIP box and a −10 promoter motif were identified in the promoter regions of almost all HrpX-activated genes. Bioinformatic analyses and reverse transcription-PCR experiments revealed novel HrpX-dependent genes, among them a NUDIX hydrolase gene and several genes with a predicted role in the degradation of the plant cell wall. We conclude that HrpX is the most downstream component of the hrp regulatory cascade, which is proposed to directly activate most genes of the hrpX regulon via binding to corresponding PIP boxes. PMID:16936021

  11. Sequence and expression analysis of the hrpB pathogenicity operon of Xanthomonas campestris pv. vesicatoria which encodes eight proteins with similarity to components of the Hrp, Ysc, Spa, and Fli secretion systems.

    PubMed

    Fenselau, S; Bonas, U

    1995-01-01

    In this paper we describe the molecular characterization of hrpB, the largest operon in the Xanthomonas campestris pv. vesicatoria hrp cluster. The hrpB region encompasses 6 kb and encodes eight putative proteins, seven of which were expressed in Escherichia coli. The HrpB3 protein is the only one carrying a signal peptide sequence at the N-terminus and is a putative lipoprotein localized in the outer membrane of X. campestris pv. vesicatoria. The HrpB4 and HrpB8 proteins contain one and five putative transmembrane domains, respectively, and are most likely associated with the inner membrane. The HrpB3, HrpB5, HrpB6, and HrpB8 proteins show sequence similarity to putative components of different type III protein secretion pathways in bacteria. Examples include Hrp proteins from other plant pathogens, YscJ, YscN, YscL, and YscT of Yersinia spp., and MxiJ, Spa47, adn Spa29 of Shigella flexneri. The transcription start site and the hrpB promoter was identified. The minimal hrpB promoter region of 90 bp contains a novel sequence motif, the PIP-box, which might play a role in transcription activation of the hrpB operon and possibly other plant-induced genes of X. campestris pv. vesicatoria.

  12. Type III-Dependent Translocation of HrpB2 by a Nonpathogenic hpaABC Mutant of the Plant-Pathogenic Bacterium Xanthomonas campestris pv. vesicatoria

    PubMed Central

    Scheibner, Felix; Schulz, Steve; Hausner, Jens; Marillonnet, Sylvestre

    2016-01-01

    ABSTRACT The plant-pathogenic bacterium Xanthomonas campestris pv. vesicatoria employs a type III secretion (T3S) system to translocate effector proteins into plant cells. The T3S apparatus spans both bacterial membranes and is associated with an extracellular pilus and a channel-like translocon in the host plasma membrane. T3S is controlled by the switch protein HpaC, which suppresses secretion and translocation of the predicted inner rod protein HrpB2 and promotes secretion of translocon and effector proteins. We previously reported that HrpB2 interacts with HpaC and the cytoplasmic domain of the inner membrane protein HrcU (C. Lorenz, S. Schulz, T. Wolsch, O. Rossier, U. Bonas, and D. Büttner, PLoS Pathog 4:e1000094, 2008, http://dx.doi.org/10.1371/journal.ppat.1000094). However, the molecular mechanisms underlying the control of HrpB2 secretion are not yet understood. Here, we located a T3S and translocation signal in the N-terminal 40 amino acids of HrpB2. The results of complementation experiments with HrpB2 deletion derivatives revealed that the T3S signal of HrpB2 is essential for protein function. Furthermore, interaction studies showed that the N-terminal region of HrpB2 interacts with the cytoplasmic domain of HrcU, suggesting that the T3S signal of HrpB2 contributes to substrate docking. Translocation of HrpB2 is suppressed not only by HpaC but also by the T3S chaperone HpaB and its secreted regulator, HpaA. Deletion of hpaA, hpaB, and hpaC leads to a loss of pathogenicity but allows the translocation of fusion proteins between the HrpB2 T3S signal and effector proteins into leaves of host and non-host plants. IMPORTANCE The T3S system of the plant-pathogenic bacterium Xanthomonas campestris pv. vesicatoria is essential for pathogenicity and delivers effector proteins into plant cells. T3S depends on HrpB2, which is a component of the predicted periplasmic inner rod structure of the secretion apparatus. HrpB2 is secreted during the early stages of the

  13. Resistance in Lycopersicon esculentum Intraspecific Crosses to Race T1 Strains of Xanthomonas campestris pv. vesicatoria Causing Bacterial Spot of Tomato.

    PubMed

    Yang, Wencai; Sacks, Erik J; Lewis Ivey, Melanie L; Miller, Sally A; Francis, David M

    2005-05-01

    ABSTRACT We used molecular markers to identify quantitative trait loci (QTL) that confer resistance in the field to Xanthomonas campestris pv. vesicatoria race T1, a causal agent of bacterial spot of tomato. An F(2) population derived from a cross between Hawaii 7998 (H 7998) and an elite breeding line, Ohio 88119, was used for the initial identification of an association between molecular markers and resistance as measured by bacterial populations in individual plants in the greenhouse. Polymorphism in this cross between a Lycopersicon esculentum donor of resistance and an elite L. esculentum parent was limited. The targeted use of a core set of 148 polymerase chain reaction-based markers that were identified as polymorphic in L. esculentum x L. esculentum crosses resulted in the identification of 37 markers that were polymorphic for the cross of interest. Previous studies using an H 7998 x L. pennellii wide cross implicated three loci, Rx1, Rx2, and Rx3, in the hypersensitive response to T1 strains. Markers that we identified were linked to the Rx1 and Rx3 loci, but no markers were identified in the region of chromosome 1 where Rx2 is located. Single marker-trait analysis suggested that chromosome 5, near the Rx3 locus, contributed to reduced bacterial populations in lines carrying the locus from H 7998. The locus on chromosome 5 explained 25% of the phenotypic variation in bacterial populations developing in infected plants. An advanced backcross population and subsequent inbred backcross lines developed using Ohio 88119 as a recurrent parent were used to confirm QTL associations detected in the F(2) population. Markers on chromosome 5 explained 41% of the phenotypic variation for resistance in replicated field trials. In contrast, the Rx1 locus on chromosome 1 did not play a role in resistance to X. campestris pv. vesicatoria race T1 strains as measured by bacterial populations in the greenhouse or symptoms in the field. A locus from H 7998 on chromosome 4 was

  14. Expression of peroxidase-like genes, H2O2 production, and peroxidase activity during the hypersensitive response to Xanthomonas campestris pv. vesicatoria in Capsicum annuum.

    PubMed

    Do, Hyun Mee; Hong, Jeum Kyu; Jung, Ho Won; Kim, Sang Hee; Ham, Jong Hyun; Hwang, Byung Kook

    2003-03-01

    Pepper ascorbate peroxidase-like (CAPOA1), thioredoxin peroxidase-like (CAPOT1), and peroxidase-like (CAPO1) clones were isolated from pepper leaves inoculated with avirulent strain Bv5-4a of Xanthomonas campestris pv. vesicatoria. CAPOA1, CAPOT1, and CAPO1 mRNA disappeared 18 to 30 h after the bacterial infection when the hypersensitive response (HR) was visible. In contrast, peroxidase activity reached a peak at 18 h after infection and then declined at 24 and 30 h when H2O2 accumulation level was maximal. These results suggest that the striking accumulation of H2O2 and strong decrease in peroxidase activity during the programmed cell death may be due to the strong suppression of CAPOA1, CAPOT1, and CAPO1 gene expression. Infection by Phytophthora capsici or Colletotricum gloeosporioides also induced the expression of the three putative peroxidase genes in pepper tissues. CAPOA1 mRNAs were in situ localized in phloem areas of vascular bundles in pepper tissues infected by Colletotricum. coccodes, P. capsici, or C. gloeosporioides. Exogenous treatment with H2O2 strongly induced the CAPOA1 and CAPOT1 transcription 1 h after treatment, while the CAPO1 transcripts accumulated 12 h after H2O2 treatment. We suggest that pepper ascorbate peroxidase and thioredoxin peroxidase genes may function as regulators of H2O2 level and total peroxidase activity in the oxidative burst during the HR to incompatible pathogen interaction in pepper plant.

  15. The Predicted Lytic Transglycosylase HpaH from Xanthomonas campestris pv. vesicatoria Associates with the Type III Secretion System and Promotes Effector Protein Translocation

    PubMed Central

    Hausner, Jens; Hartmann, Nadine; Jordan, Michael

    2016-01-01

    ABSTRACT The pathogenicity of the Gram-negative plant-pathogenic bacterium Xanthomonas campestris pv. vesicatoria depends on a type III secretion (T3S) system, which spans both bacterial membranes and translocates effector proteins into plant cells. The assembly of the T3S system presumably involves the predicted lytic transglycosylase (LT) HpaH, which is encoded adjacent to the T3S gene cluster. Bacterial LTs degrade peptidoglycan and often promote the formation of membrane-spanning macromolecular protein complexes. In the present study, we show that HpaH localizes to the bacterial periplasm and binds to peptidoglycan as well as to components of the T3S system, including the predicted periplasmic inner rod proteins HrpB1 and HrpB2 as well as the pilus protein HrpE. In vivo translocation assays revealed that HpaH promotes the translocation of various effector proteins and of early substrates of the T3S system, suggesting a general contribution of HpaH to type III-dependent protein export. Mutant studies and the analysis of reporter fusions showed that the N-terminal region of HpaH contributes to protein function and is proteolytically cleaved. The N-terminally truncated HpaH cleavage product is secreted into the extracellular milieu by a yet-unknown transport pathway, which is independent of the T3S system. PMID:27895129

  16. The Predicted Lytic Transglycosylase HpaH from Xanthomonas campestris pv. vesicatoria Associates with the Type III Secretion System and Promotes Effector Protein Translocation.

    PubMed

    Hausner, Jens; Hartmann, Nadine; Jordan, Michael; Büttner, Daniela

    2017-02-01

    The pathogenicity of the Gram-negative plant-pathogenic bacterium Xanthomonas campestris pv. vesicatoria depends on a type III secretion (T3S) system, which spans both bacterial membranes and translocates effector proteins into plant cells. The assembly of the T3S system presumably involves the predicted lytic transglycosylase (LT) HpaH, which is encoded adjacent to the T3S gene cluster. Bacterial LTs degrade peptidoglycan and often promote the formation of membrane-spanning macromolecular protein complexes. In the present study, we show that HpaH localizes to the bacterial periplasm and binds to peptidoglycan as well as to components of the T3S system, including the predicted periplasmic inner rod proteins HrpB1 and HrpB2 as well as the pilus protein HrpE. In vivo translocation assays revealed that HpaH promotes the translocation of various effector proteins and of early substrates of the T3S system, suggesting a general contribution of HpaH to type III-dependent protein export. Mutant studies and the analysis of reporter fusions showed that the N-terminal region of HpaH contributes to protein function and is proteolytically cleaved. The N-terminally truncated HpaH cleavage product is secreted into the extracellular milieu by a yet-unknown transport pathway, which is independent of the T3S system. Copyright © 2017 American Society for Microbiology.

  17. The SlMKK2 and SlMPK2 genes play a role in tomato disease resistance to Xanthomonas campestris pv. vesicatoria.

    PubMed

    Melech-Bonfil, Shiri; Sessa, Guido

    2011-01-01

    Xanthomonas campestris pv. vesicatoria (Xcv) is the causal agent of bacterial spot disease in tomato (Solanum lycopersicum) plants. We recently identified a MAPKKK gene, SlMAPKKKε, which is required for tomato resistance to Xcv strains and encodes a positive regulator of cell death. We also provided evidence that the MEK2 MAPKK, and the WIPK and SIPK MAPKs act downstream to MAPKKKε in Nicotiana benthamiana plants. Here, we used the virus-induced gene silencing technique to assess whether tomato homologs of MEK2 (SlMKK2), SIPK (SlMPK1 and SlMPK2), WIPK (SlMPK3), and other components of MAP kinase cascades (SlNPK1, SlMEK1 and SlNTF6), which were previously implicated in plant immunity, are involved in disease resistance to Xcv. Silencing of none of the tested genes caused the appearance of disease symptoms in tomato leaves challenged with an avirulent Xcv strain. However, bacterial populations were significantly higher in leaves of plants silenced for SlMKK2 and SlMPK2, as compared to control plants, suggesting that these two genes contribute to disease resistance to Xcv. It remains to be established whether SlMKK2 and SlMPK2 are activated by SlMAPKKKε directly or through a distinct MAPKKK.  

  18. Functional and proteomic analyses reveal that wxcB is involved in virulence, motility, detergent tolerance, and biofilm formation in Xanthomonas campestris pv. vesicatoria.

    PubMed

    Park, Hye-Jee; Jung, Ho Won; Han, Sang-Wook

    2014-09-26

    The bacterial envelope possesses diverse functions, including protection against environmental stress and virulence factors for host infection. Here, we report the function of wxcB in Xanthomonas campestris pv. vesicatoria (Xcv), a causal agent of bacterial leaf spot disease in tomato and pepper. To characterize roles of wxcB, we generated a knockout mutant (XcvΔwxcB) and found that the virulence of the mutant was weaker than that of the wild type in tomato plants. To predict the mechanism affected by wxcB, we compared protein expressions between the wild type and the mutant. Expression of 152 proteins showed a greater than 2-fold difference. Proteins involved in motility and cell wall/membrane were the most abundant. Through phenotypic assays, we further demonstrated that the mutant displayed reduced motility and tolerance to treatment, but it showed increased biofilm formation. Interestingly, the LPS profile was unchanged. These results lead to new insights into the functions of wxcB that is associated with cell wall/membrane functions, which contributes to pathogen virulence.

  19. Real time expression of ACC oxidase and PR-protein genes mediated by Methylobacterium spp. in tomato plants challenged with Xanthomonas campestris pv. vesicatoria.

    PubMed

    Yim, W J; Kim, K Y; Lee, Y W; Sundaram, S P; Lee, Y; Sa, T M

    2014-07-15

    Biotic stress like pathogenic infection increases ethylene biosynthesis in plants and ethylene inhibitors are known to alleviate the severity of plant disease incidence. This study aimed to reduce the bacterial spot disease incidence in tomato plants caused by Xanthomonas campestris pv. vesicatoria (XCV) by modulating stress ethylene with 1-aminocyclopropane-1-carboxylate (ACC) deaminase activity of Methylobacterium strains. Under greenhouse condition, Methylobacterium strains inoculated and pathogen challenged tomato plants had low ethylene emission compared to pathogen infected ones. ACC accumulation and ACC oxidase (ACO) activity with ACO related gene expression increased in XCV infected tomato plants over Methylobacterium strains inoculated plants. Among the Methylobacterium spp., CBMB12 resulted lowest ACO related gene expression (1.46 Normalized Fold Expression), whereas CBMB20 had high gene expression (3.42 Normalized Fold Expression) in pathogen challenged tomato. But a significant increase in ACO gene expression (7.09 Normalized Fold Expression) was observed in the bacterial pathogen infected plants. In contrast, Methylobacterium strains enhanced β-1,3-glucanase and phenylalanine ammonia-lyase (PAL) enzyme activities in pathogen challenged tomato plants. The respective increase in β-1,3-glucanase related gene expressions due to CBMB12, CBMB15, and CBMB20 strains were 66.3, 25.5 and 10.4% higher over pathogen infected plants. Similarly, PAL gene expression was high with 0.67 and 0.30 Normalized Fold Expression, in pathogen challenged tomato plants inoculated with CBMB12 and CBMB15 strains. The results suggest that ethylene is a crucial factor in bacterial spot disease incidence and that methylobacteria with ACC deaminase activity can reduce the disease severity with ultimate pathogenesis-related protein increase in tomato. Copyright © 2014 Elsevier GmbH. All rights reserved.

  20. Chloroplast-generated reactive oxygen species play a major role in localized cell death during the non-host interaction between tobacco and Xanthomonas campestris pv. vesicatoria.

    PubMed

    Zurbriggen, Matias D; Carrillo, Néstor; Tognetti, Vanesa B; Melzer, Michael; Peisker, Martin; Hause, Bettina; Hajirezaei, Mohammad-Reza

    2009-12-01

    Attempted infection of plants by pathogens elicits a complex defensive response. In many non-host and incompatible host interactions it includes the induction of defence-associated genes and a form of localized cell death (LCD), purportedly designed to restrict pathogen advance, collectively known as the hypersensitive response (HR). It is preceded by an oxidative burst, generating reactive oxygen species (ROS) that are proposed to cue subsequent deployment of the HR, although neither the origin nor the precise role played by ROS in the execution of this response are completely understood. We used tobacco plants expressing cyanobacterial flavodoxin to address these questions. Flavodoxin is an electron shuttle present in prokaryotes and algae that, when expressed in chloroplasts, specifically prevents ROS formation in plastids during abiotic stress episodes. Infiltration of tobacco wild-type leaves with high titres of Xanthomonas campestris pv. vesicatoria (Xcv), a non-host pathogen, resulted in ROS accumulation in chloroplasts, followed by the appearance of localized lesions typical of the HR. In contrast, chloroplast ROS build-up and LCD were significantly reduced in Xcv-inoculated plants expressing plastid-targeted flavodoxin. Metabolic routes normally inhibited by pathogens were protected in the transformants, whereas other aspects of the HR, including the induction of defence-associated genes and synthesis of salicylic and jasmonic acid, proceeded as in inoculated wild-type plants. Therefore, ROS generated in chloroplasts during this non-host interaction are essential for the progress of LCD, but do not contribute to the induction of pathogenesis-related genes or other signalling components of the response.

  1. Characterization of HrpB2 from Xanthomonas campestris pv. vesicatoria identifies protein regions that are essential for type III secretion pilus formation.

    PubMed

    Hartmann, Nadine; Schulz, Steve; Lorenz, Christian; Fraas, Simone; Hause, Gerd; Büttner, Daniela

    2012-05-01

    The Gram-negative plant-pathogenic bacterium Xanthomonas campestris pv. vesicatoria employs a type III secretion (T3S) system to translocate effector proteins into plant cells. T3S depends on HrpB2, which is essential for assembly of the extracellular T3S pilus and is itself weakly secreted. To characterize the role of HrpB2, we used a transposon mutagenesis approach, which led to the insertion of pentapeptide-encoding sequences into hrpB2. Complementation studies with HrpB2 mutant derivatives revealed that the N-terminal region of HrpB2 tolerates pentapeptide insertions, whereas insertions in the regions spanning amino acids 60-74 and 93-130, respectively, resulted in a loss of bacterial pathogenicity and T3S, including secretion of HrpB2 itself. The C-terminal region (amino acids 93-130) of HrpB2 contains a conserved VxTLxK amino acid motif that is also present in predicted inner rod proteins from animal-pathogenic bacteria and is required for the contribution of HrpB2 to pilus assembly and T3S. Electron microscopy and fractionation studies revealed that HrpB2 is not a component of the extracellular pilus structure but localizes to the bacterial periplasm and the outer membrane. We therefore propose that the essential contribution of HrpB2 to T3S and pilus assembly is linked to its possible function as a periplasmic component of the T3S system at the base of the pilus.

  2. Chloroplast Redox Status Modulates Genome-Wide Plant Responses during the Non-host Interaction of Tobacco with the Hemibiotrophic Bacterium Xanthomonas campestris pv. vesicatoria

    PubMed Central

    Pierella Karlusich, Juan J.; Zurbriggen, Matias D.; Shahinnia, Fahimeh; Sonnewald, Sophia; Sonnewald, Uwe; Hosseini, Seyed A.; Hajirezaei, Mohammad-Reza; Carrillo, Néstor

    2017-01-01

    Non-host resistance is the most ample and durable form of plant resistance against pathogen infection. It includes induction of defense-associated genes, massive metabolic reprogramming, and in many instances, a form of localized cell death (LCD) at the site of infection, purportedly designed to limit the spread of biotrophic and hemibiotrophic microorganisms. Reactive oxygen species (ROS) have been proposed to act as signals for LCD orchestration. They are produced in various cellular compartments including chloroplasts, mitochondria and apoplast. We have previously reported that down-regulation of ROS build-up in chloroplasts by expression of a plastid-targeted flavodoxin (Fld) suppressed LCD in tobacco leaves inoculated with the non-host bacterium Xanthomonas campestris pv. vesicatoria (Xcv), while other defensive responses were unaffected, suggesting that chloroplast ROS and/or redox status play a major role in the progress of LCD. To better understand these effects, we compare here the transcriptomic alterations caused by Xcv inoculation on leaves of Fld-expressing tobacco plants and their wild-type siblings. About 29% of leaf-expressed genes were affected by Xcv and/or Fld. Surprisingly, 5.8% of them (1,111 genes) were regulated by Fld in the absence of infection, presumably representing pathways responsive to chloroplast ROS production and/or redox status during normal growth conditions. While the majority (∼75%) of pathogen-responsive genes were not affected by Fld, many Xcv responses were exacerbated, attenuated, or regulated in opposite direction by expression of this protein. Particularly interesting was a group of 384 genes displaying Xcv responses that were already triggered by Fld in the absence of infection, suggesting that the transgenic plants had a larger and more diversified suite of constitutive defenses against the attacking microorganism compared to the wild type. Fld modulated many genes involved in pathogenesis, signal transduction

  3. Chloroplast Redox Status Modulates Genome-Wide Plant Responses during the Non-host Interaction of Tobacco with the Hemibiotrophic Bacterium Xanthomonas campestris pv. vesicatoria.

    PubMed

    Pierella Karlusich, Juan J; Zurbriggen, Matias D; Shahinnia, Fahimeh; Sonnewald, Sophia; Sonnewald, Uwe; Hosseini, Seyed A; Hajirezaei, Mohammad-Reza; Carrillo, Néstor

    2017-01-01

    Non-host resistance is the most ample and durable form of plant resistance against pathogen infection. It includes induction of defense-associated genes, massive metabolic reprogramming, and in many instances, a form of localized cell death (LCD) at the site of infection, purportedly designed to limit the spread of biotrophic and hemibiotrophic microorganisms. Reactive oxygen species (ROS) have been proposed to act as signals for LCD orchestration. They are produced in various cellular compartments including chloroplasts, mitochondria and apoplast. We have previously reported that down-regulation of ROS build-up in chloroplasts by expression of a plastid-targeted flavodoxin (Fld) suppressed LCD in tobacco leaves inoculated with the non-host bacterium Xanthomonas campestris pv. vesicatoria (Xcv), while other defensive responses were unaffected, suggesting that chloroplast ROS and/or redox status play a major role in the progress of LCD. To better understand these effects, we compare here the transcriptomic alterations caused by Xcv inoculation on leaves of Fld-expressing tobacco plants and their wild-type siblings. About 29% of leaf-expressed genes were affected by Xcv and/or Fld. Surprisingly, 5.8% of them (1,111 genes) were regulated by Fld in the absence of infection, presumably representing pathways responsive to chloroplast ROS production and/or redox status during normal growth conditions. While the majority (∼75%) of pathogen-responsive genes were not affected by Fld, many Xcv responses were exacerbated, attenuated, or regulated in opposite direction by expression of this protein. Particularly interesting was a group of 384 genes displaying Xcv responses that were already triggered by Fld in the absence of infection, suggesting that the transgenic plants had a larger and more diversified suite of constitutive defenses against the attacking microorganism compared to the wild type. Fld modulated many genes involved in pathogenesis, signal transduction

  4. Regulation of resistance to copper in Xanthomonas axonopodis pv. vesicatoria.

    PubMed

    Voloudakis, Andreas E; Reignier, Therese M; Cooksey, Donald A

    2005-02-01

    Copper-resistant strains of Xanthomonas axonopodis pv. vesicatoria were previously shown to carry plasmid-borne copper resistance genes related to the cop and pco operons of Pseudomonas syringae and Escherichia coli, respectively. However, instead of the two-component (copRS and pcoRS) systems determining copper-inducible expression of the operons in P. syringae and E. coli, a novel open reading frame, copL, was found to be required for copper-inducible expression of the downstream multicopper oxidase copA in X. axonopodis. copL encodes a predicted protein product of 122 amino acids that is rich in histidine and cysteine residues, suggesting a possible direct interaction with copper. Deletions or frameshift mutations within copL, as well as an amino acid substitution generated at the putative start codon of copL, caused a loss of copper-inducible transcriptional activation of copA. A nonpolar insertion of a kanamycin resistance gene in copL resulted in copper sensitivity in the wild-type strain. However, repeated attempts to complement copL mutations in trans failed. Analysis of the genomic sequence databases shows that there are copL homologs upstream of copAB genes in X. axonopodis pv. citri, X. campestris pv. campestris, and Xylella fastidiosa. The cloned promoter area upstream of copA in X. axonopodis pv. vesicatoria did not function in Pseudomonas syringae or in E. coli, nor did the P. syringae cop promoter function in Xanthomonas. However, a transcriptional fusion of the Xanthomonas cop promoter with the Pseudomonas copABCDRS was able to confer resistance to copper in Xanthomonas, showing divergence in the mechanisms of regulation of the resistance to copper in phytopathogenic bacteria.

  5. The N-Glycan Cluster from Xanthomonas campestris pv. campestris

    PubMed Central

    Dupoiron, Stéphanie; Zischek, Claudine; Ligat, Laetitia; Carbonne, Julien; Boulanger, Alice; Dugé de Bernonville, Thomas; Lautier, Martine; Rival, Pauline; Arlat, Matthieu; Jamet, Elisabeth; Lauber, Emmanuelle; Albenne, Cécile

    2015-01-01

    N-Glycans are widely distributed in living organisms but represent only a small fraction of the carbohydrates found in plants. This probably explains why they have not previously been considered as substrates exploited by phytopathogenic bacteria during plant infection. Xanthomonas campestris pv. campestris, the causal agent of black rot disease of Brassica plants, possesses a specific system for GlcNAc utilization expressed during host plant infection. This system encompasses a cluster of eight genes (nixE to nixL) encoding glycoside hydrolases (GHs). In this paper, we have characterized the enzymatic activities of these GHs and demonstrated their involvement in sequential degradation of a plant N-glycan using a N-glycopeptide containing two GlcNAcs, three mannoses, one fucose, and one xylose (N2M3FX) as a substrate. The removal of the α-1,3-mannose by the α-mannosidase NixK (GH92) is a prerequisite for the subsequent action of the β-xylosidase NixI (GH3), which is involved in the cleavage of the β-1,2-xylose, followed by the α-mannosidase NixJ (GH125), which removes the α-1,6-mannose. These data, combined to the subcellular localization of the enzymes, allowed us to propose a model of N-glycopeptide processing by X. campestris pv. campestris. This study constitutes the first evidence suggesting N-glycan degradation by a plant pathogen, a feature shared with human pathogenic bacteria. Plant N-glycans should therefore be included in the repertoire of molecules putatively metabolized by phytopathogenic bacteria during their life cycle. PMID:25586188

  6. [A virulence gene from Xanthomonas campestris pv. campestris homologous to the avrBs2 locus is recognized in race-specific reaction by two different resistance genes in Brassica plant species].

    PubMed

    Ignatov, A N; Monakhos, G F; Dzhalilov, F S; Pozmogova, G V

    2002-12-01

    Race-specific interaction between the Brassica plants and Xanthomonas compestris pv. campestris bacteria follows the "gene-for-gene" rule. Expression of the avirulence genes recognized by two dominant resistance genes of Brassica, Rxc1 in plants with the BB genome, and Rxc3 in the CC plants, was lost after bacterial mutation in planta. The mutation was distinguished by the elongation of CGCGC pentanucleotide repeat in the gene, which was designated as avrRxc1/3. This gene displayed strong structural similarity to the avrBs2 locus from the related species X. vesicatoria. Thus, it is the first description of the avrRxc1/3 avirulence gene conferring race-specific interaction between X. campestris pv. campestris and Brassica plants. Structural homologues of the avrBs2 are found in many Xanthomonas species, but in all cases except X. vesicatoria, their function remains unknown.

  7. Genome Sequences of Three Atypical Xanthomonas campestris pv. campestris Strains, CN14, CN15, and CN16

    PubMed Central

    Bolot, Stéphanie; Roux, Brice; Carrere, Sébastien; Jiang, Bo-Le; Tang, Ji-Liang; Arlat, Matthieu

    2013-01-01

    Xanthomonas campestris pv. campestris is the causal agent of black rot on Brassicaceae. The draft genome sequences of three strains (CN14, CN15, and CN16) that are highly aggressive on Arabidopsis have been determined. These genome sequences present an unexpected genomic diversity in X. campestris pv. campestris, which will be valuable for comparative analyses. PMID:23846270

  8. The Plant Pathogen Xanthomonas campestris pv. campestris Exploits N-Acetylglucosamine during Infection

    PubMed Central

    Boulanger, Alice; Zischek, Claudine; Lautier, Martine; Jamet, Stevie; Rival, Pauline; Carrère, Sébastien; Arlat, Matthieu

    2014-01-01

    ABSTRACT N-Acetylglucosamine (GlcNAc), the main component of chitin and a major constituent of bacterial peptidoglycan, is present only in trace amounts in plants, in contrast to the huge amount of various sugars that compose the polysaccharides of the plant cell wall. Thus, GlcNAc has not previously been considered a substrate exploited by phytopathogenic bacteria during plant infection. Xanthomonas campestris pv. campestris, the causal agent of black rot disease of Brassica plants, expresses a carbohydrate utilization system devoted to GlcNAc exploitation. In addition to genes involved in GlcNAc catabolism, this system codes for four TonB-dependent outer membrane transporters (TBDTs) and eight glycoside hydrolases. Expression of all these genes is under the control of GlcNAc. In vitro experiments showed that X. campestris pv. campestris exploits chitooligosaccharides, and there is indirect evidence that during the early stationary phase, X. campestris pv. campestris recycles bacterium-derived peptidoglycan/muropeptides. Results obtained also suggest that during plant infection and during growth in cabbage xylem sap, X. campestris pv. campestris encounters and metabolizes plant-derived GlcNAc-containing molecules. Specific TBDTs seem to be preferentially involved in the consumption of all these plant-, fungus- and bacterium-derived GlcNAc-containing molecules. This is the first evidence of GlcNAc consumption during infection by a phytopathogenic bacterium. Interestingly, N-glycans from plant N-glycosylated proteins are proposed to be substrates for glycoside hydrolases belonging to the X. campestris pv. campestris GlcNAc exploitation system. This observation extends the range of sources of GlcNAc metabolized by phytopathogenic bacteria during their life cycle. PMID:25205095

  9. Identification and Origin of Xanthomonas campestris pv. campestris Races and Related Pathovars.

    PubMed

    Vicente, J G; Conway, J; Roberts, S J; Taylor, J D

    2001-05-01

    ABSTRACT One hundred sixty-four isolates of Xanthomonas campestris pv. campestris and other X. campestris pathovars known to infect cruciferous hosts (X. campestris pvs. aberrans, raphani, armoraciae, and incanae) were inoculated onto a differential series of Brassica spp. to determine both pathogenicity to brassicas and race. Of these, 144 isolates were identified as X. campestris pv. campestris and grouped into six races, with races 1 (62%) and 4 (32%) being predominant. Other races were rare. The remaining 20 isolates from brassicas and other cruciferous hosts were either nonpathogenic or very weakly pathogenic on the differential series and could not be race-typed. Five of these isolates, from the ornamental crucifers wallflower (Cheiranthus cheiri), stock (Matthiola incana) and candytuft (Iberis sp.), showed clear evidence of pathovar-like specificity to the hosts of origin. A gene-for-gene model based on the interaction of four avirulence genes in X. campestris pv. campestris races and four matching resistance genes in the differential hosts is proposed. Knowledge of the race structure and worldwide distribution of races is fundamental to the search for sources of resistance and for the establishment of successful resistance breeding programs.

  10. Functional characterization and transcriptional analysis of icd2 gene encoding an isocitrate dehydrogenase of Xanthomonas campestris pv. campestris.

    PubMed

    Chiang, Ying-Chuan; Liao, Chao-Tsai; Du, Shin-Chiao; Hsiao, Yi-Min

    2017-08-01

    Isocitrate dehydrogenase (IDH) catalyzes the oxidative decarboxylation of isocitrate to alpha-ketoglutarate. In the genome of Xanthomonas campestris pv. campestris, the phytopathogen that causes black rot in cruciferous plants, two putative IDH genes, icd1 and icd2, have been annotated. Their physiological roles in X. campestris pv. campestris are unclear. In this study, the icd2 gene from X. campestris pv. campestris was characterized in detail. We demonstrated genetically that icd2 gene encodes a functional IDH, and is involved in virulence as well as bacterial attachment. Furthermore, the icd2 transcription initiation site was mapped at nucleotide G, 127 nucleotide upstream of the icd2 translation start codon. In addition, promoter analysis revealed that icd2 expression exhibits a distinct expression profile under different culture conditions, is subjected to catabolite repression, and is affected by acetate. This is the first time that the function and transcription of icd2 have been characterized in the crucifer pathogen X. campestris pv. campestris.

  11. The plant pathogen Xanthomonas campestris pv. campestris exploits N-acetylglucosamine during infection.

    PubMed

    Boulanger, Alice; Zischek, Claudine; Lautier, Martine; Jamet, Stevie; Rival, Pauline; Carrère, Sébastien; Arlat, Matthieu; Lauber, Emmanuelle

    2014-09-09

    N-Acetylglucosamine (GlcNAc), the main component of chitin and a major constituent of bacterial peptidoglycan, is present only in trace amounts in plants, in contrast to the huge amount of various sugars that compose the polysaccharides of the plant cell wall. Thus, GlcNAc has not previously been considered a substrate exploited by phytopathogenic bacteria during plant infection. Xanthomonas campestris pv. campestris, the causal agent of black rot disease of Brassica plants, expresses a carbohydrate utilization system devoted to GlcNAc exploitation. In addition to genes involved in GlcNAc catabolism, this system codes for four TonB-dependent outer membrane transporters (TBDTs) and eight glycoside hydrolases. Expression of all these genes is under the control of GlcNAc. In vitro experiments showed that X. campestris pv. campestris exploits chitooligosaccharides, and there is indirect evidence that during the early stationary phase, X. campestris pv. campestris recycles bacterium-derived peptidoglycan/muropeptides. Results obtained also suggest that during plant infection and during growth in cabbage xylem sap, X. campestris pv. campestris encounters and metabolizes plant-derived GlcNAc-containing molecules. Specific TBDTs seem to be preferentially involved in the consumption of all these plant-, fungus- and bacterium-derived GlcNAc-containing molecules. This is the first evidence of GlcNAc consumption during infection by a phytopathogenic bacterium. Interestingly, N-glycans from plant N-glycosylated proteins are proposed to be substrates for glycoside hydrolases belonging to the X. campestris pv. campestris GlcNAc exploitation system. This observation extends the range of sources of GlcNAc metabolized by phytopathogenic bacteria during their life cycle. Despite the central role of N-acetylglucosamine (GlcNAc) in nature, there is no evidence that phytopathogenic bacteria metabolize this compound during plant infection. Results obtained here suggest that Xanthomonas

  12. Lettuce cultivar influences Xanthomonas campestris pv. vitians population levels

    USDA-ARS?s Scientific Manuscript database

    Bacterial Leaf Spot, caused by Xanthomonas campestris pv. vitians (Xcv), is a widespread and economically important disease of lettuce. Cultivars with resistance to Xcv have been identified, but mechanisms for resistance in this pathosystem have not been investigated. We hypothesized that susceptibl...

  13. Xanthan Pyruvilation Is Essential for the Virulence of Xanthomonas campestris pv. campestris.

    PubMed

    Bianco, María Isabel; Toum, Laila; Yaryura, Pablo Marcelo; Mielnichuk, Natalia; Gudesblat, Gustavo Eduardo; Roeschlin, Roxana; Marano, María Rosa; Ielpi, Luis; Vojnov, Adrián A

    2016-09-01

    Xanthan, the main exopolysaccharide (EPS) synthesized by Xanthomonas spp., contributes to bacterial stress tolerance and enhances attachment to plant surfaces by helping in biofilm formation. Therefore, xanthan is essential for successful colonization and growth in planta and has also been proposed to be involved in the promotion of pathogenesis by calcium ion chelation and, hence, in the suppression of the plant defense responses in which this cation acts as a signal. The aim of this work was to study the relationship between xanthan structure and its role as a virulence factor. We analyzed four Xanthomonas campestris pv. campestris mutants that synthesize structural variants of xanthan. We found that the lack of acetyl groups that decorate the internal mannose residues, ketal-pyruvate groups, and external mannose residues affects bacterial adhesion and biofilm architecture. In addition, the mutants that synthesized EPS without pyruvilation or without the external mannose residues did not develop disease symptoms in Arabidopsis thaliana. We also observed that the presence of the external mannose residues and, hence, pyruvilation is required for xanthan to suppress callose deposition as well as to interfere with stomatal defense. In conclusion, pyruvilation of xanthan seems to be essential for Xanthomonas campestris pv. campestris virulence.

  14. Transfer of resistance to Xanthomonas campestris pv campestris into Brassica oleracea L. by protoplast fusion.

    PubMed

    Hansen, L N; Earle, E D

    1995-12-01

    Black rot caused by the bacterium Xanthomonas campestris pv campestris is one of the most serious diseases of Brassica oleracea. Since sources of resistance to the disease within B. oleracea are insufficient and control means are limited, the development of resistant breeding lines is extremely desirable. Certain lines of B. napus contain very high resistance controlled by a dominant gene, but crossing the two species sexually is very difficult. Therefore, somatic hybrids were produced by protoplast fusion between rapid cycling B. oleracea and a B. napus line highly resistant to X. campestris pv campestris. Hybrid identity was confirmed by morphological studies, flow cytometric estimation of nuclear DNA content, and analysis of random amplified polymorphic DNA (RAPD). Inoculations with the pathogen identified four somatic hybrids with high resistance. The resistant hybrid plants were fertile and set seed when selfed or crossed reciprocally to the bridge line '15' (Quazi 1988). Direct crosses to B. oleracea were unsuccessful, but embryo rescue facilitated the production of a first-backcross generation. The BC1 plants were resistant to the pathogen. Progeny from the crosses to 'line 15' were all susceptible. Embryo rescue techniques were not obligatory for the development of a second-backcross generation, and several resistant BC2 plants were obtained.

  15. Inheritance of Race-Specific Resistance to Xanthomonas campestris pv. campestris in Brassica Genomes.

    PubMed

    Vicente, J G; Taylor, J D; Sharpe, A G; Parkin, I A P; Lydiate, D J; King, G J

    2002-10-01

    ABSTRACT The inheritance of resistance to three Xanthomonas campestris pv. campestris races was studied in crosses between resistant and susceptible lines of Brassica oleracea (C genome), B. carinata (BC genome), and B. napus (AC genome). Resistance to race 3 in the B. oleracea doubled haploid line BOH 85c and in PI 436606 was controlled by a single dominant locus (Xca3). Resistance to races 1 and 3 in the B. oleracea line Badger Inbred-16 was quantitative and recessive. Strong resistance to races 1 and 4 was controlled by a single dominant locus (Xca1) in the B. carinata line PI 199947. This resistance probably originates from the B genome. Resistance to race 4 in three B. napus lines, cv. Cobra, the rapid cycling line CrGC5, and the doubled haploid line N-o-1, was controlled by a single dominant locus (Xca4). A set of doubled haploid lines, selected from a population used previously to develop a restriction fragment length polymorphism map, was used to map this locus. Xca4 was positioned on linkage group N5 of the B. napus A genome, indicating that this resistance originated from B. rapa. Xca4 is the first major locus to be mapped that controls race-specific resistance to X. campestris pv. campestris in Brassica spp.

  16. Genome Sequences of the Race 1 and Race 4 Xanthomonas campestris pv. campestris Strains CFBP 1869 and CFBP 5817.

    PubMed

    Bolot, Stéphanie; Cerutti, Aude; Carrère, Sébastien; Arlat, Matthieu; Fischer-Le Saux, Marion; Portier, Perrine; Poussier, Stéphane; Jacques, Marie-Agnes; Noël, Laurent D

    2015-09-17

    Xanthomonas campestris pv. campestris is the causal agent of black rot on Brassicaceae. The draft genome sequences of strains CFBP 1869 and CFBP 5817 have been determined and are the first ones corresponding to race 1 and race 4 strains, which have a predominant agronomic and economic impact on cabbage cultures worldwide. Copyright © 2015 Bolot et al.

  17. Genome Sequences of the Race 1 and Race 4 Xanthomonas campestris pv. campestris Strains CFBP 1869 and CFBP 5817

    PubMed Central

    Bolot, Stéphanie; Cerutti, Aude; Carrère, Sébastien; Arlat, Matthieu; Fischer-Le Saux, Marion; Portier, Perrine; Poussier, Stéphane; Jacques, Marie-Agnes

    2015-01-01

    Xanthomonas campestris pv. campestris is the causal agent of black rot on Brassicaceae. The draft genome sequences of strains CFBP 1869 and CFBP 5817 have been determined and are the first ones corresponding to race 1 and race 4 strains, which have a predominant agronomic and economic impact on cabbage cultures worldwide. PMID:26383661

  18. Natural genetic variation of Xanthomonas campestris pv. campestris pathogenicity on arabidopsis revealed by association and reverse genetics.

    PubMed

    Guy, Endrick; Genissel, Anne; Hajri, Ahmed; Chabannes, Matthieu; David, Perrine; Carrere, Sébastien; Lautier, Martine; Roux, Brice; Boureau, Tristan; Arlat, Matthieu; Poussier, Stéphane; Noël, Laurent D

    2013-06-04

    ABSTRACT The pathogenic bacterium Xanthomonas campestris pv. campestris, the causal agent of black rot of Brassicaceae, manipulates the physiology and the innate immunity of its hosts. Association genetic and reverse-genetic analyses of a world panel of 45 X. campestris pv. campestris strains were used to gain understanding of the genetic basis of the bacterium's pathogenicity to Arabidopsis thaliana. We found that the compositions of the minimal predicted type III secretome varied extensively, with 18 to 28 proteins per strain. There were clear differences in aggressiveness of those X. campestris pv. campestris strains on two Arabidopsis natural accessions. We identified 3 effector genes (xopAC, xopJ5, and xopAL2) and 67 amplified fragment length polymorphism (AFLP) markers that were associated with variations in disease symptoms. The nature and distribution of the AFLP markers remain to be determined, but we observed a low linkage disequilibrium level between predicted effectors and other significant markers, suggesting that additional genetic factors make a meaningful contribution to pathogenicity. Mutagenesis of type III effectors in X. campestris pv. campestris confirmed that xopAC functions as both a virulence and an avirulence gene in Arabidopsis and that xopAM functions as a second avirulence gene on plants of the Col-0 ecotype. However, we did not detect the effect of any other effector in the X. campestris pv. campestris 8004 strain, likely due to other genetic background effects. These results highlight the complex genetic basis of pathogenicity at the pathovar level and encourage us to challenge the agronomical relevance of some virulence determinants identified solely in model strains. IMPORTANCE The identification and understanding of the genetic determinants of bacterial virulence are essential to be able to design efficient protection strategies for infected plants. The recent availability of genomic resources for a limited number of pathogen

  19. Natural Genetic Variation of Xanthomonas campestris pv. campestris Pathogenicity on Arabidopsis Revealed by Association and Reverse Genetics

    PubMed Central

    Guy, Endrick; Genissel, Anne; Hajri, Ahmed; Chabannes, Matthieu; David, Perrine; Carrere, Sébastien; Lautier, Martine; Roux, Brice; Boureau, Tristan; Arlat, Matthieu; Poussier, Stéphane; Noël, Laurent D.

    2013-01-01

    ABSTRACT The pathogenic bacterium Xanthomonas campestris pv. campestris, the causal agent of black rot of Brassicaceae, manipulates the physiology and the innate immunity of its hosts. Association genetic and reverse-genetic analyses of a world panel of 45 X. campestris pv. campestris strains were used to gain understanding of the genetic basis of the bacterium’s pathogenicity to Arabidopsis thaliana. We found that the compositions of the minimal predicted type III secretome varied extensively, with 18 to 28 proteins per strain. There were clear differences in aggressiveness of those X. campestris pv. campestris strains on two Arabidopsis natural accessions. We identified 3 effector genes (xopAC, xopJ5, and xopAL2) and 67 amplified fragment length polymorphism (AFLP) markers that were associated with variations in disease symptoms. The nature and distribution of the AFLP markers remain to be determined, but we observed a low linkage disequilibrium level between predicted effectors and other significant markers, suggesting that additional genetic factors make a meaningful contribution to pathogenicity. Mutagenesis of type III effectors in X. campestris pv. campestris confirmed that xopAC functions as both a virulence and an avirulence gene in Arabidopsis and that xopAM functions as a second avirulence gene on plants of the Col-0 ecotype. However, we did not detect the effect of any other effector in the X. campestris pv. campestris 8004 strain, likely due to other genetic background effects. These results highlight the complex genetic basis of pathogenicity at the pathovar level and encourage us to challenge the agronomical relevance of some virulence determinants identified solely in model strains. PMID:23736288

  20. Xanthomonas campestris pv. campestris gum Mutants: Effects on Xanthan Biosynthesis and Plant Virulence

    PubMed Central

    Katzen, Federico; Ferreiro, Diego U.; Oddo, Cristian G.; Ielmini, M. Verónica; Becker, Anke; Pühler, Alfred; Ielpi, Luis

    1998-01-01

    Xanthan is an industrially important exopolysaccharide produced by the phytopathogenic, gram-negative bacterium Xanthomonas campestris pv. campestris. It is composed of polymerized pentasaccharide repeating units which are assembled by the sequential addition of glucose-1-phosphate, glucose, mannose, glucuronic acid, and mannose on a polyprenol phosphate carrier (L. Ielpi, R. O. Couso, and M. A. Dankert, J. Bacteriol. 175:2490–2500, 1993). A cluster of 12 genes in a region designated xpsI or gum has been suggested to encode proteins involved in the synthesis and polymerization of the lipid intermediate. However, no experimental evidence supporting this suggestion has been published. In this work, from the biochemical analysis of a defined set of X. campestris gum mutants, we report experimental data for assigning functions to the products of the gum genes. We also show that the first step in the assembly of the lipid-linked intermediate is severely affected by the combination of certain gum and non-gum mutations. In addition, we provide evidence that the C-terminal domain of the gumD gene product is sufficient for its glucosyl-1-phosphate transferase activity. Finally, we found that alterations in the later stages of xanthan biosynthesis reduce the aggressiveness of X. campestris against the plant. PMID:9537354

  1. The roles of peroxide protective regulons in protecting Xanthomonas campestris pv. campestris from sodium hypochlorite stress.

    PubMed

    Charoenlap, Nisanart; Sornchuer, Phornphan; Piwkam, Anong; Srijaruskul, Kriangsuk; Mongkolsuk, Skorn; Vattanaviboon, Paiboon

    2015-05-01

    The exposure of Xanthomonas campestris pv. campestris to sublethal concentrations of a sodium hypochlorite (NaOCl) solution induced the expression of genes that encode peroxide scavenging enzymes within the OxyR and OhrR regulons. Sensitivity testing in various X. campestris mutants indicated that oxyR, katA, katG, ahpC, and ohr contributed to protection against NaOCl killing. The pretreatment of X. campestris cultures with oxidants, such as hydrogen peroxide (H2O2), t-butyl hydroperoxide, and the superoxide generator menadione, protected the bacteria from lethal concentrations of NaOCl in an OxyR-dependent manner. Treating the bacteria with a low concentration of NaOCl resulted in the adaptive protection from NaOCl killing and also provided cross-protection from H2O2 killing. Taken together, the results suggest that the toxicity of NaOCl is partially mediated by the generation of peroxides and other reactive oxygen species that are removed by primary peroxide scavenging enzymes, such as catalases and AhpC, as a part of an overall strategy that protects the bacteria from the lethal effects of NaOCl.

  2. Comprehensive analysis of the extracellular proteins from Xanthomonas campestris pv. campestris B100.

    PubMed

    Watt, Steven Alexander; Wilke, Andreas; Patschkowski, Thomas; Niehaus, Karsten

    2005-01-01

    The extracellular proteome of Xanthomonas campestris pv. campestris (Xcc) cultivated in minimal medium was isolated from the cell-free culture supernatant and separated by two-dimensional gel electrophoresis. This technique resolved 97 clearly visible protein spots, which were excised, digested with trypsin and identified on the basis of their peptide mass fingerprints generated by matrix assisted laser desorption/ionisation-time of flight-mass spectrometry. Using this approach 87 different proteins could be distinguished. The Signal P software predicted putative signal peptides for 53% of the extracellular proteins. These proteins are probably transported over the inner membrane and are localized in the periplasm, the outer membrane or secreted into the extracellular space. Among the secreted proteins are 11 degradative enzymes, which are involved in pathogenesis of Xcc. The proteins without obvious secretion signals are known to serve functions in the cytosol. How the cytosolic proteins are delivered to the extracellular space remains unclear.

  3. In vivo proteome analysis of Xanthomonas campestris pv. campestris in the interaction with the host plant Brassica oleracea.

    PubMed

    Andrade, Aretusa E; Silva, Luciano P; Pereira, Jackeline L; Noronha, Eliane F; Reis, Fabio B; Bloch, Carlos; dos Santos, Marise F; Domont, Gilberto B; Franco, Octávio L; Mehta, Angela

    2008-04-01

    The genus Xanthomonas is composed of several species that cause severe crop losses around the world. In Latin America, one of the most relevant species is Xanthomonas campestris pv. campestris, which is responsible for black rot in cruciferous plants. This pathogen causes yield losses in several cultures, including cabbage, cauliflower and broccoli. Although the complete structural genome of X. campestris pv. campestris has been elucidated, little is known about the protein expression of this pathogen in close interaction with the host plant. Recently, a method for in vivo analysis of Xanthomonas axonopodis pv. citri was developed. In the present study, this technique was employed for the characterization of the protein expression of X. campestris pv. campestris in close interaction with the host plant Brassica oleracea. The bacterium was infiltrated into leaves of the susceptible cultivar and later recovered for proteome analysis. Recovered cells were used for protein extraction and separated by two-dimensional electrophoresis. Proteins were analysed by peptide mass fingerprinting or de novo sequencing and identified by searches in public databases. The approach used in this study may be extremely useful in further analyses in order to develop novel strategies to control this important plant pathogen.

  4. hxc2, an Arabidopsis mutant with an altered hypersensitive response to Xanthomonas campestris pv. campestris.

    PubMed

    Godard, F; Lummerzheim, M; Saindrenan, P; Balagué, C; Roby, D

    2000-12-01

    A chemical mutagenized population of Arabidopsis Col-0-gl plants was screened for an altered hypersensitive response (HR) after spray inoculation with an HR-inducing isolate of Xanthomonas campestris pv. campestris (strain 147). Three classes of mutant were identified: those exhibiting an HR- phenotype or partial loss of HR; hyper-responsive mutants showing necrotic lesions rapidly leading to the collapse of leaves; and susceptible mutants. One mutant belonging to the susceptible class, hxc-2, was extensively characterized. The compatible phenotype observed several days after initiation of the interaction was confirmed by measurement of in planta bacterial growth and use of bacterial strains constitutively expressing the GUS reporter gene. In the same way, accumulation of autofluorescent compounds, salicylic acid production and defence gene expression in the mutant were found to be similar to that displayed by the susceptible ecotype. Inoculation of hxc-2 with different avirulent bacteria suggests that the mutation is specific for the interaction with the Xcc 147 strain, although the mutation has been shown to affect a single dominant locus, different from the resistance locus defined by genetic analysis of resistance to Xcc 147. Genetic mapping of the mutation indicated that it is located on chromosome III, defining a previously unknown resistance function in response to X. c. campestris.

  5. A Xanthomonas campestris pv. campestris protein similar to catabolite activation factor is involved in regulation of phytopathogenicity.

    PubMed Central

    de Crecy-Lagard, V; Glaser, P; Lejeune, P; Sismeiro, O; Barber, C E; Daniels, M J; Danchin, A

    1990-01-01

    A DNA fragment from Xanthomonas campestris pv. campestris that partially restored the carbohydrate fermentation pattern of a cya crp Escherichia coli strain was cloned and expressed in E. coli. The nucleotide sequence of this fragment revealed the presence of a 700-base-pair open reading frame that coded for a protein highly similar to the catabolite activation factor (CAP) of E. coli (accordingly named CLP for CAP-like protein). An X. campestris pv. campestris clp mutant was constructed by reverse genetics. This strain was not affected in the utilization of various carbon sources but had strongly reduced pathogenicity. Production of xanthan gum, pigment, and extracellular enzymes was either increased or decreased, suggesting that CLP plays a role in the regulation of phytopathogenicity. PMID:2170330

  6. Mutations of ferric uptake regulator (fur) impair iron homeostasis, growth, oxidative stress survival, and virulence of Xanthomonas campestris pv. campestris.

    PubMed

    Jittawuttipoka, Thichakorn; Sallabhan, Ratiboot; Vattanaviboon, Paiboon; Fuangthong, Mayuree; Mongkolsuk, Skorn

    2010-05-01

    Iron is essential in numerous cellular functions. Intracellular iron homeostasis must be maintained for cell survival and protection against iron's toxic effects. Here, we characterize the roles of Xanthomonas campestris pv. campestris (Xcc) fur, which encodes an iron sensor and a transcriptional regulator that acts in iron homeostasis, oxidative stress, and virulence. Herein, we isolated spontaneous Xcc fur mutants that had high intracellular iron concentrations due to constitutively high siderophore levels and increased expression of iron transport genes. These mutants also had reduced aerobic plating efficiency and resistance to peroxide killing. Moreover, one fur mutant was attenuated on a host plant, thus indicating that fur has important roles in the virulence of X. campestris pv. campestris.

  7. Genes for hydrogen peroxide detoxification and adaptation contribute to protection against heat shock in Xanthomonas campestris pv. campestris.

    PubMed

    Buranajitpakorn, Sarinya; Piwkam, Anong; Charoenlap, Nisanart; Vattanaviboon, Paiboon; Mongkolsuk, Skorn

    2011-04-01

    Xanthomonas campestris pv. campestris, a soil-borne plant-pathogenic bacterium, is exposed to multiple stresses in the environment and during interaction with a host plant. The roles of hydrogen peroxide (H(2) O(2) )-protective genes (katA, katG, and ahpC) and a peroxide sensor/transcription regulator (oxyR) in the viability of X. campestris pv. campestris at an elevated temperature were evaluated. The single katA and katG mutants showed moderate decreased survival after the heat treatment, while the double katA-katG and oxyR mutants were the most vulnerable to the heat treatment compared with a wild-type strain. However, ahpC provided no protective function against the heat treatment. Flow cytometric analysis revealed an increased accumulation of peroxide in cells treated with heat. Altogether, the data revealed a crucial role of genes in the H(2) O(2) detoxification system for protection against lethal heat shock in X. campestris pv. campestris.

  8. [Identification of a new type III effector XC3176 in Xanthomonas campestris pv. campestris].

    PubMed

    Yang, Lichao; Su, Hua; Yang, Feng; Jian, Huahua; Zhou, Min; Jiang, Wei; Jiang, Bole

    2015-10-04

    Type III secretion system (T3SS) is essential for many phytopathogenic bacteria to cause disease in susceptible host plants and to elicit a hypersensitive response in resistant host and non-host plants. Xanthomonas campestris pv. campestris (Xcc) uses T3SS to deliver T3SS effectors (T3SEs) directly into host cells, where they play important roles in pathogenesis. The aim of this study was to identify a new T3SE in Xcc. To validate if XC3176 is a T3S effector translocated into plant cells, the promoter and signal region of XC3176 were fused to the plasmid pLJB of harboring HR-inducing AvrBs1 C-terminal domain lack of 58 N-terminal amino acid residues. The recombinant plasmid pLJB3176 was introduced by triparental conjugation into ΔavrBs1 and ΔhrcV. Hypersensitive response induced by the obtained strains ΔavrBs1/pLJB3176 and ΔhrcV/pLJB3176 were examined on the pepper ECW-10R. To determine transcription of XC3176, GUS fusion report strains were constructed. The virulence of Xcc strains was investigated on the Chinese radish by the leaf-clipping method. Hypersensitive response was elicited on the pepper ECW-10R by the strain ΔavrBs1/pLJB3176, but not ΔhrcV/pLJB3176. The GUS activities in the mutant strains ΔhrpX and ΔhrpG were significantly lower than that in the wild type Xcc strain. The mutant of XC3176 reduced virulence significantly and the complementary strain C3176 could restore the virulence as the wild-type strain. XC3176 is a T3SS-dependent effector of Xanthomonas campestris pv. campestris. The expression of XC3176 is regulated by hrpG and hrpX. XC3176 is required for the full virulence of Xcc 8004.

  9. Evaluation of rhizospheric Pseudomonas and Bacillus as biocontrol tool for Xanthomonas campestris pv campestris.

    PubMed

    Mishra, Shruti; Arora, Naveen K

    2012-02-01

    Xanthomonas campestris pv campestris (Xcc), causing black rot, is one of the most yield-limiting and destructive pathogens of cruciferous crops. The intention of this study was to evaluate the potential of rhizobacteria in black rot management. Fifty-four isolates from rhizosphere soil of Brassica campestris were screened against Xcc. Two isolates namely, KA19 and SE, with inhibition radius >11 mm were selected. The combined use of them produced an average inhibition zone of 18.1 ± 1.4 mm radius (P < 0.05). 16S rRNA gene sequencing and phylogenetic analysis identified KA19 and SE as the nearest homologs (>99.4%) of Pseudomonas aeruginosa and Bacillus thuringiensis, respectively. In greenhouse study, both isolates were effective (P < 0.05) in reducing black rot lesions compared to untreated control involving either a foliar spray or the combined seed soak and soil drench. However, the combined strains (KA19 + SE) were significantly more effective (P < 0.05) when the mode of application was combined seed and soil drench. The lipid content of seeds increased significantly with the application of these strains, especially with SE alone and in combination. After 9 weeks, the Xcc population was significantly lower in soil treated with combined strains (P < 0.05). KA19 produced extracellular siderophores, influenced by various carbon sources and identified as 4-hydroxy-2-nonyl-quinoline by NMR. In Bacillus SE, two antibacterial factors corresponding to autolysins (β-N-acetylglucosaminidase) and AHL-lactonases were established. This study would strengthen our understanding for application of different rhizobacteria with various active principles like Pseudomonas and Bacillus as ingredients of a biocontrol mixture.

  10. In vivo and in vitro effects of secondary metabolites against Xanthomonas campestris pv. campestris.

    PubMed

    Velasco, Pablo; Lema, Margarita; Francisco, Marta; Soengas, Pilar; Cartea, María Elena

    2013-09-11

    Brassica rapa is a crucifer that is grown worldwide, mainly as a vegetable. The quality of B. rapa crops is highly affected by the disease caused by the bacteria Xanthomonas campestris pv. campestris (Xcc). Glucosinolates and phenolic compounds can confer resistance to Brassica crops against pests and diseases, but few works have been done to evaluate their role in Xcc resistance. The objectives of this work were: (1) to evaluate the in vivo and in vitro antibacterial effect of gluconapin, its isothiocyanate and the methanolic extracts of B. rapa against the type 4 of Xcc, and (2) to test if there is induced resistance mediated by glucosinolates or phenolic compounds in two varieties of B. rapa. Gluconapin and its ITC varieties had an antibacterial effect on the development of Xanthomonas and this effect was strongly dependent on the concentration applied. Methanolic extracts from B. rapa, containing glucosinolates and phenolic compounds, inhibited the growth of these bacteria. Concentration of gluconapin is higher in resistant plants than in the susceptible ones and there is an induction of gluconapin, some flavonoids and sinapic acid 48 to 72 h after inoculation. Gluconapin plays a role in the constitutive resistance to Xcc, while gluconapin, some flavonoids and hydroxycinnamic acids are induced by a Xcc infection but it is not clear if this induction confers resistance to this disease.

  11. Establishment of an inducing medium for type III effector secretion in Xanthomonas campestris pv. campestris

    PubMed Central

    Jiang, Guo-Feng; Jiang, Bo-Le; Yang, Mei; Liu, San; Liu, Jiao; Liang, Xiao-Xia; Bai, Xian-Fang; Tang, Dong-Jie; Lu, Guang-Tao; He, Yong-Qiang; Yu, Di-Qiu; Tang, Ji-Liang

    2013-01-01

    It is well known that the type III secretion system (T3SS) and type III (T3) effectors are essential for the pathogenicity of most bacterial phytopathogens and that the expression of T3SS and T3 effectors is suppressed in rich media but induced in minimal media and plants. To facilitate in-depth studies on T3SS and T3 effectors, it is crucial to establish a medium for T3 effector expression and secretion. Xanthomonas campestris pv. campestris (Xcc) is a model bacterium for studying plant-pathogen interactions. To date no medium for Xcc T3 effector secretion has been defined. Here, we compared four minimal media (MME, MMX, XVM2, and XOM2) which are reported for T3 expression induction in Xanthomonas spp. and found that MME is most efficient for expression and secretion of Xcc T3 effectors. By optimization of carbon and nitrogen sources and pH value based on MME, we established XCM1 medium, which is about 3 times stronger than MME for Xcc T3 effectors secretion. We further optimized the concentration of phosphate, calcium, and magnesium in XCM1 and found that XCM1 with a lower concentration of magnesium (renamed as XCM2) is about 10 times as efficient as XCM1 (meanwhile, about 30 times stronger than MME). Thus, we established an inducing medium XCM2 which is preferred for T3 effector secretion in Xcc. PMID:24516463

  12. Identification of genes differentially expressed in cauliflower associated with resistance to Xanthomonas campestris pv. campestris.

    PubMed

    Jiang, Hanmin; Song, Wenqin; Li, Ai; Yang, Xiao; Sun, Deling

    2011-01-01

    Black rot, caused by Xanthomonas campestris pv. campestris (Pammel) Dowson (Xcc), is one of the most damaging diseases of cauliflower and other crucifers. In order to investigate the molecular resistance mechanisms and to find the genes related to black rot resistance in cauliflower, a suppression subtractive hybridization (SSH) cDNA library was constructed using resistant line C712 and its susceptible near-isogenic line C731 as tester and driver, respectively. A total of 280 clones were obtained from the library by reverse northern blotting. Sequencing analysis and homology searching showed that these clones represent 202 unique sequences. The library included many defense/disease-resistant related genes, such as plant defensin gene PDF1.2, lipid transfer protein, thioredoxin h. Gene expression profiles of 12 genes corresponding to different functional categories were monitored by real-time RT-PCR. The results showed that the expression induction of these genes in the susceptible line C712 in response to Xcc was quicker and more intense, while in C731 the reaction was delayed and limited. Our results imply that these up-regulated genes might be involved in cauliflower responses against Xcc infection. Information obtained from this study could be used to understand the molecular mechanisms of disease response in cauliflower under Xcc stress.

  13. Response regulator, VemR, positively regulates the virulence and adaptation of Xanthomonas campestris pv. campestris.

    PubMed

    Tao, Jun; He, Chaozu

    2010-03-01

    Xanthomonas campestris pv. campestris (Xcc) is the causal agent of black rot disease in cruciferous plants. The synthesis of known virulence factors in this organism, such as extracellular enzymes and biofilm, is strictly regulated in response to environmental stimuli. Two-component signal transduction systems sense environmental signals and alter bacterial behavior by regulating gene expression. Here, we identified a response regulator, VemR, that regulates Xcc pathogenesis. The vemR gene encodes an atypical response regulator that only contains a receiver domain. Deletion of vemR resulted in decreased virulence, exopolysaccharide production and motility of Xcc. The vemR gene is located in an operon flanked by genes fleQ and rpoN2. Genetic analysis indicated that deletion of fleQ does not affect motility significantly. However, a double mutant DeltavemR/DeltafleQ reversed the phenotype of DeltavemR, indicating that fleQ is epistatic to vemR in the regulation of virulence and adaptation.

  14. Sources and Origin of Resistance to Xanthomonas campestris pv. campestris in Brassica Genomes.

    PubMed

    Taylor, J D; Conway, J; Roberts, S J; Astley, D; Vicente, J G

    2002-01-01

    ABSTRACT Two hundred and seventy-six accessions of mainly Brassica spp. were screened for resistance to Xanthomonas campestris pv. campestris races. In Brassica oleracea (C genome), the majority of accessions were susceptible to all races, but 43% showed resistance to one or more of the rare races (2, 3, 5, and 6) and a single accession showed partial resistance to races 1, 3, 5, and 6. Further searches for resistance to races 1 and 4, currently the most important races worldwide, and race 6, the race with the widest host range, were made in accessions representing the A and B genomes. Strong resistance to race 4 was frequent in B. rapa (A genome) and B. napus (AC genome), indicating an A genome origin. Resistance to races 1 and 4 was present in a high proportion of B. nigra (B genome) and B. carinata (BC genome) accessions, indicating a B genome origin. B. juncea (AB genome) was the most resistant species, showing either strong resistance to races 1 and 4 or quantitative resistance to all races. Potentially race-nonspecific resistance was also found, but at a lower frequency, in B. rapa, B. nigra, and B. carinata. The combination of race-specific and race-nonspecific resistance could provide durable control of black rot of crucifers.

  15. Functional characterization of copA gene encoding multicopper oxidase in Xanthomonas campestris pv. campestris.

    PubMed

    Hsiao, Yi-Min; Liu, Yu-Fan; Lee, Pei-Yu; Hsu, Pei-Chi; Tseng, Szu-Yu; Pan, Yu-Chien

    2011-09-14

    The gram-negative plant pathogenic Xanthomonas campestris pv. campestris (Xcc) is the causative agent of black rot in crucifers, a disease causing tremendous loss in agriculture. Copper-containing bactericides have been widely used to control this disease for many years, possibly leading to the development of copper resistance in Xcc. Homologues of copper resistance genes copLAB are present in the Xcc genome, but none has been characterized. In this study, mutations in copL, copA, and copB decreased Xcc copper tolerance. Among them, the copA mutant displayed the most significant reduction. The copA mutant also resulted in a reduction in virulence on the host cabbage. Sequence and mutational analysis demonstrated that copA encodes a multicopper oxidase and that CopA is able to catalyze the oxidation of 2,6-dimethoxyphenol. Alanine substitutions in each of the putative copper binding residues (H538, H583, C584, and H585) of CopA caused a loss of function including copper tolerance and oxidase activity. Furthermore, reporter assays showed that copA transcription is inducible in the presence of copper, subject to catabolite repression, and repressed under conditions of high osmolarity, nitrogen starvation, or oxygen limitation. This is the first time that multicopper oxidase has been characterized in the crucifer pathogen Xcc.

  16. Identification of quantitative trait loci for resistance to Xanthomonas campestris pv. campestris in Brassica rapa.

    PubMed

    Soengas, P; Hand, P; Vicente, J G; Pole, J M; Pink, D A C

    2007-02-01

    Resistance to six known races of black rot in crucifers caused by Xanthomonas campestris pv. campestris (Pammel) Dowson is absent or very rare in Brassica oleracea (C genome). However, race specific and broad-spectrum resistance (to type strains of all six races) does appear to occur frequently in other brassica genomes including B. rapa (A genome). Here, we report the genetics of broad spectrum resistance in the B. rapa Chinese cabbage accession B162, using QTL analysis of resistance to races 1 and 4 of the pathogen. A B. rapa linkage map comprising ten linkage groups (A01-A10) with a total map distance of 664 cM was produced, based on 223 AFLP bands and 23 microsatellites from a F(2) population of 114 plants derived from a cross between the B. rapa susceptible inbred line R-o-18 and B162. Interaction phenotypes of 125 F(2) plants were assessed using two criteria: the percentage of inoculation sites in which symptoms developed, and the severity of symptoms per plant. Resistance to both races was correlated and a cluster of highly significant QTL that explained 24-64% of the phenotypic variance was located on A06. Two additional QTLs for resistance to race 4 were found on A02 and A09. Markers closely linked to these QTL could assist in the transference of the resistance into different B. rapa cultivars or into B. oleracea.

  17. Establishment of an inducing medium for type III effector secretion in Xanthomonas campestris pv. campestris.

    PubMed

    Jiang, Guo-Feng; Jiang, Bo-Le; Yang, Mei; Liu, San; Liu, Jiao; Liang, Xiao-Xia; Bai, Xian-Fang; Tang, Dong-Jie; Lu, Guang-Tao; He, Yong-Qiang; Yu, Di-Qiu; Tang, Ji-Liang

    2013-01-01

    It is well known that the type III secretion system (T3SS) and type III (T3) effectors are essential for the pathogenicity of most bacterial phytopathogens and that the expression of T3SS and T3 effectors is suppressed in rich media but induced in minimal media and plants. To facilitate in-depth studies on T3SS and T3 effectors, it is crucial to establish a medium for T3 effector expression and secretion. Xanthomonas campestris pv. campestris (Xcc) is a model bacterium for studying plant-pathogen interactions. To date no medium for Xcc T3 effector secretion has been defined. Here, we compared four minimal media (MME, MMX, XVM2, and XOM2) which are reported for T3 expression induction in Xanthomonas spp. and found that MME is most efficient for expression and secretion of Xcc T3 effectors. By optimization of carbon and nitrogen sources and pH value based on MME, we established XCM1 medium, which is about 3 times stronger than MME for Xcc T3 effectors secretion. We further optimized the concentration of phosphate, calcium, and magnesium in XCM1 and found that XCM1 with a lower concentration of magnesium (renamed as XCM2) is about 10 times as efficient as XCM1 (meanwhile, about 30 times stronger than MME). Thus, we established an inducing medium XCM2 which is preferred for T3 effector secretion in Xcc.

  18. Characterization of genes encoding proteins containing HD-related output domain in Xanthomonas campestris pv. campestris.

    PubMed

    Lee, Hsien-Ming; Liao, Chao-Tsai; Chiang, Ying-Chuan; Chang, Yu-Yin; Yeh, Yu-Tzu; Du, Shin-Chiao; Hsiao, Yi-Min

    2016-04-01

    The Gram-negative plant pathogen Xanthomonas campestris pv. campestris (Xcc) is the causative agent of black rot in crucifers. The production of Xcc virulence factors is regulated by Clp and RpfF. HD-related output domain (HDOD) is a protein domain of unknown biochemical function. The genome of Xcc encodes three proteins (GsmR, HdpA, and HdpB) with an HDOD. The GsmR has been reported to play a role in the general stress response and cell motility and its expression is positively regulated by Clp. Here, the function and transcription of hdpA and hdpB were characterized. Mutation of hdpA resulted in enhanced bacterial attachment. In addition, the expression of hdpA was positively regulated by RpfF but not by Clp, subject to catabolite repression and affected by several stress conditions. However, mutational analysis and reporter assay showed that hdpB had no effect on the production of a range of virulence factors and its expression was independent of Clp and RpfF. The results shown here not only extend the previous work on RpfF regulation to show that it influences the expression of hdpA in Xcc, but also expand knowledge of the function of the HDOD containing proteins in bacteria.

  19. Biophysical and biochemical studies of a major endoglucanase secreted by Xanthomonas campestris pv. campestris.

    PubMed

    Rosseto, Flávio Rodolfo; Manzine, Livia Regina; de Oliveira Neto, Mario; Polikarpov, Igor

    2016-09-01

    Endoglucanases are the main cellulolytic enzymes secreted by the bacterium Xanthomonas campestris pv. campestris (Xcc). The major endoglucanase exported by this bacterium into an external milieu is an enzyme XccCel5A, which belongs to GH5 family subfamily 1 and is encoded by the gene engXCA. We purified XccCel5A using ammonium sulfate precipitation followed by size exclusion chromatography and identified it by zymogram analysis. Circular dichroism and fluorescence spectroscopy studies showed that XccCel5A is stable in a wide pH range and up to about 55°C and denatures at the higher temperatures. The optimal conditions for enzyme activity were identified as T=45°C and pH=7.0. Under the optimum conditions the catalytic efficiency (kcat/KM) of the enzyme was determined as 5.16×10(4)s(-1)M(-1) using carboxymethylcellulose (CMC) as a substrate. Our SAXS studies revealed extended tadpole-shape molecular assembly, typical for cellulases, and allowed to determine an overall shape of the enzyme and a relative position of the catalytic and cellulose binding domains.

  20. Light signaling mediated by PAS domain-containing proteins in Xanthomonas campestris pv. campestris.

    PubMed

    Mao, Daqing; Tao, Jun; Li, Chunxia; Luo, Chao; Zheng, Linlin; He, Chaozu

    2012-01-01

    Per-ARNT-Sim (PAS) domains are important signalling modules that possibly monitor changes in various stimuli such as light. For the majority of PAS domains that have been identified by sequence similarity, the biological function of the signalling pathways has not yet been experimentally investigated.Thirty-three PAS proteins were discovered in Xanthomonas campestris pv. campestris(Xcc) by genome/proteome analysis. Thirteen PAS proteins were identified as contributing to light signalling and Xcc growth, motility or virulence using molecular genetics and bioinformatics methods. The PAS domains played important roles in light signalling to regulate the growth, motility and virulence of Xcc. They might be regulated by not only light quality (wavelength)but also quantity (intensity) as potential light-signalling components. Evaluating the light wavelength, three light-signalling types of PAS proteins in Xcc were shown to be involved in blue light signalling, tricolour (blue, red and far red)signalling or red/far-red signalling. This showed that Xcc had evolved a complicated light-signalling system to adapt to a complex environment.

  1. Transcriptional analysis of pmeA gene encoding a pectin methylesterase in Xanthomonas campestris pv. campestris.

    PubMed

    Hsiao, Yi-Min; Liu, Yu-Fan; Huang, Yi-Ling; Lee, Pei-Yu

    2011-04-01

    Exopolysaccharides and several extracellular enzymes of Xanthomonas campestris pv. campestris (Xcc), the causative agent of black rot in crucifers, are virulence determinants. It is known that Clp (cAMP receptor protein-like protein) and RpfF (an enoyl-CoA hydratase homologue required for the synthesis of the diffusible signal factor, DSF) regulate production of these factors. In this study, plate assay revealed that Xcc possesses pectin methylesterase activity and that its expression is controlled by Clp and RpfF. Mutational analysis has demonstrated that pmeA encodes a pectin methylesterase. Using the 5' RACE method, the pmeA transcription initiation site was mapped. Transcriptional fusion assays showed that pmeA transcription is positively regulated by Clp and RpfF, subject to catabolite repression which is independent of Clp or RpfF, and repressed under conditions of high osmolarity or oxygen limitation. This study not only extends previous work on Clp and RpfF regulation by showing that they both influence the expression of pmeA in Xcc, but also, for the first time, characterizes pectin methylesterase gene expression in Xanthomonas.

  2. Sensitive and specific detection of Xanthomonas campestris pv campestris by PCR using species-specific primers based on hrpF gene sequences.

    PubMed

    Park, Young Jin; Lee, Byoung Moo; Ho-Hahn, Jang; Lee, Gil Bok; Park, Dong Suk

    2004-01-01

    A sensitive and specific assay was developed to detect bacterial black rot of crucifers caused by Xanthomonas campestris pv. campestris (X. c. pv. campestris), in cabbage seed and plant. Primers XCF and XCR from hrpF homologous to nolX, host recognition protein, were used to amplify a 525 bp DNA fragment. PCR technique was applied to detect the pathogen in naturally infected seed and plant of cabbage. The PCR product was only produced from X. c. pv. campestris among 40 isolates of Xanthomonas strains, Escherichia coli (O157:H7), Pectobacterium carotovorum subsp. carotovorum, and other reference bacteria.

  3. Xanthomonas campestris pv. campestris requires a functional pigB for epiphytic survival and host infection.

    PubMed

    Poplawsky, A R; Chun, W

    1998-06-01

    When cauliflower plants (Brassica oleraceae) were misted with bacterial suspensions of Xanthomonas campestris pv. campestris (causal agent of black rot of cruciferous plants), two separate populations of the pathogen were associated with the leaves. Initially, bacteria removable by sonication and sensitive to sodium hypochlorite treatment predominated (easily removable epiphytic bacteria, EREB). However, after 2 weeks, bacteria not removable by sonication and insensitive to sodium hypochlorite treatment were dominant. Although the exact location of this second population of the pathogen was not determined, evidence is presented to support its location in protected sites on the leaf surface, pigB of this pathogen is required for production of extracellular polysaccharide (EPS), xanthomonadin pigments, and the diffusible signal molecule, DF (diffusible factor). DF can extracellularly restore EPS and xanthomonadin production to pigB mutant strains. Parent strain B-24 and pigB mutant strain B24-B2 were identical for in planta growth and symptomatology after artificial infection by injection in leaf mid-veins. Subsequently, X. campestris pv. campestris parent strain B-24, Tn3HoHo1 pigB insertion mutation strain B24-B2, chromosomally restored pigB mutation strain B24-B2R, and strain B24-79 with a Tn3HoHo1 insertion in an unrelated part of the genome were compared for epiphytic survival on, and natural infection of, cauliflower. After application, strains B-24, B24-B2R, and B24-79 all maintained leaf EREB populations of between approximately 3 and 6 (log [1 + CFU per g of fresh weight]) over a 3-week period, whereas B24-B2 populations fell to nearly undetectable levels. Plants sprayed with strains B-24, B24-B2R, and B24-79 averaged between 1.0 and 1.2 lesions, whereas those sprayed with B24-B2 averaged only 0.03 lesions per plant after 3 weeks. Differences in EREB population levels did not explain the observed differences in host infection frequencies, and the results

  4. Identification of proteins in susceptible and resistant Brassica oleracea responsive to Xanthomonas campestris pv. campestris infection.

    PubMed

    Villeth, Gabriela R C; Carmo, Lílian S T; Silva, Luciano Paulino; Santos, Mateus Figueiredo; de Oliveira Neto, Osmundo Brilhante; Grossi-de-Sá, Maria Fátima; Ribeiro, Igor Sousa; Dessaune, Suelen Nogueira; Fragoso, Rodrigo Rocha; Franco, Octávio L; Mehta, Angela

    2016-06-30

    Cruciferous plants are important edible vegetables widely consumed around the world, including cabbage, cauli-flower and broccoli. The main disease that affects crucifer plants is black rot, caused by Xanthomonas campestris pv. campestris (Xcc). In order to better understand this specific plant-pathogen interaction, proteins responsive to Xcc infection in resistant (União) and susceptible (Kenzan) Brassica oleracea cultivars were investigated by 2-DE followed by mass spectrometry. A total of 47 variable spots were identified and revealed that in the susceptible interaction there is a clear reduction in the abundance of proteins involved in energetic metabolism and defense. It was interesting to observe that in the resistant interaction, these proteins showed an opposite behavior. Based on our results, we conclude that resistance is correlated with the ability of the plant to keep sufficient photosynthesis metabolism activity to provide energy supplies necessary for an active defense. As a follow-up study, qRT-PCR analysis of selected genes was performed and revealed that most genes showed an up-regulation trend from 5 to 15days after inoculation (DAI), showing highest transcript levels at 15DAI. These results revealed the gradual accumulation of transcripts providing a more detailed view of the changes occurring during different stages of the plant-pathogen interaction. In this study we have compared cultivars of Brassica oleracea (cabbage), susceptible and resistant to black rot, by using the classical 2-DE approach. We have found that resistance is correlated with the ability of the plant to keep sufficient photosynthesis metabolism activity to provide energy supplies necessary for an active defense. Copyright © 2016 Elsevier B.V. All rights reserved.

  5. Function-related positioning of the type II secretion ATPase of Xanthomonas campestris pv. campestris.

    PubMed

    Chen, Yih-Lin; Hu, Nien-Tai

    2013-01-01

    Gram-negative bacteria use the type II secretion (T2S) system to secrete exoproteins for attacking animal or plant cells or to obtain nutrients from the environment. The system is unique in helping folded proteins traverse the outer membrane. The secretion machine comprises multiple proteins spanning the cell envelope and a cytoplasmic ATPase. Activity of the ATPase, when copurified with the cytoplasmic domain of an interactive ATPase partner, is stimulated by an acidic phospholipid, suggesting the membrane-associated ATPase is actively engaged in secretion. How the stimulated ATPase activity is terminated when secretion is complete is unclear. We fused the T2S ATPase of Xanthomonas campestris pv. campestris, the causal agent of black rot in the crucifers, with fluorescent protein and found that the ATPase in secretion-proficient cells was mainly diffused in cytoplasm. Focal spots at the cell periphery were detectable only in a few cells. The discrete foci were augmented in abundance and intensity when the secretion channel was depleted and the exoprotein overproduced. The foci abundance was inversely related to secretion efficiency of the secretion channel. Restored function of the secretion channel paralleled reduced ATPase foci abundance. The ATPase foci colocalized with the secretion channel. The ATPase may be transiently associated with the T2S machine by alternating between a cytoplasmic and a machine-associated state in a secretion-dependent manner. This provides a logical means for terminating the ATPase activity when secretion is completed. Function-related dynamic assembly may be the essence of the T2S machine.

  6. Real Time Live Imaging of Phytopathogenic Bacteria Xanthomonas campestris pv. campestris MAFF106712 in ‘Plant Sweet Home’

    PubMed Central

    Akimoto-Tomiyama, Chiharu; Furutani, Ayako; Ochiai, Hirokazu

    2014-01-01

    Xanthomonas is one of the most widespread phytobacteria, causing diseases on a variety of agricultural plants. To develop novel control techniques, knowledge of bacterial behavior inside plant cells is essential. Xanthomonas campestris pv. campestris, a vascular pathogen, is the causal agent of black rot on leaves of Brassicaceae, including Arabidopsis thaliana. Among the X. campestris pv. campestris stocks in the MAFF collection, we selected XccMAFF106712 as a model compatible pathogen for the A. thaliana reference ecotype Columbia (Col-0). Using modified green fluorescent protein (AcGFP) as a reporter, we observed real time XccMAFF106712 colonization in planta with confocal microscopy. AcGFP-expressing bacteria colonized the inside of epidermal cells and the apoplast, as well as the xylem vessels of the vasculature. In the case of the type III mutant, bacteria colonization was never detected in the xylem vessel or apoplast, though they freely enter the xylem vessel through the wound. After 9 days post inoculation with XccMAFF106712, the xylem vessel became filled with bacterial aggregates. This suggests that Xcc colonization can be divided into main four steps, (1) movement in the xylem vessel, (2) movement to the next cell, (3) adhesion to the host plant cells, and (4) formation of bacterial aggregates. The type III mutant abolished at least steps (1) and (2). Better understanding of Xcc colonization is essential for development of novel control techniques for black rot. PMID:24736478

  7. NagZ is required for beta-lactamase expression and full pathogenicity in Xanthomonas campestris pv. campestris str. 17.

    PubMed

    Yang, Tsuey-Ching; Chen, Tzu-Fan; Tsai, Jeffrey J P; Hu, Rouh-Mei

    2014-10-01

    Xanthomonas campestris pv. campestris expresses a chromosomally encoded class A β-lactamase Blaxc. Basal expression and induction of blaxc require the transcriptional factor AmpRxc and the peptidoglycan-monomers permease AmpGxc. NagZ is a β-GlcNAcase which cleaves GlcNAc-anhMurNAc peptides (peptidoglycan-monomers) to generate anhMurNAc-peptides. In many bacteria, anhMurNAc-peptides act as activation ligands for AmpR. Nevertheless, the role of NagZ in β-lactamase induction differs among species. In this paper, we studied the roles of nagZxc in the regulation of blaxc and pathogenicity in X. campestris pv. campestris. Our data showed that cells lacking nagZxc dramatically reduced the basal expression and induction of blaxc, suggesting that anhMurNAc-peptides, products of NagZxc, are required for blaxc expression regardless of the presence or absence of inducers. Expression of blaxc is regulated via an ampG-nagZ-ampR pathway. Pathogenicity assay demonstrated that an ampGxc mutant excited more severe symptoms than the wild-type; on the contrary, the nagZxc mutant became less virulent. To our knowledge, this is the first demonstration of a link between the ampG or nagZ defects and the pathogenicity in a plant pathogen.

  8. Evaluation of the virulence of Xanthomonas campestris pv. campestris mutant strains lacking functional genes in the OxyR regulon.

    PubMed

    Charoenlap, Nisanart; Buranajitpakorn, Sarinya; Duang-Nkern, Jintana; Namchaiw, Poommaree; Vattanaviboon, Paiboon; Mongkolsuk, Skorn

    2011-08-01

    Xanthomonas campestris pv. campestris causes black rot in cruciferous crops. Hydrogen peroxide (H(2)O(2)) production and accumulation is an important initial response in plant defense against invading microbes. The role of genes involved in the bacterial H(2)O(2) protection system in pathogenicity was evaluated. Mutants of katA (encoding a monofunctional catalase) and, to a lesser extent, katG (encoding a catalase-peroxidase) and oxyR (encoding a H(2)O(2) sensor and a transcription regulator), are hypersensitive to H(2)O(2) treatments that mimic the plant H(2)O(2) burst. These data correlate with the results of pathogenicity testing that show katA, katG, and oxyR mutants are avirulent on a compatible plant. Moreover, exposure to H(2)O(2) (1, 2, and 4 mM) highly induces the expression of genes in the OxyR regulon, including katA, katG, and ahpC. The avirulent phenotype of the oxyR mutant is partly because of its inability to mount an adaptive response upon exposure to an H(2)O(2) burst. Our data provide insights into important roles of a transcription regulator and other genes involved in peroxide stress protection in the virulence of X. campestris pv. campestris.

  9. Real time live imaging of phytopathogenic bacteria Xanthomonas campestris pv. campestris MAFF106712 in 'plant sweet home'.

    PubMed

    Akimoto-Tomiyama, Chiharu; Furutani, Ayako; Ochiai, Hirokazu

    2014-01-01

    Xanthomonas is one of the most widespread phytobacteria, causing diseases on a variety of agricultural plants. To develop novel control techniques, knowledge of bacterial behavior inside plant cells is essential. Xanthomonas campestris pv. campestris, a vascular pathogen, is the causal agent of black rot on leaves of Brassicaceae, including Arabidopsis thaliana. Among the X. campestris pv. campestris stocks in the MAFF collection, we selected XccMAFF106712 as a model compatible pathogen for the A. thaliana reference ecotype Columbia (Col-0). Using modified green fluorescent protein (AcGFP) as a reporter, we observed real time XccMAFF106712 colonization in planta with confocal microscopy. AcGFP-expressing bacteria colonized the inside of epidermal cells and the apoplast, as well as the xylem vessels of the vasculature. In the case of the type III mutant, bacteria colonization was never detected in the xylem vessel or apoplast, though they freely enter the xylem vessel through the wound. After 9 days post inoculation with XccMAFF106712, the xylem vessel became filled with bacterial aggregates. This suggests that Xcc colonization can be divided into main four steps, (1) movement in the xylem vessel, (2) movement to the next cell, (3) adhesion to the host plant cells, and (4) formation of bacterial aggregates. The type III mutant abolished at least steps (1) and (2). Better understanding of Xcc colonization is essential for development of novel control techniques for black rot.

  10. Identification of Crucifer Accessions from the NC-7 and NE-9 Plant Introduction Collections that are Resistant to Black Rot (Xanthomonas campestris pv. campestris) Races 1 and 4

    USDA-ARS?s Scientific Manuscript database

    Black rot, caused by Xanthomonas campestris pv. campestris (Pam.) Dawson (Xcc), is a serious disease of vegetable crucifers worldwide. The USDA NC-7 and NE-9 regional plant introduction stations maintain vegetable, mustard and oilseed crucifers, of which 4084 accessions were available for testing, ...

  11. The AvrB_AvrC domain of AvrXccC of Xanthomonas campestris pv. campestris is required to elicit plant defense responses and manipulate ABA homeostasis.

    PubMed

    Ho, Yi-Ping; Tan, Choon Meng; Li, Meng-Ying; Lin, Hong; Deng, Wen-Ling; Yang, Jun-Yi

    2013-04-01

    Plant disease induced by Xanthomonas campestris pv. campestris depends on type III effectors but the molecular basis is poorly understood. Here, AvrXccC8004 was characterized, and it was found that the AvrB_AvrC domain was essential and sufficient to elicit defense responses in an Arabidopsis-resistant ecotype (Col-0). An upregulation of genes in responding to the AvrB_AvrC domain of AvrXccC8004 was shown in a profile of host gene expression. The molecular changes were correlated with morphological changes observed in phenotypic and ultrastructural characterizations. Interestingly, the abscisic acid (ABA)-signaling pathway was also a prominent target for the AvrB_AvrC domain of AvrXccC8004. The highly elicited NCED5, encoding a key enzyme of ABA biosynthesis, was increased in parallel with ABA levels in AvrXccC8004 transgenic plants. Consistently, the X. campestris pv. campestris 8004 ΔavrXccC mutant was severely impaired in the ability to manipulate the accumulation of ABA and induction of ABA-related genes in challenged leaves. Moreover, exogenous application of ABA also enhanced the susceptibility of Arabidopsis to the X. campestris pv. campestris strains. These results indicate that the AvrB_AvrC domain of AvrXccC8004 alone has the activity to manipulate ABA homeostasis, which plays an important role in regulating the interactions of X. campestris pv. campestris and Arabidopsis.

  12. The galU gene of Xanthomonas campestris pv. campestris is involved in bacterial attachment, cell motility, polysaccharide synthesis, virulence, and tolerance to various stresses.

    PubMed

    Liao, Chao-Tsai; Du, Shin-Chiao; Lo, Hsueh-Hsia; Hsiao, Yi-Min

    2014-10-01

    Uridine triphosphate (UTP)-glucose-1-phosphate uridylyltransferase (GalU; EC 2.7.7.9) is an enzyme that catalyzes the formation of uridine diphosphate (UDP)-glucose from UTP and glucose-1-phosphate. GalU is involved in virulence in a number of animal-pathogenic bacteria since its product, UDP-glucose, is indispensable for the biosynthesis of virulence factors such as lipopolysaccharide and exopolysaccharide. However, its function in Xanthomonas campestris pv. campestris, the phytopathogen that causes black rot in cruciferous plants, is unclear. Here, we characterized a galU mutant of X. campestris pv. campestris and showed that the X. campestris pv. campestris galU mutant resulted in a reduction in virulence on the host cabbage. We also demonstrated that galU is involved in bacterial attachment, cell motility, and polysaccharide synthesis. Furthermore, the galU mutant showed increased sensitivity to various stress conditions including copper sulfate, hydrogen peroxide, and sodium dodecyl sulfate. In addition, mutation of galU impairs the expression of the flagellin gene fliC as well as the attachment-related genes xadA, fhaC, and yapH. In conclusion, our results indicate involvement of galU in the virulence factor production and pathogenicity in X. campestris pv. campestris, and a role for galU in stress tolerance of this crucifer pathogen.

  13. Synergistic Activation of the Pathogenicity-Related Proline Iminopeptidase Gene in Xanthomonas campestris pv. campestris by HrpX and a LuxR Homolog

    PubMed Central

    Zhang, Jingxi; Kan, Jinhong; Zhang, Jieqiong; Guo, Ping; Chen, Xiaoying

    2012-01-01

    Xanthomonas campestris pv. campestris strain 8004 contains an orphan quorum-sensing (QS) locus, xccR-pipXcc, in which the proline iminopeptidase (pipXcc) gene (where “Xcc” indicates that the pip gene is from X. campestris pv. campestris) is positively regulated by the LuxR homologue XccR by binding to the luxXc box of the pipXcc promoter. The disruption of pipXcc significantly attenuated the virulence of X. campestris pv. campestris. An imperfect plant-inducible promoter (PIP) box is located in the upstream region of the pipXcc promoter, which is the putative binding site of the transcriptional activator HrpX. To explore whether the expression of the pipXcc gene is regulated by HrpX, the expression level of a pipXcc promoter-gusA fusion gene was assayed in an hrpX disruption mutant. The results showed that the lack of HrpX dramatically decreased the β-glucuronidase (GUS) activity. Further analyses using an electrophoretic mobility shift assay (EMSA) and chromatin immunoprecipitation (ChIP)-PCR indicated that the imperfect PIP box in X. campestris pv. campestris is specifically bound to HrpX. These data demonstrated that the pipXcc gene belongs to the hrp regulon and that the imperfect PIP box of the pipXcc promoter could be a cis element for the HrpX protein. We further showed in a pulldown assay that XccR can bind HrpX, suggesting that these two regulatory proteins coactivate the virulence factor by binding to the different cis elements of the pipXcc gene and adapt to the host environment during X. campestris pv. campestris infection. PMID:22865058

  14. Synergistic activation of the pathogenicity-related proline iminopeptidase gene in Xanthomonas campestris pv. campestris by HrpX and a LuxR homolog.

    PubMed

    Zhang, Jingxi; Kan, Jinhong; Zhang, Jieqiong; Guo, Ping; Chen, Xiaoying; Fang, Rongxiang; Jia, Yantao

    2012-10-01

    Xanthomonas campestris pv. campestris strain 8004 contains an orphan quorum-sensing (QS) locus, xccR-pip(Xcc), in which the proline iminopeptidase (pip(Xcc)) gene (where "Xcc" indicates that the pip gene is from X. campestris pv. campestris) is positively regulated by the LuxR homologue XccR by binding to the luxXc box of the pip(Xcc) promoter. The disruption of pip(Xcc) significantly attenuated the virulence of X. campestris pv. campestris. An imperfect plant-inducible promoter (PIP) box is located in the upstream region of the pip(Xcc) promoter, which is the putative binding site of the transcriptional activator HrpX. To explore whether the expression of the pip(Xcc) gene is regulated by HrpX, the expression level of a pip(Xcc) promoter-gusA fusion gene was assayed in an hrpX disruption mutant. The results showed that the lack of HrpX dramatically decreased the β-glucuronidase (GUS) activity. Further analyses using an electrophoretic mobility shift assay (EMSA) and chromatin immunoprecipitation (ChIP)-PCR indicated that the imperfect PIP box in X. campestris pv. campestris is specifically bound to HrpX. These data demonstrated that the pip(Xcc) gene belongs to the hrp regulon and that the imperfect PIP box of the pip(Xcc) promoter could be a cis element for the HrpX protein. We further showed in a pulldown assay that XccR can bind HrpX, suggesting that these two regulatory proteins coactivate the virulence factor by binding to the different cis elements of the pip(Xcc) gene and adapt to the host environment during X. campestris pv. campestris infection.

  15. Insights into the extracytoplasmic stress response of Xanthomonas campestris pv. campestris: role and regulation of {sigma}E-dependent activity.

    PubMed

    Bordes, Patricia; Lavatine, Laure; Phok, Kounthéa; Barriot, Roland; Boulanger, Alice; Castanié-Cornet, Marie-Pierre; Déjean, Guillaume; Lauber, Emmanuelle; Becker, Anke; Arlat, Matthieu; Gutierrez, Claude

    2011-01-01

    Xanthomonas campestris pv. campestris is an epiphytic bacterium that can become a vascular pathogen responsible for black rot disease of crucifers. To adapt gene expression in response to ever-changing habitats, phytopathogenic bacteria have evolved signal transduction regulatory pathways, such as extracytoplasmic function (ECF) σ factors. The alternative sigma factor σ(E), encoded by rpoE, is crucial for envelope stress response and plays a role in the pathogenicity of many bacterial species. Here, we combine different approaches to investigate the role and mechanism of σ(E)-dependent activation in X. campestris pv. campestris. We show that the rpoE gene is organized as a single transcription unit with the anti-σ gene rseA and the protease gene mucD and that rpoE transcription is autoregulated. rseA and mucD transcription is also controlled by a highly conserved σ(E)-dependent promoter within the σ(E) gene sequence. The σ(E)-mediated stress response is required for stationary-phase survival, resistance to cadmium, and adaptation to membrane-perturbing stresses (elevated temperature and ethanol). Using microarray technology, we started to define the σ(E) regulon of X. campestris pv. campestris. These genes encode proteins belonging to different classes, including periplasmic or membrane proteins, biosynthetic enzymes, classical heat shock proteins, and the heat stress σ factor σ(H). The consensus sequence for the predicted σ(E)-regulated promoter elements is GGAACTN(15-17)GTCNNA. Determination of the rpoH transcription start site revealed that rpoH was directly regulated by σ(E) under both normal and heat stress conditions. Finally, σ(E) activity is regulated by the putative regulated intramembrane proteolysis (RIP) proteases RseP and DegS, as previously described in many other bacteria. However, our data suggest that RseP and DegS are not only dedicated to RseA cleavage and that the proteolytic cascade of RseA could involve other proteases.

  16. Mutagenesis of all eight avr genes in Xanthomonas campestris pv. campestris had no detected effect on pathogenicity, but one avr gene affected race specificity.

    PubMed

    Castañeda, Adriana; Reddy, Joseph D; El-Yacoubi, Basma; Gabriel, Dean W

    2005-12-01

    Suppression subtractive hybridization (SSH) was used to identify genes present in the systemic crucifer black rot pathogen Xanthomonas campestris pv. campestris 528T but missing from the nonsystemic crucifer leaf spot pathogen, X. campestris pv. armoraciae 417. Among the DNA fragments unique to 528T was Xcc2109, one of eight putative avr genes identified in the published 528T genome (NC_003902). Individual and sequential deletion, insertion mutations, or both of all eight 528T avr gene loci were made, but no change in pathogenicity was observed with any combination of avr mutations, including a strain with all eight avr genes deleted. However, insertion or deletion mutants affecting the Xcc2109 locus lost avirulence (i.e., became virulent) on Florida Mustard, an X. campestris pv. campestris race-determining, differential host. The Xcc2109 open reading frame as annotated was cloned and found to be nonfunctional. A longer gene, encompassing Xcc2109 and here designated avrXccFM, was cloned and found to complement the Xcc2109 mutants and to confer avirulence to two additional wild-type X. campestris pv. campestris strains, thereby changing their races. Resistance in Florida Mustard to 528T strains carrying avrXccFM occurred without a typical hypersensitive response (HR) on leaves, although a vascular HR was observed in seedlings.

  17. Crystallization and preliminary X-ray diffraction analysis of a new xyloglucanase from Xanthomonas campestris pv. campestris

    PubMed Central

    de Araújo, Evandro Ares; Tomazini, Atílio; Kadowaki, Marco Antonio Seiki; Murakami, Mário Tyago; Polikarpov, Igor

    2013-01-01

    Xyloglucanases (Xghs) are important enzymes involved in xyloglucan modification and degradation. Xanthomonas campestris pv. campestris (Xcc) is a phytopathogenic bacterium which produces a large number of glycosyl hydrolases (GH), but has only one family 74 GH (Xcc-Xgh). This enzyme was overexpressed in Escherichia coli, purified and crystallized. Diffraction data sets were collected for the native enzyme and its complex with glucose to maximum resolutions of 2.0 and 2.1 Å, respectively. The data were indexed in a hexagonal crystal system with unit-cell parameters a = b = 153.4, c = 84.9 Å. As indicated by molecular-replacement solution, the crystals belonged to space group P61. PMID:23722852

  18. Crystallization and preliminary diffraction analysis of the catalytic domain of major extracellular endoglucanase from Xanthomonas campestris pv. campestris

    PubMed Central

    Rosseto, Flávio R.; Puhl, Ana C.; Andrade, Maxuel O.; Polikarpov, Igor

    2013-01-01

    Cellulases, such as endoglucanases, exoglucanases and β-glucosidases, are important enzymes used in the process of enzymatic hydrolysis of plant biomass. The bacteria Xanthomonas campestris pv. campestris expresses a large number of hydrolases and the major endoglucanase (XccEG), a member of glycoside hydrolase family 5 (GH5), is the most strongly secreted extracellularly. In this work, the native XccEG was purified from the extracellular extract and crystallization assays were performed on its catalytic domain. A complete data set was collected on an in-house X-ray source. The crystal diffracted to 2.7 Å resolution and belonged to space group C2, with unit-cell parameters a = 174.66, b = 141.53, c = 108.00 Å, β = 110.49°. The Matthews coefficient suggests a solvent content of 70.1% and the presence of four protein subunits in the asymmetric unit. PMID:23385754

  19. Induction of a secretable beta-lactamase requires a long lag time in Xanthomonas campestris pv. campestris str. 17.

    PubMed

    Yang, Tsuey-Ching; Tsai, Mei-Jung; Tsai, Jeffrey J P; Hu, Rouh-Mei

    2011-12-01

    Xanthomonas campestris pv. campestris (Xcc) constitutively expresses penicillinase activity in the absence of an inducer. An ampR-bla module is required for the antibiotic resistance phenotype. In this study, we demonstrate that AmpR negatively autoregulates its own expression in a β-lactam-independent manner. In the absence of inducer, bla is expressed at a high basal level. Expression of bla is inducible by β-lactam, however, with a period. AmpR protein and the LysR-motif located upstream of bla promoter are essential for basal expression and induction of bla. Most β-lactamase activity is present in the culture medium, suggesting that Bla protein can be secreted by Xcc into the environment.

  20. Crystallization and preliminary diffraction analysis of the catalytic domain of major extracellular endoglucanase from Xanthomonas campestris pv. campestris.

    PubMed

    Rosseto, Flávio R; Puhl, Ana C; Andrade, Maxuel O; Polikarpov, Igor

    2013-02-01

    Cellulases, such as endoglucanases, exoglucanases and β-glucosidases, are important enzymes used in the process of enzymatic hydrolysis of plant biomass. The bacteria Xanthomonas campestris pv. campestris expresses a large number of hydrolases and the major endoglucanase (XccEG), a member of glycoside hydrolase family 5 (GH5), is the most strongly secreted extracellularly. In this work, the native XccEG was purified from the extracellular extract and crystallization assays were performed on its catalytic domain. A complete data set was collected on an in-house X-ray source. The crystal diffracted to 2.7 Å resolution and belonged to space group C2, with unit-cell parameters a = 174.66, b = 141.53, c = 108.00 Å, β = 110.49°. The Matthews coefficient suggests a solvent content of 70.1% and the presence of four protein subunits in the asymmetric unit.

  1. Crystallization and preliminary X-ray diffraction analysis of a new xyloglucanase from Xanthomonas campestris pv. campestris.

    PubMed

    de Araújo, Evandro Ares; Tomazini, Atílio; Kadowaki, Marco Antonio Seiki; Murakami, Mário Tyago; Polikarpov, Igor

    2013-06-01

    Xyloglucanases (Xghs) are important enzymes involved in xyloglucan modification and degradation. Xanthomonas campestris pv. campestris (Xcc) is a phytopathogenic bacterium which produces a large number of glycosyl hydrolases (GH), but has only one family 74 GH (Xcc-Xgh). This enzyme was overexpressed in Escherichia coli, purified and crystallized. Diffraction data sets were collected for the native enzyme and its complex with glucose to maximum resolutions of 2.0 and 2.1 Å, respectively. The data were indexed in a hexagonal crystal system with unit-cell parameters a = b = 153.4, c = 84.9 Å. As indicated by molecular-replacement solution, the crystals belonged to space group P6(1).

  2. XC_0531 encodes a c-type cytochrome biogenesis protein and is required for pathogenesis in Xanthomonas campestris pv. campestris.

    PubMed

    Chen, Lei; Wang, Mingpeng; Huang, Li; Zhang, Zhaojie; Liu, Fanghua; Lu, Guangtao

    2017-06-27

    The phytopathogenic Xanthomonas campestris pv.campestris is a gram-negative bacterium and the causal agent of black-rot disease of cruciferous crops. Many gram-negative bacteria possess a family of proteins, called Dsbs, which are involved in disulfide bond formation in certain periplasmic proteins. In our preliminary screening of the virulence to the plants we identified that gene XC_0531 which annotated gene dsbD of Xanthomonas campestris pv. campestris (Xcc) is related to the virulence to the host plants. Here, we found XC_0531 encoded a DsbD like protein. Its deletion is sensitive to DTT and copper, decreased accumulation of free thiols in periplasm. Its deletion also affected heme synthesis, position of Soret band and the production of peak c550. This suggests that XC_0531 is related to c-type cytochromes biogenesis. XC_0531 mutation decreased the utilization of different carbon sources (such as galactose, xylose, maltose, saccharose and glucose), reduced extracellular polysaccharide (EPS) production, decreased extracellular enzyme activities (protease, cellulose and amylase), slowed down growth rate of Xcc and weakened virulence to the plants. These results suggest that these phenotypes caused by XC_0531 mutation is possibly due to deficient biosynthesis of c-type cytochromes in respiration chain and the formation of disulfide bonds. Our work confirmed the function of XC_0531 and provide theory basis for scientists working on molecular mechanisms of cytochrome c biogenesis, pathogenesis of Xcc, development of EPS commercial values and protecting plant from black rot. We confirmed the function of gene XC_0531, which encodes a DsbD like protein, a protein correlated with c-type cytochrome biogenesis. This gene is related to the virulence to plants by affecting funtion of cytochromes c and probably disulfide bonds modification of proteins in type II secretion system (T2SS).

  3. The apo structure of sucrose hydrolase from Xanthomonas campestris pv. campestris shows an open active-site groove.

    PubMed

    Champion, Elise; Remaud-Simeon, Magali; Skov, Lars K; Kastrup, Jette S; Gajhede, Michael; Mirza, Osman

    2009-12-01

    Glycoside hydrolase family 13 (GH-13) mainly contains starch-degrading or starch-modifying enzymes. Sucrose hydrolases utilize sucrose instead of amylose as the primary glucosyl donor. Here, the catalytic properties and X-ray structure of sucrose hydrolase from Xanthomonas campestris pv. campestris are reported. Sucrose hydrolysis catalyzed by the enzyme follows Michaelis-Menten kinetics, with a K(m) of 60.7 mM and a k(cat) of 21.7 s(-1). The structure of the enzyme was solved at a resolution of 1.9 A in the resting state with an empty active site. This represents the first apo structure from subfamily 4 of GH-13. Comparisons with structures of the highly similar sucrose hydrolase from X. axonopodis pv. glycines most notably showed that residues Arg516 and Asp138, which form a salt bridge in the X. axonopodis sucrose complex and define part of the subsite -1 glucosyl-binding determinants, are not engaged in salt-bridge formation in the resting X. campestris enzyme. In the absence of the salt bridge an opening is created which gives access to subsite -1 from the ;nonreducing' end. Binding of the glucosyl moiety in subsite -1 is therefore likely to induce changes in the conformation of the active-site cleft of the X. campestris enzyme. These changes lead to salt-bridge formation that shortens the groove. Additionally, this finding has implications for understanding the molecular mechanism of the closely related subfamily 4 glucosyl transferase amylosucrase, as it indicates that sucrose could enter the active site from the ;nonreducing' end during the glucan-elongation cycle.

  4. The N-Glycan cluster from Xanthomonas campestris pv. campestris: a toolbox for sequential plant N-glycan processing.

    PubMed

    Dupoiron, Stéphanie; Zischek, Claudine; Ligat, Laetitia; Carbonne, Julien; Boulanger, Alice; Dugé de Bernonville, Thomas; Lautier, Martine; Rival, Pauline; Arlat, Matthieu; Jamet, Elisabeth; Lauber, Emmanuelle; Albenne, Cécile

    2015-03-06

    N-Glycans are widely distributed in living organisms but represent only a small fraction of the carbohydrates found in plants. This probably explains why they have not previously been considered as substrates exploited by phytopathogenic bacteria during plant infection. Xanthomonas campestris pv. campestris, the causal agent of black rot disease of Brassica plants, possesses a specific system for GlcNAc utilization expressed during host plant infection. This system encompasses a cluster of eight genes (nixE to nixL) encoding glycoside hydrolases (GHs). In this paper, we have characterized the enzymatic activities of these GHs and demonstrated their involvement in sequential degradation of a plant N-glycan using a N-glycopeptide containing two GlcNAcs, three mannoses, one fucose, and one xylose (N2M3FX) as a substrate. The removal of the α-1,3-mannose by the α-mannosidase NixK (GH92) is a prerequisite for the subsequent action of the β-xylosidase NixI (GH3), which is involved in the cleavage of the β-1,2-xylose, followed by the α-mannosidase NixJ (GH125), which removes the α-1,6-mannose. These data, combined to the subcellular localization of the enzymes, allowed us to propose a model of N-glycopeptide processing by X. campestris pv. campestris. This study constitutes the first evidence suggesting N-glycan degradation by a plant pathogen, a feature shared with human pathogenic bacteria. Plant N-glycans should therefore be included in the repertoire of molecules putatively metabolized by phytopathogenic bacteria during their life cycle.

  5. AmpG is required for BlaXc beta-lactamase expression in Xanthomonas campestris pv. campestris str. 17.

    PubMed

    Yang, Tsuey-Ching; Chen, Tzu-Fan; Tsai, Jeffrey J P; Hu, Rouh-Mei

    2013-03-01

    The chromosomal ampR(Xc) -bla(Xc) module is essential for the β-lactam resistance of Xanthomonas campestris pv. campestris. Bla(Xc) β-lactamase is expressed at a high basal level in the absence of an inducer and its expression can be further induced by β-lactam. In enterobacteria, ampG encodes an inner membrane facilitator involved in the recycling of murein degradation compounds. An isogenic ampG mutant (XcampG) of X. campestris pv. campestris str. 17 (Xc17) was constructed to investigate the link between murein recycling and bla(Xc) expression. Our data demonstrate that (1) XcampG is susceptible to β-lactam antibiotics; (2) AmpG(Xc) is essential for expression of bla(Xc) ; (3) AmpGs of Xc17, Stenotrophomonas maltophilia KJ (SmKJ) and Escherichia coli DH5α can complement the defect of XcampG; (4) overexpression of AmpG(X) (c) significantly increased bla(Xc) expression; and (5) AmpG(Xc) from Xc17 is able to restore β-lactamase induction of the ampN(Xc) -ampG(Xc) double mutant of SmKJ. In Xc17, ampG(Xc) can be expressed from the promoter residing in the intergenic region of ampN(Xc) -ampG(Xc) and the expression is independent of β-lactam induction. AmpN, which is required for β-lactamases induction in SmKJ, is not required for the β-lactam antibiotic resistance of Xc17.

  6. Host Genotype and Hypersensitive Reaction Influence Population Levels of Xanthomonas campestris pv. vitians in Lettuce.

    PubMed

    Bull, Carolee T; Gebben, Samantha J; Goldman, Polly H; Trent, Mark; Hayes, Ryan J

    2015-03-01

    Dynamics of population sizes of Xanthomonas campestris pv. vitians inoculated onto or into lettuce leaves were monitored on susceptible and resistant cultivars. In general, population growth was greater for susceptible (Clemente, Salinas 88, Vista Verde) than resistant (Batavia Reine des Glaces, Iceberg, Little Gem) cultivars. When spray-inoculated or infiltrated, population levels of X. campestris pv. vitians were consistently significantly lower on Little Gem than on susceptible cultivars, while differences in the other resistant cultivars were not consistently statistically significant. Populations increased at an intermediate rate on cultivars Iceberg and Batavia Reine des Glaces. There were significant positive correlations between bacterial concentration applied and disease severity for all cultivars, but bacterial titer had a significantly greater influence on disease severity in the susceptible cultivars than in Little Gem and an intermediate influence in Iceberg and Batavia Reine des Glaces. Infiltration of X. campestris pv. vitians strains into leaves of Little Gem resulted in an incompatible reaction, whereas compatible reactions were observed in all other cultivars. It appears that the differences in the relationship between population dynamics for Little Gem and the other cultivars tested were due to the hypersensitive response in cultivar Little Gem. These findings have implications for disease management and lettuce breeding because X. campestris pv. vitians interacts differently with cultivars that differ for resistance mechanisms.

  7. Host genotype and hypersensitive reaction influence population levels of Xanthomonas campestris pv. vitians in lettuce

    USDA-ARS?s Scientific Manuscript database

    Population dynamics of Xanthomonas campestris pv. vitians spray inoculated on or infiltrated into lettuce leaves were monitored on cultivars that were well characterized for resistance or susceptibility to the pathogen. In general, population growth was greater for susceptible (Clemente, Salinas 88,...

  8. Genetic and pathogenic variability of Indian strains of Xanthomonas campestris pv. campestris causing black rot disease in crucifers.

    PubMed

    Singh, Dinesh; Dhar, Shri; Yadava, D K

    2011-12-01

    Xanthomonas campestris pv. campestris (Pammel) Dowson (Xcc) causing black rot of crucifers is a serious disease in India and causes >50% crop losses in favorable environmental conditions. Pathogenic variability of Xcc, X. oryzae pv. oryzae (Xoo), and X. axonopodis pv. citri (Xac) were tested on 19 cultivars of cruciferae including seven Brassica spp. viz., B. campestris, B. carinata, B. juncea, B. napus, B. nigra, B. oleracea and B. rapa, and Raphanus sativus for two consecutive years viz., 2007-2008 and 2008-2009 under field conditions at Indian Agricultural Research Institute, New Delhi. Xcc (22 strains) and other species of Xanthomonas (2 strains), they formed three distinct groups of pathogenic variability i.e., Group 1, 2, and 3 under 50% minimum similarity coefficient. All strains of Xcc clustered under Groupl except Xcc-C20. The strains of Xcc further clustered in 6 subgroups viz., A, B, C, D, E, and F based on diseases reaction on host. Genetic variability of 22 strains of Xcc was studied by using Rep-PCR (REP-, BOX- and ERIC-PCR) and 10 strains for hrp (hypersensitive reaction and pathogenecity) gene sequence analysis. Xcc strains comprised in cluster 1, Xac under cluster 2, while Xoo formed separate cluster 3 based on >50% similarity coefficient. Cluster 1 was further divided into 8 subgroups viz., A, B, C, D, E, F, G, and H at 75% similarity coefficient. The hrpF gene sequence analysis also showed distinctness of Xcc strains from other Xanthomonads. In this study, genetic and pathogenic variability in Indian strains of Xcc were established, which will be of immense use in the development of resistant genotypes against this bacterial pathogen.

  9. Evaluation of the role of recA protein in plant virulence with recA mutants of Xanthomonas campestris pv. campestris.

    PubMed

    Martínez, S; Martínez-Salazar, J; Camas, A; Sánchez, R; Soberón-Chávez, G

    1997-09-01

    Xanthomonas campestris pv. campestris NRRL B1459 recA mutants were isolated by recombination with an interrupted Rhizobium etli recA gene and selection of double recombinants. The mutants were impaired in homologous genetic recombination and in DNA repair as judged by their sensitivity to methyl-methane-sulfonate and to UV irradiation; these defects are complemented in trans by the R. etli recA gene. Virulence of X. campestris pv. campestris NRRL B1459 to cabbage is considerably diminished by the recA mutation. The recA mutation is not correlated with the frequency of occurrence of a genetic rearrangement that affects chemotaxis, plant virulence, and xanthan gum production.

  10. Xanthoferrin, the α-hydroxycarboxylate-type siderophore of Xanthomonas campestris pv. campestris, is required for optimum virulence and growth inside cabbage.

    PubMed

    Pandey, Sheo Shankar; Patnana, Pradeep Kumar; Rai, Rikky; Chatterjee, Subhadeep

    2016-06-27

    Xanthomonas campestris pv. campestris causes black rot, a serious disease of crucifers. Xanthomonads encode a siderophore biosynthesis and uptake gene cluster xss (Xanthomonas siderophore synthesis) involved in the production of a vibrioferrin-type siderophore. However, little is known about the role of the siderophore in the iron uptake and virulence of X. campestris pv. campestris. In this study, we show that X. campestris pv. campestris produces an α-hydroxycarboxylate-type siderophore (named xanthoferrin), which is required for growth under low-iron conditions and for optimum virulence. A mutation in the siderophore synthesis xssA gene causes deficiency in siderophore production and growth under low-iron conditions. In contrast, the siderophore utilization ΔxsuA mutant is able to produce siderophore, but exhibits a defect in the utilization of the siderophore-iron complex. Our radiolabelled iron uptake studies confirm that the ΔxssA and ΔxsuA mutants exhibit defects in ferric iron (Fe(3+) ) uptake. The ΔxssA mutant is able to utilize and transport the exogenous xanthoferrin-Fe(3+) complex; in contrast, the siderophore utilization or uptake mutant ΔxsuA exhibits defects in siderophore uptake. Expression analysis of the xss operon using a chromosomal gusA fusion indicates that the xss operon is expressed during in planta growth and under low-iron conditions. Furthermore, exogenous iron supplementation in cabbage leaves rescues the in planta growth deficiency of ΔxssA and ΔxsuA mutants. Our study reveals that the siderophore xanthoferrin is an important virulence factor of X. campestris pv. campestris which promotes in planta growth by the sequestration of Fe(3+) .

  11. Gene expression in Brassica campestris showing a hypersensitive response to the incompatible pathogen Xanthomonas campestris pv. vitians.

    PubMed

    Collinge, D B; Milligan, D E; Dow, J M; Scofield, G; Daniels, M J

    1987-09-01

    Xanthomonas campestris pv. vitians, a pathogen of lettuce, elicits a hypersensitive response within 12 hours of inoculation into Brassica leaves, characterized by tissue collapse, loss of membrane integrity, vein blockage and melanin production. In contrast, the compatible pathogen, X. c. pv. campestris, has no visible effects on leaves for 48 hours, after which inoculated areas show chlorosis which eventually spreads, followed by rotting.mRNA was prepared from leaves inoculated with suspensions of both pathovars or with sterile medium up to 24 hours following inoculation. In vitro translation of total and poly A(+) RNA in rabbit reticulocyte lysate in the presence of (35)S methionine followed by separation of the polypeptide products by 2D-PAGE, allowed comparison of the effects of these treatments on plant gene expression. Major changes in gene expression were observed as a consequence of the inoculation technique. In addition, after inoculation with X. c. vitians, up to fifteen additional major polypeptides appeared or greatly increased by four hours. Some of these had disappeared by nine hours and several more had appeared. No major polypeptides disappeared or decreased greatly in intensity following inoculation with X. c. vitians.

  12. Comparative proteome analysis of Xanthomonas campestris pv. campestris in the interaction with the susceptible and the resistant cultivars of Brassica oleracea.

    PubMed

    Villeth, Gabriela R; Reis, Fabio B; Tonietto, Angela; Huergo, Luciano; de Souza, Emanuel M; Pedrosa, Fabio O; Franco, Octávio L; Mehta, Angela

    2009-09-01

    Black rot of cruciferous plants, caused by Xanthomonas campestris pv. campestris, causes severe losses in agriculture around the world. This disease affects several cultures, including cabbage and broccoli, among others. Proteome studies of this bacterium have been reported; however, most of them were performed using the bacterium grown under culture media conditions. Recently, we have analyzed the proteome of X. campestris pv. campestris during the interaction with the susceptible cultivar of Brassica oleracea and several proteins were identified. The objective of the present study was to analyze the expressed proteins of X. campestris pv. campestris during the interaction with the resistant cultivar of B. oleracea. The bacterium was infiltrated in the leaves of the resistant plant and recovered for protein extraction and two-dimensional electrophoresis. The protein profile was compared with that of the bacterium isolated from the susceptible host and the results obtained revealed a group of proteins exclusive to the resistant interaction. Among the proteins identified in this study were plant and bacterium proteins, some of which were exclusively expressed during the resistant interaction.

  13. Genome-scale mutagenesis and phenotypic characterization of two-component signal transduction systems in Xanthomonas campestris pv. campestris ATCC 33913.

    PubMed

    Qian, Wei; Han, Zhong-Ji; Tao, Jun; He, Chaozu

    2008-08-01

    The gram-negative bacterium Xanthomonas campestris pv. campestris is the causal agent of black rot disease of cruciferous plants. Its genome encodes a large repertoire of two-component signal transduction systems (TCSTSs), which consist of histidine kinases and response regulators (RR) to monitor and respond to environmental stimuli. To investigate the biological functions of these TCSTS genes, we aimed to inactivate all 54 RR genes in X. campestris pv. campestris ATCC 33913, and successfully generated 51 viable mutants using the insertion inactivation method. Plant inoculation identified two novel response regulator genes (XCC1958 and XCC3107) that are involved in virulence of this strain. Genetic complementation demonstrated that XCC3107, designated as vgrR (virulence and growth regulator), also affects bacterial growth and activity of extracellular proteases. In addition, we assessed the survival of these mutants under various stresses, including osmotic stress, high sodium concentration, heat shock, and sodium dodecyl sulfate exposure, and identified a number of genes that may be involved in the general stress response of X. campestris pv. campestris. Mutagenesis and phenotypic characterization of RR genes in this study will facilitate future studies on signaling networks in this important phytopathogenic bacterium.

  14. Transcriptome profiling of Xanthomonas campestris pv. campestris grown in minimal medium MMX and rich medium NYG.

    PubMed

    Liu, Wei; Yu, Yan-Hua; Cao, Shi-Yuan; Niu, Xiang-Na; Jiang, Wei; Liu, Guo-Fang; Jiang, Bo-Le; Tang, Dong-Jie; Lu, Guang-Tao; He, Yong-Qiang; Tang, Ji-Liang

    2013-06-01

    Xanthomonas campestris pathovar campestris (Xcc) is the causal agent of black rot disease in cruciferous plants worldwide. Although the complete genomes of several Xcc strains have been determined, the gene expression and regulation mechanisms in this pathogen are far from clear. In this work, transcriptome profiling of Xcc 8004 grown in MMX medium (minimal medium for Xanthomonas campestris) and NYG medium (peptone yeast glycerol medium) were investigated by RNA-Seq. Using the Illumina HiSeq 2000 platform, a total of 26,514,630 reads (90 nt in average) were generated, of which 15,708,478 reads mapped uniquely to coding regions of Xcc 8004 genome. Of the 4273 annotated protein-coding genes of Xcc 8004, 629 were found differentially expressed in Xcc grown in MMX and NYG. Of the differentially expressed genes, 495 were up-regulated and 134 were down-regulated in MMX. The MMX-induced genes are mainly involved in amino acid metabolism, transport systems, atypical condition adaptation and pathogenicity, especially the type III secretion system, while the MMX-repressed genes are mainly involved in chemotaxis and degradation of small molecules. The global transcriptome analyzes of Xcc 8004 grown in MMX and NYG might facilitate the gene functional characterization of this phytopathogenic bacterium.

  15. Identification of six type III effector genes with the PIP box in Xanthomonas campestris pv. campestris and five of them contribute individually to full pathogenicity.

    PubMed

    Jiang, Wei; Jiang, Bo-Le; Xu, Rong-Qi; Huang, Jun-Ding; Wei, Hong-Yu; Jiang, Guo-Feng; Cen, Wei-Jian; Liu, Jiao; Ge, Ying-Ying; Li, Guang-Hua; Su, Li-Li; Hang, Xiao-Hong; Tang, Dong-Jie; Lu, Guang-Tao; Feng, Jia-Xun; He, Yong-Qiang; Tang, Ji-Liang

    2009-11-01

    Xanthomonas campestris pv. campestris is the pathogen of black rot of cruciferous plants. The pathogenicity of the pathogen depends on the type III secretion system (T3SS) that translocates directly effector proteins into plant cells, where they play important roles in the molecular interaction between the pathogen and its hosts. The T3SS of Xanthomonas spp. is encoded by a cluster of hypersensitive response and pathogenicity (hrp) genes. It has been demonstrated that the expression of hrp genes and some type III secreted (T3S)-effector genes is coactivated by the key hrp regulatory protein HrpX. The regulation by HrpX can be mediated by the binding of HrpX protein to a cis-regulatory element named the plant-inducible promoter (PIP) box present in the promoter region of HrpX-regulated genes. A genome screen revealed that X. campestris pv. campestris 8004 possesses 56 predicted genes with the PIP box. Nine of these genes have been shown to encode T3S effectors, Hrp, and Hrp-associated proteins. In this study, we employed an established T3S effector translocation assay with the hypersensitive-reaction-inducing domain of X. campestris pv. campestris AvrBs1 as a reporter to characterize the remaining 47 genes with the PIP box and showed that 6 of them, designated as XopXccE1, XopXccP, XopXccQ, XopXccR1, XopXccLR, and AvrXccB, harbor a functional translocation signal in their N-terminal regions, indicating that they are T3S effectors of X. campestris pv. campestris. We provided evidence to demonstrate that all these effectors are expressed in an HrpX-dependent manner and their translocation into plant cells relies on the translocon protein HrpF and the chaperone HpaB. Mutational analyses demonstrated that all these effectors, except AvrXccB, are individually required for full virulence and growth of X. campestris pv. campestris in the host plant Chinese radish.

  16. Antibacterial activity of metabolite produced by Paenibacillus polymyxa strain HKA-15 against Xanthomonas campestris pv. phaseoli.

    PubMed

    Mageshwaran, V; Walia, Suresh; Govindasamy, V; Annapurna, K

    2011-03-01

    An antibacterial metabolite extracted from Paenibacillus polymyxa HKA-15 showed strong inhibition against Xanthomonas campestris pv. phaseoli strains CP-1-1 and M-5. Minimum inhibitory concentration (MIC) of crude extract against strains CP-1-1 and M-5 was found to be 1.7 mg/ml and 1.52 mg/ml, respectively. In UV-Vis range, the absorption peak of crude extract was maximum at 240 nm. The compound is resilience to wide range of temperature, pH, surfactants and organic solvents. The complete loss of activity was observed when crude metabolite was treated with pepsin (400 unit/ml). Characterization of crude metabolite suggested its hydrophobic and peptide nature. Inhibition of Xanthomonas campestris pv. phaseoli by peptide like metabolite produced by Paenibacillus polymyxa strain HKA-15 under in vitro conditions showed ecological and biotechnological potential of strain HKA-15 to control common blight disease in beans.

  17. Diffusible signal factor family signals provide a fitness advantage to Xanthomonas campestris pv. campestris in interspecies competition.

    PubMed

    Deng, Yinyue; Wu, Jien; Yin, Wenfang; Li, Peng; Zhou, Jianuan; Chen, Shaohua; He, Fei; Cai, Jun; Zhang, Lian-Hui

    2016-05-01

    Diffusible signal factor (DSF) represents a new class of widely conserved quorum sensing signals, which regulates various biological functions through intra- or interspecies signaling. The previous studies identified that there is an antagonistic interaction between Xanthomonas and Bacillus species bacteria in natural ecosystem, but the detailed molecular mechanism of interspecies competition is not clear. This study showed that Xanthomonas campestris pv. campestris (Xcc) interfered with morphological transition and sporulation of Bacillus thuringiensis in mixed cultures, whereas abrogation of the DSF synthase RpfF reduced the interference. DSF inhibited B. thuringiensis cell division and sporulation through modulation of ftsZ, which encodes an important cell division protein in bacterial cells. In addition, RpfF is essential for production of six DSF-family signals in Xcc, which employ the same signaling pathways to regulate biological functions in Xcc and play similar effects on reduction of cell division, sporulation and antibiotic resistance of B. thuringiensis. Furthermore, abrogation of RpfF decreased the competitive capability of Xcc against B. thuringiensis on the surface of Chinese cabbage leaves. Our findings provide new insights into the role of DSF-family signals in interspecies competition and depict molecular mechanisms with which Xcc competes with B. thuringiensis. © 2016 Society for Applied Microbiology and John Wiley & Sons Ltd.

  18. Identification of a novel type III secretion-associated outer membrane-bound protein from Xanthomonas campestris pv. campestris

    PubMed Central

    Li, Lei; Li, Rui-Fang; Ming, Zhen-Hua; Lu, Guang-Tao; Tang, Ji-Liang

    2017-01-01

    Many bacterial pathogens employ the type III secretion system (T3SS) to translocate effector proteins into eukaryotic cells to overcome host defenses. To date, most of our knowledge about the T3SS molecular architecture comes from the studies on animal pathogens. In plant pathogens, nine Hrc proteins are believed to be structural components of the T3SS, of which HrcC and HrcJ form the outer and inner rings of the T3SS, respectively. Here, we demonstrated that a novel outer membrane-bound protein (HpaM) of Xanthomonas campestris pv. campestris is critical for the type III secretion and is structurally and functionally conserved in phytopathogenic Xanthomonas spp. We showed that the C-terminus of HpaM extends into the periplasm to interact physically with HrcJ and the middle part of HpaM interacts physically with HrcC. It is clear that the outer and inner rings compose the main basal body of the T3SS apparatus in animal pathogens. Therefore, we presume that HpaM may act as a T3SS structural component, or play a role in assisting assembling or affecting the stability of the T3SS apparatus. HpaM is a highly prevalent and specific protein in Xanthomonas spp., suggesting that the T3SS of Xanthomonas is distinctive in some aspects from other pathogens. PMID:28198457

  19. Analysis of outer membrane vesicle associated proteins isolated from the plant pathogenic bacterium Xanthomonas campestris pv. campestris.

    PubMed

    Sidhu, Vishaldeep K; Vorhölter, Frank-Jörg; Niehaus, Karsten; Watt, Steven A

    2008-06-02

    Outer membrane vesicles (OMVs) are released from the outer membrane of many Gram-negative bacteria. These extracellular compartments are known to transport compounds involved in cell-cell signalling as well as virulence associated proteins, e.g. the cytolysine from enterotoxic E. coli. We have demonstrated that Xanthomonas campestris pv. campestris (Xcc) releases OMVs into the culture supernatant during growth. A proteome study identified 31 different proteins that associate with the OMV fraction of which half are virulence-associated. A comparison with the most abundant outer membrane (OM) proteins revealed that some proteins are enriched in the OMV fraction. This may be connected to differences in the LPS composition between the OMVs and the OM. Furthermore, a comparison of the OMV proteomes from two different culture media indicated that the culture conditions have an impact on the protein composition. Interestingly, the proteins that are common to both culture conditions are mainly involved in virulence. Outer membrane vesicles released from the OM of Xcc contain membrane- and virulence-associated proteins. Future experiments will prove whether these structures can serve as "vehicles" for the transport of virulence factors into the host membrane.

  20. pigB determines a diffusible factor needed for extracellular polysaccharide slime and xanthomonadin production in Xanthomonas campestris pv. campestris.

    PubMed

    Poplawsky, A R; Chun, W

    1997-01-01

    Seven xanthomonadin transcriptional units (pigA through pigG) were identified by transposon saturation mutagenesis within an 18.6-kbp portion of the previously identified 25.4-kbp pig region from Xanthomonas campestris pv. campestris (strain B-24). Since marker exchange mutant strains with insertions in one 3.7-kbp portion of pig could not be obtained, mutations in this region may be lethal to the bacterium. Complementation analyses with different insertion mutations further defined and confirmed the seven transcriptional units. Insertional inactivation of one of the transcriptional units, pigB, resulted in greatly reduced levels of both xanthomonadins and extracellular polysaccharide slime, and a pigB-encoding plasmid restored both traits to these strains. pigB mutant strains could also be restored extracellularly by growth adjacent to strains with insertion mutations in any of the other six xanthomonadin transcriptional units, the parent strain (B-24), or strains of five different species of Xanthomonas. Strain B-24 produced a nontransforming diffusible factor (DF), which could be restored to pigB mutants by the pigB-encoding plasmid. Several lines of evidence indicate that DF is a novel bacterial pheromone, different from the known signal molecules of Vibrio, Agrobacterium, Erwinia, Pseudomonas, and Burkholderia spp.

  1. Genetic and proteomic analyses of a Xanthomonas campestris pv. campestris purC mutant deficient in purine biosynthesis and virulence.

    PubMed

    Yuan, Zhihui; Wang, Li; Sun, Shutao; Wu, Yao; Qian, Wei

    2013-09-20

    Bacterial proliferation in hosts requires activation of a number of housekeeping pathways, including purine de novo biosynthesis. Although inactivation of purine biosynthesis genes can attenuate virulence, it is unclear which biochemical or virulence factors are associated with the purine biosynthesis pathway in vivo. We report that inactivation of purC, a gene encoding phosphoribosylaminoimidazole-succinocarboxamide synthase, caused complete loss of virulence in Xanthomonas campestris pv. campestris, the causal agent of black rot disease of cruciferous plants. The purC mutant was a purine auxotroph; it could not grow on minimal medium, whereas addition of purine derivatives, such as hypoxanthine or adenine plus guanine, restored growth of the mutant. The purC mutation also significantly enhanced the production of an unknown purine synthesis associated pigment and extracellular polysaccharides by the bacterium. In addition, comparative proteomic analyses of bacteria grown on rich and minimal media revealed that the purC mutation affected the expression levels of diverse proteins involved in purine and pyrimidine synthesis, carbon and energy metabolisms, iron uptake, proteolysis, protein secretion, and signal transduction. These results provided clues to understanding the contributions of purine synthesis to bacterial virulence and interactions with host immune systems.

  2. Identification of a novel type III secretion-associated outer membrane-bound protein from Xanthomonas campestris pv. campestris.

    PubMed

    Li, Lei; Li, Rui-Fang; Ming, Zhen-Hua; Lu, Guang-Tao; Tang, Ji-Liang

    2017-02-15

    Many bacterial pathogens employ the type III secretion system (T3SS) to translocate effector proteins into eukaryotic cells to overcome host defenses. To date, most of our knowledge about the T3SS molecular architecture comes from the studies on animal pathogens. In plant pathogens, nine Hrc proteins are believed to be structural components of the T3SS, of which HrcC and HrcJ form the outer and inner rings of the T3SS, respectively. Here, we demonstrated that a novel outer membrane-bound protein (HpaM) of Xanthomonas campestris pv. campestris is critical for the type III secretion and is structurally and functionally conserved in phytopathogenic Xanthomonas spp. We showed that the C-terminus of HpaM extends into the periplasm to interact physically with HrcJ and the middle part of HpaM interacts physically with HrcC. It is clear that the outer and inner rings compose the main basal body of the T3SS apparatus in animal pathogens. Therefore, we presume that HpaM may act as a T3SS structural component, or play a role in assisting assembling or affecting the stability of the T3SS apparatus. HpaM is a highly prevalent and specific protein in Xanthomonas spp., suggesting that the T3SS of Xanthomonas is distinctive in some aspects from other pathogens.

  3. Transcriptional reprogramming and phenotypical changes associated with growth of Xanthomonas campestris pv. campestris in cabbage xylem sap.

    PubMed

    Dugé de Bernonville, Thomas; Noël, Laurent D; SanCristobal, Magali; Danoun, Saida; Becker, Anke; Soreau, Paul; Arlat, Matthieu; Lauber, Emmanuelle

    2014-09-01

    Xylem sap (XS) is the first environment that xylem phytopathogens meet in planta during the early infection steps. Xanthomonas campestris pv. campestris (Xcc), the causative agent of Brassicaceae black rot, colonizes the plant xylem vessels to ensure its multiplication and dissemination. Besides suppression of plant immunity, Xcc has to adapt its metabolism to exploit plant-derived nutrients present in XS. To study Xcc behaviour in the early infection steps, we used cabbage XS to analyse bacterial growth. Mineral and organic composition of XS were determined. Significant growth of Xcc in XS was allowed by the rapid catabolism of amino acids, sugars and organic acids, and it was accompanied by the formation of biofilm-like structures. Transcriptome analysis of Xcc cultivated in XS using cDNA microarrays revealed a XS-specific transcriptional reprogramming compared to minimal or rich media. More specifically, up-regulation of genes encoding transporters such as TonB-dependent transporters (TBDTs), that could be associated with nutrient acquisition and detoxification, was observed. In agreement with the aggregation phenotype, expression of genes important for twitching motility and adhesion was up-regulated in XS. Taken together, our data show specific responses of Xcc to colonization of cabbage XS that could be important for the pathogenesis process and establish XS as a model medium to study mechanisms important for the early infection events.

  4. Transcriptional analysis and functional characterization of XCC1294 gene encoding a GGDEF domain protein in Xanthomonas campestris pv. campestris.

    PubMed

    Hsiao, Yi-Min; Song, Wan-Ling; Liao, Chao-Tsai; Lin, I-Hsuan; Pan, Mei-Ying; Lin, Ching-Fen

    2012-04-01

    The nucleotide cyclic di-GMP is a second messenger in bacteria that regulates a range of cellular functions including the virulence of pathogens. GGDEF is a protein domain involved in the synthesis of cyclic di-GMP. The genome of the crucifer pathogen Xanthomonas campestris pv. campestris (Xcc) encodes 21 proteins with a GGDEF domain. Clp, a homolog of the model transcription factor Crp of Escherichia coli, is a global regulator in Xcc. The aim of this study is to identify genes encoding GGDEF domain proteins whose expression is regulated by Clp. Results of reporter assay and RT-PCR analysis suggested that Clp regulates the expression of a set of genes encoding proteins harboring GGDEF domain. The transcription initiation site of XCC1294, one of the Clp regulated gene encoding a GGDEF domain protein, was mapped. Promoter analysis and gel retardation assay indicated that the transcription of XCC1294 is positively and directly regulated by Clp. Furthermore, transcription of XCC1294 was subject to catabolite repression and affected by several stress conditions. We also showed that mutation of XCC1294 results in enhanced surface attachment. In addition, transcription of three putative adhesin genes (xadA, fhaC, and yapH) was increased in the XCC1294 mutant. Taken together, the data presented here indicate that Clp positively regulates expression of XCC1294, and that XCC1294 serves a regulator of bacterial attachment and regulates different adhesin genes expression.

  5. pigB determines a diffusible factor needed for extracellular polysaccharide slime and xanthomonadin production in Xanthomonas campestris pv. campestris.

    PubMed Central

    Poplawsky, A R; Chun, W

    1997-01-01

    Seven xanthomonadin transcriptional units (pigA through pigG) were identified by transposon saturation mutagenesis within an 18.6-kbp portion of the previously identified 25.4-kbp pig region from Xanthomonas campestris pv. campestris (strain B-24). Since marker exchange mutant strains with insertions in one 3.7-kbp portion of pig could not be obtained, mutations in this region may be lethal to the bacterium. Complementation analyses with different insertion mutations further defined and confirmed the seven transcriptional units. Insertional inactivation of one of the transcriptional units, pigB, resulted in greatly reduced levels of both xanthomonadins and extracellular polysaccharide slime, and a pigB-encoding plasmid restored both traits to these strains. pigB mutant strains could also be restored extracellularly by growth adjacent to strains with insertion mutations in any of the other six xanthomonadin transcriptional units, the parent strain (B-24), or strains of five different species of Xanthomonas. Strain B-24 produced a nontransforming diffusible factor (DF), which could be restored to pigB mutants by the pigB-encoding plasmid. Several lines of evidence indicate that DF is a novel bacterial pheromone, different from the known signal molecules of Vibrio, Agrobacterium, Erwinia, Pseudomonas, and Burkholderia spp. PMID:8990296

  6. Copper ions potentiate organic hydroperoxide and hydrogen peroxide toxicity through different mechanisms in Xanthomonas campestris pv. campestris.

    PubMed

    Patikarnmonthon, Nisa; Nawapan, Sirikan; Buranajitpakorn, Sarinya; Charoenlap, Nisanart; Mongkolsuk, Skorn; Vattanaviboon, Paiboon

    2010-12-01

    Copper (Cu)-based biocides are important chemical controls for both fungal and bacterial diseases in crop fields. Here, we showed that Cu ions at a concentration of 100 μM enhanced t-butyl hydroperoxide (tBOOH) and hydrogen peroxide (H(2) O(2) ) killing of Xanthomonas campestris pv. campestris through different mechanisms. The addition of an antilipid peroxidation agent (α-tocopherol) and hydroxyl radical scavengers (glycerol and dimethyl sulphoxide) partially protected the bacteria from the Cu-enhanced tBOOH and H(2) O(2) killing, respectively. Inactivation of the alkyl hydroperoxide reductase gene rendered the mutant vulnerable to lethal doses of copper sulphate, which could be alleviated by the addition of an H(2) O(2) scavenger (pyruvate) and α-tocopherol. Taken together, the data suggest that Cu ions influence the killing effect of tBOOH through the stimulation of lipid peroxidation, while hydroxyl radical production is the underlying mechanism responsible for the Cu-ion-enhanced H(2) O(2) killing effects.

  7. Comparative and functional genomic analyses of the pathogenicity of phytopathogen Xanthomonas campestris pv. campestris

    PubMed Central

    Qian, Wei; Jia, Yantao; Ren, Shuang-Xi; He, Yong-Qiang; Feng, Jia-Xun; Lu, Ling-Feng; Sun, Qihong; Ying, Ge; Tang, Dong-Jie; Tang, Hua; Wu, Wei; Hao, Pei; Wang, Lifeng; Jiang, Bo-Le; Zeng, Shenyan; Gu, Wen-Yi; Lu, Gang; Rong, Li; Tian, Yingchuan; Yao, Zhijian; Fu, Gang; Chen, Baoshan; Fang, Rongxiang; Qiang, Boqin; Chen, Zhu; Zhao, Guo-Ping; Tang, Ji-Liang; He, Chaozu

    2005-01-01

    Xanthomonas campestris pathovar campestris (Xcc) is the causative agent of crucifer black rot disease, which causes severe losses in agricultural yield world-wide. This bacterium is a model organism for studying plant-bacteria interactions. We sequenced the complete genome of Xcc 8004 (5,148,708 bp), which is highly conserved relative to that of Xcc ATCC 33913. Comparative genomics analysis indicated that, in addition to a significant genomic-scale rearrangement cross the replication axis between two IS1478 elements, loss and acquisition of blocks of genes, rather than point mutations, constitute the main genetic variation between the two Xcc strains. Screening of a high-density transposon insertional mutant library (16,512 clones) of Xcc 8004 against a host plant (Brassica oleraceae) identified 75 nonredundant, single-copy insertions in protein-coding sequences (CDSs) and intergenic regions. In addition to known virulence factors, full virulence was found to require several additional metabolic pathways and regulatory systems, such as fatty acid degradation, type IV secretion system, cell signaling, and amino acids and nucleotide metabolism. Among the identified pathogenicity-related genes, three of unknown function were found in Xcc 8004-specific chromosomal segments, revealing a direct correlation between genomic dynamics and Xcc virulence. The present combination of comparative and functional genomic analyses provides valuable information about the genetic basis of Xcc pathogenicity, which may offer novel insight toward the development of efficient methods for prevention of this important plant disease. PMID:15899963

  8. Comparative and functional genomic analyses of the pathogenicity of phytopathogen Xanthomonas campestris pv. campestris.

    PubMed

    Qian, Wei; Jia, Yantao; Ren, Shuang-Xi; He, Yong-Qiang; Feng, Jia-Xun; Lu, Ling-Feng; Sun, Qihong; Ying, Ge; Tang, Dong-Jie; Tang, Hua; Wu, Wei; Hao, Pei; Wang, Lifeng; Jiang, Bo-Le; Zeng, Shenyan; Gu, Wen-Yi; Lu, Gang; Rong, Li; Tian, Yingchuan; Yao, Zhijian; Fu, Gang; Chen, Baoshan; Fang, Rongxiang; Qiang, Boqin; Chen, Zhu; Zhao, Guo-Ping; Tang, Ji-Liang; He, Chaozu

    2005-06-01

    Xanthomonas campestris pathovar campestris (Xcc) is the causative agent of crucifer black rot disease, which causes severe losses in agricultural yield world-wide. This bacterium is a model organism for studying plant-bacteria interactions. We sequenced the complete genome of Xcc 8004 (5,148,708 bp), which is highly conserved relative to that of Xcc ATCC 33913. Comparative genomics analysis indicated that, in addition to a significant genomic-scale rearrangement cross the replication axis between two IS1478 elements, loss and acquisition of blocks of genes, rather than point mutations, constitute the main genetic variation between the two Xcc strains. Screening of a high-density transposon insertional mutant library (16,512 clones) of Xcc 8004 against a host plant (Brassica oleraceae) identified 75 nonredundant, single-copy insertions in protein-coding sequences (CDSs) and intergenic regions. In addition to known virulence factors, full virulence was found to require several additional metabolic pathways and regulatory systems, such as fatty acid degradation, type IV secretion system, cell signaling, and amino acids and nucleotide metabolism. Among the identified pathogenicity-related genes, three of unknown function were found in Xcc 8004-specific chromosomal segments, revealing a direct correlation between genomic dynamics and Xcc virulence. The present combination of comparative and functional genomic analyses provides valuable information about the genetic basis of Xcc pathogenicity, which may offer novel insight toward the development of efficient methods for prevention of this important plant disease.

  9. Characterization of the pyrophosphate-dependent 6-phosphofructokinase from Xanthomonas campestris pv. campestris.

    PubMed

    Frese, Marcel; Schatschneider, Sarah; Voss, Julia; Vorhölter, Frank-Jörg; Niehaus, Karsten

    2014-03-15

    Xanthomonads are plant pathogenic proteobacteria that produce the polysaccharide xanthan. They are assumed to catabolize glucose mainly via the Entner-Doudoroff pathway. Whereas previous studies have demonstrated no phosphofructokinase (PFK) activity in xanthomonads, detailed genome analysis revealed in Xanthomonas campestris pathovar campestris (Xcc) genes for all Embden-Meyerhof-Parnas pathway (glycolysis) enzymes, including a conserved pfkA gene similar to 6-phosphofructokinase genes. To address this discrepancy between genetic and physiological properties, the pfkA gene of Xcc strain B100 was cloned into the expression vector pET28a+. The 45-kDa pfkA gene product exhibited no conventional PFK activity. Bioinformatic analysis of the Xcc PfkA amino acid sequence suggested utilization of pyrophosphate as an alternative cosubstrate. Pyrophosphate-dependent PFK activity was shown in an in vitro enzyme assay for purified Xcc PfkA, as well as in the Xcc B100 crude protein extract. Kinetic constants were determined for the forward and reverse reactions. Primary structure conservation indicates the global presence of similar enzymes among Xanthomonadaceae.

  10. New Culture Medium to Xanthan Production by Xanthomonas campestris pv. campestris.

    PubMed

    Carignatto, Cíntia Regina Rodrigues; Oliveira, Kassandra Sussi Mustafé; de Lima, Valéria Marta Gomes; de Oliva Neto, Pedro

    2011-07-01

    Xanthan is a important biopolymer for commercial purpose and it is produced in two stages by Xanthomonas campestris. In the first one, the bacterium is cultivated in the complex medium enriched in nitrogen and the biomass produced is used as inoculum for the next stage in which the gum is produced in another medium. In this work a new medium for the first stage is proposed in place of currently used YM medium. Different formulated growth media were studied and the correspondent biomass produced was analysed as inoculum for the second stage. The inoculum and gum were produced by batch process in shaker at 27°C in pH 6.0 and at 30°C in pH 7.0, respectively. The gum was precipitated with ethanol (3:1 v/v). The dryed biomass and xathan gum produced were determined by drying in oven at 105 and 40°C, respectively. The viscosity of the fermentation broth and 1% gum solution in water were determined in Brookfield viscometer. The formulated medium presented the increase in gum production (30%), broth (136%) and 1% gum solution viscosity (60%) compared to YM, besides the inferior cost. The results showed the importance of the quality of the inoculum from the first stage of the culture which influenced on the gum viscosity in the second stage.

  11. [Putative promoter region of type III effector gene avrAC(Xcc8004) in Xanthomonas campestris pv. campestris].

    PubMed

    Jiang, Guofeng; Wu, Qiuju; Liang, Xiaoxia; Yang, Lichao; Yang, Liyan; Wang, Lin; Wu, Xiaojian; Jiang, Bole

    2014-02-04

    Xanthomonas campestris pv. campestris (Xcc) is the cause agent of black rot of crucifers. Xcc uses type III secretion system (T3SS) to deliver T3SS effectors (T3SEs) directly into host cells, where they play important roles in pathogenesis. Many identified T3SEs genes contain plant-inducible promoter(PIP) box and -10 box in their promoter regions. However, the relation among PIP-box, -10 box and -10 region, -35 region of the classic promoter is unclear, and the conservative characteristic of -10 box sequence is hardly reported. The aim of this study was to analyze the putative promoter region of T3SE gene avrAC(Xcc8004). Through 5' RACE, the transcriptional start site of avrAC(Xcc8004) was identified. Fusion PCR was introduced to generate the site-mutagenesis of -10 box for constructing the GUS fusion report strains. The 5' RACE results indicate that the transcription start site was A. After analysis, we found that -35 region was located 8 bp downstream of PIP-box, and -10 box was exactly overlapped with -10 region. The whole motif of PIP-box, -35 region, and -10 box was then counted as: TTCAC-N15-TTCGC-N8-TTGATG-N18-TACGTT. The GUS assay results demonstrate that the site-mutagenesis of -10 box caused a higher expression of avrAC(Xcc8004). The GUS activities in the mutant strains delta hrpX and delta hrpG were significantly lower than that in the wild type Xcc strains. PIP-box is tandem with -35 region, -10 box is just the same as -10 region, -10 box is important for the transcription of avrAC(Xcc8004), and HrpG and HrpX activate the expression of avrAC(X8004), despite of -10 box site-mutagenesis.

  12. The bifunctional effector AvrXccC of Xanthomonas campestris pv. campestris requires plasma membrane-anchoring for host recognition.

    PubMed

    Wang, Lifeng; Tang, Xiaoyan; He, Chaozu

    2007-07-01

    Bacterial pathogens use type III secretion systems (TTSS) to deliver effector proteins into eukaryotic cells for pathogenesis. In bacterial-plant interactions, one effector may function as an avirulence factor to betray the pathogen to the plant surveillance system and induce the hypersensitive response (HR) in the resistant host carrying a corresponding resistance (R) gene. However, the same effector can also sustain the growth of the pathogen by acting as a virulence factor to modulate plant physiology in the susceptible host lacking the corresponding R gene. Here, we identified and characterized a bifunctional TTSS effector AvrXccC belonging to the AvrB effector family in Xanthomonas campestris pv. campestris 8004. This effector is required for full bacterial virulence in the susceptible host cabbage (Brassica oleracea) and avirulence in the resistant host mustard (Brassica napiformis L.H. Baily). Expressing avrXccC in mustard-virulent strain Xcc HRI 3849A converts its virulence to avirulence. The effector AvrXccC is anchored to the plant plasma membrane, and the N-terminal myristoylation site (amino acids 2-7: GLcaSK) is essential for its localization. In addition, the avirulence function of AvrXccC for host recognition depends on its plasma membrane localization. Promoter activity assays showed that the expression of avrXccC is hrpG/hrpX-dependent. Moreover, the secretion of AvrXccC displayed hrp-dependency and the core sequence for AvrXccC translocation was defined to the N-terminal 40 amino acids.

  13. Two isocitrate dehydrogenases from a plant pathogen Xanthomonas campestris pv. campestris 8004. Bioinformatic analysis, enzymatic characterization, and implication in virulence.

    PubMed

    Lv, Changqi; Wang, Peng; Wang, Wencai; Su, Ruirui; Ge, Yadong; Zhu, Youming; Zhu, Guoping

    2016-09-01

    Isocitrate dehydrogenase (IDH) is a key enzyme in the tricarboxylate (TCA) cycle, which may play an important role in the virulence of pathogenic bacteria. Here, two structurally different IDHs from a plant pathogen Xanthomonas campestris pv. campestris 8004 (XccIDH1 and XccIDH2) were characterized in detail. The recombinant XccIDH1 forms homodimer in solution, while the recombinant XccIDH2 is a typical monomer. Phylogenetic analysis showed that XccIDH1 belongs to the type I IDH subfamily and XccIDH2 groups into the monomeric IDH clade. Kinetic characterization demonstrated that XccIDH1's specificity towards NAD(+) was 110-fold greater than NADP(+) , while XccIDH2's specificity towards NADP(+) was 353-fold greater than NAD(+) . The putative coenzyme discriminating amino acids (Asp268, Ile269 and Ala275 for XccIDH1, and Lys589, His590 and Arg601 for XccIDH2) were studied by site-directed mutagenesis. The coenzyme specificities of the two mutants, mXccIDH1 and mXccIDH2, were completely reversed from NAD(+) to NADP(+) , and NADP(+) to NAD(+) , respectively. Furthermore, Ser80 of XccIDH1, and Lys256 and Tyr421 of XccIDH2, were the determinants for the substrate binding. The detailed biochemical properties, such as optimal pH and temperature, thermostability, and metal ion effects, of XccIDH1 and XccIDH2 were further investigated. The possibility of taking the two IDHs into consideration as the targets for drug development to control the plant diseases caused by Xcc 8004 were described and discussed thoroughly.

  14. Identification and Regulation of the N-Acetylglucosamine Utilization Pathway of the Plant Pathogenic Bacterium Xanthomonas campestris pv. campestris ▿ †

    PubMed Central

    Boulanger, Alice; Déjean, Guillaume; Lautier, Martine; Glories, Marie; Zischek, Claudine; Arlat, Matthieu; Lauber, Emmanuelle

    2010-01-01

    Xanthomonas campestris pv. campestris, the causal agent of black rot disease of brassicas, is known for its ability to catabolize a wide range of plant compounds. This ability is correlated with the presence of specific carbohydrate utilization loci containing TonB-dependent transporters (CUT loci) devoted to scavenging specific carbohydrates. In this study, we demonstrate that there is an X. campestris pv. campestris CUT system involved in the import and catabolism of N-acetylglucosamine (GlcNAc). Expression of genes belonging to this GlcNAc CUT system is under the control of GlcNAc via the LacI family NagR and GntR family NagQ regulators. Analysis of the NagR and NagQ regulons confirmed that GlcNAc utilization involves NagA and NagB-II enzymes responsible for the conversion of GlcNAc-6-phosphate to fructose-6-phosphate. Mutants with mutations in the corresponding genes are sensitive to GlcNAc, as previously reported for Escherichia coli. This GlcNAc sensitivity and analysis of the NagQ and NagR regulons were used to dissect the X. campestris pv. campestris GlcNAc utilization pathway. This analysis revealed specific features, including the fact that uptake of GlcNAc through the inner membrane occurs via a major facilitator superfamily transporter and the fact that this amino sugar is phosphorylated by two proteins belonging to the glucokinase family, NagK-IIA and NagK-IIB. However, NagK-IIA seems to play a more important role in GlcNAc utilization than NagK-IIB under our experimental conditions. The X. campestris pv. campestris GlcNAc NagR regulon includes four genes encoding TonB-dependent active transporters (TBDTs). However, the results of transport experiments suggest that GlcNAc passively diffuses through the bacterial envelope, an observation that calls into question whether GlcNAc is a natural substrate for these TBDTs and consequently is the source of GlcNAc for this nonchitinolytic plant-associated bacterium. PMID:20081036

  15. An oxidoreductase is involved in cercosporin degradation by the bacterium Xanthomonas campestris pv. zinniae.

    PubMed

    Taylor, Tanya V; Mitchell, Thomas K; Daub, Margaret E

    2006-09-01

    The polyketide toxin cercosporin plays a key role in pathogenesis by fungal species of the genus Cercospora. The bacterium Xanthomonas campestris pv. zinniae is able to rapidly degrade this toxin. Growth of X. campestris pv. zinniae strains in cercosporin-containing medium leads to the breakdown of cercosporin and to the formation of xanosporic acid, a nontoxic breakdown product. Five non-cercosporin-degrading mutants of a strain that rapidly degrades cercosporin (XCZ-3) were generated by ethyl methanesulfonate mutagenesis and were then transformed with a genomic library from the wild-type strain. All five mutants were complemented with the same genomic clone, which encoded a putative transcriptional regulator and an oxidoreductase. Simultaneous expression of these two genes was necessary to complement the mutant phenotype. Sequence analysis of the mutants showed that all five mutants had point mutations in the oxidoreductase gene and no mutations in the regulator. Quantitative reverse transcription-PCR (RT-PCR) showed that the expression of both of these genes in the wild-type strain is upregulated after exposure to cercosporin. Both the oxidoreductase and transcriptional regulator genes were transformed into three non-cercosporin-degrading bacteria to determine if they are sufficient for cercosporin degradation. Quantitative RT-PCR analysis confirmed that the oxidoreductase was expressed in all transconjugants. However, none of the transconjugants were able to degrade cercosporin, suggesting that additional factors are required for cercosporin degradation. Further study of cercosporin degradation in X. campestris pv. zinniae may allow for the engineering of Cercospora-resistant plants by using a suite of genes.

  16. Influence of agitation and aeration in xanthan production by Xanthomonas campestris pv pruni strain 101.

    PubMed

    Borges, C D; da Moreira, A S; Vendruscolo, C T; Ayub, M A Z

    2008-01-01

    Production, viscosity, and chemical composition of xanthan synthesized by bacterium Xanthomonas campestris pv pruni strain 101 were evaluated in bioreactor systems. During the process, the volumetric oxygen mass transfer coefficient (k(L)a) and the biomass were determined and the pH was monitored. The cultures were grown in a 3 I bioreactor, with aeration and agitation varying as follows: conditions (A) 300 rpm, 3 vvm and (B) 200 rpm, 2 vvm, at 28 degrees C. Our results showed that gum production was dependent on k(L)a, with a maximum yield of 8.15 g/l at 300 rpm, 3 vvm, 54 h of fermentation, k(L)a 21.4/h, while biomass was not affected. All aqueous solutions of 3% (w/v) xanthans synthesized showed a pseudoplastic behavior. The highest viscosity was reached under the strongest aeration/agitation conditions. All xanthan samples contained glucose, mannose, rhamnose, and glucuronic acid as their main components. The highest agitation and aeration rates used under condition A (300 rpm and 3 vvm) favorably influenced the yield and viscosity of the xanthan produced by bacterium X. campestris pv pruni 101 at different fermentation times.

  17. The inheritance of resistance to bacterial leaf spot of lettuce caused by Xanthomonas campestris pv. vitians in three lettuce cultivars

    USDA-ARS?s Scientific Manuscript database

    Lettuce yields can be reduced by the disease bacterial leaf spot (BLS) caused by the pathogen Xanthomonas campestris pv. vitians (Xcv) and host resistance is the most feasible method to reduce disease losses. The cultivars La Brillante, Pavane, and Little Gem express an incompatible host-pathogen in...

  18. Baby leaf lettuce germplasm enhancement: developing diverse populations with resistance to bacterial leaf spot caused by Xanthomonas campestris pv. vitians

    USDA-ARS?s Scientific Manuscript database

    Baby leaf lettuce cultivars with resistance to bacterial leaf spot (BLS) caused by Xanthomonas campestris pv. vitians (Xcv) are needed to reduce crop losses. The objectives of this research were to assess the genetic diversity for BLS resistance in baby leaf lettuce cultivars and to select early gen...

  19. Genetic Diversity of Lettuce (Lactuca sativa) for Resistance to Bacterial Leaf Spot Caused by Xanthomonas campestris pv. vitians.

    USDA-ARS?s Scientific Manuscript database

    Lettuce plants were artificially inoculated with three isolates of Xanthomonas campestris pv. vitians in field and greenhouse evaluations for genetic variation in resistance to bacterial leaf spot. The cultivar Little Gem had the least amount of disease, whether evaluated for disease severity or dis...

  20. Insights into the Extracytoplasmic Stress Response of Xanthomonas campestris pv. campestris: Role and Regulation of σE-Dependent Activity ▿ ‡

    PubMed Central

    Bordes, Patricia; Lavatine, Laure; Phok, Kounthéa; Barriot, Roland; Boulanger, Alice; Castanié-Cornet, Marie-Pierre; Déjean, Guillaume; Lauber, Emmanuelle; Becker, Anke; Arlat, Matthieu; Gutierrez, Claude

    2011-01-01

    Xanthomonas campestris pv. campestris is an epiphytic bacterium that can become a vascular pathogen responsible for black rot disease of crucifers. To adapt gene expression in response to ever-changing habitats, phytopathogenic bacteria have evolved signal transduction regulatory pathways, such as extracytoplasmic function (ECF) σ factors. The alternative sigma factor σE, encoded by rpoE, is crucial for envelope stress response and plays a role in the pathogenicity of many bacterial species. Here, we combine different approaches to investigate the role and mechanism of σE-dependent activation in X. campestris pv. campestris. We show that the rpoE gene is organized as a single transcription unit with the anti-σ gene rseA and the protease gene mucD and that rpoE transcription is autoregulated. rseA and mucD transcription is also controlled by a highly conserved σE-dependent promoter within the σE gene sequence. The σE-mediated stress response is required for stationary-phase survival, resistance to cadmium, and adaptation to membrane-perturbing stresses (elevated temperature and ethanol). Using microarray technology, we started to define the σE regulon of X. campestris pv. campestris. These genes encode proteins belonging to different classes, including periplasmic or membrane proteins, biosynthetic enzymes, classical heat shock proteins, and the heat stress σ factor σH. The consensus sequence for the predicted σE-regulated promoter elements is GGAACTN15-17GTCNNA. Determination of the rpoH transcription start site revealed that rpoH was directly regulated by σE under both normal and heat stress conditions. Finally, σE activity is regulated by the putative regulated intramembrane proteolysis (RIP) proteases RseP and DegS, as previously described in many other bacteria. However, our data suggest that RseP and DegS are not only dedicated to RseA cleavage and that the proteolytic cascade of RseA could involve other proteases. PMID:20971899

  1. Characterization of the hrpF pathogenicity peninsula of Xanthomonas oryzae pv. oryzae.

    PubMed

    Sugio, Akiko; Yang, Bing; White, Frank F

    2005-06-01

    The hrp gene cluster of Xanthomonas spp. contains genes for the assembly and function of a type III secretion system (TTSS). The hrpF genes reside in a region between hpaB and the right end of the hrp cluster. The region of the hrpF gene of Xanthomonas oryzae pv. oryzae is bounded by two IS elements and also contains a homolog of hpaF of X. campestris pv. vesicatoria and two newly identified genes, hpa3 and hpa4. A comparison of the hrp gene clusters of different species of Xanthomonas revealed that the hrpF region is a constant yet more variable peninsula of the hrp pathogenicity island. Mutations in hpaF, hpa3, and hpa4 had no effect on virulence, whereas hrpF mutants were severely reduced in virulence on susceptible rice cultivars. The hrpF genes from X. campestris pv. vesicatoria, X. campestris pv. campestris, and X. axonopodis pv. citri each were capable of restoring virulence to the hrpF mutant of X. oryzae pv. oryzae. Correspondingly, none of the Xanthomonas pathovars with hrpF from X. oryzae pv. oryzae elicited a hypersensitive reaction in their respective hosts. Therefore, no evidence was found for hrpF as a host-specialization factor. In contrast to the loss of Bs3-dependent reactions by hrpF mutants of X. campestris pv. vesicatoria, hrpF mutants of X. oryzae pv. oryzae with either avrXa10 or avrXa7 elicited hypersensitive reactions in rice cultivars with the corresponding R genes. A double hrpFxoo-hpa1 mutant also elicited an Xa10-dependent resistance reaction. Thus, loss of hrpF, hpal, or both may reduce delivery or effectiveness of type III effectors. However, the mutations did not completely prevent the delivery of effectors from X. oryzae pv. oryzae into the host cells.

  2. [RAPD-markers linked to the locus for resistance to the race 4 pathogen for black rot, Xanthomonas campestris pv. campestris (Pamm.) Dow., in Brassica rapa L].

    PubMed

    Ignatov, A N; Kuginuki, Y; Suprunova, T P; Pozmogova, G E; Seitova, A M; Dorokhov, D B; Hirai, M

    2000-03-01

    Association between the RAPD markers and the resistance to race 4 of the black rot causative agent was studied in Brassica rapa L. Experiments were carried out using doubled haploid lines, obtained via crosses between the race 4-susceptible fodder turnip and resistant pak-choi, and the F2 progeny of the crosses between the doubled haploid lines with contrasting resistance. The WE(22)980 RAPD marker inherited from the pak-choi and associated with the clubroot susceptibility was also linked to the locus responsible for the resistance to race 4 of Xanthomonas campestris pv. campestris. The two other RAPD markers were linked to susceptibility to black rot. Simultaneous association of the same DNA markers with the resistance/susceptibility to two different obligate pathogens favored the hypothesis on cluster organization of the resistance genes in plants. The markers described can be used in plant breeding and in further investigation of the genetic bases of resistance in plants.

  3. Expression of sweet pepper Hrap gene in banana enhances resistance to Xanthomonas campestris pv. musacearum.

    PubMed

    Tripathi, Leena; Mwaka, Henry; Tripathi, Jaindra Nath; Tushemereirwe, Wilberforce Kateera

    2010-11-01

    Banana Xanthomonas wilt (BXW), caused by the bacterium Xanthomonas campestris pv. musacearum, is the most devastating disease of banana in the Great Lakes region of Africa. The pathogen's rapid spread has threatened the livelihood of millions of Africans who rely on banana fruit for food security and income. The disease is very destructive, infecting all banana varieties, including both East African Highland bananas and exotic types of banana. In the absence of natural host plant resistance among banana cultivars, the constitutive expression of the hypersensitivity response-assisting protein (Hrap) gene from sweet pepper (Capsicum annuum) was evaluated for its ability to confer resistance to BXW. Transgenic lines expressing the Hrap gene under the regulation of the constitutive CaMV35S promoter were generated using embryogenic cell suspensions of two banana cultivars: 'Sukali Ndiizi' and 'Mpologoma'. These lines were characterized by molecular analysis, and were challenged with Xanthomonas campestris pv. musacearum to analyse the efficacy of the Hrap gene against BXW. The majority of transgenic lines (six of eight) expressing Hrap did not show any symptoms of infection after artificial inoculation of potted plants in the screenhouse, whereas control nontransgenic plants showed severe symptoms resulting in complete wilting. This study demonstrates that the constitutive expression of the sweet pepper Hrap gene in banana results in enhanced resistance to BXW. We describe the development of transgenic banana varieties resistant to BXW, which will boost the arsenal available to fight this epidemic disease and save livelihoods in the Great Lakes region of East and Central Africa. © 2010 IITA/NARO. Molecular Plant Pathology © 2010 BSPP and Blackwell Publishing Ltd.

  4. The genome of Xanthomonas campestris pv. campestris B100 and its use for the reconstruction of metabolic pathways involved in xanthan biosynthesis.

    PubMed

    Vorhölter, Frank-Jörg; Schneiker, Susanne; Goesmann, Alexander; Krause, Lutz; Bekel, Thomas; Kaiser, Olaf; Linke, Burkhard; Patschkowski, Thomas; Rückert, Christian; Schmid, Joachim; Sidhu, Vishaldeep Kaur; Sieber, Volker; Tauch, Andreas; Watt, Steven Alexander; Weisshaar, Bernd; Becker, Anke; Niehaus, Karsten; Pühler, Alfred

    2008-03-20

    The complete genome sequence of the Xanthomonas campestris pv. campestris strain B100 was established. It consisted of a chromosome of 5,079,003bp, with 4471 protein-coding genes and 62 RNA genes. Comparative genomics showed that the genes required for the synthesis of xanthan and xanthan precursors were highly conserved among three sequenced X. campestris pv. campestris genomes, but differed noticeably when compared to the remaining four Xanthomonas genomes available. For the xanthan biosynthesis genes gumB and gumK earlier translational starts were proposed, while gumI and gumL turned out to be unique with no homologues beyond the Xanthomonas genomes sequenced. From the genomic data the biosynthesis pathways for the production of the exopolysaccharide xanthan could be elucidated. The first step of this process is the uptake of sugars serving as carbon and energy sources wherefore genes for 15 carbohydrate import systems could be identified. Metabolic pathways playing a role for xanthan biosynthesis could be deduced from the annotated genome. These reconstructed pathways concerned the storage and metabolization of the imported sugars. The recognized sugar utilization pathways included the Entner-Doudoroff and the pentose phosphate pathway as well as the Embden-Meyerhof pathway (glycolysis). The reconstruction indicated that the nucleotide sugar precursors for xanthan can be converted from intermediates of the pentose phosphate pathway, some of which are also intermediates of glycolysis or the Entner-Doudoroff pathway. Xanthan biosynthesis requires in particular the nucleotide sugars UDP-glucose, UDP-glucuronate, and GDP-mannose, from which xanthan repeat units are built under the control of the gum genes. The updated genome annotation data allowed reconsidering and refining the mechanistic model for xanthan biosynthesis.

  5. Characterization of Xanthomonas campestris pv. campestris heat shock protein A (HspA), which possesses an intrinsic ability to reactivate inactivated proteins.

    PubMed

    Lin, Ching-Hsuan; Lee, Chia-Ni; Lin, Juey-Wen; Tsai, Wan-Ju; Wang, Szu-Wen; Weng, Shu-Fen; Tseng, Yi-Hsiung

    2010-10-01

    hspA encodes a small heat shock protein (sHSP) in Xanthomonas campestris pv. campestris, the causative agent of black rot in cruciferous plants. In this study, two-dimensional gel electrophoresis, promoter activity assays, and Northern hybridization results revealed that HspA expression was induced by heat shock but not by other stresses, although low-level expression was detectable by reverse transcription-polymerase chain reaction (RT-PCR) under normal culture conditions. An hspA mutant exhibited reduced tolerance to heat, especially in the presence of MgSO4, but no change in pathogenicity. Results of size-exclusion chromatography indicated that purified HspA(his), containing six C-terminal histidine residues, formed two different size classes of oligomeric complexes--410 and 820 kDa. In contrast, HspA(ter), the unmodified protein translated from the original hspA gene, formed only the 820-kDa complex. These results suggest that the C-terminus of HspA is important for oligomerization. Both HspA820(his) and HspA410(his) were able to partially protect luciferase against heat-induced aggregation. Unlike other reported sHSPs that commonly capture denaturing proteins in refoldable states until refolded by adenosine triphosphate-dependent chaperone systems, HspA(his) alone was capable of reactivating heat-inactivated EcoRI. Thus, Xanthomonas campestris pv. campestris HspA has potential application as a protective agent during the storage of proteins.

  6. Amplified fragment length polymorphism and multilocus sequence analysis-based genotypic relatedness among pathogenic variants of Xanthomonas citri pv. citri and Xanthomonas campestris pv. bilvae.

    PubMed

    Bui Thi Ngoc, Lan; Vernière, Christian; Jouen, Emmanuel; Ah-You, Nathalie; Lefeuvre, Pierre; Chiroleu, Frédéric; Gagnevin, Lionel; Pruvost, Olivier

    2010-03-01

    Three pathogenic variants (i.e. pathotypes) have been described within Xanthomonas citri pv. citri, the causal agent of Asiatic citrus canker. Pathotype A strains naturally infect a wide range of Citrus species and members of some related genera. In contrast, pathotypes A* and A(w) have narrow host ranges within the genus Citrus and have been isolated from Mexican lime (Citrus aurantifolia L.) and from Mexican lime and alemow (Citrus macrophylla L.), respectively. We used amplified fragment length polymorphism (AFLP) and multilocus sequence analysis (MLSA) based on four partial housekeeping gene sequences (atpD, dnaK, efp and gyrB ) for the genotypic classification of Xanthomonas citri pv. citri and the poorly characterized citrus pathogen Xanthomonas campestris pv. bilvae. A Mantel test showed that genetic distances derived from AFLP and MLSA were highly correlated. X. campestris pv. bilvae showed a close relatedness to the type strain of X. citri, indicating that this pathovar should be reclassified as X. citri pv. bilvae. All pathotype A* and A(w) strains were most closely related to X. citri pv. citri strains with a wide host range (pathotype A), confirming previous DNA-DNA hybridization data. Pathotype A(w) should be considered a junior synonym of pathotype A* on the basis of pathogenicity tests, AFLP, MLSA and PCR using pathovar-specific primers. Evolutionary genome divergences computed from AFLP data suggested that pathotype A* (including A(w) strains) is a group of strains that shows a wider genetic diversity than pathotype A.

  7. Structure of proline iminopeptidase from Xanthomonas campestris pv. citri: a prototype for the prolyl oligopeptidase family.

    PubMed

    Medrano, F J; Alonso, J; García, J L; Romero, A; Bode, W; Gomis-Rüth, F X

    1998-01-02

    The proline iminopeptidase from Xanthomonas campestris pv. citri is a serine peptidase that catalyses the removal of N-terminal proline residues from peptides with high specificity. We have solved its three-dimensional structure by multiple isomorphous replacement and refined it to a crystallographic R-factor of 19.2% using X-ray data to 2.7 A resolution. The protein is folded into two contiguous domains. The larger domain shows the general topology of the alpha/beta hydrolase fold, with a central eight-stranded beta-sheet flanked by two helices and the 11 N-terminal residues on one side, and by four helices on the other side. The smaller domain is placed on top of the larger domain and essentially consists of six helices. The active site, located at the end of a deep pocket at the interface between both domains, includes a catalytic triad of Ser110, Asp266 and His294. Cys269, located at the bottom of the active site very close to the catalytic triad, presumably accounts for the inhibition by thiol-specific reagents. The overall topology of this iminopeptidase is very similar to that of yeast serine carboxypeptidase. The striking secondary structure similarity to human lymphocytic prolyl oligopeptidase and dipeptidyl peptidase IV makes this proline iminopeptidase structure a suitable model for the three-dimensional structure of other peptidases of this family.

  8. Structure of proline iminopeptidase from Xanthomonas campestris pv. citri: a prototype for the prolyl oligopeptidase family.

    PubMed Central

    Medrano, F J; Alonso, J; García, J L; Romero, A; Bode, W; Gomis-Rüth, F X

    1998-01-01

    The proline iminopeptidase from Xanthomonas campestris pv. citri is a serine peptidase that catalyses the removal of N-terminal proline residues from peptides with high specificity. We have solved its three-dimensional structure by multiple isomorphous replacement and refined it to a crystallographic R-factor of 19.2% using X-ray data to 2.7 A resolution. The protein is folded into two contiguous domains. The larger domain shows the general topology of the alpha/beta hydrolase fold, with a central eight-stranded beta-sheet flanked by two helices and the 11 N-terminal residues on one side, and by four helices on the other side. The smaller domain is placed on top of the larger domain and essentially consists of six helices. The active site, located at the end of a deep pocket at the interface between both domains, includes a catalytic triad of Ser110, Asp266 and His294. Cys269, located at the bottom of the active site very close to the catalytic triad, presumably accounts for the inhibition by thiol-specific reagents. The overall topology of this iminopeptidase is very similar to that of yeast serine carboxypeptidase. The striking secondary structure similarity to human lymphocytic prolyl oligopeptidase and dipeptidyl peptidase IV makes this proline iminopeptidase structure a suitable model for the three-dimensional structure of other peptidases of this family. PMID:9427736

  9. Fatty acid DSF binds and allosterically activates histidine kinase RpfC of phytopathogenic bacterium Xanthomonas campestris pv. campestris to regulate quorum-sensing and virulence.

    PubMed

    Cai, Zhen; Yuan, Zhi-Hui; Zhang, Huan; Pan, Yue; Wu, Yao; Tian, Xiu-Qi; Wang, Fang-Fang; Wang, Li; Qian, Wei

    2017-04-01

    As well as their importance to nutrition, fatty acids (FA) represent a unique group of quorum sensing chemicals that modulate the behavior of bacterial population in virulence. However, the way in which full-length, membrane-bound receptors biochemically detect FA remains unclear. Here, we provide genetic, enzymological and biophysical evidences to demonstrate that in the phytopathogenic bacterium Xanthomonas campestris pv. campestris, a medium-chain FA diffusible signal factor (DSF) binds directly to the N-terminal, 22 amino acid-length sensor region of a receptor histidine kinase (HK), RpfC. The binding event remarkably activates RpfC autokinase activity by causing an allosteric change associated with the dimerization and histidine phosphotransfer (DHp) and catalytic ATP-binding (CA) domains. Six residues were found essential for sensing DSF, especially those located in the region adjoining to the inner membrane of cells. Disrupting direct DSF-RpfC interaction caused deficiency in bacterial virulence and biofilm development. In addition, two amino acids within the juxtamembrane domain of RpfC, Leu172 and Ala178, are involved in the autoinhibition of the RpfC kinase activity. Replacements of them caused constitutive activation of RpfC-mediated signaling regardless of DSF stimulation. Therefore, our results revealed a biochemical mechanism whereby FA activates bacterial HK in an allosteric manner, which will assist in future studies on the specificity of FA-HK recognition during bacterial virulence regulation and cell-cell communication.

  10. Conformational changes associated with the binding of zinc acetate at the putative active site of XcTcmJ, a cupin from Xanthomonas campestris pv. campestris

    PubMed Central

    Axelrod, Herbert L.; Kozbial, Piotr; McMullan, Daniel; Krishna, S. Sri; Miller, Mitchell D.; Abdubek, Polat; Acosta, Claire; Astakhova, Tamara; Carlton, Dennis; Caruthers, Jonathan; Chiu, Hsiu-Ju; Clayton, Thomas; Deller, Marc C.; Duan, Lian; Elias, Ylva; Feuerhelm, Julie; Grzechnik, Slawomir K.; Grant, Joanna C.; Han, Gye Won; Jaroszewski, Lukasz; Jin, Kevin K.; Klock, Heath E.; Knuth, Mark W.; Kumar, Abhinav; Marciano, David; Morse, Andrew T.; Murphy, Kevin D.; Nigoghossian, Edward; Okach, Linda; Oommachen, Silvya; Paulsen, Jessica; Reyes, Ron; Rife, Christopher L.; Tien, Henry J.; Trout, Christina V.; van den Bedem, Henry; Weekes, Dana; White, Aprilfawn; Xu, Qingping; Zubieta, Chloe; Hodgson, Keith O.; Wooley, John; Elsliger, Marc-André; Deacon, Ashley M.; Godzik, Adam; Lesley, Scott A.; Wilson, Ian A.

    2010-01-01

    In the plant pathogen Xanthomonas campestris pv. campestris, the product of the tcmJ gene, XcTcmJ, encodes a protein belonging to the RmlC family of cupins. XcTcmJ was crystallized in a monoclinic space group (C2) in the presence of zinc acetate and the structure was determined to 1.6 Å resolution. Previously, the apo structure has been reported in the absence of any bound metal ion [Chin et al. (2006 ▶), Proteins, 65, 1046–1050]. The most significant difference between the apo structure and the structure of XcTcmJ described here is a reorganization of the binding site for zinc acetate, which was most likely acquired from the crystallization solution. This site is located in the conserved metal ion-binding domain at the putative active site of XcTcmJ. In addition, an acetate was also bound within coordination distance of the zinc. In order to accommodate this binding, rearrangement of a conserved histidine ligand is required as well as several nearby residues within and around the putative active site. These observations indicate that binding of zinc serves a functional role in this cupin protein. PMID:20944231

  11. Conformational changes associated with the binding of zinc acetate at the putative active site of XcTcmJ, a cupin from Xanthomonas campestris pv. campestris.

    PubMed

    Axelrod, Herbert L; Kozbial, Piotr; McMullan, Daniel; Krishna, S Sri; Miller, Mitchell D; Abdubek, Polat; Acosta, Claire; Astakhova, Tamara; Carlton, Dennis; Caruthers, Jonathan; Chiu, Hsiu Ju; Clayton, Thomas; Deller, Marc C; Duan, Lian; Elias, Ylva; Feuerhelm, Julie; Grzechnik, Slawomir K; Grant, Joanna C; Han, Gye Won; Jaroszewski, Lukasz; Jin, Kevin K; Klock, Heath E; Knuth, Mark W; Kumar, Abhinav; Marciano, David; Morse, Andrew T; Murphy, Kevin D; Nigoghossian, Edward; Okach, Linda; Oommachen, Silvya; Paulsen, Jessica; Reyes, Ron; Rife, Christopher L; Tien, Henry J; Trout, Christina V; van den Bedem, Henry; Weekes, Dana; White, Aprilfawn; Xu, Qingping; Zubieta, Chloe; Hodgson, Keith O; Wooley, John; Elsliger, Marc André; Deacon, Ashley M; Godzik, Adam; Lesley, Scott A; Wilson, Ian A

    2010-10-01

    In the plant pathogen Xanthomonas campestris pv. campestris, the product of the tcmJ gene, XcTcmJ, encodes a protein belonging to the RmlC family of cupins. XcTcmJ was crystallized in a monoclinic space group (C2) in the presence of zinc acetate and the structure was determined to 1.6 Å resolution. Previously, the apo structure has been reported in the absence of any bound metal ion [Chin et al. (2006), Proteins, 65, 1046-1050]. The most significant difference between the apo structure and the structure of XcTcmJ described here is a reorganization of the binding site for zinc acetate, which was most likely acquired from the crystallization solution. This site is located in the conserved metal ion-binding domain at the putative active site of XcTcmJ. In addition, an acetate was also bound within coordination distance of the zinc. In order to accommodate this binding, rearrangement of a conserved histidine ligand is required as well as several nearby residues within and around the putative active site. These observations indicate that binding of zinc serves a functional role in this cupin protein.

  12. Fatty acid DSF binds and allosterically activates histidine kinase RpfC of phytopathogenic bacterium Xanthomonas campestris pv. campestris to regulate quorum-sensing and virulence

    PubMed Central

    Zhang, Huan; Pan, Yue; Wu, Yao; Tian, Xiu-Qi; Wang, Fang-Fang; Wang, Li

    2017-01-01

    As well as their importance to nutrition, fatty acids (FA) represent a unique group of quorum sensing chemicals that modulate the behavior of bacterial population in virulence. However, the way in which full-length, membrane-bound receptors biochemically detect FA remains unclear. Here, we provide genetic, enzymological and biophysical evidences to demonstrate that in the phytopathogenic bacterium Xanthomonas campestris pv. campestris, a medium-chain FA diffusible signal factor (DSF) binds directly to the N-terminal, 22 amino acid-length sensor region of a receptor histidine kinase (HK), RpfC. The binding event remarkably activates RpfC autokinase activity by causing an allosteric change associated with the dimerization and histidine phosphotransfer (DHp) and catalytic ATP-binding (CA) domains. Six residues were found essential for sensing DSF, especially those located in the region adjoining to the inner membrane of cells. Disrupting direct DSF-RpfC interaction caused deficiency in bacterial virulence and biofilm development. In addition, two amino acids within the juxtamembrane domain of RpfC, Leu172 and Ala178, are involved in the autoinhibition of the RpfC kinase activity. Replacements of them caused constitutive activation of RpfC-mediated signaling regardless of DSF stimulation. Therefore, our results revealed a biochemical mechanism whereby FA activates bacterial HK in an allosteric manner, which will assist in future studies on the specificity of FA-HK recognition during bacterial virulence regulation and cell-cell communication. PMID:28369120

  13. Differential accumulation of Xanthomonas campestris pv. campestris proteins during the interaction with the host plant: Contributions of an in vivo system.

    PubMed

    Santos, Cristiane; Maximiano, Mariana R; Ribeiro, Daiane G; Oliveira-Neto, Osmundo B; Murad, André M; Franco, Octávio L; Mehta, Angela

    2017-06-01

    Xanthomonas campestris pv. campestris (Xcc) is the causal agent of black rot, a highly destructive disease that affects all brassicas. This work aimed to study the interaction Xcc-Brassica oleracea using an in vivo system in an attempt to identify proteins involved in pathogenicity. We used label-free shotgun 2D-nanoUPLC/MS(E) to analyze Xcc proteins in three conditions: in the interaction with susceptible (REK) and resistant (REU) plants and in culture medium (control condition). A model of Xcc-susceptible host interaction is proposed and shows that Xcc increases the abundance of several crucial proteins for infection and cell protection. In this study, we also confirmed the differential expression by qPCR analysis of selected genes. This is the first report showing a large-scale identification of proteins in an in vivo host plant condition. Considering that most studies involving phytopathogens are in vitro (growth in culture medium or in plant extract), this work contributes with relevant information related to the plant-pathogen interaction in planta. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  14. A proline iminopeptidase gene upregulated in planta by a LuxR homologue is essential for pathogenicity of Xanthomonas campestris pv. campestris.

    PubMed

    Zhang, Lili; Jia, Yantao; Wang, Li; Fang, Rongxiang

    2007-07-01

    Expression of bacterial genes is often regulated by complex mechanisms, some of which involve host cues. Analysis of the Xanthomonas campestris pv. campestris (Xcc) genome sequence revealed the presence of an xccR/pip locus. The upstream gene xccR is a luxR homologue, while pip codes for a proline iminopeptidase. A lux box-like element, named luxXc box, locates in the pip promoter region. In this work, we show that disruption of either xccR or pip resulted in significantly attenuated virulence of Xcc. Under medium culture conditions, the pip expression was significantly enhanced by overexpression of XccR and the luxXc box is necessary for this enhancement. We further show that expression of a pip promoter-gusA fusion either inserted in the bacterial chromosome or resided in a plasmid was markedly induced when the bacteria grew in planta. Disruption of either xccR or the luxXc box abolished the in planta induction, while disruption of pip enhanced the induction. Taken together, these data demonstrate that pip is indispensable for Xcc virulence and suggest a model for Xcc-host interaction in which the pathogen senses some host factor(s) to activate XccR that subsequently interacts with the luxXc box to induce the expression of pip for facilitating Xcc infection.

  15. Functional characterization and transcriptome analysis reveal multiple roles for prc in the pathogenicity of the black rot pathogen Xanthomonas campestris pv. campestris.

    PubMed

    Liao, Chao-Tsai; Liu, Yu-Fan; Chiang, Ying-Chuan; Lo, Hsueh-Hsia; Du, Shin-Chiao; Hsu, Pei-Chi; Hsiao, Yi-Min

    2016-05-01

    Gram-negative phytopathogenic Xanthomonas campestris pv. campestris (Xcc) is the causal agent of black rot in crucifers. The ability of Xcc to incite this disease in plants depends on a number of factors, including exopolysaccharides, extracellular enzymes and biofilm production. In this study, transposon mutagenesis led to identification of the prc gene, encoding a tail-specific protease, which plays a role in Xcc pathogenesis. Mutation of prc resulted in decreased virulence, extracellular protease production and bacterial attachment, with restoration to the levels of wild type by the intact prc gene. From subsequent quantitative RT-PCR analysis and reporter assay, the major extracellular protease gene prt1, biofilm-related gene galE encoding a UDP-galactose 4-epimerase and two putative adhesin genes (yapH and XC_4290 encoding autotransporter-like protein H and hemagglutinin, respectively) were found to be reduced in the prc mutant. Results of transcriptome profiling of Xcc wild type and prc mutant by RNA sequencing (RNA-Seq) showed that mutation of prc in Xcc leads to alteration in the transcriptional levels (more than twofold) of 91 genes. These differentially expressed genes were associated with a wide range of biological functions such as carbohydrate transport and metabolism, cell wall/membrane biogenesis, posttranslational modification, protein turnover and chaperones, inorganic ion transport and metabolism and signal transduction mechanisms. The results of this study facilitate the functional understanding of and provide new information about the regulatory role of prc.

  16. Functional characterization and transcriptional analysis of galE gene encoding a UDP-galactose 4-epimerase in Xanthomonas campestris pv. campestris.

    PubMed

    Li, Chien-Te; Liao, Chao-Tsai; Du, Shin-Chiao; Hsiao, Yu-Ping; Lo, Hsueh-Hsia; Hsiao, Yi-Min

    2014-01-01

    The Gram-negative plant pathogen Xanthomonas campestris pv. campestris (Xcc) is the causative agent of black rot in crucifers, a disease that causes tremendous agricultural loss. In this study, the Xcc galE gene was characterized. Sequence and mutational analysis demonstrated that the Xcc galE encodes a UDP-galactose 4-epimerase (EC 5.1.3.2), which catalyzes the interconversion of UDP-galactose and UDP-glucose. Alanine substitution of the putative catalytic residues (Ser124, Tyr147, and Lys151) of GalE caused loss of epimerase activity. Further study showed that the Xcc galE mutant had reduced biofilm formation ability. Furthermore, reporter assays revealed that galE transcription exhibits a distinct expression profile under different culture conditions, is subject to catabolite repression, and is positively regulated by Clp and RpfF. In addition, the galE transcription initiation site was mapped. This is the first time that UDP-galactose 4-epimerase has been characterized in the crucifer pathogen Xcc.

  17. Systematic mutagenesis of all predicted gntR genes in Xanthomonas campestris pv. campestris reveals a GntR family transcriptional regulator controlling hypersensitive response and virulence.

    PubMed

    An, Shi-Qi; Lu, Guang-Tao; Su, Hui-Zhao; Li, Rui-Fang; He, Yong-Qiang; Jiang, Bo-Le; Tang, Dong-Jie; Tang, Ji-Liang

    2011-09-01

    The GntR family is one of the most abundant and widely distributed groups of helix-turn-helix transcriptional regulators in bacteria. Six open reading frames in the genome of the plant pathogen Xanthomonas campestris pv. campestris were predicted to encode GntR regulators. All six of the predicted GntR-encoding genes were individually mutagenized and mutants from five of them were successfully obtained. Plant disease response assays revealed that one, whose product belongs to the YtrA subfamily and has been named HpaR1, is involved in the hypersensitive response (HR) and virulence. Electrophoretic mobility shift assays and in vitro transcription assays revealed that HpaR1 could repress its own transcription level through binding to its promoter sequence, indicating an autoregulatory feedback inhibition mechanism for HpaR1 expression. Promoter-gusA reporter and reverse-transcription polymerase chain reaction analyses revealed that HpaR1 positively and negatively affects the expression of HR and pathogenicity (hrp) genes in host plant and standard media, respectively. Constitutive expression of the key hrp regulator, hrpG, in the hpaR1 mutant could bypass the requirement of HpaR1 for the induction of wild-type HR, suggesting that HpaR1 regulates the expression of hrp genes that encode the type III secretion system via hrpG.

  18. Establishment, in silico analysis, and experimental verification of a large-scale metabolic network of the xanthan producing Xanthomonas campestris pv. campestris strain B100.

    PubMed

    Schatschneider, Sarah; Persicke, Marcus; Watt, Steven Alexander; Hublik, Gerd; Pühler, Alfred; Niehaus, Karsten; Vorhölter, Frank-Jörg

    2013-08-20

    The γ-proteobacterium Xanthomonas campestris pv. campestris (Xcc) B100 synthesizes the polysaccharide xanthan, a commercially important viscosifier. Since the complete genome of Xcc B100 is available, systems biology tools were applied to obtain a deeper understanding of the metabolism involved in xanthan biosynthesis. A large-scale metabolic network was reconstructed and manually curated. The reconstructed network included 352 genes, 437 biochemical reactions, 10 transport reactions, and 338 internal metabolites. To use this network for flux balance analysis, the biomass composition of Xcc B100 was determined. The comprehensive model obtained was applied for in silico analyses to predict biomass generation and gene essentiality. Predictions were extensively validated by analyzing batch culture performance and by carbon balancing including xanthan production. Single gene deletion mutants causing deficiencies in the central carbohydrate metabolism were constructed to enforce major flux redistributions. The impact of xanthan production was studied in vivo and in silico, comparing the physiology of a gumD mutant, negative in xanthan production, with the original strain. The results indicate a redistribution of resources from xanthan to biomass, rather than a reduction in carbon uptake. With this high quality metabolic model, both systems biology analyses and synthetic biology reengineering of Xcc gained an important tool.

  19. The xylan utilization system of the plant pathogen Xanthomonas campestris pv campestris controls epiphytic life and reveals common features with oligotrophic bacteria and animal gut symbionts.

    PubMed

    Déjean, Guillaume; Blanvillain-Baufumé, Servane; Boulanger, Alice; Darrasse, Armelle; Dugé de Bernonville, Thomas; Girard, Anne-Laure; Carrére, Sébastien; Jamet, Stevie; Zischek, Claudine; Lautier, Martine; Solé, Magali; Büttner, Daniela; Jacques, Marie-Agnès; Lauber, Emmanuelle; Arlat, Matthieu

    2013-05-01

    Xylan is a major structural component of plant cell wall and the second most abundant plant polysaccharide in nature. Here, by combining genomic and functional analyses, we provide a comprehensive picture of xylan utilization by Xanthomonas campestris pv campestris (Xcc) and highlight its role in the adaptation of this epiphytic phytopathogen to the phyllosphere. The xylanolytic activity of Xcc depends on xylan-deconstruction enzymes but also on transporters, including two TonB-dependent outer membrane transporters (TBDTs) which belong to operons necessary for efficient growth in the presence of xylo-oligosaccharides and for optimal survival on plant leaves. Genes of this xylan utilization system are specifically induced by xylo-oligosaccharides and repressed by a LacI-family regulator named XylR. Part of the xylanolytic machinery of Xcc, including TBDT genes, displays a high degree of conservation with the xylose-regulon of the oligotrophic aquatic bacterium Caulobacter crescentus. Moreover, it shares common features, including the presence of TBDTs, with the xylan utilization systems of Bacteroides ovatus and Prevotella bryantii, two gut symbionts. These similarities and our results support an important role for TBDTs and xylan utilization systems for bacterial adaptation in the phyllosphere, oligotrophic environments and animal guts.

  20. Crystallization and preliminary X-ray characterization of the full-length bacteriophytochrome from the plant pathogen Xanthomonas campestris pv. campestris.

    PubMed

    Klinke, Sebastián; Otero, Lisandro H; Rinaldi, Jimena; Sosa, Santiago; Guimarães, Beatriz G; Shepard, William E; Goldbaum, Fernando A; Bonomi, Hernán R

    2014-12-01

    Phytochromes give rise to the largest photosensor family known to date. However, they are underrepresented in the Protein Data Bank. Plant, cyanobacterial, fungal and bacterial phytochromes share a canonical architecture consisting of an N-terminal photosensory module (PAS2-GAF-PHY domains) and a C-terminal variable output module. The bacterium Xanthomonas campestris pv. campestris, a worldwide agricultural pathogen, codes for a single bacteriophytochrome (XccBphP) that has this canonical architecture, bearing a C-terminal PAS9 domain as the output module. Full-length XccBphP was cloned, expressed and purified to homogeneity by nickel-NTA affinity and size-exclusion chromatography and was then crystallized at room temperature bound to its cofactor biliverdin. A complete native X-ray diffraction data set was collected to a maximum resolution of 3.25 Å. The crystals belonged to space group P43212, with unit-cell parameters a = b = 103.94, c = 344.57 Å and a dimer in the asymmetric unit. Refinement is underway after solving the structure by molecular replacement.

  1. Two non-consensus Clp binding sites are involved in upregulation of the gum operon involved in xanthan polysaccharide synthesis in Xanthomonas campestris pv. campestris.

    PubMed

    Chen, Chih-Hua; Lin, Nien-Tsung; Hsiao, Yi-Min; Yang, Chiou-Ying; Tseng, Yi-Hsiung

    2010-09-01

    Biosynthesis of xanthan polysaccharide, a virulence factor of phytopathogenic Xanthomonas campestris pv. campestris (Xcc), involves the gum operon and the cyclic AMP receptor protein (CRP) homologue Clp. Clp was shown to have the same DNA binding specificity as the CRP at positions 5, 6, and 7 (GTG motif) of the left arm. Therefore, Clp binding sites (CBSs) have typically been identified by pattern searching of the Xcc genome using the consensus CRP binding sequence. Here, results of a reporter assay and electrophoretic mobility shift assay suggest that Clp upregulates the gum operon by binding to two non-consensus sites, in which a more conserved right arm may compensate for the lack of conservation in the left arm, a high GC content in the central region (6 bp) may be important for binding, and binding may be enhanced if the GC-rich central region is palindromic. These suggest that atypical CBSs exist in Xcc promoters and that Clp, while retaining the capacity to bind typical CBSs, has evolved to bind atypical CBS because: 1) Clp shares only moderate homology with the CRP and is modulated by cyclic di-GMP; and 2) Xcc has a higher GC content (65%) than Escherichia coli (50%).

  2. Dynamic protein phosphorylation during the growth of Xanthomonas campestris pv. campestris B100 revealed by a gel-based proteomics approach.

    PubMed

    Musa, Yaarub Raji; Bäsell, Katrin; Schatschneider, Sarah; Vorhölter, Frank-Jörg; Becher, Dörte; Niehaus, Karsten

    2013-08-20

    Xanthomonas campestris pv. campestris (Xcc) synthesizes huge amounts of the exopolysaccharide xanthan and is a plant pathogen affecting Brassicaceae, among them the model plant Arabidopsis thaliana. Xanthan is produced as a thickening agent at industrial scale by fermentation of Xcc. In an approach based on 2D gel electrophoresis, protein samples from different growth phases were characterized to initialize analysis of the Xanthomonas phosphoproteome. The 2D gels were stained with Pro-Q Diamond phosphoprotein stain to identify putatively phosphorylated proteins. Spots of putatively phosphorylated proteins were excised from the gel and analyzed by mass spectrometry. Three proteins were confirmed to be phosphorylated, the phosphoglucomutase/phosphomannomutase XanA that is important for xanthan and lipopolysaccharide biosynthesis, the phosphoenolpyruvate synthase PspA that is involved in gluconeogenesis, and an anti-sigma factor antagonist RsbR that was so far uncharacterized in xanthomonads. The growth phase in which the samples were collected had an influence on protein phosphorylation in Xcc, particular distinct in case of RsbR, which was phosphorylated during the transition from the late exponential growth phase to the stationary phase.

  3. Loss of chloroplast localized protein phosphatase 2Cs in Arabidopsis thaliana leads to enhancement of the plant immunity and resistance to Xanthomonas campestris pv. campestris infection.

    PubMed

    Akimoto-Tomiyama, Chiharu; Tanabe, Shigeru; Kajiwara, Hideyuki; Minami, Eiichi; Ochiai, Hirokazu

    2017-08-16

    Protein phosphatases (PPs) counteract kinases in reversible phosphorylation events during numerous signal transduction pathways in eukaryotes. PP2Cs, one of the four major classes of serine/threonine specific PPs family is greatly expanded in plants. Thus PP2C is thought to have a specific role in a signal transduction pathway. Some rice PP2Cs classified to subgroup K were responsive to the infection by the compatible Xanthomonas oryzae pv. oryzae, the causal agent of bacterial blight. In A. thaliana, orthologous PP2C genes (AtPP2C62 and AtPP2C26) classified to subgroup K were also responsive to Xanthomonas campestris pv. campestris (Xcc, causal agent of the Black rot) infection. To elucidate the function of these subgroup K PP2Cs, atpp2c62 and atpp2c26 deficient A. thaliana mutants were characterized. A double mutant plant which was inoculated with a compatible Xcc showed reduced lesion development as well as the suppression of bacterial multiplication. AtPP2C62 and AtPP2C26 localized to the chloroplast. Furthermore, photosynthesis related proteins, chaperonin-60 was indicated as the potent candidate for de-phosphorylated substrate catalyzed by AtPP2C62 and AtPP2C26 using the 2D-IDF-SDS-PAGE method. Taken together, AtPP2C62 and AtPP2C26 are suggested to be involved in both photosynthesis and suppression of plant immune system. These results imply the occurrence of the crosstalk between photosynthesis and the plant defense system to control the productivity under pathogen infection. This article is protected by copyright. All rights reserved. © 2017 BSPP and John Wiley & Sons Ltd.

  4. The type III effector AvrXccB in Xanthomonas campestris pv. campestris targets putative methyltransferases and suppresses innate immunity in Arabidopsis.

    PubMed

    Liu, Lijuan; Wang, Yanping; Cui, Fuhao; Fang, Anfei; Wang, Shanzhi; Wang, Jiyang; Wei, Chao; Li, Shuai; Sun, Wenxian

    2016-05-31

    Xanthomonas campestris pv. campestris (Xcc) causes black rot, one of the most important diseases of brassica crops worldwide. The type III effector inventory plays important roles in the virulence and pathogenicity of the pathogen. However, little is known about the virulence function(s) of the putative type III effector AvrXccB in Xcc. Here, we investigated the immune suppression ability of AvrXccB and the possible underlying mechanisms. AvrXccB was demonstrated to be secreted in a type III secretion system-dependent manner. AvrXccB tagged with green fluorescent protein is localized to the plasma membrane in Arabidopsis, and the putative N-myristoylation motif is essential for its localization. Chemical-induced expression of AvrXccB suppresses flg22-triggered callose deposition and the oxidative burst, and promotes the in planta growth of Xcc and Pseudomonas syringae pv. tomato in transgenic Arabidopsis plants. The putative catalytic triad and plasma membrane localization of AvrXccB are required for its immunosuppressive activity. Furthermore, it was demonstrated that AvrXccB interacts with the Arabidopsis S-adenosyl-l-methionine-dependent methyltransferases SAM-MT1 and SAM-MT2. Interestingly, SAM-MT1 is not only self-associated, but also associated with SAM-MT2 in vivo. SAM-MT1 and SAM-MT2 expression is significantly induced upon stimulation of microbe-associated molecular patterns and bacterial infection. Collectively, these findings indicate that AvrXccB targets a putative methyltransferase complex and suppresses plant immunity.

  5. Interaction of Xanthomonas campestris with Arabidopsis thaliana: characterization of a gene from X. c. pv. raphani that confers avirulence to most A. thaliana accessions.

    PubMed

    Parker, J E; Barber, C E; Fan, M J; Daniels, M J

    1993-01-01

    Infiltration of leaves of Arabidopsis thaliana accession Columbia with Xanthomonas campestris pathovar campestris leads to bacterial growth and disease symptoms reminiscent of those incited in Brassica plants inoculated under the same conditions. A search among A. thaliana accessions for variation in the reaction phenotype to strains of X. campestris pathovars campestris, aberrans, and raphani showed that there were no clear differential responses between plant accessions to the individual bacterial strains tested. X. c. pv. raphani strain 1067 was avirulent to all A. thaliana accessions tested. A gene was cloned from X. c. pv. raphani 1067 which, when transferred into the virulent X. c. pv. campestris strain 8004, strongly reduced symptom development and bacterial growth in A. thaliana Columbia plants but did not affect virulence to Brassica plants. The gene (denoted avrXca) interacted with all A. thaliana accessions tested except one, Kas-1, which developed disease symptoms and supported growth of the transconjugant to levels similar to those with X. c. pv. campestris 8004 alone. Sequence analysis of avrXca revealed a probable open reading frame encoding a protein of 66,566 Da that has no homology with other known sequences. A sequence motif conserved among hrp genes was identified in the 5' noncoding region of avrXca, and features characteristic of a signal peptide were found in the N-terminal portion of the presumed AvrXca protein. DNA from different phytopathogenic bacteria contained sequences hybridizing with avrXca in related X. campestris pathovars but not in Erwinia or Pseudomonas strains.

  6. Association of the Cytoplasmic Membrane Protein XpsN with the Outer Membrane Protein XpsD in the Type II Protein Secretion Apparatus of Xanthomonas campestris pv. Campestris

    PubMed Central

    Lee, Hsien-Ming; Wang, Kuan-Cheng; Liu, Yi-Ling; Yew, Hsin-Yan; Chen, Ling-Yun; Leu, Wei-Ming; Chen, David Chanhen; Hu, Nien-Tai

    2000-01-01

    An xps gene cluster composed of 11 open reading frames is required for the type II protein secretion in Xanthomonas campestris pv. campestris. Immediately upstream of the xpsD gene, which encodes an outer membrane protein that serves as the secretion channel by forming multimers, there exists an open reading frame (previously designated ORF2) that could encode a protein of 261 amino acid residues. Its N-terminal hydrophobic region is a likely membrane-anchoring sequence. Antibody raised against this protein could detect in the wild-type strain of X. campestris pv. campestris a protein band with an apparent molecular mass of 36 kDa by Western blotting. Its aberrant slow migration in sodium dodecyl sulfate-polyacrylamide gels might be due to its high proline content. We designated this protein XpsN. By constructing a mutant strain with an in-frame deletion of the chromosomal xpsN gene, we demonstrated that it is required for the secretion of extracellular enzyme by X. campestris pv. campestris. Subcellular fractionation studies indicated that the XpsN protein was tightly associated with the membrane. Sucrose gradient sedimentation followed by immunoblot analysis revealed that it primarily appeared in the cytoplasmic membrane fractions. Immune precipitation experiments indicated that the XpsN protein was coprecipitated with the XpsD protein. In addition, the XpsN protein was co-eluted with the (His)6-tagged XpsD protein from the metal affinity chromatography column. All observations suggested that the XpsN protein forms a stable complex with the XpsD protein. In addition, immune precipitation analysis of the XpsN protein with various truncated XpsD proteins revealed that the C-terminal region of the XpsD protein between residues 650 and 759 was likely to be involved in complex formation between the two. PMID:10692359

  7. Genome scale analysis of diffusible signal factor regulon in Xanthomonas campestris pv. campestris: identification of novel cell-cell communication-dependent genes and functions.

    PubMed

    He, Ya-Wen; Xu, Min; Lin, Kui; Ng, Yu-Jin Alvin; Wen, Chao-Ming; Wang, Lian-Hui; Liu, Zi-Duo; Zhang, Hai-Bao; Dong, Yi-Hu; Dow, J Maxwell; Zhang, Lian-Hui

    2006-01-01

    The bacterial pathogen Xanthomonas campestris pv. campestris (Xcc) recruits a diffusible signal factor (DSF), which has recently been structurally characterized as cis-11-methyl-2-dodecenoic acid, as a cell-cell communication signal to synchronize virulence gene expression and biofilm dispersal. In this study, we showed that despite the existance of phenotype variations in different Xcc isolates, the DSF-mediated functions were in general conserved. To investigate the genomic profiles of DSF regulation, we designed and conducted oligomicroarray analysis by comparison of the gene expression patterns of wild-type strain XC1 and its DSF-deficient mutant XC1dF, as well as those of XC1dF in the presence or absence of DSF signals. The analyses led to identification of 165 genes, whose expression was significantly influenced by DSF signals. These genes encode proteins and enzymes belonging to at least 12 functional groups. In addition to those previously known DSF-dependent activities such as production of extracellular enzymes and extracellular polysaccharides, microarray analyses also revealed new functions mediated by DSF, such as flagellum synthesis, resistance to toxins and oxidative stress, and aerobic respiration. Phenotype analyses confirmed that DSF signalling contributed to resistance to toxin acriflavin and hydrogen peroxide, and to the survival of bacterial cells at different temperatures. We conclude that DSF cell-cell signalling is not only essential for co-ordinating the expression of virulence genes but also plays a vital role in keeping up the general competence of the pathogen in ecosystems.

  8. XerR, a negative regulator of XccR in Xanthomonas campestris pv. campestris, relieves its repressor function in planta.

    PubMed

    Wang, Li; Zhang, Lili; Geng, Yunfeng; Xi, Wei; Fang, Rongxiang; Jia, Yantao

    2011-07-01

    We previously reported that XccR, a LuxR-type regulator of Xanthomonas campestris pv. campestris (Xcc), activates the downstream proline iminopeptidase virulence gene (pip) in response to certain host plant factor(s). In this report, we further show that the expression of the xccR gene was repressed in the culture medium by an NtrC-type response regulator, which we named XerR (XccR expression-related, repressor), and that this repression was relieved when the bacteria were grown in planta. Such a regulatory mechanism is reinforced by the observations that XerR directly bound to the xccR promoter in vitro, and that mutations at the phosphorylation-related residues of XerR resulted in the loss of its repressor function. Furthermore, the expression level of xccR increased even in XerR-overexpressing Xcc cells when they were vacuum infiltrated into cabbage plants. We also preliminarily characterized the host factor(s) involved in the above mentioned interactions between Xcc and the host plant, showing that a plant material(s) with molecular weight(s) less than 1 kDa abolished the binding of XerR to the xccR promoter, while the same material enhanced the binding of XccR to the luxXc box in the pip promoter. Taken together, our results implicate XerR in a new layer of the regulatory mechanism controlling the expression of the virulence-related xccR/pip locus and provide clues to the identification of plant signal molecules that interact with XerR and XccR to enhance the virulence of Xcc.

  9. XerR, a negative regulator of XccR in Xanthomonas campestris pv. campestris, relieves its repressor function in planta

    PubMed Central

    Wang, Li; Zhang, Lili; Geng, Yunfeng; Xi, Wei; Fang, Rongxiang; Jia, Yantao

    2011-01-01

    We previously reported that XccR, a LuxR-type regulator of Xanthomonas campestris pv. campestris (Xcc), activates the downstream proline iminopeptidase virulence gene (pip) in response to certain host plant factor(s). In this report, we further show that the expression of the xccR gene was repressed in the culture medium by an NtrC-type response regulator, which we named XerR (XccR expression-related, repressor), and that this repression was relieved when the bacteria were grown in planta. Such a regulatory mechanism is reinforced by the observations that XerR directly bound to the xccR promoter in vitro, and that mutations at the phosphorylation-related residues of XerR resulted in the loss of its repressor function. Furthermore, the expression level of xccR increased even in XerR-overexpressing Xcc cells when they were vacuum infiltrated into cabbage plants. We also preliminarily characterized the host factor(s) involved in the above mentioned interactions between Xcc and the host plant, showing that a plant material(s) with molecular weight(s) less than 1 kDa abolished the binding of XerR to the xccR promoter, while the same material enhanced the binding of XccR to the luxXc box in the pip promoter. Taken together, our results implicate XerR in a new layer of the regulatory mechanism controlling the expression of the virulence-related xccR/pip locus and provide clues to the identification of plant signal molecules that interact with XerR and XccR to enhance the virulence of Xcc. PMID:21483448

  10. Mutation of the gene encoding a major outer-membrane protein in Xanthomonas campestris pv. campestris causes pleiotropic effects, including loss of pathogenicity.

    PubMed

    Chen, Yih-Yuan; Wu, Chieh-Hao; Lin, Juey-Wen; Weng, Shu-Fen; Tseng, Yi-Hsiung

    2010-09-01

    Xanthomonas campestris pv. campestris (Xcc) is the phytopathogen that causes black rot in crucifers. The xanthan polysaccharide and extracellular enzymes produced by this organism are virulence factors, the expression of which is upregulated by Clp (CRP-like protein) and DSF (diffusible signal factor), which is synthesized by RpfF. It is also known that biofilm formation/dispersal, regulated by the effect of controlled synthesis of DSF on cell-cell signalling, is required for virulence. Furthermore, a deficiency in DSF causes cell aggregation with concomitant production of a gum-like substance that can be dispersed by addition of DSF or digested by exogenous endo-beta-1,4-mannanase expressed by Xcc. In this study, Western blotting of proteins from a mopB mutant (XcMopB) showed Xcc MopB to be the major outer-membrane protein (OMP); Xcc MopB shared over 97 % identity with homologues from other members of Xanthomonas. Similarly to the rpfF mutant, XcMopB formed aggregates with simultaneous production of a gummy substance, but these aggregates could not be dispersed by DSF or endo-beta-1,4-mannanase, indicating that different mechanisms were involved in aggregation. In addition, XcMopB showed surface deformation, altered OMP composition, impaired xanthan production, increased sensitivity to stressful conditions including SDS, elevated temperature and changes in pH, reduced adhesion and motility and defects in pathogenesis. The finding that the major OMP is required for pathogenicity is unprecedented in phytopathogenic bacteria.

  11. Genome wide transcription start sites analysis of Xanthomonas campestris pv. campestris B100 with insights into the gum gene cluster directing the biosynthesis of the exopolysaccharide xanthan.

    PubMed

    Alkhateeb, Rabeaa S; Vorhölter, Frank-Jörg; Rückert, Christian; Mentz, Almut; Wibberg, Daniel; Hublik, Gerd; Niehaus, Karsten; Pühler, Alfred

    2016-05-10

    Xanthomonas campestris pv. campestris (Xcc) is the major producer of the exopolysaccharide xanthan, the commercially most important natural polysaccharide of microbial origin. The current work provides deeper insights into the yet uncharacterized transcriptomic features of the xanthan producing strain Xcc-B100. Towards this goal, RNA sequencing of a library based on the selective enrichment of the 5' ends of native transcripts was performed. This approach resulted in the genome wide identification of 3067 transcription start sites (TSSs) that were further classified based on their genomic positions. Among them, 1545 mapped upstream of an actively transcribed CDS and 1363 were classified as novel TSSs representing antisense, internal, and TSSs belonging to previously unidentified genomic features. Analyzing the transcriptional strength of primary and antisense TSSs revealed that in some instances antisense transcription seemed to be initiated at a higher level than its sense counterpart. Mapping the exact positions of TSSs aided in the identification of promoter consensus motifs, ribosomal binding sites, and enhanced the genome annotation of 159 in silico predicted translational start (TLS) sites. The global view on length distribution of the 5' untranslated regions (5'-UTRs) deduced from the data pointed to the occurrence of leaderless transcripts and transcripts with unusually long 5'-UTRs, in addition to identifying seven putative riboswitch elements for Xcc-B100. Concerning the biosynthesis of xanthan, we focused on the transcriptional organization of the gum gene cluster. Under the conditions tested, we present evidence for a complex transcription pattern of the gum genes with multiple TSSs and an obvious considerable role of antisense transcription. The gene gumB, encoding an outer membrane xanthan exporter, is presented here as an example for genes that possessed a strong antisense TSS.

  12. Detection of the plant pathogenic bacterium Xanthomonas campestris pv. Campestris in seed extracts of Brassica sp. Applying fluorescent antibodies and flow cytometry.

    PubMed

    Chitarra, L G; Langerak, C J; Bergervoet, J H W; van den Bulk, R W

    2002-02-01

    Xanthomonas campestris pv. campestris (Xcc) is a seed-transmitted plant pathogenic bacterium that causes black rot of crucifers. Seed lots and plants are screened for contamination with this pathogen using plating or serological assays. These methods, however, are time consuming and not very sensitive, respectively. Therefore, flow cytometry (FCM) was evaluated as a tool for the rapid detection and quantification of Xcc cells labeled with a mixture of specific fluorescein isothicyanate (FITC)-monoclonal antibodies (mAb) in pure culture, in mixed cultures of Xcc with either the common saprophyte Pseudomonas fluorescens (Psf) or a nonpathogenic X. campestris isolate (Xc), and in crude seed extracts. The mAb 18G12, conjugated with FITC, was tested at dilutions of 1:50, 1:100, 1:200, and 1:400. For mixed suspensions of Xcc and Psf, mAb 18G12 was used at a dilution of 1:100. The combination of mAbs 18G12, 2F4, and 20H6, all conjugated with FITC, was used at a dilution of 1:100 for the detection and quantification of Xcc cells in mixed suspensions containing Xcc and Xc and in crude seed extracts. The analyses were performed with a Coulter EPICS XL-MCL flow cytometer, at low flow rate during 2 min. Using FCM, Xcc cells labeled with FITC-conjugated mAbs (18G12, 2F4, and 20H6) were detected and quantified rapidly at low numbers, i.e., 10(3) colony-forming units per milliliter in pure and in mixed cultures with Psf. The presence of the nonpathogenic Xc in the seed extracts did not interfere with the FCM results. Xcc cells were distinguished from the cells of other organisms and from small particles present in the seed extract based on the high-intensity fluorescence of the labeled cells. The application of FCM in combination with FITC-conjugated mAbs appears to be a promising technique for the detection and quantification of Xcc cells in seed extracts of crucifers. Copyright 2002 Wiley-Liss, Inc.

  13. Xanthomonas campestris pv. campestris (cause of black rot of crucifers) in the genomic era is still a worldwide threat to brassica crops.

    PubMed

    Vicente, Joana G; Holub, Eric B

    2013-01-01

    Xanthomonas campestris pv. campestris (Xcc) (Pammel) Dowson is a Gram-negative bacterium that causes black rot, the most important disease of vegetable brassica crops worldwide. Intensive molecular investigation of Xcc is gaining momentum and several whole genome sequences are available. Bacteria; Phylum Proteobacteria; Class Gammaproteobacteria; Order Xanthomonadales; Family Xanthomonadacea; Genus Xanthomonas; Species X. campestris. Xcc can cause disease in a large number of species of Brassicaceae (ex-Cruciferae), including economically important vegetable Brassica crops and a number of other cruciferous crops, ornamentals and weeds, including the model plant Arabidopsis thaliana. Black rot is a systemic vascular disease. Typical disease symptoms include V-shaped yellow lesions starting from the leaf margins and blackening of the veins. Collections of Xcc isolates have been differentiated into physiological races based on the response of several brassica species lines. Black rot is a seed-borne disease. The disease is favoured by warm, humid conditions and can spread rapidly from rain dispersal and irrigation water. The control of black rot is difficult and relies on the use of pathogen-free planting material and the elimination of other potential inoculum sources (infected crop debris and cruciferous weeds). Major gene resistance is very rare in B. oleracea (brassica C genome). Resistance is more readily available in other species, including potentially useful sources of broad-spectrum resistance in B. rapa and B. carinata (A and BC genomes, respectively) and in the wild relative A. thaliana. The reference genomes of three isolates have been released. The genome consists of a single chromosome of approximately 5 100 000 bp, with a GC content of approximately 65% and an average predicted number of coding DNA sequences (CDS) of 4308. Three different secretion systems have been identified and studied in Xcc. The gene clusters xps and xcs encode a type II

  14. Involvement of bacterial TonB-dependent signaling in the generation of an oligogalacturonide damage-associated molecular pattern from plant cell walls exposed to Xanthomonas campestris pv. campestris pectate lyases.

    PubMed

    Vorhölter, Frank-Jörg; Wiggerich, Heinrich-Günter; Scheidle, Heiko; Sidhu, Vishaldeep Kaur; Mrozek, Kalina; Küster, Helge; Pühler, Alfred; Niehaus, Karsten

    2012-10-19

    Efficient perception of attacking pathogens is essential for plants. Plant defense is evoked by molecules termed elicitors. Endogenous elicitors or damage-associated molecular patterns (DAMPs) originate from plant materials upon injury or pathogen activity. While there are comparably well-characterized examples for DAMPs, often oligogalacturonides (OGAs), generated by the activity of fungal pathogens, endogenous elicitors evoked by bacterial pathogens have been rarely described. In particular, the signal perception and transduction processes involved in DAMP generation are poorly characterized. A mutant strain of the phytopathogenic bacterium Xanthomonas campestris pv. campestris deficient in exbD2, which encodes a component of its unusual elaborate TonB system, had impaired pectate lyase activity and caused no visible symptoms for defense on the non-host plant pepper (Capsicum annuum). A co-incubation of X. campestris pv. campestris with isolated cell wall material from C. annuum led to the release of compounds which induced an oxidative burst in cell suspension cultures of the non-host plant. Lipopolysaccharides and proteins were ruled out as elicitors by polymyxin B and heat treatment, respectively. After hydrolysis with trifluoroacetic acid and subsequent HPAE chromatography, the elicitor preparation contained galacturonic acid, the monosaccharide constituent of pectate. OGAs were isolated from this crude elicitor preparation by HPAEC and tested for their biological activity. While small OGAs were unable to induce an oxidative burst, the elicitor activity in cell suspension cultures of the non-host plants tobacco and pepper increased with the degree of polymerization (DP). Maximal elicitor activity was observed for DPs exceeding 8. In contrast to the X. campestris pv. campestris wild type B100, the exbD2 mutant was unable to generate elicitor activity from plant cell wall material or from pectin. To our knowledge, this is the second report on a DAMP generated

  15. Involvement of bacterial TonB-dependent signaling in the generation of an oligogalacturonide damage-associated molecular pattern from plant cell walls exposed to Xanthomonas campestris pv. campestris pectate lyases

    PubMed Central

    2012-01-01

    Background Efficient perception of attacking pathogens is essential for plants. Plant defense is evoked by molecules termed elicitors. Endogenous elicitors or damage-associated molecular patterns (DAMPs) originate from plant materials upon injury or pathogen activity. While there are comparably well-characterized examples for DAMPs, often oligogalacturonides (OGAs), generated by the activity of fungal pathogens, endogenous elicitors evoked by bacterial pathogens have been rarely described. In particular, the signal perception and transduction processes involved in DAMP generation are poorly characterized. Results A mutant strain of the phytopathogenic bacterium Xanthomonas campestris pv. campestris deficient in exbD2, which encodes a component of its unusual elaborate TonB system, had impaired pectate lyase activity and caused no visible symptoms for defense on the non-host plant pepper (Capsicum annuum). A co-incubation of X. campestris pv. campestris with isolated cell wall material from C. annuum led to the release of compounds which induced an oxidative burst in cell suspension cultures of the non-host plant. Lipopolysaccharides and proteins were ruled out as elicitors by polymyxin B and heat treatment, respectively. After hydrolysis with trifluoroacetic acid and subsequent HPAE chromatography, the elicitor preparation contained galacturonic acid, the monosaccharide constituent of pectate. OGAs were isolated from this crude elicitor preparation by HPAEC and tested for their biological activity. While small OGAs were unable to induce an oxidative burst, the elicitor activity in cell suspension cultures of the non-host plants tobacco and pepper increased with the degree of polymerization (DP). Maximal elicitor activity was observed for DPs exceeding 8. In contrast to the X. campestris pv. campestris wild type B100, the exbD2 mutant was unable to generate elicitor activity from plant cell wall material or from pectin. Conclusions To our knowledge, this is the

  16. Identification of Isolates that Cause a Leaf Spot Disease of Brassicas as Xanthomonas campestris pv. raphani and Pathogenic and Genetic Comparison with Related Pathovars.

    PubMed

    Vicente, J G; Everett, B; Roberts, S J

    2006-07-01

    ABSTRACT Twenty-five Xanthomonas isolates, including some isolates received as either X. campestris pv. armoraciae or pv. raphani, caused discrete leaf spot symptoms when spray-inoculated onto at least one Brassica oleracea cultivar. Twelve of these isolates and four other Xanthomonas isolates were spray- and pin-inoculated onto 21 different plant species/cultivars including horseradish (Armoracia rusticana), radish (Raphanus sativus), and tomato (Lycopersicon esculentum). The remaining 13 leaf spot isolates were spray-inoculated onto a subset of 10 plant species/cultivars. The leaf spot isolates were very aggressive on several Brassica spp., radish, and tomato causing leaf spots and dark sunken lesions on the middle vein, petiole, and stem. Based on the differential reactions of several Brassica spp. and radish cultivars, the leaf spot isolates were divided into three races, with races 1 and 3 predominating. A differential series was established to determine the race-type of isolates and a gene-for-gene model based on the interaction of two avirulence genes in the pathogen races and two matching resistance genes in the differential hosts is proposed. Repetitive-DNA polymerase chain reaction-based fingerprinting was used to assess the genetic diversity of the leaf spot isolates and isolates of closely related Xanthomonas pathovars. Although there was variability within each race, the leaf spot isolates were clustered separately from the X. campestris pv. campestris isolates. We propose that X. campestris isolates that cause a nonvascular leaf spot disease on Brassica spp. should be identified as pv. raphani and not pv. armoraciae. Race-type strains and a neopathotype strain for X. campestris pv. raphani are proposed.

  17. Genome-enabled determination of amino acid biosynthesis in Xanthomonas campestris pv. campestris and identification of biosynthetic pathways for alanine, glycine, and isoleucine by 13C-isotopologue profiling.

    PubMed

    Schatschneider, Sarah; Vorhölter, Frank-Jörg; Rückert, Christian; Becker, Anke; Eisenreich, Wolfgang; Pühler, Alfred; Niehaus, Karsten

    2011-10-01

    To elucidate the biosynthetic pathways for all proteinogenic amino acids in Xanthomonas campestris pv. campestris, this study combines results obtained by in silico genome analysis and by (13)C-NMR-based isotopologue profiling to provide a panoramic view on a substantial section of bacterial metabolism. Initially, biosynthesis pathways were reconstructed from an improved annotation of the complete genome of X. campestris pv. campestris B100. This metabolic reconstruction resulted in the unequivocal identification of biosynthesis routes for 17 amino acids in total: arginine, asparagine, aspartate, cysteine, glutamate, glutamine, histidine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, and valine. Ambiguous pathways were reconstructed from the genome data for alanine, glycine, and isoleucine biosynthesis. (13)C-NMR analyses supported the identification of the metabolically active pathways. The biosynthetic routes for these amino acids were derived from the precursor molecules pyruvate, serine, and pyruvate, respectively. By combining genome analysis and isotopologue profiling, a comprehensive set of biosynthetic pathways covering all proteinogenic amino acids was unraveled for this plant pathogenic bacterium, which plays an important role in biotechnology as a producer of the exopolysaccharide xanthan. The data obtained lay ground for subsequent functional analyses in post-genomics and biotechnology, while the innovative combination of in silico and wet lab technology described here is promising as a general approach to elucidate metabolic pathways.

  18. The influence of a modified lipopolysaccharide O-antigen on the biosynthesis of xanthan in Xanthomonas campestris pv. campestris B100.

    PubMed

    Steffens, Tim; Vorhölter, Frank-Jörg; Giampà, Marco; Hublik, Gerd; Pühler, Alfred; Niehaus, Karsten

    2016-05-23

    The exopolysaccharide xanthan is a natural product which is extensively used in industry. It is a thickening agent in many fields, from oil recovery to the food sector. Xanthan is produced by the Gram negative bacterium Xanthomonas campestris pv. campestris (Xcc). We analyzed the lipopolysaccharide (LPS) of three mutant strains of the Xcc wild type B100 to distinguish if the xanthan production can be increased when LPS biosynthesis is affected. The Xcc B100 O-antigen (OA) is composed of a linear main chain of rhamnose residues with N-acetylfucosamine (FucNAc) side branches at every second rhamnose. It is the major LPS constituent. The O-antigen was missing completely in the mutant strain H21012 (deficient in wxcB), since neither rhamnose nor FucNAc could be detected as part of the LPS by MALDI-TOF-MS, and only a slight amount of rhamnose and no FucNAc was found by GC analysis. The LPS of two other mutants was analyzed, Xcc H28110 (deficient in wxcK) and H20110 (wxcN). In both of them no FucNAc could be detected in the LPS fraction, while the rhamnose moieties were more abundant than in wild type LPS. The measurements were carried out by GC and confirmed by MALDI-TOF-MS analyses that indicated an altered OA in which the branches are missing, while the rhamnan main chain seemed longer than in the wild type. Quantification of xanthan confirmed our hypothesis that a missing OA can lead to an increased production of the extracellular polysaccharide. About 6.3 g xanthan per g biomass were produced by the Xcc mutant H21012 (wxcB), as compared to the wild type production of approximately 5 g xanthan per g biomass. In the two mutant strains with modified OA however, Xcc H28110 (wxcK) and Xcc H20110 (wxcN), the xanthan production of 5.5 g and 5.3 g, respectively, was not significantly increased. Mutations affecting LPS biosynthesis can be beneficial for the production of the extracellular polysaccharide xanthan. However, only complete inhibition of the OA resulted in

  19. Reclassification of Xanthomonas campestris pv. citri (ex Hasse 1915) Dye 1978 forms A, B/C/D, and E as X. smithii subsp. citri (ex Hasse) sp. nov. nom. rev. comb. nov., X. fuscans subsp. aurantifolii (ex Gabriel 1989) sp. nov. nom. rev. comb. nov., and X. alfalfae subsp. citrumelo (ex Riker and Jones) Gabriel et al., 1989 sp. nov. nom. rev. comb. nov.; X. campestris pv malvacearum (ex smith 1901) Dye 1978 as X. smithii subsp. smithii nov. comb. nov. nom. nov.; X. campestris pv. alfalfae (ex Riker and Jones, 1935) dye 1978 as X. alfalfae subsp. alfalfae (ex Riker et al., 1935) sp. nov. nom. rev.; and "var. fuscans" of X. campestris pv. phaseoli (ex Smith, 1987) Dye 1978 as X. fuscans subsp. fuscans sp. nov.

    PubMed

    Schaad, Norman W; Postnikova, Elena; Lacy, George H; Sechler, Aaron; Agarkova, Irina; Stromberg, Paul E; Stromberg, Verlyn K; Vidaver, Anne K

    2005-08-01

    Bacterial canker of citrus is a serious disease of citrus worldwide. Five forms of the disease have been described, cankers "A", "B", "C", "D", and "E". Although considerable genetic diversity has been described among the causal agents of the five forms of citrus canker and supports multiple taxons, the causal agents currently are classified as pathovars citri ("A"), aurantifolii ("B/C/D") and citrumelo ("E") of a single species, Xanthomonas campestris pv. citri (or X. axonopodis pv. citri). To determine the taxonomic relatedness among strains of X. campestris pv. citri, we conducted DNA-DNA relatedness assays, sequenced the 16S-23S intergenic spacer (ITS) regions, and performed amplified fragment length polymorphism (AFLP) analysis, using 44 strains representative of the five recognized forms of citrus canker. Under stringent DNA reassociation conditions (Tm - 15 degrees C), three distinct genotypes of citrus pathogens were revealed: taxon I included all "A" strains; taxon II contained all "B", "C", and "D" strains; and taxon III contained all "E" strains. The three citrus taxa showed less than 50% (mean) DNA-DNA relatedness to each other and less than 30% (mean) to X. campestris pv. campestris and X. axonopodis pv. axonopodis. Taxa I and II strains share over 70% DNA relatedness to X. campestris pv. malvacearum and X. campestris pv. phaseoli var. fuscans, respectively (at Tm - 15 degrees C). Taxon III strains share 70% relatedness to X. campestris pv. alfalfae. Previous and present phenotypic data support these DNA reassociation data. Taxon II strains grow more slowly on agar media than taxa I and III strains. Taxa I and III strains utilize maltose, and liquefy gelatin whereas taxon II strains do not. Taxon I strains hydrolyze pectate (pH 7.0) whereas Taxon II strains do not. Taxon III strains utilize raffinose whereas Taxon I strains do not. Each taxon can be differentiated by serology and pathogenicity. We propose taxa I, II, and III citrus strains be named

  20. Ecological and genetic analysis of copper and streptomycin resistance in Pseudomonas syringae pv. syringae.

    PubMed Central

    Sundin, G W; Bender, C L

    1993-01-01

    Strains of Pseudomonas syringae pv. syringae resistant to copper, streptomycin, or both compounds were recovered from symptomless and diseased tissue of four woody hosts in three nurseries in Oklahoma. In strains resistant to copper and streptomycin (Cur Smr), resistance to both compounds was cotransferred with a single plasmid which was either 68, 190, or 220 kilobase pairs (kb). All Cus Smr strains contained a 68-kb conjugative plasmid. Cur Sms strains contained one plasmid which varied in size from 60 to 73 kb. All conjugative plasmids which transferred streptomycin resistance contained sequences homologous to the strA and strB Smr genes from the broad-host-range plasmid RSF1010. The Smr determinant was subsequently cloned from a 68-kb Cur Smr plasmid designated pPSR1. A restriction map detailing the organization of the homologous Smr genes from pPSR1 and RSF1010 and cloned Smr genes from P. syringae pv. papulans and Xanthomonas campestris pv. vesicatoria revealed the conservation of all sites studied. The Cur genes cloned from P. syringae pv. tomato PT23 and X. campestris pv. vesicatoria XV10 did not hybridize to the Cur plasmids identified in the present study, indicating that copper resistance in these P. syringae pv. syringae strains may be conferred by a distinct genetic determinant. Images PMID:8476279

  1. Isolation and partial characterization of antibacterial lipopeptide produced by Paenibacillus polymyxa HKA-15 against phytopathogen Xanthomonas campestris pv. phaseoli M-5.

    PubMed

    Mageshwaran, Vellaichamy; Walia, Suresh; Annapurna, Kannepalli

    2012-03-01

    An antibacterial metabolite was isolated from Paenibacillus polymyxa HKA-15, a soybean bacterial endophyte. The purification of the crude metabolite from Paenibacillus polymyxa HKA-15 was done by column chromatography. In TLC, a spot with an R ( f ) value of 0.86 (±0.02) from the purified fraction showed bioactivity against Xanthomonas campestris pv. phaseoli M-5. In SDS-PAGE, the purified antibiotic was separated in the molecular weight range of 3.5 kDa. The exact molecular weight of the active compound was identified as 1,347.7 Da using MS-MS analysis. Infra red spectrum and (1)H NMR analysis showed the presence of amino acids and fatty acids in the active compound. The characterization of the antibacterial compound revealed its lipopeptide nature. In an agar diffusion assay, the crude metabolite showed a broad spectrum of activity, being able to inhibit the growth of the fungal pathogen, Rhizoctonia bataticola, Macrophomina phaseolina and Fusarium udum. A stronger inhibition was observed against bacterial pathogens viz., X. campestris pv.phaseoli M-5, X. campestris pv. phaseoli CP-1-1, Xanthomonas oryzae, Ralstonia solanacearum and Micrococcus luteus.

  2. Restoration of pathogenicity of avirulent Xanthomonas oryzae pv. oryzae and X. campestris pathovars by reciprocal complementation with the hrpXo and hrpXc genes and identification of HrpX function by sequence analyses.

    PubMed Central

    Kamdar, H V; Kamoun, S; Kado, C I

    1993-01-01

    The molecular basis of pathogenesis by Xanthomonas oryzae pv. oryzae has been partly elucidated by the identification of a gene, hrpXo, required for bacterial blight on rice. A mutation in hrpXo results in the loss of pathogenicity on rice and the loss of hypersensitivity on nonhosts such as Datura stramonium and radishes. Pathogenicity and its ability to cause the hypersensitive reaction is restored by complementing the mutant with the heterologous hrpXc gene derived from X. campestris pv. campestris. Conversely, hrpXo complements nonpathogenic mutants of X. campestris pv. campestris and X. campetstris pv, armoraciae. Mutants bearing the heterologous hrpX gene are restored in their abilities to cause diseases typical of their chromosomal background and not the hypersensitive reaction on their respective hosts. The hrpXo and hrpXc genes are therefore functionally equivalent, and this functional equivalence extends into X. campestris pv. armoraciae and possibly into other X. campestris pathovars, since this gene is highly conserved among eight other pathovars tested. Sequence analyses of hrpXo revealed an open reading frame of 1,452 bp with a coding capacity for a protein of 52.3 kDa. The protein contains a consensus domain for possible protein myristoylation whose consequence may result in a loss of recognition by host defense and surveillance systems. Images PMID:8458844

  3. Metabolic flux pattern of glucose utilization by Xanthomonas campestris pv. campestris: prevalent role of the Entner-Doudoroff pathway and minor fluxes through the pentose phosphate pathway and glycolysis.

    PubMed

    Schatschneider, Sarah; Huber, Claudia; Neuweger, Heiko; Watt, Tony Francis; Pühler, Alfred; Eisenreich, Wolfgang; Wittmann, Christoph; Niehaus, Karsten; Vorhölter, Frank-Jörg

    2014-10-01

    The well-studied plant pathogenic bacterium Xanthomonas campestris pv. campestris (Xcc) synthesizes the biotechnologically important polysaccharide xanthan gum, which is also regarded as a virulence factor in plant interactions. In Xcc, sugars like glucose are utilized as a source to generate energy and biomass for growth and pathogenicity. In this study, we used [1-(13)C]glucose as a tracer to analyze the fluxes in the central metabolism of the bacterium growing in a minimal medium. (13)C-Metabolic flux analysis based on gas chromatography-mass spectrometry (GC-MS) confirmed the prevalent catabolic role of the Entner-Doudoroff pathway. Comparative nuclear magnetic resonance (NMR)-based isotopologue profiling of a mutant deficient in glycolysis gave evidence for a moderate flux via glycolysis in the wild-type. In addition to reconfirming the Entner-Doudoroff pathway as a catabolic main route, this approach affirmed a numerically minor but important flux via the pentose phosphate pathway.

  4. Cloning and molecular characterization of hrpX from Xanthomonas axonopodis pv. citri.

    PubMed

    Iwamoto, M; Oku, T

    2000-01-01

    The hrpX gene of plant pathogenic Xanthomonas species is essential for pathogenicity on host plants and to cause hypersensitive reaction on non-host plants. We cloned and analyzed a hrpX homologue, designated hrpXct, of X. axonopodis pv. citri, a pathogen of citrus canker. The open reading frame of hrpXct has 1431 bp in nucleotides which has a coding capacity of 476 amino acid residues with a molecular mass of 52.4 kDa. The predicted amino acid sequence of HrpXct has 90% identity to the AraC family type transcriptional activator protein HrpXc of X. campestris pv. campestris, 95% to HrpXo of X. oryzae pv. oryzae and 97% to X. vesicatoria. These findings clearly indicate and confirm that the structure of the hrpX genes in plant pathogenic Xanthomonas species is highly conserved.

  5. Comparative genomic analysis of Xanthomonas axonopodis pv. citrumelo F1, which causes citrus bacterial spot disease, and related strains provides insights into virulence and host specificity.

    PubMed

    Jalan, Neha; Aritua, Valente; Kumar, Dibyendu; Yu, Fahong; Jones, Jeffrey B; Graham, James H; Setubal, João C; Wang, Nian

    2011-11-01

    Xanthomonas axonopodis pv. citrumelo is a citrus pathogen causing citrus bacterial spot disease that is geographically restricted within the state of Florida. Illumina, 454 sequencing, and optical mapping were used to obtain a complete genome sequence of X. axonopodis pv. citrumelo strain F1, 4.9 Mb in size. The strain lacks plasmids, in contrast to other citrus Xanthomonas pathogens. Phylogenetic analysis revealed that this pathogen is very close to the tomato bacterial spot pathogen X. campestris pv. vesicatoria 85-10, with a completely different host range. We also compared X. axonopodis pv. citrumelo to the genome of citrus canker pathogen X. axonopodis pv. citri 306. Comparative genomic analysis showed differences in several gene clusters, like those for type III effectors, the type IV secretion system, lipopolysaccharide synthesis, and others. In addition to pthA, effectors such as xopE3, xopAI, and hrpW were absent from X. axonopodis pv. citrumelo while present in X. axonopodis pv. citri. These effectors might be responsible for survival and the low virulence of this pathogen on citrus compared to that of X. axonopodis pv. citri. We also identified unique effectors in X. axonopodis pv. citrumelo that may be related to the different host range as compared to that of X. axonopodis pv. citri. X. axonopodis pv. citrumelo also lacks various genes, such as syrE1, syrE2, and RTX toxin family genes, which were present in X. axonopodis pv. citri. These may be associated with the distinct virulences of X. axonopodis pv. citrumelo and X. axonopodis pv. citri. Comparison of the complete genome sequence of X. axonopodis pv. citrumelo to those of X. axonopodis pv. citri and X. campestris pv. vesicatoria provides valuable insights into the mechanism of bacterial virulence and host specificity.

  6. Comparative Genomic Analysis of Xanthomonas axonopodis pv. citrumelo F1, Which Causes Citrus Bacterial Spot Disease, and Related Strains Provides Insights into Virulence and Host Specificity ▿ #

    PubMed Central

    Jalan, Neha; Aritua, Valente; Kumar, Dibyendu; Yu, Fahong; Jones, Jeffrey B.; Graham, James H.; Setubal, João C.; Wang, Nian

    2011-01-01

    Xanthomonas axonopodis pv. citrumelo is a citrus pathogen causing citrus bacterial spot disease that is geographically restricted within the state of Florida. Illumina, 454 sequencing, and optical mapping were used to obtain a complete genome sequence of X. axonopodis pv. citrumelo strain F1, 4.9 Mb in size. The strain lacks plasmids, in contrast to other citrus Xanthomonas pathogens. Phylogenetic analysis revealed that this pathogen is very close to the tomato bacterial spot pathogen X. campestris pv. vesicatoria 85-10, with a completely different host range. We also compared X. axonopodis pv. citrumelo to the genome of citrus canker pathogen X. axonopodis pv. citri 306. Comparative genomic analysis showed differences in several gene clusters, like those for type III effectors, the type IV secretion system, lipopolysaccharide synthesis, and others. In addition to pthA, effectors such as xopE3, xopAI, and hrpW were absent from X. axonopodis pv. citrumelo while present in X. axonopodis pv. citri. These effectors might be responsible for survival and the low virulence of this pathogen on citrus compared to that of X. axonopodis pv. citri. We also identified unique effectors in X. axonopodis pv. citrumelo that may be related to the different host range as compared to that of X. axonopodis pv. citri. X. axonopodis pv. citrumelo also lacks various genes, such as syrE1, syrE2, and RTX toxin family genes, which were present in X. axonopodis pv. citri. These may be associated with the distinct virulences of X. axonopodis pv. citrumelo and X. axonopodis pv. citri. Comparison of the complete genome sequence of X. axonopodis pv. citrumelo to those of X. axonopodis pv. citri and X. campestris pv. vesicatoria provides valuable insights into the mechanism of bacterial virulence and host specificity. PMID:21908674

  7. The inheritance of resistance to bacterial leaf spot of lettuce caused by Xanthomonas campestris pv. vitians in three lettuce cultivars

    PubMed Central

    Hayes, Ryan J; Trent, Mark A; Truco, Maria Jose; Antonise, Rudie; Michelmore, Richard W; Bull, Carolee T

    2014-01-01

    Lettuce yields can be reduced by the disease bacterial leaf spot (BLS) caused by the pathogen Xanthomonas campestris pv. vitians (Xcv) and host resistance is the most feasible method to reduce disease losses. The cultivars La Brillante, Pavane and Little Gem express an incompatible host–pathogen interaction as a hypersensitive response (HR) to California strains of Xcv resulting in resistance. Little was known about the inheritance of resistance; however, resistance to other lettuce pathogens is often determined by resistance gene candidates (RGCs) encoding nucleotide-binding leucine-rich repeat (NB-LRR) proteins. Therefore, we determined the inheritance of BLS resistance in the cultivars La Brillante, Little Gem and Pavane and mapped it relative to RGCs. The reaction to Xcv was analyzed in nine F1, F2 and recombinant inbred line populations of lettuce from HR×compatible or HR×HR crosses. The HR in La Brillante, Pavane and Little Gem is conditioned by single dominant genes, which are either allelic or closely linked genes. The resistance gene in La Brillante was designated Xanthomonas resistance 1 (Xar1) and mapped to lettuce linkage group 2. Xar1 is present in a genomic region that contains numerous NB-LRR encoding RGCs and functional pathogen resistance loci in the RGC2 family. The Xar1 gene confers a high level of BLS resistance in the greenhouse and field that can be introgressed into commercial lettuce cultivars to reduce BLS losses using molecular markers. PMID:26504558

  8. Differential expression of peach ERF transcriptional activators in response to signaling molecules and inoculation with Xanthomonas campestris pv. pruni.

    PubMed

    Sherif, S; El-Sharkawy, I; Paliyath, G; Jayasankar, S

    2012-05-01

    Ethylene response factors (ERFs) are a large family of transcription factors (TFs) that have diverse functions in plant development and immunity. However, very little is known about the molecular regulation of these TFs in stone fruits during disease incidence. In the present study, we describe the identification of five peach ERFs (Pp-ERFs), aiming to elucidate their potential roles in defense against Xanthomonas campestris pv. pruni (Xcp), the causal agent of bacterial spot disease. The phylogenetic analysis along with sequence comparisons indicated that all Pp-ERFs are transcriptional activators belonging to groups IX and IIV ERFs. The transactivation capacity of these proteins was verified in vivo where they all induced the expression of the GUS reporter gene and in a GCC-dependent manner. The nuclear localization was also confirmed for two of these proteins, Pp-ERF2.b and Pp-ERF2.c, after their transient expression in onion epidermal cells. The induction kinetics of Pp-ERFs after inoculation with Xcp was determined by qRT-PCR. Except for Pp-ERF2.b, transcript levels of Pp-ERFs increased strongly and rapidly in the resistant 'Venture' compared to the susceptible 'BabyGold 5' cultivar after infection with Xcp. In contrast, the expression of Pp-ERF2.b was several-fold higher in the susceptible cultivar after bacterial infection. The expression of Pp-ERFs was also monitored after treating with signaling compounds; salicylic acid (SA) (1 mM), ethephon (1 mM) and methyl jasmonate (MeJA) (50 μM). Although the results generally emphasize the role of ethylene/jasmonic acid (ET/JA) signaling pathways in regulating the expression of Pp-ERFs, there was a coordination of the timing of ET/JA responses, suggesting compensatory rather than synergistic interactions between these pathways during defense against Xcp. Copyright © 2012 Elsevier GmbH. All rights reserved.

  9. Identification of a putative cognate sensor kinase for the two-component response regulator HrpG, a key regulator controlling the expression of the hrp genes in Xanthomonas campestris pv. campestris.

    PubMed

    Li, Rui-Fang; Lu, Guang-Tao; Li, Lei; Su, Hui-Zhao; Feng, Guo-fang; Chen, Ya; He, Yong-Qiang; Jiang, Bo-Le; Tang, Dong-Jie; Tang, Ji-Liang

    2014-07-01

    The bacterial phytopathogen Xanthomonas campestris pv. campestris (Xcc) relies on the hrp (hypersensitive response and pathogenicity) genes to cause disease and induce hypersensitive response (HR). The hrp genes of bacterial phytopathogens are divided into two groups. Xcc hrp genes belong to group II. It has long been known that the group II hrp genes are activated by an AraC-type transcriptional regulator whose expression is controlled by a two-component system (TCS) response regulator (named HrpG in Xcc). However, no cognate sensor kinase has yet been identified. Here, we present evidence showing that the Xcc open-reading frame XC_3670 encodes a TCS sensor kinase (named HpaS). Mutation of hpaS almost completely abolished the HR induction and virulence. Bacterial two-hybrid and protein pull-down assays revealed that HpaS physically interacted with HrpG. Phos-tag™ SDS-PAGE analysis showed that mutation in hpaS reduced markedly the phosphorylation of HrpG in vivo. These data suggest that HpaS and HrpG are most likely to form a TCS. We also showed that XC_3669 (named hpaR2), which is adjacent to hpaS and encodes a putative TCS response regulator, is required for full virulence but not HR induction. HpaR2 also physically interacted with HpaS, suggesting that HpaS may also form another TCS with HpaR2.

  10. Cloning and characterization of katA, encoding the major monofunctional catalase from Xanthomonas campestris pv. phaseoli and characterization of the encoded catalase KatA.

    PubMed

    Chauvatcharin, Nopmanee; Vattanaviboon, Paiboon; Switala, Jack; Loewen, Peter C; Mongkolsuk, Skorn

    2003-02-01

    The first cloning and characterization of the gene katA, encoding the major catalase (KatA), from Xanthomonas is reported. A reverse genetic approach using a synthesized katA-specific DNA probe to screen a X. campestris pv. phaseoli genomic library was employed. A positively hybridizing clone designated pKat29 that contained a full-length katA was isolated. Analysis of the nucleotide sequence revealed an open reading frame of 1,521 bp encoding a 507-amino acid protein with a theoretical molecular mass of 56 kDa. The deduced amino acid sequence of KatA revealed 84% and 78% identity to CatF of Pseudomonas syringae and KatB of P. aeruginosa, respectively. Phylogenetic analysis places Xanthomonas katA in the clade I group of bacterial catalases. Unexpectedly, expression of katA in a heterologous Escherichia coli host resulted in a temperature-sensitive expression. The KatA enzyme was purified from an overproducing mutant of X. campestris and was characterized. It has apparent K(m) and V(max) values of 75 m M [H(2)O(2)] and 2.55 x 10(5) micromol H(2)O(2) micromol heme(-1) s(-1), respectively. The enzyme is highly sensitive to 3-amino-1,2,4-triazole and NaN(3), has a narrower optimal pH range than other catalases, and is more sensitive to heat inactivation.

  11. Transgenic expression of the rice Xa21 pattern recognition receptor in banana (Musa sp.) confers resistance to Xanthomonas campestris pv. musacearum

    PubMed Central

    Tripathi, Jaindra Nath; Lorenzen, Jim; Bahar, Ofir; Ronald, Pamela; Tripathi, Leena

    2014-01-01

    Summary Banana Xanthomonas wilt (BXW), caused by the bacterium Xanthomonas campestris pv. musacearum (Xcm), is the most devastating disease of banana in east and central Africa. The spread of BXW threatens the livelihood of millions of African farmers who depend on banana for food security and income. There are no commercial chemicals, bio-control agents or resistant cultivars available to control BXW. Here we take advantage of the robust resistance conferred by the rice pattern recognition receptor (PRR), XA21, to the rice pathogen Xanthomonas oryzae pv. oryzae (Xoo). We identified a set of genes required for activation of Xa21 mediated immunity (rax) that were conserved in both Xoo and Xcm. Based on the conservation, we hypothesized that intergeneric transfer of Xa21 would confer resistance to Xcm. We evaluated 25 transgenic lines of the banana cultivar ‘Gonja manjaya’ (AAB) using a rapid bioassay and 12 transgenic plants in the glass house for resistance against Xcm. About fifty percent of the transgenic lines showed complete resistance to Xcm in both assays. In contrast, all of the non-transgenic control plants showed severe symptoms that progressed to complete wilting. These results indicate that the constitutive expression of the rice Xa21 gene in banana results in enhanced resistance against Xcm. Furthermore this work demonstrates the feasibility of PRR gene transfer between monocotyledonous species and provides a valuable new tool for controlling the BXW pandemic of banana, a staple food for 100 million people in east Africa. PMID:24612254

  12. Transgenic expression of the rice Xa21 pattern-recognition receptor in banana (Musa sp.) confers resistance to Xanthomonas campestris pv. musacearum.

    PubMed

    Tripathi, Jaindra N; Lorenzen, Jim; Bahar, Ofir; Ronald, Pamela; Tripathi, Leena

    2014-08-01

    Banana Xanthomonas wilt (BXW), caused by the bacterium Xanthomonas campestris pv. musacearum (Xcm), is the most devastating disease of banana in east and central Africa. The spread of BXW threatens the livelihood of millions of African farmers who depend on banana for food security and income. There are no commercial chemicals, biocontrol agents or resistant cultivars available to control BXW. Here, we take advantage of the robust resistance conferred by the rice pattern-recognition receptor (PRR), XA21, to the rice pathogen Xanthomonas oryzae pv. oryzae (Xoo). We identified a set of genes required for activation of Xa21-mediated immunity (rax) that were conserved in both Xoo and Xcm. Based on the conservation, we hypothesized that intergeneric transfer of Xa21 would confer resistance to Xcm. We evaluated 25 transgenic lines of the banana cultivar 'Gonja manjaya' (AAB) using a rapid bioassay and 12 transgenic lines in the glasshouse for resistance against Xcm. About 50% of the transgenic lines showed complete resistance to Xcm in both assays. In contrast, all of the nontransgenic control plants showed severe symptoms that progressed to complete wilting. These results indicate that the constitutive expression of the rice Xa21 gene in banana results in enhanced resistance against Xcm. Furthermore, this work demonstrates the feasibility of PRR gene transfer between monocotyledonous species and provides a valuable new tool for controlling the BXW pandemic of banana, a staple food for 100 million people in east Africa. © 2014 Society for Experimental Biology, Association of Applied Biologists and John Wiley & Sons Ltd.

  13. Isolation of rhizospheric and roots endophytic actinomycetes from Leguminosae plant and their activities to inhibit soybean pathogen, Xanthomonas campestris pv. glycine.

    PubMed

    Mingma, Ratchanee; Pathom-aree, Wasu; Trakulnaleamsai, Savitr; Thamchaipenet, Arinthip; Duangmal, Kannika

    2014-01-01

    In this study, actinomycetes from roots and rhizospheric soils of leguminous plants were isolated using starch casein agar supplemented with antifungal and antibacterial antibiotics. Three hundred and seventeen actinomycetes were isolated with 77 isolates obtained from plant roots and 240 isolates from rhizospheric soils. Analysis of whole-organism hydrolysates showed that 289 strains were rich in the LL-isomer of diaminopimelic acid, a result consistent with their assignment to the streptomycetes. The remaining 28 strains were assigned to non-streptomycetes based on the presence of meso-isomer of diaminopimelic acid in cell wall. Sixty-four isolates (20.2%) showed antagonistic activity against soybean pathogen Xanthomonas campestris pv. glycine by agar overlay method. Isolate RM 365 showed the highest activity with an inhibition ratio of 3.79, with no inhibitory activity on the growth of Rhizobium japonicum TISTR 079, Rhizobium sp. TISTR 061 and Rhizobium sp. TISTR 063. The 16S rRNA gene sequence analysis revealed that isolate RM 365 shared 99.28% similarity to Streptomyces caeruleatus GIMN4(T) (GQ329712). In addition, isolates which contained meso-DAP were also identified by 16S rRNA gene sequence analysis. The results showed that they were members of the genus Amycolatopsis, Isoptericola, Micromonospora, Microbispora, Nocardia, Nonomuraea, Promicromonospora and Pseudonocardia.

  14. Salicylic Acid Mediated by the Oxidative Burst Is a Key Molecule in Local and Systemic Responses of Cotton Challenged by an Avirulent Race of Xanthomonas campestris pv malvacearum

    PubMed Central

    Martinez, Christelle; Baccou, Jean-Claude; Bresson, Estelle; Baissac, Yves; Daniel, Jean-François; Jalloul, Aïda; Montillet, Jean-Luc; Geiger, Jean-Paul; Assigbetsé, Komi; Nicole, Michel

    2000-01-01

    We analyzed the production of reactive oxygen species, the accumulation of salicylic acid (SA), and peroxidase activity during the incompatible interaction between cotyledons of the cotton (Gossypium hirsutum) cv Reba B50/Xanthomonas campestris pv malvacearum (Xcm) race 18. SA was detected in petioles of cotyledons 6 h after infection and 24 h post inoculation in cotyledons and untreated leaves. The first peak of SA occurred 3 h after generation of superoxide (O2·−), and was inhibited by infiltration of catalase. Peroxidase activity and accumulation of SA increased in petioles of cotyledons and leaves following H2O2 infiltration of cotyledons from 0.85 to 1 mm. Infiltration of 2 mm SA increased peroxidase activity in treated cotyledons and in the first leaves, but most of the infiltrated SA was rapidly conjugated within the cotyledons. When increasing concentrations of SA were infiltrated 2.5 h post inoculation at the beginning of the oxidative burst, the activity of the apoplastic cationic O2·−-generating peroxidase decreased in a dose-dependent manner. We have shown that during the cotton hypersensitive response to Xcm, H2O2 is required for local and systemic accumulation of SA, which may locally control the generation of O2·−. Detaching cotyledons at intervals after inoculation demonstrated that the signal leading to systemic accumulation of SA was emitted around 3 h post inoculation, and was associated with the oxidative burst. SA produced 6 h post infection at HR sites was not the primary mobile signal diffusing systemically from infected cotyledons. PMID:10712539

  15. Xanthomonas campestris pv. musacearum: a major constraint to banana, plantain and enset production in central and east Africa over the past decade.

    PubMed

    Nakato, Valentine; Mahuku, George; Coutinho, Teresa

    2017-07-05

    Bacteria; Phylum Proteobacteria; Class Gammaproteobacteria; Order Xanthomonadales; Family Xanthomonadaceae; Genus Xanthomonas; currently classified as X. campestris pv. musacearum (Xcm). However, fatty acid methyl ester analysis and genetic and genomic evidence suggest that this pathogen is X. vasicola and resides in a separate pathovar. Xcm can be isolated on yeast extract peptone glucose agar (YPGA), cellobiose cephalexin agar and yeast extract tryptone sucrose agar (YTSA) complemented with 5-fluorouracil, cephalexin and cycloheximide to confer semi-selectivity. Xcm can also be identified using direct antigen coating enzyme-linked immunosorbent assay (DAC-ELISA), species-specific polymerase chain reaction (PCR) using GspDm primers and lateral flow devices that detect latent infections. Causes Xanthomonas wilt on plants belonging to the Musaceae, primarily banana (Musa acuminata), plantain (M. acuminata × balbisiana) and enset (Ensete ventricosum). There is a high level of genetic homogeneity within Xcm, although genome sequencing has revealed two major sublineages. Yellowing and wilting of leaves, premature fruit ripening and dry rot, bacterial exudate from cut stems. Xcm has only been found in African countries, namely Burundi, Ethiopia, Democratic Republic of the Congo, Kenya, Rwanda, Tanzania and Uganda. Xcm is transmitted by insects, bats, birds and farming implements. Long-distance dispersal of the pathogen is by the transportation of latently infected plants into new areas. The management of Xcm has relied on cultural practices that keep the pathogen population at tolerable levels. Biotechnology programmes have been successful in producing resistant banana plants. However, the deployment of such genetic material has not as yet been achieved in farmers' fields, and the sustainability of transgenic resistance remains to be addressed. © 2017 BSPP AND JOHN WILEY & SONS LTD.

  16. Salicylic acid mediated by the oxidative burst is a key molecule in local and systemic responses of cotton challenged by an avirulent race of Xanthomonas campestris pv malvacearum.

    PubMed

    Martinez, C; Baccou, J C; Bresson, E; Baissac, Y; Daniel, J F; Jalloul, A; Montillet, J L; Geiger, J P; Assigbetsé, K; Nicole, M

    2000-03-01

    We analyzed the production of reactive oxygen species, the accumulation of salicylic acid (SA), and peroxidase activity during the incompatible interaction between cotyledons of the cotton (Gossypium hirsutum) cv Reba B50/Xanthomonas campestris pv malvacearum (Xcm) race 18. SA was detected in petioles of cotyledons 6 h after infection and 24 h post inoculation in cotyledons and untreated leaves. The first peak of SA occurred 3 h after generation of superoxide (O(2)(.-)), and was inhibited by infiltration of catalase. Peroxidase activity and accumulation of SA increased in petioles of cotyledons and leaves following H(2)O(2) infiltration of cotyledons from 0.85 to 1 mM. Infiltration of 2 mM SA increased peroxidase activity in treated cotyledons and in the first leaves, but most of the infiltrated SA was rapidly conjugated within the cotyledons. When increasing concentrations of SA were infiltrated 2. 5 h post inoculation at the beginning of the oxidative burst, the activity of the apoplastic cationic O(2)(.-)-generating peroxidase decreased in a dose-dependent manner. We have shown that during the cotton hypersensitive response to Xcm, H(2)O(2) is required for local and systemic accumulation of SA, which may locally control the generation of O(2)(.-). Detaching cotyledons at intervals after inoculation demonstrated that the signal leading to systemic accumulation of SA was emitted around 3 h post inoculation, and was associated with the oxidative burst. SA produced 6 h post infection at HR sites was not the primary mobile signal diffusing systemically from infected cotyledons.

  17. ERIC-PCR-generated genomic fingerprints and their relationship with pathogenic variability of Xanthomonas campestris pv. punicae, the incitant of bacterial blight of pomegranate.

    PubMed

    Mondal, Kalyan K; Mani, C

    2009-12-01

    Bacterial blight caused by Xanthomonas campestris pv. punicae (Xcp) has emerged as a potential threat in pomegranate (Punica granatum) cultivation in India. Here, we report the genomic fingerprints and their correlation with virulence pattern of Xcp isolates from Maharashtra and Delhi. The genomic fingerprints of Xcp isolates were generated using enterobacterial repetitive intergenic consensus (ERIC) sequence-based primers, and virulence level was based on their reaction upon infiltration to susceptible pomegranate cultivar. Maharashtra isolate PGM1 showed only 50% similarity with Delhi isolate PGD8 forming a distinct genotype, whereas the Delhi isolates PGD5 and PGD6 form a cluster with Maharashtra isolates PGM2 and PGM4. The isolates PGM2, PGM4, PGD5, and PGD6 showing mean disease score of 7.47 were marked as group A or highly virulent. The moderately virulent or group B isolates PGM3 and PGD7 produced mean disease score of 4.19, whereas less virulent or group C isolates PGD8 and PGM1 gave mean disease intensity of 1.91. A correlation between genotypic groups based on ERIC fingerprints and pathogenicity of the isolates was established. The highly virulent isolates PGM2, PGM4, PGD5, and PGD6 formed a single cluster. A unique 900 bp amplicon present in all highly virulent isolates has been identified that can be used as genetic marker to screen isolates for virulence. The less virulent isolates PGD8 and PGM1 formed single cluster at 50% similarity coefficient. This seems to be the first report to establish a correlation between ERIC-PCR fingerprints and their corresponding virulence pattern of the pomegranate bacterial blight pathogen.

  18. Lipopolysaccharide from Xanthomonas campestris induces defense-related gene expression in Brassica campestris.

    PubMed

    Newman, M A; Daniels, M J; Dow, J M

    1995-01-01

    Purified lipopolysaccharide (LPS) from Xanthomonas campestris pv. campestris induced accumulation of transcript for beta-1,3-glucanase in turnip at concentrations of 1 micrograms/ml. The lipid A-inner core structure was required for activity but the O-antigen had no role. We suggest that release of LPS in planta triggers expression of at least some defense-related genes.

  19. The exbD2 gene as well as the iron-uptake genes tonB, exbB and exbD1 of Xanthomonas campestris pv. campestris are essential for the induction of a hypersensitive response on pepper (Capsicum annuum).

    PubMed

    Wiggerich, H G; Pühler, A

    2000-05-01

    The tonB, exbB and exbD1 genes of Xanthomonas campestris pv. campestris are essential for ferric iron uptake. In contrast, the exbD2 gene located in the same gene cluster is not essential. Mutational analysis revealed that the ferric-iron-uptake genes tonB, exbB and exbD1 are necessary for the induction of a hypersensitive response (HR) on the nonhost plant pepper (Capsicum annuum) and the induction of typical black rot symptoms on the host plant cauliflower (Brassica oleracea). Again, the exbD2 gene behaved differently. It was found to play a role only in the induction of the HR in pepper but not in the induction of black rot symptoms in cauliflower. Due to the low iron concentration in the plant tissue, the titre of viable bacteria of the ferric-iron-uptake mutants tonB, exbB and exbD1 decreased after leaf infiltration of pepper. The exbD2 mutant, however, which is not impaired in ferric iron uptake, multiplied in the pepper leaf tissue and grew even better than the wild-type strain, probably due to its failure to induce the HR. Nevertheless, the tonB, exbB and exbD1 mutant strains were able to spread systemically in cauliflower.

  20. Low molecular weight plant extract induces metabolic changes and the secretion of extracellular enzymes, but has a negative effect on the expression of the type-III secretion system in Xanthomonas campestris pv. campestris.

    PubMed

    Watt, Tony Francis; Vucur, Mihael; Baumgarth, Birgit; Watt, Steven Alexander; Niehaus, Karsten

    2009-03-10

    Xanthomonas campestris pathovar campestris (Xcc) is a plant pathogenic bacterium and as such has to adapt to a variety of environments. During the course of disease, Xcc colonizes the surface of its host, infects the xylem in the early stages, and develops a fully saprophytic life-style, aided by secreted degradative enzymes, in the late stages. To get some insight into this complex regulation, Xcc was cultivated in the presence of low molecular weight host plant extract (<10 kDa). From this experiments it could be observed, that malate and sucrose are taken up preferably in such an environment. Furthermore, it was demonstrated, that the plant extract has a negative effect on the gene expression of the hrp-gene cluster, although the activator hrpG was induced. Also, the secretion of degradative enzymes was shown to be upregulated. These observations indicate, that a low molecular weight plant extract (<10 kDa) is a sufficient signal to regulate metabolic pathways and the secretion of enzymes relevant for the development of virulence in Xanthomonas, but has a negative effect on the expression of genes involved in type-III secretion.

  1. Contribution of Phe-7 to Tat-Dependent Export of β-Lactamase in Xanthomonas campestris

    PubMed Central

    Lee, Chen-Wei; Tseng, Yi-Hsuan; Deng, Fu-Seng; Lin, Juey-Wen

    2012-01-01

    Strains of Xanthomonas campestris pv. campestris isolated in Taiwan are commonly resistant to ampicillin owing to the constitutive expression of a chromosomally encoded β-lactamase that is secreted into the periplasm. In this study, we found that levels of β-lactamase vary among X. campestris pv. campestris strains, a difference that can be attributed to amino acid substitutions at least at positions 7 and 206, with the former having the major impact. Bioinformatic and PCR analyses indicated that X. campestris pv. campestris possesses tatABC genes and that the signal peptide of X. campestris pv. campestris pre-Bla contains the typical twin-arginine motif (N-R-R-Q-F-L at amino acid residues 3 to 8 in strain X. campestris pv. campestris strain 11), suggesting that Bla is secreted via the Tat pathway. To assess the importance of Phe7 in the efficient export of X. campestris pv. campestris Bla, we prepared mutant constructs containing amino acid substitutions and monitored their expression by measuring enzyme activity and detecting Bla protein by Western blotting. The results indicate that replacement of Phe7 with Leu severely inhibited Bla export whereas replacement with Pro almost abolished it. Although a change to Arg caused moderate inhibition of export, replacement with Tyr had no effect. These results suggest that for efficient export of Bla by X. campestris pv. campestris, the aromatic-aromatic interactions and stability of protein structure around the twin-arginine motif are important, since only proteins that can attain a folded state in the cytoplasm are competent for export via the Tat pathway. PMID:22526303

  2. Contribution of Phe-7 to Tat-dependent export of β-lactamase in Xanthomonas campestris.

    PubMed

    Lee, Chen-Wei; Tseng, Yi-Hsuan; Deng, Fu-Seng; Lin, Juey-Wen; Tseng, Yi-Hsiung; Weng, Shu-Fen

    2012-07-01

    Strains of Xanthomonas campestris pv. campestris isolated in Taiwan are commonly resistant to ampicillin owing to the constitutive expression of a chromosomally encoded β-lactamase that is secreted into the periplasm. In this study, we found that levels of β-lactamase vary among X. campestris pv. campestris strains, a difference that can be attributed to amino acid substitutions at least at positions 7 and 206, with the former having the major impact. Bioinformatic and PCR analyses indicated that X. campestris pv. campestris possesses tatABC genes and that the signal peptide of X. campestris pv. campestris pre-Bla contains the typical twin-arginine motif (N-R-R-Q-F-L at amino acid residues 3 to 8 in strain X. campestris pv. campestris strain 11), suggesting that Bla is secreted via the Tat pathway. To assess the importance of Phe(7) in the efficient export of X. campestris pv. campestris Bla, we prepared mutant constructs containing amino acid substitutions and monitored their expression by measuring enzyme activity and detecting Bla protein by Western blotting. The results indicate that replacement of Phe(7) with Leu severely inhibited Bla export whereas replacement with Pro almost abolished it. Although a change to Arg caused moderate inhibition of export, replacement with Tyr had no effect. These results suggest that for efficient export of Bla by X. campestris pv. campestris, the aromatic-aromatic interactions and stability of protein structure around the twin-arginine motif are important, since only proteins that can attain a folded state in the cytoplasm are competent for export via the Tat pathway.

  3. Comparative and functional genomics reveals genetic diversity and determinants of host specificity among reference strains and a large collection of Chinese isolates of the phytopathogen Xanthomonas campestris pv. campestris

    PubMed Central

    He, Yong-Qiang; Zhang, Liang; Jiang, Bo-Le; Zhang, Zheng-Chun; Xu, Rong-Qi; Tang, Dong-Jie; Qin, Jing; Jiang, Wei; Zhang, Xia; Liao, Jie; Cao, Jin-Ru; Zhang, Sui-Sheng; Wei, Mei-Liang; Liang, Xiao-Xia; Lu, Guang-Tao; Feng, Jia-Xun; Chen, Baoshan; Cheng, Jing; Tang, Ji-Liang

    2007-01-01

    Background Xanthomonas campestris pathovar campestris (Xcc) is the causal agent of black rot disease of crucifers worldwide. The molecular genetic diversity and host specificity of Xcc are poorly understood. Results We constructed a microarray based on the complete genome sequence of Xcc strain 8004 and investigated the genetic diversity and host specificity of Xcc by array-based comparative genome hybridization analyses of 18 virulent strains. The results demonstrate that a genetic core comprising 3,405 of the 4,186 coding sequences (CDSs) spotted on the array are conserved and a flexible gene pool with 730 CDSs is absent/highly divergent (AHD). The results also revealed that 258 of the 304 proved/presumed pathogenicity genes are conserved and 46 are AHD. The conserved pathogenicity genes include mainly the genes involved in type I, II and III secretion systems, the quorum sensing system, extracellular enzymes and polysaccharide production, as well as many other proved pathogenicity genes, while the AHD CDSs contain the genes encoding type IV secretion system (T4SS) and type III-effectors. A Xcc T4SS-deletion mutant displayed the same virulence as wild type. Furthermore, three avirulence genes (avrXccC, avrXccE1 and avrBs1) were identified. avrXccC and avrXccE1 conferred avirulence on the hosts mustard cultivar Guangtou and Chinese cabbage cultivar Zhongbai-83, respectively, and avrBs1 conferred hypersensitive response on the nonhost pepper ECW10R. Conclusion About 80% of the Xcc CDSs, including 258 proved/presumed pathogenicity genes, is conserved in different strains. Xcc T4SS is not involved in pathogenicity. An efficient strategy to identify avr genes determining host specificity from the AHD genes was developed. PMID:17927820

  4. Comparative and functional genomics reveals genetic diversity and determinants of host specificity among reference strains and a large collection of Chinese isolates of the phytopathogen Xanthomonas campestris pv. campestris.

    PubMed

    He, Yong-Qiang; Zhang, Liang; Jiang, Bo-Le; Zhang, Zheng-Chun; Xu, Rong-Qi; Tang, Dong-Jie; Qin, Jing; Jiang, Wei; Zhang, Xia; Liao, Jie; Cao, Jin-Ru; Zhang, Sui-Sheng; Wei, Mei-Liang; Liang, Xiao-Xia; Lu, Guang-Tao; Feng, Jia-Xun; Chen, Baoshan; Cheng, Jing; Tang, Ji-Liang

    2007-01-01

    Xanthomonas campestris pathovar campestris (Xcc) is the causal agent of black rot disease of crucifers worldwide. The molecular genetic diversity and host specificity of Xcc are poorly understood. We constructed a microarray based on the complete genome sequence of Xcc strain 8004 and investigated the genetic diversity and host specificity of Xcc by array-based comparative genome hybridization analyses of 18 virulent strains. The results demonstrate that a genetic core comprising 3,405 of the 4,186 coding sequences (CDSs) spotted on the array are conserved and a flexible gene pool with 730 CDSs is absent/highly divergent (AHD). The results also revealed that 258 of the 304 proved/presumed pathogenicity genes are conserved and 46 are AHD. The conserved pathogenicity genes include mainly the genes involved in type I, II and III secretion systems, the quorum sensing system, extracellular enzymes and polysaccharide production, as well as many other proved pathogenicity genes, while the AHD CDSs contain the genes encoding type IV secretion system (T4SS) and type III-effectors. A Xcc T4SS-deletion mutant displayed the same virulence as wild type. Furthermore, three avirulence genes (avrXccC, avrXccE1 and avrBs1) were identified. avrXccC and avrXccE1 conferred avirulence on the hosts mustard cultivar Guangtou and Chinese cabbage cultivar Zhongbai-83, respectively, and avrBs1 conferred hypersensitive response on the nonhost pepper ECW10R. About 80% of the Xcc CDSs, including 258 proved/presumed pathogenicity genes, is conserved in different strains. Xcc T4SS is not involved in pathogenicity. An efficient strategy to identify avr genes determining host specificity from the AHD genes was developed.

  5. Effector-triggered innate immunity contributes Arabidopsis resistance to Xanthomonas campestris.

    PubMed

    Rong, Wei; Feng, Feng; Zhou, Jianmin; He, Chaozu

    2010-11-01

    Xanthomonas campestris pv. campestris, the causal agent of black rot disease, depends on its type III secretion system (TTSS) to infect cruciferous plants, including Brassica oleracea, B. napus and Arabidopsis. Previous studies on the Arabidopsis-Pseudomonas syringae model pathosystem have indicated that a major function of TTSS from virulent bacteria is to suppress host defences triggered by pathogen-associated molecular patterns. Similar analyses have not been made for the Arabidopsis-X. campestris pv. campestris pathosystem. In this study, we report that X. campestris pv. campestris strain 8004, which is modestly pathogenic on Arabidopsis, induces strong defence responses in Arabidopsis in a TTSS-dependent manner. Furthermore, the induction of defence responses and disease resistance to X. campestris pv. campestris strain 8004 requires NDR1 (NON-RACE-SPECIFIC DISEASE RESISTANCE1), RAR1 (required for Mla12 resistance) and SGT1b (suppressor of G2 allele of skp1), suggesting that effector-triggered immunity plays a large role in resistance to this strain. Consistent with this notion, AvrXccC, an X. campestris pv. campestris TTSS effector protein, induces PR1 expression and confers resistance in Arabidopsis in a RAR1- and SGT1b-dependent manner. In rar1 and sgt1b mutants, AvrXccC acts as a virulence factor, presumably because of impaired resistance gene function. © 2010 The Authors. Molecular Plant Pathology © 2010 BSPP and Blackwell Publishing Ltd.

  6. Identification of novel type III secretion effectors in Xanthomonas oryzae pv. oryzae.

    PubMed

    Furutani, Ayako; Takaoka, Minako; Sanada, Harumi; Noguchi, Yukari; Oku, Takashi; Tsuno, Kazunori; Ochiai, Hirokazu; Tsuge, Seiji

    2009-01-01

    Many gram-negative bacteria secrete so-called effector proteins via a type III secretion (T3S) system. Through genome screening for genes encoding potential T3S effectors, 60 candidates were selected from rice pathogen Xanthomonas oryzae pv. oryzae MAFF311018 using these criteria: i) homologs of known T3S effectors in plant-pathogenic bacteria, ii) genes with expression regulated by hrp regulatory protein HrpX, or iii) proteins with N-terminal amino acid patterns associated with T3S substrates of Pseudomonas syringae. Of effector candidates tested with the Bordetella pertussis calmodulin-dependent adenylate cyclase reporter for translocation into plant cells, 16 proteins were translocated in a T3S system-dependent manner. Of these 16 proteins, nine were homologs of known effectors in other plant-pathogenic bacteria and seven were not. Most of the effectors were widely conserved in Xanthomonas spp.; however, some were specific to X. oryzae. Interestingly, all these effectors were expressed in an HrpX-dependent manner, suggesting coregulation of effectors and the T3S system. In X. campestris pv. vesicatoria, HpaB and HpaC (HpaP in X. oryzae pv. oryzae) have a central role in recruiting T3S substrates to the secretion apparatus. Secretion of all but one effector was reduced in both HpaB() and HpaP() mutant strains, indicating that HpaB and HpaP are widely involved in efficient secretion of the effectors.

  7. The 9-lipoxygenase GhLOX1 gene is associated with the hypersensitive reaction of cotton Gossypium hirsutum to Xanthomonas campestris pv malvacearum.

    PubMed

    Marmey, Philippe; Jalloul, Aïda; Alhamdia, Majd; Assigbetse, Komi; Cacas, Jean-Luc; Voloudakis, Andreas E; Champion, Antony; Clerivet, Alain; Montillet, Jean-Luc; Nicole, Michel

    2007-08-01

    Hypersensitive reaction (HR) cell death of cotton to the incompatible race 18 from Xanthomonas campestris pathovar malvacearum (Xcm) is associated with 9S-lipoxygenase activity (LOX) responsible for lipid peroxidation. Here, we report the cloning of cotton (Gossypium hirsutum L.) LOX gene (GhLOX1) and the sequencing of its promoter. GhLOX1 was found to be highly expressed during Xcm induced HR. Sequence analysis showed that GhLOX1 is a putative 9-LOX, and GhLOX1 promoter contains SA and JA responsive elements. Investigation on LOX signalisation on cotyledons infiltrated with salicylic acid (SA), or incubated with methyl-jasmonate (MeJA) revealed that both treatments induced LOX activity and GhLOX1 gene expression. HR-like symptoms were observed when LOX substrates were then injected in treated (MeJA and SA) cotyledons or when Xcm compatible race 20 was inoculated on MeJA treated cotyledons. Together these results support the fact that GhLOX1 encodes a 9 LOX whose activity would be involved in cell death during cotton HR.

  8. Diversity of Xanthomonas campestris Isolates from Symptomatic Crucifers in New York State.

    PubMed

    Lange, H W; Tancos, M A; Carlson, M O; Smart, C D

    2016-02-01

    To assess the diversity of Xanthomonas campestris spp. infecting crucifers in New York, 154 isolates were collected over 10 years across the state. The goal was to determine if isolates of the pathogen were overwintering in New York and serving as primary inoculum in subsequent years, or if novel isolates were entering the state each year. Pure cultures of isolates were characterized using multilocus sequence analysis (MLSA), a greenhouse pathogenicity assay, repetitive element-polymerase chain reaction (Rep-PCR) using the BOX-A1R primer, and enzyme-linked immunosorbent assay. The MLSA scheme proved to be more efficient than Rep-PCR for a large sample population and for comparison with global isolates. X. campestris isolated from crucifers in New York comprised of X. campestris pv. campestris and X. campestris pv. raphani, with X. campestris pv. raphani being predominately isolated from transplants. Evidence for unique haplotypes persisting on the same farm for several years due to improper seedbed rotations was documented in addition to novel haplotypes being spread throughout states through infected transplants and seed. Rep-PCR confirmed the high diversity of X. campestris and was used to generate 15 unique fingerprint patterns from isolates collected in the first 5 years. A worldwide comparison of isolates suggests that the X. campestris pv. campestris population appears to be very homogenous with dominant haplotypes persisting for extended periods and being globally disseminated.

  9. A hot pepper gene encoding WRKY transcription factor is induced during hypersensitive response to Tobacco mosaic virus and Xanthomonas campestris.

    PubMed

    Park, Chang-Jin; Shin, Yun-Chul; Lee, Boo-Ja; Kim, Ki-Jeong; Kim, Jeong-Kook; Paek, Kyung-Hee

    2006-01-01

    Plant WRKY transcription factors were previously implicated in the alteration of gene expression in response to various pathogens. The WRKY proteins constitute a large family of plant transcription factors, whose precise functions have yet to be elucidated. Using a domain-specific differential display procedure, we isolated a WRKY gene, which is rapidly induced during an incompatible interaction between hot pepper and Tobacco mosaic virus (TMV) or Xanthomonas campestris pv . vesicatoria (Xcv). The full-length cDNA of CaWRKY-a (Capsicum annuum WRKY-a) encodes a putative polypeptide of 546 amino acids, containing two WRKY domains with a zinc finger motif. The expression of CaWRKY-a could be rapidly induced by not only chemical elicitor such as salicylic acid (SA) or ethephon but also wounding treatments. The nuclear localization of CaWRKY-a was determined in transient expression system using tobacco BY-2 cells by polyethylene glycol (PEG)-mediated transformation experiment. With oligonucleotide molecules containing the putative W-box sequences as a probe, we confirmed that CaWRKY-a protein had W-box-binding activity. These results suggest that CaWRKY-a might be involved as a transcription factor in plant defense-related signal transduction pathways.

  10. Unconventional membrane lipid biosynthesis in Xanthomonas campestris.

    PubMed

    Aktas, Meriyem; Narberhaus, Franz

    2015-09-01

    All bacteria are surrounded by at least one bilayer membrane mainly composed of phospholipids (PLs). Biosynthesis of the most abundant PLs phosphatidylethanolamine (PE), phosphatidylglycerol (PG) and cardiolipin (CL) is well understood in model bacteria such as Escherichia coli. It recently emerged, however, that the diversity of bacterial membrane lipids is huge and that not yet explored biosynthesis pathways exist, even for the common PLs. A good example is the plant pathogen Xanthomonas campestris pv. campestris. It contains PE, PG and CL as major lipids and small amounts of the N-methylated PE derivatives monomethyl PE and phosphatidylcholine (PC = trimethylated PE). Xanthomonas campestris uses a repertoire of canonical and non-canonical enzymes for the synthesis of its membrane lipids. In this minireview, we briefly recapitulate standard pathways and integrate three recently discovered pathways into the overall picture of bacterial membrane biosynthesis.

  11. AvrACXcc8004, a Type III Effector with a Leucine-Rich Repeat Domain from Xanthomonas campestris Pathovar campestris Confers Avirulence in Vascular Tissues of Arabidopsis thaliana Ecotype Col-0▿ †

    PubMed Central

    Xu, Rong-Qi; Blanvillain, Servane; Feng, Jia-Xun; Jiang, Bo-Le; Li, Xian-Zhen; Wei, Hong-Yu; Kroj, Thomas; Lauber, Emmanuelle; Roby, Dominique; Chen, Baoshan; He, Yong-Qiang; Lu, Guang-Tao; Tang, Dong-Jie; Vasse, Jacques; Arlat, Matthieu; Tang, Ji-Liang

    2008-01-01

    Xanthomonas campestris pathovar campestris causes black rot, a vascular disease on cruciferous plants, including Arabidopsis thaliana. The gene XC1553 from X. campestris pv. campestris strain 8004 encodes a protein containing leucine-rich repeats (LRRs) and appears to be restricted to strains of X. campestris pv. campestris. LRRs are found in a number of type III-secreted effectors in plant and animal pathogens. These prompted us to investigate the role of the XC1553 gene in the interaction between X. campestris pv. campestris and A. thaliana. Translocation assays using the hypersensitive-reaction-inducing domain of X. campestris pv. campestris AvrBs1 as a reporter revealed that XC1553 is a type III effector. Infiltration of Arabidopsis leaf mesophyll with bacterial suspensions showed no differences between the wild-type strain and an XC1553 gene mutant; both strains induced disease symptoms on Kashmir and Col-0 ecotypes. However, a clear difference was observed when bacteria were introduced into the vascular system by piercing the central vein of leaves. In this case, the wild-type strain 8004 caused disease on the Kashmir ecotype, but not on ecotype Col-0; the XC1553 gene mutant became virulent on the Col-0 ecotype and still induced disease on the Kashmir ecotype. Altogether, these data show that the XC1553 gene, which was renamed avrACXcc8004, functions as an avirulence gene whose product seems to be recognized in vascular tissues. PMID:17951377

  12. The Host Plant Metabolite Glucose Is the Precursor of Diffusible Signal Factor (DSF) Family Signals in Xanthomonas campestris

    PubMed Central

    Liu, Xiaoling; Wu, Ji'en; Lee, Jasmine; Chen, Shaohua; Cheng, Yingying; Zhang, Chunyan

    2015-01-01

    Plant pathogen Xanthomonas campestris pv. campestris produces cis-11-methyl-2-dodecenoic acid (diffusible signal factor [DSF]) as a cell-cell communication signal to regulate biofilm dispersal and virulence factor production. Previous studies have demonstrated that DSF biosynthesis is dependent on the presence of RpfF, an enoyl-coenzyme A (CoA) hydratase, but the DSF synthetic mechanism and the influence of the host plant on DSF biosynthesis are still not clear. We show here that exogenous addition of host plant juice or ethanol extract to the growth medium of X. campestris pv. campestris could significantly boost DSF family signal production. It was subsequently revealed that X. campestris pv. campestris produces not only DSF but also BDSF (cis-2-dodecenoic acid) and another novel DSF family signal, which was designated DSF-II. BDSF was originally identified in Burkholderia cenocepacia to be involved in regulation of motility, biofilm formation, and virulence in B. cenocepacia. Functional analysis suggested that DSF-II plays a role equal to that of DSF in regulation of biofilm dispersion and virulence factor production in X. campestris pv. campestris. Furthermore, chromatographic separation led to identification of glucose as a specific molecule stimulating DSF family signal biosynthesis in X. campestris pv. campestris. 13C-labeling experiments demonstrated that glucose acts as a substrate to provide a carbon element for DSF biosynthesis. The results of this study indicate that X. campestris pv. campestris could utilize a common metabolite of the host plant to enhance DSF family signal synthesis and therefore promote virulence. PMID:25681189

  13. The host plant metabolite glucose is the precursor of diffusible signal factor (DSF) family signals in Xanthomonas campestris.

    PubMed

    Deng, Yinyue; Liu, Xiaoling; Wu, Ji'en; Lee, Jasmine; Chen, Shaohua; Cheng, Yingying; Zhang, Chunyan; Zhang, Lian-Hui

    2015-04-01

    Plant pathogen Xanthomonas campestris pv. campestris produces cis-11-methyl-2-dodecenoic acid (diffusible signal factor [DSF]) as a cell-cell communication signal to regulate biofilm dispersal and virulence factor production. Previous studies have demonstrated that DSF biosynthesis is dependent on the presence of RpfF, an enoyl-coenzyme A (CoA) hydratase, but the DSF synthetic mechanism and the influence of the host plant on DSF biosynthesis are still not clear. We show here that exogenous addition of host plant juice or ethanol extract to the growth medium of X. campestris pv. campestris could significantly boost DSF family signal production. It was subsequently revealed that X. campestris pv. campestris produces not only DSF but also BDSF (cis-2-dodecenoic acid) and another novel DSF family signal, which was designated DSF-II. BDSF was originally identified in Burkholderia cenocepacia to be involved in regulation of motility, biofilm formation, and virulence in B. cenocepacia. Functional analysis suggested that DSF-II plays a role equal to that of DSF in regulation of biofilm dispersion and virulence factor production in X. campestris pv. campestris. Furthermore, chromatographic separation led to identification of glucose as a specific molecule stimulating DSF family signal biosynthesis in X. campestris pv. campestris. (13)C-labeling experiments demonstrated that glucose acts as a substrate to provide a carbon element for DSF biosynthesis. The results of this study indicate that X. campestris pv. campestris could utilize a common metabolite of the host plant to enhance DSF family signal synthesis and therefore promote virulence.

  14. The Xanthomonas campestris type III effector XopJ proteolytically degrades proteasome subunit RPT6.

    PubMed

    Üstün, Suayib; Börnke, Frederik

    2015-05-01

    Many animal and plant pathogenic bacteria inject type III effector (T3E) proteins into their eukaryotic host cells to suppress immunity. The Yersinia outer protein J (YopJ) family of T3Es is a widely distributed family of effector proteins found in both animal and plant pathogens, and its members are highly diversified in virulence functions. Some members have been shown to possess acetyltransferase activity; however, whether this is a general feature of YopJ family T3Es is currently unknown. The T3E Xanthomonas outer protein J (XopJ), a YopJ family effector from the plant pathogen Xanthomonas campestris pv vesicatoria, interacts with the proteasomal subunit Regulatory Particle AAA-ATPase6 (RPT6) in planta to suppress proteasome activity, resulting in the inhibition of salicylic acid-related immune responses. Here, we show that XopJ has protease activity to specifically degrade RPT6, leading to reduced proteasome activity in the cytoplasm as well as in the nucleus. Proteolytic degradation of RPT6 was dependent on the localization of XopJ to the plasma membrane as well as on its catalytic triad. Mutation of the Walker B motif of RPT6 prevented XopJ-mediated degradation of the protein but not XopJ interaction. This indicates that the interaction of RPT6 with XopJ is dependent on the ATP-binding activity of RPT6, but proteolytic cleavage additionally requires its ATPase activity. Inhibition of the proteasome impairs the proteasomal turnover of Nonexpressor of Pathogenesis-Related1 (NPR1), the master regulator of salicylic acid responses, leading to the accumulation of ubiquitinated NPR1, which likely interferes with the full induction of NPR1 target genes. Our results show that YopJ family T3Es are not only highly diversified in virulence function but also appear to possess different biochemical activities. © 2015 American Society of Plant Biologists. All Rights Reserved.

  15. The Xanthomonas campestris Type III Effector XopJ Proteolytically Degrades Proteasome Subunit RPT61[OPEN

    PubMed Central

    2015-01-01

    Many animal and plant pathogenic bacteria inject type III effector (T3E) proteins into their eukaryotic host cells to suppress immunity. The Yersinia outer protein J (YopJ) family of T3Es is a widely distributed family of effector proteins found in both animal and plant pathogens, and its members are highly diversified in virulence functions. Some members have been shown to possess acetyltransferase activity; however, whether this is a general feature of YopJ family T3Es is currently unknown. The T3E Xanthomonas outer protein J (XopJ), a YopJ family effector from the plant pathogen Xanthomonas campestris pv vesicatoria, interacts with the proteasomal subunit Regulatory Particle AAA-ATPase6 (RPT6) in planta to suppress proteasome activity, resulting in the inhibition of salicylic acid-related immune responses. Here, we show that XopJ has protease activity to specifically degrade RPT6, leading to reduced proteasome activity in the cytoplasm as well as in the nucleus. Proteolytic degradation of RPT6 was dependent on the localization of XopJ to the plasma membrane as well as on its catalytic triad. Mutation of the Walker B motif of RPT6 prevented XopJ-mediated degradation of the protein but not XopJ interaction. This indicates that the interaction of RPT6 with XopJ is dependent on the ATP-binding activity of RPT6, but proteolytic cleavage additionally requires its ATPase activity. Inhibition of the proteasome impairs the proteasomal turnover of Nonexpressor of Pathogenesis-Related1 (NPR1), the master regulator of salicylic acid responses, leading to the accumulation of ubiquitinated NPR1, which likely interferes with the full induction of NPR1 target genes. Our results show that YopJ family T3Es are not only highly diversified in virulence function but also appear to possess different biochemical activities. PMID:25739698

  16. Induction of Hydrolytic Enzymes in Brassica campestris in Response to Pathovars of Xanthomonas campestris.

    PubMed

    Conrads-Strauch, J; Dow, J M; Milligan, D E; Parra, R; Daniels, M J

    1990-05-01

    Inoculation of mature leaves of turnip (Brassica campestris) with the incompatible Xanthomonas campestris pv vitians resulted in the induction of beta-1,3-glucanase and chitinase/lysozyme (CHL) activity. No increase in the basal activity of beta-1,3-glucanase was observed after inoculation of leaves with heat- or rifampicin-killed X. c. vitians, Escherichia coli, or sterile water. Inoculation with the compatible X. campestris pv campestris resulted in a slower induction of glucanase than that seen with X. c. vitians. In contrast, all bacteria caused an induction of CHL activity. One major beta-1,3-glucanase (molecular mass 36.5 kilodaltons, isoelectric point [pl] ~8.5) was purified from both inoculated and untreated leaves by ion-exchange chromatography. The enzyme degraded laminarin by an endo-glycolytic mechanism. Two major CHL isozymes (CHL 1 and CHL 2, molecular mass 30 kilodaltons and pl 9.4 and 10.2, respectively) were purified from X. c. vitians inoculated leaves by affinity chromatography on a chitin column followed by ion-exchange chromatography. Both enzymes degraded chitin by an endo-glycolytic mechanism although the ratio of lysozyme to chitinase specific activities for CHL 1 and CHL2 were different. The induction of CHL 1 was associated with the hypersensitive reaction caused by X. c. vitians whereas all other treatments induced largely CHL 2.

  17. Differential expression of conserved protease genes in crucifer-attacking pathovars of Xanthomonas campestris.

    PubMed

    Dow, J M; Fan, M J; Newman, M A; Daniels, M J

    1993-12-01

    Strains of Xanthomonas campestris pathovars armoraciae and raphani, which cause leaf spotting diseases in brassicas, produce a major extracellular protease in liquid culture which was partially purified. The protease (PRT 3) was a zinc-requiring metalloenzyme and was readily distinguishable from the two previously characterized proteases (PRT 1 and PRT 2) of X. campestris pv. campestris by the pattern of degradation of beta-casein and sensitivity to inhibitors. PRT 3 was produced at a low level in the vascular brassica pathogen X. campestris pv. campestris (five strains tested), in which PRT 1 and PRT 2 predominate. In contrast, expression of PRT 1, a serine protease, could not be detected in the six tested strains of the leaf spotting mesophyll pathogens. However, all these strains had DNA fragments which hybridized to a prtA probe and which probably carry a functional prtA (the structural gene for PRT 1). The structural gene for PRT 3 (prtC) was cloned by screening a genomic library of X. campestris pv. raphani in a protease-deficient X. campestris pv. campestris strain. Subcloning and Tn5 mutagenesis located the structural gene to 1.2 kb of DNA. DNA fragments which hybridized to the structural gene were found in all strains of the crucifer-attacking X. campestris pathovars tested as well as in a number of other pathovars. Experiments in which the pattern of protease production of the pathovars was manipulated by introduction of cloned genes into heterologous pathovars suggested that no determinative relationship exists between the pattern of protease gene expression and the (vascular or mesophyllic) mode of pathogenesis.

  18. [The extracellular proteases of the phytopathogenic bacterium Xanthomonas campestris].

    PubMed

    Kalashnikova, E E; Chernyshova, M P; Ignatov, V V

    2003-01-01

    The culture liquids of three Xanthomonas campestris pv. campestris strains were found to possess proteolytic activity. The culture liquid of strain B-611 with the highest proteolytic activity was fractionated by salting-out with ammonium sulfate, gel filtration, and ion-exchange chromatography. The electrophoretic analysis of active fractions showed the presence of two proteases in the culture liquid of strain B-611, the major of which being serine protease. The treatment of cabbage seedlings with the proteases augmented the activity of peroxidase in the cabbage roots by 28%.

  19. A metalloprotease from Xanthomonas campestris that specifically degrades proline/hydroxyproline-rich glycoproteins of the plant extracellular matrix.

    PubMed

    Dow, J M; Davies, H A; Daniels, M J

    1998-11-01

    Culture supernatants of Xanthomonas campestris pv. campestris contain an enzymic activity capable of degrading gp120, a proline-rich glycoprotein associated with the extracellular matrix of the vascular bundles in petioles of turnip (Brassica campestris). This activity did not reside in any of the three previously characterized proteases of X. campestris pv. campestris that were identified by their action against the model substrate beta-casein. The novel enzyme was purified by ion-exchange and size-exclusion high-performance liquid chromatography (HPLC). The enzyme, which has no activity against beta-casein, is active against some plant glycoproteins of the hydroxyproline-rich class such as extensin from potato and tomato and gpS-3, a glycoprotein induced in B. campestris petioles by wounding. Other hydroxyproline-rich glycoproteins, such as the solanaceous lectins, were not substrates however. Studies of the products released upon degradation of tomato extensin suggested that the degradative mechanism was proteolysis. Inhibitor studies suggested that the enzyme was a zinc-requiring metalloprotease. Extracellular matrix glycoproteins of the proline-rich and hydroxyproline-rich classes have been implicated in plant resistance to microbial attack, hence their degradation by X. campestris pv. campestris may have considerable significance for black rot pathogenesis.

  20. Characterization of Xanthomonas campestris Pathovars by rRNA Gene Restriction Patterns

    PubMed Central

    Berthier, Yvette; Verdier, Valérie; Guesdon, Jean-Luc; Chevrier, Danièle; Denis, Jean-Baptiste; Decoux, Guy; Lemattre, Monique

    1993-01-01

    Genomic DNA of 191 strains of the family Pseudomonadaceae, including 187 strains of the genus Xanthomonas, was cleaved by EcoRI endonuclease. After hybridization of Southern transfer blots with 2-acetylamino-fluorene-labelled Escherichia coli 16+23S rRNA probe, 27 different patterns were obtained. The strains are clearly distinguishable at the genus, species, and pathovar levels. The variability of the rRNA gene restriction patterns was determined for four pathovars of Xanthomonas campestris species. The 16 strains of X. campestris pv. begoniae analyzed gave only one pattern. The variability of rRNA gene restriction patterns of X. campestris pv. manihotis strains could be related to ecotypes. In contrast, the variability of patterns observed for X. campestris pv. malvacearum was not correlated with pathogenicity or with the geographical origins of the strains. The highest degree of variability of DNA fingerprints was observed within X. campestris pv. dieffenbachiae, which is pathogenic to several hosts of the Araceae family. In this case, variability was related to both host plant and pathogenicity. Images PMID:16348894

  1. Synergistic Antimycobacterial Actions of Knowltonia vesicatoria (L.f) Sims

    PubMed Central

    Labuschagné, Antoinette; Hussein, Ahmed A.; Rodríguez, Benjamín; Lall, Namrita

    2012-01-01

    Euclea natalensis A.DC., Knowltonia vesicatoria (L.f) Sims, and Pelargonium sidoides DC. are South African plants traditionally used to treat tuberculosis. Extracts from these plants were used in combination with isoniazid (INH) to investigate the possibility of synergy with respect to antimycobacterial activity. The ethanol extract of K. vesicatoria was subjected to fractionation to identify the active compounds. The activity of the Knowltonia extract remained superior to the fractions with a minimum inhibitory concentration (MIC) of 625.0 μg/mL against Mycobacterium smegmatis and an MIC of 50.00 μg/mL against M. tuberculosis. The K. vesicatoria extract was tested against two different drug-resistant strains of M. tuberculosis, which resulted in an MIC of 50.00 μg/mL on both strains. The combination of K. vesicatoria with INH exhibited the best synergistic antimycobacterial activity with a fractional inhibitory concentration index of 0.25 (a combined concentration of 6.28 μg/mL). A fifty percent inhibitory concentration of this combination against U937 cells was 121.0 μg/mL. Two compounds, stigmasta-5,23-dien-3-ol (1) and 5-(hydroxymethyl)furan-2(5H)-one (2), were isolated from K. vesicatoria as the first report of isolation for both compounds from this plant and the first report of antimycobacterial activity. Compound (1) was active against drug-sensitive M. tuberculosis with an MIC of 50.00 μg/mL. PMID:22611433

  2. The Xanthomonas campestris type III effector XopJ targets the host cell proteasome to suppress salicylic-acid mediated plant defence.

    PubMed

    Üstün, Suayib; Bartetzko, Verena; Börnke, Frederik

    2013-01-01

    The phytopathogenic bacterium Xanthomonas campestris pv. vesicatoria (Xcv) requires type III effector proteins (T3Es) for virulence. After translocation into the host cell, T3Es are thought to interact with components of host immunity to suppress defence responses. XopJ is a T3E protein from Xcv that interferes with plant immune responses; however, its host cellular target is unknown. Here we show that XopJ interacts with the proteasomal subunit RPT6 in yeast and in planta to inhibit proteasome activity. A C235A mutation within the catalytic triad of XopJ as well as a G2A exchange within the N-terminal myristoylation motif abolishes the ability of XopJ to inhibit the proteasome. Xcv ΔxopJ mutants are impaired in growth and display accelerated symptom development including tissue necrosis on susceptible pepper leaves. Application of the proteasome inhibitor MG132 restored the ability of the Xcv ΔxopJ to attenuate the development of leaf necrosis. The XopJ dependent delay of tissue degeneration correlates with reduced levels of salicylic acid (SA) and changes in defence- and senescence-associated gene expression. Necrosis upon infection with Xcv ΔxopJ was greatly reduced in pepper plants with reduced expression of NPR1, a central regulator of SA responses, demonstrating the involvement of SA-signalling in the development of XopJ dependent phenotypes. Our results suggest that XopJ-mediated inhibition of the proteasome interferes with SA-dependent defence response to attenuate onset of necrosis and to alter host transcription. A central role of the proteasome in plant defence is discussed.

  3. The Xanthomonas campestris Type III Effector XopJ Targets the Host Cell Proteasome to Suppress Salicylic-Acid Mediated Plant Defence

    PubMed Central

    Börnke, Frederik

    2013-01-01

    The phytopathogenic bacterium Xanthomonas campestris pv. vesicatoria (Xcv) requires type III effector proteins (T3Es) for virulence. After translocation into the host cell, T3Es are thought to interact with components of host immunity to suppress defence responses. XopJ is a T3E protein from Xcv that interferes with plant immune responses; however, its host cellular target is unknown. Here we show that XopJ interacts with the proteasomal subunit RPT6 in yeast and in planta to inhibit proteasome activity. A C235A mutation within the catalytic triad of XopJ as well as a G2A exchange within the N-terminal myristoylation motif abolishes the ability of XopJ to inhibit the proteasome. Xcv ΔxopJ mutants are impaired in growth and display accelerated symptom development including tissue necrosis on susceptible pepper leaves. Application of the proteasome inhibitor MG132 restored the ability of the Xcv ΔxopJ to attenuate the development of leaf necrosis. The XopJ dependent delay of tissue degeneration correlates with reduced levels of salicylic acid (SA) and changes in defence- and senescence-associated gene expression. Necrosis upon infection with Xcv ΔxopJ was greatly reduced in pepper plants with reduced expression of NPR1, a central regulator of SA responses, demonstrating the involvement of SA-signalling in the development of XopJ dependent phenotypes. Our results suggest that XopJ-mediated inhibition of the proteasome interferes with SA-dependent defence response to attenuate onset of necrosis and to alter host transcription. A central role of the proteasome in plant defence is discussed. PMID:23785289

  4. The cloning, crystallization and preliminary X-ray analysis of XC2113, a YaeQ protein from Xanthomonas campestris

    SciTech Connect

    Chio, Kuo-Cheng; Chin, Ko-Hsin; Gao, Fei Philip; Lyu, Ping-Chiang; Shr, Hui-Lin; Wang, Andrew H.-J.; Chou, Shan-Ho

    2006-10-01

    A YaeQ protein from the plant pathogen X. campestris pv. campestris has been overexpressed in E. coli, purified and crystallized. The crystals diffracted well to a resolution of 1.28 Å. Xanthomonas campestris is a Gram-negative bacterium that is phytopathogenic to cruciferous plants and causes worldwide agricultural loss. It is therefore important to identify potential pathogenic factors involved in this plant disease. Here, the cloning, expression, crystallization and preliminary X-ray analysis of XC2113, a YaeQ protein possibly involved in the production of virulence factors in Xanthomonas campestris pathovar campestris, are reported. The XC2113 crystals diffracted well to a resolution of at least 1.28 Å. They are orthorhombic and belong to space group P2{sub 1}2{sub 1}2{sub 1}, with unit-cell parameters a = 32.86, b = 62.69, c = 79.96 Å.

  5. An adenosine kinase exists in Xanthomonas campestris pathovar campestris and is involved in extracellular polysaccharide production, cell motility, and virulence.

    PubMed

    Lu, Guang-Tao; Tang, Yong-Qin; Li, Cai-Yue; Li, Rui-Fang; An, Shi-Qi; Feng, Jia-Xun; He, Yong-Qiang; Jiang, Bo-Le; Tang, Dong-Jie; Tang, Ji-Liang

    2009-06-01

    Adenosine kinase (ADK) is a purine salvage enzyme and a typical housekeeping enzyme in eukaryotes which catalyzes the phosphorylation of adenosine to form AMP. Since prokaryotes synthesize purines de novo and no endogenous ADK activity is detectable in Escherichia coli, ADK has long been considered to be rare in bacteria. To date, only two prokaryotes, both of which are gram-positive bacteria, have been reported to contain ADK. Here we report that the gram-negative bacterium Xanthomonas campestris pathovar campestris, the causal agent of black rot of crucifers, possesses a gene (designated adk(Xcc)) encoding an ADK (named ADK(Xcc)), and we demonstrate genetically that the ADK(Xcc) is involved in extracellular polysaccharide (EPS) production, cell motility, and pathogenicity of X. campestris pv. campestris. adk(Xcc) was overexpressed as a His(6)-tagged protein in E. coli, and the purified His(6)-tagged protein exhibited ADK activity. Mutation of adk(Xcc) did not affect bacterial growth in rich and minimal media but led to an accumulation of intracellular adenosine and diminutions of intracellular ADK activity and ATP level, as well as EPS. The adk(Xcc) mutant displayed significant reductions in bacterial growth and virulence in the host plant.

  6. Defense-related gene induction in Brassica campestris in response to defined mutants of Xanthomonas campestris with altered pathogenicity.

    PubMed

    Newman, M A; Conrads-Strauch, J; Scofield, G; Daniels, M J; Dow, J M

    1994-01-01

    We have studied the induction of beta-1,3-glucanase (BGL) in turnip following inoculation with pathovars of Xanthomonas campestris and derived mutants. BGL transcript accumulated more rapidly in leaves in the incompatible interactions with X. c. pv. armoraciae and X. c. pv. raphani than in the compatible interaction with X. c. pv. campestris. No accumulation was seen in response to wounding or inoculation with water, salicylic acid, or Escherichia coli. Deletion of the hrp cluster from the X. campestris pathovars caused a reduction in the level of transcript accumulation; these effects were much more pronounced in the incompatible than in the compatible interaction, in which bacterial growth was also affected. In the compatible interaction, bacterial growth and BGL transcript accumulation were not altered by mutation of bacterial genes involved in the regulation of the synthesis of extracellular enzymes or their export from the cell, or by mutation of the structural genes for extracellular endoglucanase and serine protease. Mutation of genes involved in the synthesis of extracellular polysaccharide or lipopolysaccharide reduced bacterial survival in planta, so that the numbers were between two and three orders of magnitude lower than the number of wild-type bacteria. However, total BGL transcript accumulation after inoculation with these mutants was about 80% of that seen after inoculation with the wild-type bacteria, suggesting that one aspect of the role of extracellular polysaccharide and lipopolysaccharide in pathogenesis is to mask the presence of bacteria in the plant. Our results are discussed in the context of work on other plant-microbe interactions.

  7. Novel Roles of SoxR, a Transcriptional Regulator from Xanthomonas campestris, in Sensing Redox-Cycling Drugs and Regulating a Protective Gene That Have Overall Implications for Bacterial Stress Physiology and Virulence on a Host Plant

    PubMed Central

    Mahavihakanont, Aekkapol; Charoenlap, Nisanart; Namchaiw, Poommaree; Eiamphungporn, Warawan; Chattrakarn, Sorayut

    2012-01-01

    In Xanthomonas campestris pv. campestris, SoxR likely functions as a sensor of redox-cycling drugs and as a transcriptional regulator. Oxidized SoxR binds directly to its target site and activates the expression of xcc0300, a gene that has protective roles against the toxicity of redox-cycling compounds. In addition, SoxR acts as a noninducible repressor of its own expression. X. campestris pv. campestris requires SoxR both for protection against redox-cycling drugs and for full virulence on a host plant. The X. campestris model of the gene regulation and physiological roles of SoxR represents a novel variant of existing bacterial SoxR models. PMID:22056938

  8. Novel roles of SoxR, a transcriptional regulator from Xanthomonas campestris, in sensing redox-cycling drugs and regulating a protective gene that have overall implications for bacterial stress physiology and virulence on a host plant.

    PubMed

    Mahavihakanont, Aekkapol; Charoenlap, Nisanart; Namchaiw, Poommaree; Eiamphungporn, Warawan; Chattrakarn, Sorayut; Vattanaviboon, Paiboon; Mongkolsuk, Skorn

    2012-01-01

    In Xanthomonas campestris pv. campestris, SoxR likely functions as a sensor of redox-cycling drugs and as a transcriptional regulator. Oxidized SoxR binds directly to its target site and activates the expression of xcc0300, a gene that has protective roles against the toxicity of redox-cycling compounds. In addition, SoxR acts as a noninducible repressor of its own expression. X. campestris pv. campestris requires SoxR both for protection against redox-cycling drugs and for full virulence on a host plant. The X. campestris model of the gene regulation and physiological roles of SoxR represents a novel variant of existing bacterial SoxR models.

  9. Use of bioluminescence for detection of genetically engineered microorganisms released into the environment. [Xanthomonas campestris

    SciTech Connect

    Shaw, J.J.; Dane, F.; Geiger, D.; Kloepper, J.W. )

    1992-01-01

    The persistence and movement of strain JS414 of Xanthomonas campestris pv. campestris, which was genetically engineered to bioluminesce, were monitored during a limited field introduction. Bioluminescence and traditional dilution plate counts were determined. Strain JS414 was applied to cabbage plants and surrounding soil by mist inoculation, by wound inoculation, by scattering infested debris among plants, and by incorporating bacteria into the soil. Bioluminescent X. campestris pv. campestris was detected in plant samples and in the rhizosphere up to 6 weeks after inoculation. Movement to uninoculated plants was detected on one occasion, but movement from the immediate release area was not detected. Strain JS414 was detected in soil samples beneath mist- and wound-inoculated plants only at intentionally infested locations and in aerial samples only on the day of inoculation. The authors bioluminescence methods proved to be as sensitive as plating methods for detecting the genetically engineered microorganisms in environmental samples. Their results demonstrate that transgenic incorporation of the luxCDABE operon provides a non-labor-intensive, sensitive detection method for monitoring genetically engineered microorganisms in nature.

  10. The structure of the lipooligosaccharide from Xanthomonas oryzae pv. Oryzae: the causal agent of the bacterial leaf blight in rice.

    PubMed

    Di Lorenzo, Flaviana; Palmigiano, Angelo; Silipo, Alba; Desaki, Yoshitake; Garozzo, Domenico; Lanzetta, Rosa; Shibuya, Naoto; Molinaro, Antonio

    2016-06-02

    The structure of the lipooligosaccharide (LOS) from the rice pathogen Xanthomonas oryzae pv. oryzae has been elucidated. The characterization of the core oligosaccharide structure was obtained by the employment of two chemical degradation protocols and by analysis of the products via NMR spectroscopy. The structure of the lipid A portion was achieved by MALDI mass spectrometry analysis on purified lipid A. The LOS from Xanthomonas oryzae pv. oryzae revealed to possess the same core structure of Xanthomonas campestris pv. campestris and interesting novel features on its lipid A domain. The evaluation of the biological activity of both LOS and isolated lipid A was also executed.

  11. Nutritional Similarity between Leaf-Associated Nonpathogenic Bacteria and the Pathogen Is Not Predictive of Efficacy in Biological Control of Bacterial Spot of Tomato

    PubMed Central

    Dianese, Alexei C.; Ji, Pingsheng; Wilson, Mark

    2003-01-01

    It has been demonstrated that for a nonpathogenic, leaf-associated bacterium, effectiveness in the control of bacterial speck of tomato is correlated with the similarity in the nutritional needs of the nonpathogenic bacterium and the pathogen Pseudomonas syringae pv. tomato. This relationship was investigated further in this study by using the pathogen Xanthomonas campestris pv. vesicatoria, the causal agent of bacterial spot of tomato, and a collection of nonpathogenic bacteria isolated from tomato foliage. The effects of inoculation of tomato plants with one of 34 nonpathogenic bacteria prior to inoculation with the pathogen X. campestris pv. vesicatoria were quantified by determining (i) the reduction in disease severity (number of lesions per square centimeter) in greenhouse assays and (ii) the reduction in leaf surface pathogen population size (log10 of the number of CFU per leaflet) in growth chamber assays. Nutritional similarity between the nonpathogenic bacteria and X. campestris pv. vesicatoria was quantified by using either niche overlap indices (NOI) or relatedness in cluster analyses based upon in vitro utilization of carbon or nitrogen sources reported to be present in tomato tissues or in Biolog GN plates. In contrast to studies with P. syringae pv. tomato, nutritional similarity between the nonpathogenic bacteria and the pathogen X. campestris pv. vesicatoria was not correlated with reductions in disease severity. Nutritional similarity was also not correlated with reductions in pathogen population size. Further, the percentage of reduction in leaf surface pathogen population size was not correlated with the percentage of reduction in disease severity, suggesting that the epiphytic population size of X. campestris pv. vesicatoria is not related to disease severity and that X. campestris pv. vesicatoria exhibits behavior in the phyllosphere prior to lesion formation that is different from that of P. syringae pv. tomato. PMID:12788754

  12. Cloning, purification crystallization and preliminary X-ray characterization of a conserved hypothetical protein XC6422 from Xanthomonas campestris

    SciTech Connect

    Yang, Chao-Yu; Chin, Ko-Hsin; Chou, Chia-Cheng; Shr, Hui-Lin; Gao, Fei Philip; Lyu, Ping-Chiang; Wang, Andrew H.-J.; Chou, Shan-Ho

    2005-07-01

    A conserved hypothetical protein XC6422 from X. campestris pv. campestris has been overexpressed in E. coli, purified and crystallized. Crystals obtained from the purified recombinant protein showed a variety of forms that diffracted to at least 1.6 Å resolution. Xanthomonas campestris pv. campestris is a Gram-negative yellow-pigmented pathogenic bacterium that causes black rot, one of the major worldwide diseases of cruciferous crops. Its genome contains approximately 4500 genes, roughly one third of which have no known structure and/or function. However, some genes of unknown function are highly conserved among several different bacterial genuses. XC6422 is one such conserved hypothetical protein and has been overexpressed in Escherichia coli, purified and crystallized in a variety of forms using the hanging-drop vapour-diffusion method. Crystals grew to approximately 2 × 1.5 × 0.4 mm in size after one week and diffracted to at least 1.6 Å resolution. They belong to the monoclinic space group C2, with one molecule per asymmetric unit and unit-cell parameters a = 75.8, b = 79.3, c = 38.2 Å, β = 109.4°. Determination of this structure may provide insights into the protein’s function.

  13. A putative polyketide-synthesis protein XC5357 from Xanthomonas campestris: heterologous expression, crystallization and preliminary X-ray analysis

    SciTech Connect

    Chu, Chiao-Li; Chin, Ko-Hsin; Lin, Fu-Yang; Chou, Chia-Cheng; Lee, Cheng-Chung; Shr, Hui-Lin; Lyu, Ping-Chiang; Wang, Andrew H.-J.; Chou, Shan-Ho

    2005-07-01

    A putative polyketide-synthesis protein XC5357 from X. campestris pv. campestris has been overexpressed in E. coli, purified and crystallized. The crystals diffracted to a resolution of at least 1.85 Å. Xanthomonas campestris pv. campestris (Xcc) is a Gram-negative yellow-pigmented bacterium and is the causative agent of black rot, one of the major worldwide diseases of cruciferous crops. It also synthesizes a variety of polyketide metabolites that lead to important antibiotics. XC5357 is a putative 12.2 kDa protein of unknown structure from Xcc that is likely to be essential for polyketide synthesis. It was overexpressed in Escherichia coli, purified and crystallized using the hanging-drop vapour-diffusion method. The crystals belong to the triclinic space group P1, with unit-cell parameters a = 43.7, b = 43.7, c = 46.5 Å, α = 65.0, β = 64.9, γ = 73.4°, and diffracted to a resolution of 1.85 Å.

  14. Cloning, purification, crystallization and preliminary X-ray analysis of XC229, a conserved hypothetical protein from Xanthomonas campestris

    SciTech Connect

    Chin, Ko-Hsin; Kuo, Wei-Tien; Chou, Chia-Cheng; Shr, Hui-Lin; Lyu, Ping-Chiang; Wang, Andrew H.-J.; Chou, Shan-Ho

    2005-07-01

    A conserved hypothetical protein XC229 from X. campestris pv. campestris has been overexpressed in E. coli, purified and crystallized. A crystal of the purified recombinant protein diffracted to a resolution of 1.80 Å. Xanthomonas campestris pv. campestris is a Gram-negative yellow-pigmented pathogenic bacterium that causes black rot, one of the major worldwide diseases of cruciferous crops. Its genome contains approximately 4500 genes, roughly one third of which have no known structure and/or function. However, some of these unknown genes are highly conserved among several different bacterial genuses. XC229 is one such protein containing 134 amino acids. It was overexpressed in Escherichia coli, purified and crystallized using the hanging-drop vapour-diffusion method. The crystal diffracted to a resolution of at least 1.80 Å. It is cubic and belongs to space group I2{sub x}3, with unit-cell parameters a = b = c = 106.8 Å. It contains one or two molecules per asymmetric unit.

  15. Preparation, crystallization and preliminary X-ray characterization of a conserved hypothetical protein XC1692 from Xanthomonas campestris

    SciTech Connect

    Chin, Ko-Hsin; Huang, Zhao-Wei; Wei, Kun-Chou; Chou, Chia-Cheng; Lee, Cheng-Chung; Shr, Hui-Lin; Gao, Fei Philip; Lyu, Ping-Chiang; Wang, Andrew H.-J.; Chou, Shan-Ho

    2005-07-01

    A conserved hypothetical protein XC1692 from X. campestris pv. campestris has been overexpressed in E. coli. The purified recombinant protein crystallized in a variety of forms and diffracted to a resolution of at least 1.45 Å. Xanthomonas campestris pv. campestris strain 17 is a Gram-negative yellow-pigmented pathogenic bacterium that causes black rot, one of the major worldwide diseases of cruciferous crops. Its genome contains approximately 4500 genes, one third of which have no known structure and/or function yet are highly conserved among several different bacterial genuses. One of these gene products is XC1692 protein, containing 141 amino acids. It was overexpressed in Escherichia coli, purified and crystallized in a variety of forms using the hanging-drop vapour-diffusion method. The crystals diffract to at least 1.45 Å resolution. They are hexagonal and belong to space group P6{sub 3}, with unit-cell parameters a = b = 56.9, c = 71.0 Å. They contain one molecule per asymmetric unit.

  16. Genomics and transcriptomics of Xanthomonas campestris species challenge the concept of core type III effectome.

    PubMed

    Roux, Brice; Bolot, Stéphanie; Guy, Endrick; Denancé, Nicolas; Lautier, Martine; Jardinaud, Marie-Françoise; Fischer-Le Saux, Marion; Portier, Perrine; Jacques, Marie-Agnès; Gagnevin, Lionel; Pruvost, Olivier; Lauber, Emmanuelle; Arlat, Matthieu; Carrère, Sébastien; Koebnik, Ralf; Noël, Laurent D

    2015-11-18

    The bacterial species Xanthomonas campestris infects a wide range of Brassicaceae. Specific pathovars of this species cause black rot (pv. campestris), bacterial blight of stock (pv. incanae) or bacterial leaf spot (pv. raphani). In this study, we extended the genomic coverage of the species by sequencing and annotating the genomes of strains from pathovar incanae (CFBP 1606R and CFBP 2527R), pathovar raphani (CFBP 5828R) and a pathovar formerly named barbareae (CFBP 5825R). While comparative analyses identified a large core ORFeome at the species level, the core type III effectome was limited to only three putative type III effectors (XopP, XopF1 and XopAL1). In Xanthomonas, these effector proteins are injected inside the plant cells by the type III secretion system and contribute collectively to virulence. A deep and strand-specific RNA sequencing strategy was adopted in order to experimentally refine genome annotation for strain CFBP 5828R. This approach also allowed the experimental definition of novel ORFs and non-coding RNA transcripts. Using a constitutively active allele of hrpG, a master regulator of the type III secretion system, a HrpG-dependent regulon of 141 genes co-regulated with the type III secretion system was identified. Importantly, all these genes but seven are positively regulated by HrpG and 56 of those encode components of the Hrp type III secretion system and putative effector proteins. This dataset is an important resource to mine for novel type III effector proteins as well as for bacterial genes which could contribute to pathogenicity of X. campestris.

  17. hpaR, a putative marR family transcriptional regulator, is positively controlled by HrpG and HrpX and involved in the pathogenesis, hypersensitive response, and extracellular protease production of Xanthomonas campestris pathovar campestris.

    PubMed

    Wei, Ke; Tang, Dong-Jie; He, Yong-Qiang; Feng, Jia-Xun; Jiang, Bo-Le; Lu, Guang-Tao; Chen, Baoshan; Tang, Ji-Liang

    2007-03-01

    The MarR family of transcriptional regulators of bacteria are involved in the regulation of many cellular processes, including pathogenesis. In this work, we have demonstrated genetically that hpaR (hpa, hrp associated), which encodes a putative MarR family regulator, is involved in the hypersensitive response (HR), pathogenicity, and extracellular protease production of the phytopathogenic bacterium Xanthomonas campestris pathovar campestris. A mutation in hpaR resulted in complete loss of virulence in the host plant cabbage, a delayed and weakened HR in the nonhost plant pepper ECW-10R, and an increase in extracellular protease production. Detection of the beta-glucuronidase activity of a plasmid-driven hpaR promoter-gusA reporter revealed that the expression of hpaR is positively controlled by HrpG and HrpX and is suppressed in rich medium while being strongly induced in minimal and hrp-inducing media and inside the host. These findings indicate that hpaR belongs to the hrpG and hrpX regulon and that HrpX regulates the extracellular protease production via hpaR in X. campestris pv. campestris.

  18. Virulence deficiency caused by a transposon insertion in the purH gene of Xanthomonas oryzae pv. oryzae.

    PubMed

    Chatterjee, Subhadeep; Sonti, Ramesh V

    2005-07-01

    Xanthomonas oryzae pv. oryzae causes bacterial leaf blight, a serious disease of rice. We have identified a Tn5-induced virulence-deficient mutant (BXO1704) of X. oryzae pv. oryzae. The BXO1704 mutant exhibited growth deficiency in minimal medium but was proficient in inducing a hypersensitive response in a non-host tomato plant. Sequence analysis of the chromosomal DNA flanking the Tn5 insertion indicated that the Tn5 insertion is in the purH gene, which is highly homologous to purH genes of other closely related plant pathogenic bacteria Xanthomonas axonopodis pv. citri and Xanthomonas campestris pv. campestris. Purine supplementation reversed the growth deficiency of BXO1704 in minimal medium. These results suggest that the virulence deficiency of BXO1704 may be due to the inability to use sufficient purine in the host.

  19. Cloning, expression, crystallization and preliminary X-ray analysis of a putative multiple antibiotic resistance repressor protein (MarR) from Xanthomonas campestris

    SciTech Connect

    Tu, Zhi-Le; Li, Juo-Ning; Chin, Ko-Hsin; Chou, Chia-Cheng; Lee, Cheng-Chung; Shr, Hui-Lin; Lyu, Ping-Chiang; Gao, Fei Philip; Wang, Andrew H.-J.; Chou, Shan-Ho

    2005-07-01

    A putative repressor for the multiple antibiotic resistance operon from a plant pathogen X. campestris pv. campestris has been overexpressed in E. coli, purified and crystallized. The crystals diffracted to 2.3 Å with good quality. The multiple antibiotic resistance operon (marRAB) is a member of the multidrug-resistance system. When induced, this operon enhances resistance of bacteria to a variety of medically important antibiotics, causing a serious global health problem. MarR is a marR-encoded protein that represses the transcription of the marRAB operon. Through binding with salicylate and certain antibiotics, however, MarR can derepress and activate the marRAB operon. In this report, the cloning, expression, crystallization and preliminary X-ray analysis of XC1739, a putative MarR repressor protein present in the Xanthomonas campestris pv. campestris, a Gram-negative bacterium causing major worldwide disease of cruciferous crops, are described. The XC1739 crystals diffracted to a resolution of at least 1.8 Å. They are orthorhombic and belong to space group P2{sub 1}2{sub 1}2{sub 1}, with unit-cell parameters a = 39.5, b = 54.2 and c = 139.5 Å, respectively. They contain two molecules in the asymmetric unit from calculation of the self-rotation function.

  20. Preparation, crystallization and preliminary X-ray analysis of XC2382, an ApaG protein of unknown structure from Xanthomonas campestris

    SciTech Connect

    Chin, Ko-Hsin; Chou, Chia-Cheng; Lee, Cheng-Chung; Shr, Hui-Lin; Lyu, Ping-Chiang; Wang, Andrew H.-J.; Chou, Shan-Ho

    2005-07-01

    A putative ApaG gene product from X. campestris pv. campestris was overexpressed in E. coli, purified and crystallized. The crystals diffracted to a resolution of at least 2.3 Å. Xanthomonas campestris pv. campestris is the causative agent of black rot, one of the major worldwide diseases of cruciferous crops. Its genome encodes approximately 4500 proteins, roughly one third of which have unknown function. XC2382 is one such protein, with a MW of 14.2 kDa. Based on a bioinformatics study, it was annotated as an ApaG gene product that serves multiple functions. The ApaG protein has been overexpressed in Escherichia coli, purified and crystallized using the hanging-drop vapour-diffusion method. The crystals diffracted to a resolution of at least 2.30 Å. They are tetragonal and belong to space group P4{sub 1/3}, with unit-cell parameters a = b = 57.6, c = 122.9 Å. There are two, three or four molecules in the asymmetric unit.

  1. Xanthomonas campestris diffusible factor is 3-hydroxybenzoic acid and is associated with xanthomonadin biosynthesis, cell viability, antioxidant activity, and systemic invasion.

    PubMed

    He, Ya-Wen; Wu, Ji'en; Zhou, Lian; Yang, Fan; He, Yong-Qiang; Jiang, Bo-Le; Bai, Linquan; Xu, Yuquan; Deng, Zixin; Tang, Ji-Liang; Zhang, Lian-Hui

    2011-08-01

    Xanthomonas campestris pv. campestris produces a membrane-bound yellow pigment called xanthomonadin. A diffusible factor (DF) has been reported to regulate xanthomonadin biosynthesis. In this study, DF was purified from bacterial culture supernatants using a combination of solvent extraction, flash chromatography, and high-performance liquid chromatography. Mass spectrometry and nuclear magnetic resonance analyses resolved the DF chemical structure as 3-hydroxybenzoic acid (3-HBA), which was further confirmed by synthetic 3-HBA. Significantly, bioassay and in silico analysis suggest that DF production is widely conserved in a range of bacterial species. Analysis of DF derivatives established the hydroxyl group and its position as the key structural features for the role of DF in xanthomonadin biosynthesis. In addition, we showed that DF is also associated with bacterial survival, H2O2 resistance, and systemic invasion. Furthermore, evidence was also presented that DF and diffusible signaling factor have overlapping functions in modulation of bacterial survival, H2O2 resistance, and virulence. Utilization of different mechanisms to modulate similar virulence traits may provide X. campestris pv. campestris with plasticity in response to various environmental cues.

  2. Cloning, purification, crystallization and preliminary X-ray crystallographic analysis of XC847, a 3′-5′ oligoribonuclease from Xanthomonas campestris

    SciTech Connect

    Wu, Yan-You; Chin, Ko-Hsin; Chou, Chia-Cheng; Lee, Cheng-Chung; Shr, Hui-Lin; Gao, Fei Philip; Lyu, Ping-Chiang; Wang, Andrew H.-J.; Chou, Shan-Ho

    2005-10-01

    A DEDDh-type 3′-5′ oligoribonuclease from the plant pathogen X. campestris pv. campestris has been overexpressed in E. coli, purified and crystallized. The crystals diffracted to 2.1 Å with good quality. Oligoribonucleases are essential components of RNA and DNA metabolism and close homologues of genes encoding them are found not only in prokaryotes but also in a wide range of eukaryotes, including yeast and humans. Inactivation of the oligoribonuclease gene (orn) can result in cellular lethality. Despite their important biological function, they have been studied little from a structural point of view. In this report, the cloning, expression, crystallization and preliminary X-ray analysis of XC847, a DEDDh-type 3′-5′ oligoribonuclease from the plant pathogen Xanthomonas campestris pv. campestris, a Gram-negative bacterium causing major worldwide disease of cruciferous crops, is described. The XC847 crystals diffracted to a resolution of at least 2.1 Å. They are tetragonal and belong to space group P4{sub 3}2{sub 1}2, with unit-cell parameters a = b = 67.5, c = 89.8 Å. One molecule is present per asymmetric unit.

  3. A SNP Haplotype Associated with a gene resistant to Xanthomonas axonopodis pv. malvacearum in Upland Cotton (Gossyium hirsutum L.)

    USDA-ARS?s Scientific Manuscript database

    An F5 population of 285 families with each tracing back to a different F2 plant , derived from a cotton bacterial blight resistant line ‘DeltaOpal’ and a susceptible line ‘DP388’, was artificially inoculated with bacterial blight race 18 (Xanthomonas campestris pv. Malvacearum) to assay their resist...

  4. The Xanthomonas Hrp type III system secretes proteins from plant and mammalian bacterial pathogens

    PubMed Central

    Rossier, Ombeline; Wengelnik, Kai; Hahn, Karoline; Bonas, Ulla

    1999-01-01

    Studies of essential pathogenicity determinants in Gram-negative bacteria have revealed the conservation of type III protein secretion systems that allow delivery of virulence factors into host cells from plant and animal pathogens. Ten of 21 Hrp proteins of the plant pathogen Xanthomonas campestris pv. vesicatoria have been suggested to be part of a type III machinery. Here, we report the hrp-dependent secretion of two avirulence proteins, AvrBs3 and AvrRxv, by X. campestris pv. vesicatoria strains that constitutively express hrp genes. Secretion occurred without leakage of a cytoplasmic marker in minimal medium containing BSA, at pH 5.4. Secretion was strictly hrp-dependent because a mutant carrying a deletion in hrcV, a conserved hrp gene, did not secrete AvrBs3 and AvrRxv. Moreover, the Hrp system of X. campestris pv. vesicatoria was able to secrete proteins from two other plant pathogens: PopA, a protein secreted via the Hrp system in Ralstonia solanacearum, and AvrB, an avirulence protein from Pseudomonas syringae pv. glycinea. Interestingly, X. campestris pv. vesicatoria also secreted YopE, a type III-secreted cytotoxin of the mammalian pathogen Yersinia pseudotuberculosis in a hrp-dependent manner. YerA, a YopE-specific chaperone, was required for YopE stability but not for secretion in X. campestris pv. vesicatoria. Our results demonstrate the functional conservation of the type III system of X. campestris for secretion of proteins from both plant and mammalian pathogens and imply recognition of their respective secretion signals. PMID:10430949

  5. Characterization of the GntR family regulator HpaR1 of the crucifer black rot pathogen Xanthomonas campestris pathovar campestris

    PubMed Central

    Su, Hui-Zhao; Wu, Liu; Qi, Yan-Hua; Liu, Guo-Fang; Lu, Guang-Tao; Tang, Ji-Liang

    2016-01-01

    The GntR family transcription regulator HpaR1 identified from Xanthomonas campestris pv. campestris has been previously shown to positively regulate the genes responsible for hypersensitive reaction and pathogenicity and to autorepress its own expression. Here, we demonstrated that HpaR1 is a global regulator that positively regulates diverse biological processes, including xanthan polysaccharide production, extracellular enzyme activity, cell motility and tolerance to various stresses. To investigate the regulatory mechanisms of HpaR1, we began with xanthan polysaccharide production, which is governed by a cluster of gum genes. These are directed by the gumB promoter. Disruption of HpaR1 significantly reduced gumB transcription and an electrophoretic mobility shift assay demonstrated that HpaR1 interacts directly with gumB promoter. DNase I footprint analysis revealed that HpaR1 and RNA polymerase were bound to the sequences extending from −21 to +10 and −41 to +29 relative to the transcription initiation site of gumB, respectively. Furthermore, in vitro transcription assays showed that HpaR1 facilitated the binding of RNA polymerase to gumB promoter, leading to an enhancement of its transcription. These results suggest that HpaR1 regulates gumB transcription via a mechanism similar but different to what was found, until now, to only be used by some MerR family transcription activators. PMID:26818230

  6. Characterization of the GntR family regulator HpaR1 of the crucifer black rot pathogen Xanthomonas campestris pathovar campestris.

    PubMed

    Su, Hui-Zhao; Wu, Liu; Qi, Yan-Hua; Liu, Guo-Fang; Lu, Guang-Tao; Tang, Ji-Liang

    2016-01-28

    The GntR family transcription regulator HpaR1 identified from Xanthomonas campestris pv. campestris has been previously shown to positively regulate the genes responsible for hypersensitive reaction and pathogenicity and to autorepress its own expression. Here, we demonstrated that HpaR1 is a global regulator that positively regulates diverse biological processes, including xanthan polysaccharide production, extracellular enzyme activity, cell motility and tolerance to various stresses. To investigate the regulatory mechanisms of HpaR1, we began with xanthan polysaccharide production, which is governed by a cluster of gum genes. These are directed by the gumB promoter. Disruption of HpaR1 significantly reduced gumB transcription and an electrophoretic mobility shift assay demonstrated that HpaR1 interacts directly with gumB promoter. DNase I footprint analysis revealed that HpaR1 and RNA polymerase were bound to the sequences extending from -21 to +10 and -41 to +29 relative to the transcription initiation site of gumB, respectively. Furthermore, in vitro transcription assays showed that HpaR1 facilitated the binding of RNA polymerase to gumB promoter, leading to an enhancement of its transcription. These results suggest that HpaR1 regulates gumB transcription via a mechanism similar but different to what was found, until now, to only be used by some MerR family transcription activators.

  7. Supported PV module assembly

    DOEpatents

    Mascolo, Gianluigi; Taggart, David F.; Botkin, Jonathan D.; Edgett, Christopher S.

    2013-10-15

    A supported PV assembly may include a PV module comprising a PV panel and PV module supports including module supports having a support surface supporting the module, a module registration member engaging the PV module to properly position the PV module on the module support, and a mounting element. In some embodiments the PV module registration members engage only the external surfaces of the PV modules at the corners. In some embodiments the assembly includes a wind deflector with ballast secured to a least one of the PV module supports and the wind deflector. An array of the assemblies can be secured to one another at their corners to prevent horizontal separation of the adjacent corners while permitting the PV modules to flex relative to one another so to permit the array of PV modules to follow a contour of the support surface.

  8. Proposal to Reject the Name Ulmus Campestris L. (Ullmaceae)

    USDA-ARS?s Scientific Manuscript database

    The name Ulmus campestris has nomenclatural priority over U. glabra and technically should be the correct name for the wych elm under the international rules of botanical nomenclature. The name Ulmus campestris has a very confused history, having been used for three different elm species in the past...

  9. Construction of a genetic linkage map for identification of molecular markers associated with resistance to Xanthomonas arboriciola pv. pruni in peach [Prunus persica (L.) Batsch

    USDA-ARS?s Scientific Manuscript database

    Bacterial spot, caused by Xanthomonas campestris pv. pruni, is a serious disease that can affect peach fruit quality and production. The molecular basis of its tolerance and susceptibility is yet to be understood. To study the genetics of the peach in response to bacterial spot, an F2 population of ...

  10. Regulation of the synthesis of cyclic glucan in Xanthomonas campestris by a diffusible signal molecule.

    PubMed

    Vojnov, A A; Slater, H; Newman, M A; Daniels, M J; Dow, J M

    2001-12-01

    The rpf gene cluster of Xanthomonas campestris pv. campestris is involved in the co-ordinate positive regulation of the production of extracellular enzymes and the extracellular polysaccharide xanthan. Several of the rpf genes are involved in a regulatory system involving the small diffusible molecule DSF (for diffusible signal factor). Synthesis of DSF requires RpfF, and a two-component sensory transduction system involving RpfC has been implicated in the perception of the signal and signal transduction. Here we show that mutations in both rpfF and rpfC lead to reductions in the levels of cyclic glucan. The levels of cyclic glucan synthetase in membrane preparations from rpfF and rpfC mutants were, however, unaltered from the wild-type. Similar alterations in the level of cyclic glucan without changes in cyclic glucan synthetase activity were seen when wild-type bacteria were exposed to osmotic stress. These results extend the range of cellular functions subject to regulation by the rpf genes and DSF system.

  11. A functional 4-hydroxybenzoate degradation pathway in the phytopathogen Xanthomonas campestris is required for full pathogenicity

    PubMed Central

    Wang, Jia-Yuan; Zhou, Lian; Chen, Bo; Sun, Shuang; Zhang, Wei; Li, Ming; Tang, Hongzhi; Jiang, Bo-Le; Tang, Ji-Liang; He, Ya-Wen

    2015-01-01

    Plants contain significant levels of natural phenolic compounds essential for reproduction and growth, as well as defense mechanisms against pathogens. Xanthomonas campestris pv. campestris (Xcc) is the causal agent of crucifers black rot. Here we showed that genes required for the synthesis, utilization, transportation, and degradation of 4-hydroxybenzoate (4-HBA) are present in Xcc. Xcc rapidly degrades 4-HBA, but has no effect on 2-hydroxybenzoate and 3-hydroxybenzoate when grown in XOLN medium. The genes for 4-HBA degradation are organized in a superoperonic cluster. Bioinformatics, biochemical, and genetic data showed that 4-HBA is hydroxylated by 4-HBA 3-hydroxylase (PobA), which is encoded by Xcc0356, to yield PCA. The resulting PCA is further metabolized via the PCA branches of the β-ketoadipate pathway, including Xcc0364, Xcc0365, and PcaFHGBDCR. Xcc0364 and Xcc0365 encode a new form of β-ketoadipate succinyl-coenzyme A transferase that is required for 4-HBA degradation. pobA expression was induced by 4-HBA via the transcriptional activator, PobR. Radish and cabbage hydrolysates contain 2-HBA, 3-HBA, 4-HBA, and other phenolic compounds. Addition of radish and cabbage hydrolysates to Xcc culture significantly induced the expression of pobA via PobR. The 4-HBA degradation pathway is required for full pathogenicity of Xcc in radish. PMID:26672484

  12. A functional 4-hydroxybenzoate degradation pathway in the phytopathogen Xanthomonas campestris is required for full pathogenicity.

    PubMed

    Wang, Jia-Yuan; Zhou, Lian; Chen, Bo; Sun, Shuang; Zhang, Wei; Li, Ming; Tang, Hongzhi; Jiang, Bo-Le; Tang, Ji-Liang; He, Ya-Wen

    2015-12-17

    Plants contain significant levels of natural phenolic compounds essential for reproduction and growth, as well as defense mechanisms against pathogens. Xanthomonas campestris pv. campestris (Xcc) is the causal agent of crucifers black rot. Here we showed that genes required for the synthesis, utilization, transportation, and degradation of 4-hydroxybenzoate (4-HBA) are present in Xcc. Xcc rapidly degrades 4-HBA, but has no effect on 2-hydroxybenzoate and 3-hydroxybenzoate when grown in XOLN medium. The genes for 4-HBA degradation are organized in a superoperonic cluster. Bioinformatics, biochemical, and genetic data showed that 4-HBA is hydroxylated by 4-HBA 3-hydroxylase (PobA), which is encoded by Xcc0356, to yield PCA. The resulting PCA is further metabolized via the PCA branches of the β-ketoadipate pathway, including Xcc0364, Xcc0365, and PcaFHGBDCR. Xcc0364 and Xcc0365 encode a new form of β-ketoadipate succinyl-coenzyme A transferase that is required for 4-HBA degradation. pobA expression was induced by 4-HBA via the transcriptional activator, PobR. Radish and cabbage hydrolysates contain 2-HBA, 3-HBA, 4-HBA, and other phenolic compounds. Addition of radish and cabbage hydrolysates to Xcc culture significantly induced the expression of pobA via PobR. The 4-HBA degradation pathway is required for full pathogenicity of Xcc in radish.

  13. Detection of Xanthomonas arboricola pv. pruni by PCR using primers based on DNA sequences related to the hrp genes.

    PubMed

    Park, So Yeon; Lee, Young Sun; Koh, Young Jin; Hur, Jae-Sun; Jung, Jae Sung

    2010-10-01

    Efficient control of Xanthomonas arboricola pv. pruni, the causal agent of bacterial spot on stone fruit, requires a sensitive and reliable diagnostic tool. A PCR detection method that utilizes primers to target the hrp gene cluster region was developed in this study. The nucleotide sequence of the PCR product amplified with primers specific for the hrp region of the xanthomonads and genomic DNA of X. arboricola pv. pruni was determined, and the sequence was aligned with that of X. campestris pv. campestris, which was obtained from the GenBank database. On the basis of the sequence of the amplified hrp region, a PCR primer set of XapF/R specific to X. arboricola pv. pruni was designed. This primer set yielded a 243-bp product from the genomic DNA of X. aboricola pv. pruni strains, but no products from other 21 strains of Xanthomonas or from two epiphytic bacterial species. Southern blot hybridization with the probe derived from the PCR product with the primer set and X. aboricola pv. pruni DNA confirmed the PCR results. The Xap primer system was successfully applied to detect the pathogen from infected peach fruits. When it was applied in field samples, the primer set was proved as a reliable diagnostic tool for specific detection of X. aboricola pv. pruni from peach orchards.

  14. [Xanthan production by Xanthomonas campestris B-1459].

    PubMed

    Lorda, G S; Pastor, M D; Balatti, A P

    1994-01-01

    The production of xantano from Xanthomonas campestris B-1459 was analyzed. The experiments were performed in shaked flasks at 250 rpm and 2.5 cm eccentricity. Using a base modified medium it was possible to achieve xantano concentration of 35 g/l in 72 h of process. The modified medium contained glucose as carbon source, and yeast extract, meat peptone, malt extract and amaranth meal as growth factors and nitrogen sources, in a KH2PO4/K2HPO4 buffer.

  15. Functional Analysis of the Ferric Uptake Regulator Gene fur in Xanthomonas vesicatoria.

    PubMed

    Liu, Huiqin; Dong, Chunling; Zhao, Tingchang; Han, Jucai; Wang, Tieling; Wen, Xiangzhen; Huang, Qi

    2016-01-01

    Iron is essential for the growth and survival of many organisms. Intracellular iron homeostasis must be maintained for cell survival and protection against iron toxicity. The ferric uptake regulator protein (Fur) regulates the high-affinity ferric uptake system in many bacteria. To investigate the function of the fur gene in Xanthomonas vesicatoria (Xv), we generated a fur mutant strain, fur-m, by site-directed mutagenesis. Whereas siderophore production increased in the Xv fur mutant, extracellular polysaccharide production, biofilm formation, swimming ability and quorum sensing signals were all significantly decreased. The fur mutant also had significantly reduced virulence in tomato leaves. The above-mentioned phenotypes significantly recovered when the Xv fur mutation allele was complemented with a wild-type fur gene. Thus, Fur either negatively or positively regulates multiple important physiological functions in Xv.

  16. Functional Analysis of the Ferric Uptake Regulator Gene fur in Xanthomonas vesicatoria

    PubMed Central

    Liu, Huiqin; Dong, Chunling; Zhao, Tingchang; Han, Jucai; Wang, Tieling; Wen, Xiangzhen; Huang, Qi

    2016-01-01

    Iron is essential for the growth and survival of many organisms. Intracellular iron homeostasis must be maintained for cell survival and protection against iron toxicity. The ferric uptake regulator protein (Fur) regulates the high-affinity ferric uptake system in many bacteria. To investigate the function of the fur gene in Xanthomonas vesicatoria (Xv), we generated a fur mutant strain, fur-m, by site-directed mutagenesis. Whereas siderophore production increased in the Xv fur mutant, extracellular polysaccharide production, biofilm formation, swimming ability and quorum sensing signals were all significantly decreased. The fur mutant also had significantly reduced virulence in tomato leaves. The above-mentioned phenotypes significantly recovered when the Xv fur mutation allele was complemented with a wild-type fur gene. Thus, Fur either negatively or positively regulates multiple important physiological functions in Xv. PMID:26910324

  17. Energy 101: Solar PV

    ScienceCinema

    None

    2016-07-12

    Solar photovoltaic (PV) systems can generate clean, cost-effective power anywhere the sun shines. This video shows how a PV panel converts the energy of the sun into renewable electricity to power homes and businesses.

  18. Energy 101: Solar PV

    SciTech Connect

    2011-01-01

    Solar photovoltaic (PV) systems can generate clean, cost-effective power anywhere the sun shines. This video shows how a PV panel converts the energy of the sun into renewable electricity to power homes and businesses.

  19. Descriptions of the pupae of Ochlerotatus aloponotum and Ochlerotatus campestris.

    PubMed

    Darsie, Richard F

    2011-09-01

    Pupae of 2 western species, Ochlerotatus aloponotum and Oc. campestris, are described and illustrated. Sources of the original descriptions and known stages are given. The relation of Oc. aloponotum to Oc. excrucians and characters to separate them are discussed.

  20. Mutants of Xanthomonas oryzae pv. oryzae deficient in general secretory pathway are virulence deficient and unable to secrete xylanase.

    PubMed

    Ray, S K; Rajeshwari, R; Sonti, R V

    2000-04-01

    Xanthomonas oryzae pv. oryzae (Xoo) causes bacterial leaf blight, a serious disease of rice. A virulence- and xylanase-deficient mutant of Xoo was isolated following ethyl methane sulfonate (EMS) mutagenesis. A cosmid clone that restored virulence and xylanase secretion was obtained from a genomic library by functional complementation. Transposon mutagenesis and marker exchange studies revealed genes on the cloned DNA that were required for xylanase production and virulence. Sequence analysis with transposon-specific primers revealed that these genes were homologues of xps F and xps D, which encode components of a protein secretion system in Xanthomonas campestris pv. campestris. Enzyme assays showed xylanase accumulation in the periplasmic space and cytoplasm of the xps F mutant and the complementing clone restored transport to the extracellular space.

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

    PubMed

    Hurabielle, M; Eberle, J; Paris, M

    1982-10-01

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

  2. Fire resistant PV shingle assembly

    DOEpatents

    Lenox, Carl J.

    2012-10-02

    A fire resistant PV shingle assembly includes a PV assembly, including PV body, a fire shield and a connection member connecting the fire shield below the PV body, and a support and inter-engagement assembly. The support and inter-engagement assembly is mounted to the PV assembly and comprises a vertical support element, supporting the PV assembly above a support surface, an upper interlock element, positioned towards the upper PV edge, and a lower interlock element, positioned towards the lower PV edge. The upper interlock element of one PV shingle assembly is inter-engageable with the lower interlock element of an adjacent PV shingle assembly. In some embodiments the PV shingle assembly may comprise a ventilation path below the PV body. The PV body may be slidably mounted to the connection member to facilitate removal of the PV body.

  3. The molybdate-binding protein (ModA) of the plant pathogen Xanthomonas axonopodis pv. citri.

    PubMed

    Balan, Andrea; Santacruz, Carolina P; Moutran, Alexandre; Ferreira, Rita C C; Medrano, Francisco J; Pérez, Carlos A; Ramos, Carlos H I; Ferreira, Luís C S

    2006-12-01

    The modABC operon of phytopathogen Xanthomonas axonopodis pv. citri (X. citri) encodes a putative ABC transporter involved in the uptake of the molybdate and tungstate anions. Sequence analyses showed high similarity values of ModA orthologs found in X. campestris pv. campestris (X. campestris) and Escherichia coli. The X. citri modA gene was cloned in pET28a and the recombinant protein, expressed in the E. coli BL21 (DE3) strain, purified by immobilized metal affinity chromatography. The purified protein remained soluble and specifically bound molybdate and tungstate with K(d) 0.29+/-0.12 microM and 0.58+/-0.14 microM, respectively. Additionally binding of molybdate drastically enhanced the thermal stability of the recombinant ModA as compared to the apoprotein. This is the first characterization of a ModA ortholog expressed by a phytopathogen and represents an important tool for functional, biochemical and structural analyses of molybdate transport in Xanthomonas species.

  4. Site-directed gene replacement of the phytopathogen Xanthomonas axonopodis pv. citri.

    PubMed

    Oshiro, Elisa E; Nepomuceno, Roberto S L; Faria, Juarez B; Ferreira, Luís C S; Ferreira, Rita C C

    2006-04-01

    In this work we defined experimental conditions for site-directed gene replacement of the Xanthomonas axonopodis pv. citri (Xac), an economically relevant pathogen of citrus plants. The procedure involved, first, optimizing the electrotransformation conditions of the Xac 306 strain and, second, constructing non-replicative suicide vectors carrying knockout copies of the target gene. Using specific experimental conditions, transformation efficiencies of Xac were at least 100 fold higher than those achieved with electroporation protocols previously designed for X. campestris transformation. Successful gene replacement events were achieved with a suicide vector derived from R6K plasmid (pWR-SS) but not with those with ColE1 replication origin. We have chosen the oppA as a target gene, encoding the binding component (OppA) of the major oligopeptide uptake system found in the genome of the Xac 306 strain, although not in X. campestris pv. campestris (Xcc). Defining the experimental conditions, which allow for the specific mutagenesis of the Xac 306 strain, represents a step in the understanding of both genetics and physiology of this economically important bacterial species.

  5. First isolation of Xanthomonas campestris from the blood of a Chinese woman.

    PubMed

    Li, Z X; Bian, Z S; Zheng, H P; Yue, Y S; Yao, J Y; Gong, Y P; Cai, M Y; Dong, X Z

    1990-05-01

    Xanthomonas campestris isolated from the blood of a patient with a fever was first reported. Xanthomonas campestris is a bacterium that can cause black rot of some vegetables, such as rape. Chinese cabbage, etc. Human infection due to X. campestris has not been reported so far. The characteristics of this organism, including morphology, staining, physiology and biochemistry were studied. We believe that X. campestris is also one of the opportunistic pathogens, which can infect compromised host.

  6. Xanthomonas campestris lipooligosaccharides trigger innate immunity and oxidative burst in Arabidopsis.

    PubMed

    Proietti, S; Giangrande, C; Amoresano, A; Pucci, P; Molinaro, A; Bertini, L; Caporale, C; Caruso, C

    2014-12-01

    Plants lack the adaptive immunity mechanisms of jawed vertebrates, so they rely on innate immune responses to defense themselves from pathogens. The plant immune system perceives the presence of pathogens by recognition of molecules known as pathogen-associated molecular patterns (PAMPs). PAMPs have several common characteristics, including highly conserved structures, essential for the microorganism but absent in host organisms. Plants can specifically recognize PAMPs using a large set of receptors and can respond with appropriate defenses by activating a multicomponent and multilayered response. Lipopolysaccharides (LPSs) and lipooligosaccharides (LOSs) are major components of the cell surface of Gram-negative bacteria with diverse roles in bacterial pathogenesis of animals and plants that include elicitation of host defenses. Little is known on the mechanisms of perception of these molecules by plants and the associated signal transduction pathways that trigger plant immunity.Here we addressed the question whether the defense signaling pathway in Arabidopsis thaliana was triggered by LOS from Xanthomonas campestris pv. campestris (Xcc), using proteomic and transcriptomic approaches. By using affinity capture strategies with immobilized LOS and LC-MS/MS analyses, we identified 8 putative LOS protein ligands. Further investigation of these interactors led to the definition that LOS challenge is able to activate a signal transduction pathway that uses nodal regulators in common with salicylic acid-mediated pathway. Moreover, we proved evidence that Xcc LOS are responsible for oxidative burst in Arabidopsis either in infiltrated or systemic leaves. In addition, gene expression studies highlighted the presence of gene network involved in reactive oxygen species transduction pathway.

  7. Xanthomonas campestris attenuates virulence by sensing light through a bacteriophytochrome photoreceptor.

    PubMed

    Bonomi, Hernán R; Toum, Laila; Sycz, Gabriela; Sieira, Rodrigo; Toscani, Andrés M; Gudesblat, Gustavo E; Leskow, Federico C; Goldbaum, Fernando A; Vojnov, Adrián A; Malamud, Florencia

    2016-11-01

    Phytochromes constitute a major photoreceptor family found in plants, algae, fungi, and prokaryotes, including pathogens. Here, we report that Xanthomonas campestris pv. campestris (Xcc), the causal agent of black rot disease which affects cruciferous crops worldwide, codes for a functional bacteriophytochrome (XccBphP). XccBphP possesses an N-terminal PAS2-GAF-PHY photosensory domain triad and a C-terminal PAS9 domain as its output module. Our results show that illumination of Xcc, prior to plant infection, attenuates its virulence in an XccBphP-dependent manner. Moreover, in response to light, XccBphP downregulates xanthan exopolysaccharide production and biofilm formation, two known Xcc virulence factors. Furthermore, the XccbphP null mutant shows enhanced virulence, similar to that of dark-adapted Xcc cultures. Stomatal aperture regulation and callose deposition, both well-established plant defense mechanisms against bacterial pathogens, are overridden by the XccbphP strain. Additionally, an RNA-Seq analysis reveals that far-red light or XccBphP overexpression produces genomewide transcriptional changes, including the inhibition of several Xcc virulence systems. Our findings indicate that Xcc senses light through XccBphP, eliciting bacterial virulence attenuation via downregulation of bacterial virulence factors. The capacity of XccBphP to respond to light both in vitro and in vivo was abolished by a mutation on the conserved Cys13 residue. These results provide evidence for a novel bacteriophytochrome function affecting an infectious process. © 2016 The Authors.

  8. PV_LIB Toolbox

    SciTech Connect

    2012-09-11

    While an organized source of reference information on PV performance modeling is certainly valuable, there is nothing to match the availability of actual examples of modeling algorithms being used in practice. To meet this need, Sandia has developed a PV performance modeling toolbox (PV_LIB) for Matlab. It contains a set of well-documented, open source functions and example scripts showing the functions being used in practical examples. This toolbox is meant to help make the multi-step process of modeling a PV system more transparent and provide the means for model users to validate and understand the models they use and or develop. It is fully integrated into Matlab’s help and documentation utilities. The PV_LIB Toolbox provides more than 30 functions that are sorted into four categories

  9. The rice bacterial pathogen Xanthomonas oryzae pv. oryzae produces 3-hydroxybenzoic acid and 4-hydroxybenzoic acid via XanB2 for use in xanthomonadin, ubiquinone, and exopolysaccharide biosynthesis.

    PubMed

    Zhou, Lian; Huang, Tin-Wei; Wang, Jia-Yuan; Sun, Shuang; Chen, Gongyou; Poplawsky, Alan; He, Ya-Wen

    2013-10-01

    Xanthomonas oryzae pv. oryzae, the causal agent of rice bacterial blight, produces membrane-bound yellow pigments, referred to as xanthomonadins. Xanthomonadins protect the pathogen from photodamage and host-induced perioxidation damage. They are also required for epiphytic survival and successful host plant infection. Here, we show that XanB2 encoded by PXO_3739 plays a key role in xanthomonadin and coenzyme Q8 biosynthesis in X. oryzae pv. oryzae PXO99A. A xanB2 deletion mutant exhibits a pleiotropic phenotype, including xanthomonadin deficiency, producing less exopolysaccharide (EPS), lower viability and H2O2 resistance, and lower virulence. We further demonstrate that X. oryzae pv. oryzae produces 3-hydroxybenzoic acid (3-HBA) and 4-hydroxybenzoic acid (4-HBA) via XanB2. 3-HBA is associated with xanthomonadin biosynthesis while 4-HBA is mainly used as a precursor for coenzyme Q (CoQ)8 biosynthesis. XanB2 is the alternative source of 4-HBA for CoQ8 biosynthesis in PXO99A. These findings suggest that the roles of XanB2 in PXO99A are generally consistent with those in X. campestris pv. campestris. The present study also demonstrated that X. oryzae pv. oryzae PXO99A has evolved several specific features in 3-HBA and 4-HBA signaling. First, our results showed that PXO99A produces less 3-HBA and 4-HBA than X. campestris pv. campestris and this is partially due to a degenerated 4-HBA efflux pump. Second, PXO99A has evolved unique xanthomonadin induction patterns via 3-HBA and 4-HBA. Third, our results showed that 3-HBA or 4-HBA positively regulates the expression of gum cluster to promote EPS production in PXO99A. Taken together, the results of this study indicate that XanB2 is a key metabolic enzyme linking xanthomonadin, CoQ, and EPS biosynthesis, which are collectively essential for X. oryzae pv. oryzae pathogenesis.

  10. Quantitative disease resistance to the bacterial pathogen Xanthomonas campestris involves an Arabidopsis immune receptor pair and a gene of unknown function.

    PubMed

    Debieu, Marilyne; Huard-Chauveau, Carine; Genissel, Anne; Roux, Fabrice; Roby, Dominique

    2016-05-01

    Although quantitative disease resistance (QDR) is a durable and broad-spectrum form of resistance in plants, the identification of the genes underlying QDR is still in its infancy. RKS1 (Resistance related KinaSe1) has been reported recently to confer QDR in Arabidopsis thaliana to most but not all races of the bacterial pathogen Xanthomonas campestris pv. campestris (Xcc). We therefore explored the genetic bases of QDR in A. thaliana to diverse races of X. campestris (Xc). A nested genome-wide association mapping approach was used to finely map the genomic regions associated with QDR to Xcc12824 (race 2) and XccCFBP6943 (race 6). To identify the gene(s) implicated in QDR, insertional mutants (T-DNA) were selected for the candidate genes and phenotyped in response to Xc. We identified two major QTLs that confer resistance specifically to Xcc12824 and XccCFBP6943. Although QDR to Xcc12824 is conferred by At5g22540 encoding for a protein of unknown function, QDR to XccCFBP6943 involves the well-known immune receptor pair RRS1/RPS4. In addition to RKS1, this study reveals that three genes are involved in resistance to Xc with strikingly different ranges of specificity, suggesting that QDR to Xc involves a complex network integrating multiple response pathways triggered by distinct pathogen molecular determinants.

  11. Preliminary X-ray analysis of XC5848, a hypothetical ORFan protein with an Sm-like motif from Xanthomonas campestris

    SciTech Connect

    Ruan, Sz-Kai; Chin, Ko-Hsin; Shr, Hui-Lin; Lyu, Ping-Chiang; Wang, Andrew H.-J.; Chou, Shan-Ho

    2007-01-01

    A conserved hypothetical protein Se-XC5848 from X. campestris pv. campestris has been overexpressed in E. coli, purified and crystallized. Crystals obtained from the purified recombinant protein diffracted to a resolution of 1.68 Å. XC5848, a hypothetical protein from the pathogenic bacterium Xanthomonas campestris that causes black rot, has been chosen as a potential target for the discovery of novel folds. It is unique to the Xanthomonas genus and has significant sequence identity mainly to corresponding proteins from the Xanthomonas genus. In this paper, the cloning, overexpression, purification and crystallization of the XC5848 protein are reported. The XC5848 crystals diffracted to a resolution of at least 1.68 Å. They belong to the orthorhombic space group P2{sub 1}2{sub 1}2{sub 1}, with unit-cell parameters a = 48.13, b = 51.62, c = 82.32 Å. Two molecules were found in each asymmetric unit. Preliminary structural studies nevertheless indicate that XC5848 belongs to the highly conserved Sm-like α-β-β-β-β fold. However, significant differences in sequence and structure were observed. It therefore represents a novel variant of the crucial Sm-like motif that is heavily involved in mRNA splicing and degradation.

  12. Controlled synthesis of the DSF cell-cell signal is required for biofilm formation and virulence in Xanthomonas campestris.

    PubMed

    Torres, Pablo S; Malamud, Florencia; Rigano, Luciano A; Russo, Daniela M; Marano, María Rosa; Castagnaro, Atilio P; Zorreguieta, Angeles; Bouarab, Kamal; Dow, John Maxwell; Vojnov, Adrián A

    2007-08-01

    Virulence of the black rot pathogen Xanthomonas campestris pv. campestris (Xcc) is regulated by cell-cell signalling involving the diffusible signal factor DSF. Synthesis and perception of DSF require products of genes within the rpf cluster (for regulation of pathogenicity factors). RpfF directs DSF synthesis whereas RpfC and RpfG are involved in DSF perception. Here we have examined the role of the rpf/DSF system in biofilm formation in minimal medium using confocal laser-scanning microscopy of GFP-labelled bacteria. Wild-type Xcc formed microcolonies that developed into a structured biofilm. In contrast, an rpfF mutant (DSF-minus) and an rpfC mutant (DSF overproducer) formed only unstructured arrangements of bacteria. A gumB mutant, defective in xanthan biosynthesis, was also unable to develop the typical wild-type biofilm. Mixed cultures of gumB and rpfF mutants formed a typical biofilm in vitro. In contrast, in mixed cultures the rpfC mutant prevented the formation of the structured biofilm by the wild-type and did not restore wild-type biofilm phenotypes to gumB or rpfF mutants. These effects on structured biofilm formation were correlated with growth and disease development by Xcc strains in Nicotiana benthamiana leaves. These findings suggest that DSF signalling is finely balanced during both biofilm formation and virulence.

  13. Deduction of upstream sequences of Xanthomonas campestris flagellar genes responding to transcription activation by FleQ

    SciTech Connect

    Hu, R.-M.; Yang, T.-C.; Yang, S.-H.; Tseng, Y.-H. . E-mail: yhtseng@chtai.ctc.edu.tw

    2005-10-07

    Xanthomonas campestris pv. campestris (Xcc), a close relative to Pseudomonas aeruginosa, is the pathogen causing black rot in cruciferous plants. In P. aeruginosa, FleQ serves as a cognate activator of {sigma}{sup 54} in transcription from several {sigma}{sup 54}-dependent promoters of flagellar genes. These P. aeruginosa promoters have been analyzed for FleQ-binding sequences; however, no consensus was deduced. Xcc, although lacks fleSR, has a fleQ homologue residing among over 40 contiguously clustered flagellar genes. A fleQ mutant, Xc17fleQ, constructed by insertional mutation is deficient in FleQ protein, non-flagellated, and immobile. Transcriptional fusion assays on six putative {sigma}{sup 54}-dependent promoters of the flagellar genes, fliE, fliQ, fliL, flgG, flgB, and flhF, indicated that each of them is also FleQ dependent. Each of these promoters has a sequence with weak consensus to 5'-gaaacCCgccgCcgctTt-3', immediately upstream of the predicted {sigma}{sup 54}-binding site, with an imperfect inverted repeat containing a GC-rich center flanked by several A and T at 5'- and 3'-ends, respectively. Replacing this region in fliE promoter with a HindIII recognition sequence abolished the transcription, indicating that this region responds to transcription activation by FleQ.

  14. Applying DNA affinity chromatography to specifically screen for sucrose-related DNA-binding transcriptional regulators of Xanthomonas campestris.

    PubMed

    Leßmeier, Lennart; Alkhateeb, Rabeaa S; Schulte, Fabian; Steffens, Tim; Loka, Tobias Pascal; Pühler, Alfred; Niehaus, Karsten; Vorhölter, Frank-Jörg

    2016-08-20

    At a molecular level, the regulation of many important cellular processes is still obscure in xanthomonads, a bacterial group of outstanding relevance as world-wide plant pathogens and important for biotechnology as producers of the polysaccharide xanthan. Transcriptome analysis indicated a sucrose-dependent regulation of 18 genes in Xanthomonas campestris pv. campestris (Xcc) B100. The expression of 12 of these genes was clearly increased in the presence of sucrose. Only part of these genes was obviously involved in sucrose utilization. To identify regulatory proteins involved in transcriptional regulation, a DNA fragment-specific pull-down approach was established for Xcc. Putative promoter regions were identified and used to isolate DNA-binding proteins, which were separated by SDS PAGE and identified by MALDI-TOF mass spectrometry. This led to the identification of four transcriptional regulators, among them the global transcriptional regulator Clp and a previously identified regulator of sucrose utilization, SuxR, plus a third DNA-binding transcriptional regulator encoded by xcc-b100_2861 and recently shown to interact with a cyclic di-GMP-binding protein. The fourth regulatory protein was encoded by xcc-b100_2791. These results indicate DNA fragment-specific pull-down experiments as promising approaches to screen for specific DNA-binding regulatory proteins in Xcc.

  15. RavA/RavR two-component system regulates Xanthomonas campestris pathogenesis and c-di-GMP turnover.

    PubMed

    Tao, Jun; Li, Chunxia; Luo, Chao; He, Chaozu

    2014-09-01

    The two-component system (TCS), consisting of a response regulator (RR) and a cognate histidine kinase (HK), responds to extra-/intercellular cues and triggers adaptive changes. The RR, RavR, has been reported to act as a positive virulence regulator and a c-di-GMP hydrolase in Xanthomonas campestris pv. campestris (Xcc). Here, we identified the cognate HK, RavA, that regulate RavR phosphorylation levels and bacterial pathogenesis. Deletion of ravA, a putative HK gene flanking ravR, dramatically attenuated Xcc virulence. Phenotypes of the double mutant ΔravR/ΔravA were similar to those of ΔravR, suggesting that RavR is a downstream component of RavA signaling. RavA interacts with RavR and positively influences the phosphorylated RavR levels. In vitro analysis suggests that RavR is a bifunctional enzyme involved in c-di-GMP synthesis and degradation. Importantly, mutation and enzyme activity assays indicate that the phosphorylation level affects RavR c-di-GMP turnover activity. These results show that RavA acts as the RavR cognate HK, which fine-tunes RavR functions and enables bacteria to adapt quickly to intracellular changes.

  16. IscR plays a role in oxidative stress resistance and pathogenicity of a plant pathogen, Xanthomonas campestris.

    PubMed

    Fuangthong, Mayuree; Jittawuttipoka, Thichakorn; Wisitkamol, Ratiphorn; Romsang, Adisak; Duang-nkern, Jintana; Vattanaviboon, Paiboon; Mongkolsuk, Skorn

    2015-01-01

    Iron-sulfur ([Fe-S]) cluster is an essential cofactor of proteins involved in various physiological processes including cellular defense against oxidative stress. In Xanthomonas campestris pv. campestris (Xcc), IscR plays a negative role in regulation of the transcription of [Fe-S] assembly genes, iscR-sufBCDS. The expression level of sufBCDS was up-regulated in an Xcc iscR mutant. In addition, the iscR promoter activity in an Xcc iscR mutant was also higher than the wild-type strain, indicating an autoregulatory circuit. Purified IscR was shown to bind at the iscR promoter region and three putative IscR binding sites were identified. The expression of iscR-suf operon was highly induced by oxidant treatments and iron limited conditions. The iscR mutant showed increased sensitivity toward hydrogen peroxide phenotype but, surprisingly, had hyper-resistant phenotype toward plumbagin compared to the wild-type strain. Most importantly, the iscR mutant was impaired in its ability to cause lesion on leaves of a compatible host plant, Chinese radish (Raphanus sativus). These results demonstrate that a transcription regulator gene, iscR, negatively regulates genes involved in [Fe-S] biosynthesis and plays a role in oxidative stress response and pathogenesis of Xcc.

  17. Xanthomonas campestris atcc 31601 and process for use

    SciTech Connect

    Weisrock, W.P.; McCarthy, E.F.

    1983-11-29

    A degenerative-resistant strain of Xanthomonas campestris has been developed and a process for using this strain to effectively overcome the problems of continuous xanthan production. This strain of X. campestris, designated X. campestris XCP-19 ATCC 31601, is capable of continuously producing xanthan at high specific productivities, i.e., 0.24 to 0.32 gm xanthan/gm cells/hr, for several hundred hours without culture degeneration from inexpensive aqueous nutrient media such as, for example, a minimal medium consisting primarily of inorganic salts, glucose, and NH4Cl. The medium may or may not also contain a yeast extract or yeast autolysate as a supplemental nitrogen source. Any medium having assimilable sources of carbon, nitrogen, and inorganic substances will serve satisfactorily for use with this new organism. 14 claims.

  18. GridPV Toolbox

    SciTech Connect

    Broderick, Robert; Quiroz, Jimmy; Grijalva, Santiago; Reno, Matthew; Coogan, Kyle

    2014-07-15

    Matlab Toolbox for simulating the impact of solar energy on the distribution grid. The majority of the functions are useful for interfacing OpenDSS and MATLAB, and they are of generic use for commanding OpenDSS from MATLAB and retrieving GridPV Toolbox information from simulations. A set of functions is also included for modeling PV plant output and setting up the PV plant in the OpenDSS simulation. The toolbox contains functions for modeling the OpenDSS distribution feeder on satellite images with GPS coordinates. Finally, example simulations functions are included to show potential uses of the toolbox functions.

  19. Molecular cloning, characterization, and mutagenesis of a pel gene from Pseudomonas syringae pv. lachyrmans encoding a member of the Erwinia chrysanthemi pelADE family of pectate lyases.

    PubMed

    Bauer, D W; Collmer, A

    1997-04-01

    The pelS gene from Pseudomonas syringae pv. lachrymans 859 was cloned by heterologous expression in nonpectolytic P. syringae pv. syringae BUVS1, using genomic DNA libraries constructed with two novel broad-host-range cosmid vectors, pCPP34 and pCPP47. Screening of P. syringae pv. syringae transconjugants for the ability to pit pectate media at pH 6.0 and 8.5 yielded several overlapping clones of the same DNA region. Ultrathin-layer isoelectric focusing gels, activity-stained with diagnostically buffered substrate overlays, revealed that this region encoded a single pectate lyase (PelS) with a pI of 9.4. pelS was subcloned from cosmid pCPP5020 and sequenced, revealing it to encode a member of the Erwinia chrysanthemi PelADE family, with highest similarity to Pseudomonas viridiflava PelV. A pelS probe hybridized at high stringency in DNA gel blots with total DNA from P. syringae pv. lachrymans strains 859 and Pla5, P. syringae pv. tabaci, P. syringae pv. phaseolicola, P. syringae pv. glycinea, P. fluorescens (marginalis), P. viridiflava, and Xanthomonas campestris pv. campestris, but not with P. syringae pv. pisi, P. syringae pv. syringae, P. syringae pv. tomato, P. syringae pv. papulans, E. chrysanthemi, or Ralstonia (Pseudomonas or Burkholderia) solanacearum. The PelS sequence revealed an N-terminal signal peptide, whose processing in Escherichia coli was confirmed by protein sequence analysis. PelS was similar to E. chrysanthemi PelE in its substrate preference and ability to reduce the viscosity of pectate and to macerate potato tuber tissue. A pelS:: omega Kmr mutation was marker-exchanged into P. syringae pv. lachrymans Pla5, pelS was also subcloned into the broad-host-range expression vector pML122 under control of the vector nptII promoter, and then transformed into P. syringae pv. lachrymans Pla5 to produce a strain overproducing PelS. Necrotic lesions developed in cotyledons following inoculation with all of the P. syringae pv. lachrymans Pla5 derivatives

  20. Grid integrated distributed PV (GridPV).

    SciTech Connect

    Reno, Matthew J.; Coogan, Kyle

    2013-08-01

    This manual provides the documentation of the MATLAB toolbox of functions for using OpenDSS to simulate the impact of solar energy on the distribution system. The majority of the functions are useful for interfacing OpenDSS and MATLAB, and they are of generic use for commanding OpenDSS from MATLAB and retrieving information from simulations. A set of functions is also included for modeling PV plant output and setting up the PV plant in the OpenDSS simulation. The toolbox contains functions for modeling the OpenDSS distribution feeder on satellite images with GPS coordinates. Finally, example simulations functions are included to show potential uses of the toolbox functions. Each function in the toolbox is documented with the function use syntax, full description, function input list, function output list, example use, and example output.

  1. Xanthomonas campestris overcomes Arabidopsis stomatal innate immunity through a DSF cell-to-cell signal-regulated virulence factor.

    PubMed

    Gudesblat, Gustavo E; Torres, Pablo S; Vojnov, Adrián A

    2009-02-01

    Pathogen-induced stomatal closure is part of the plant innate immune response. Phytopathogens using stomata as a way of entry into the leaf must avoid the stomatal response of the host. In this article, we describe a factor secreted by the bacterial phytopathogen Xanthomonas campestris pv campestris (Xcc) capable of interfering with stomatal closure induced by bacteria or abscisic acid (ABA). We found that living Xcc, as well as ethyl acetate extracts from Xcc culture supernatants, are capable of reverting stomatal closure induced by bacteria, lipopolysaccharide, or ABA. Xcc ethyl acetate extracts also complemented the infectivity of Pseudomonas syringae pv tomato (Pst) mutants deficient in the production of the coronatine toxin, which is required to overcome stomatal defense. By contrast, the rpfF and rpfC mutant strains of Xcc, which are unable to respectively synthesize or perceive a diffusible molecule involved in bacterial cell-to-cell signaling, were incapable of reverting stomatal closure, indicating that suppression of stomatal response by Xcc requires an intact rpf/diffusible signal factor system. In addition, we found that guard cell-specific Arabidopsis (Arabidopsis thaliana) Mitogen-Activated Protein Kinase3 (MPK3) antisense mutants were unresponsive to bacteria or lipopolysaccharide in promotion of stomatal closure, and also more sensitive to Pst coronatine-deficient mutants, showing that MPK3 is required for stomatal immune response. Additionally, we found that, unlike in wild-type Arabidopsis, ABA-induced stomatal closure in MPK3 antisense mutants is not affected by Xcc or by extracts from Xcc culture supernatants, suggesting that the Xcc factor might target some signaling component in the same pathway as MPK3.

  2. Atypical regulation of virulence-associated functions by a diffusible signal factor in Xanthomonas oryzae pv. oryzae.

    PubMed

    Rai, Rikky; Ranjan, Manish; Pradhan, Binod B; Chatterjee, Subhadeep

    2012-06-01

    In Xanthomonas oryzae pv. oryzae, the bacterial blight pathogen of rice, a secreted fatty acid signaling molecule known as diffusible signal factor (DSF) is required for virulence and growth on low-iron medium. To identify other virulence-associated traits that are regulated by DSF in this pathogen, we have performed microarray analysis of transcriptional changes between the wild type and DSF-deficient mutants of X. oryzae pv. oryzae. Expression of genes that encode secreted hydrolytic enzymes, motility, and chemotaxis functions are negatively regulated by DSF while functions involved in adhesion and biofilm formation are positively regulated. Enzymatic assays for hydrolytic enzymes as well as assays for chemotaxis, motility, attachment, and biofilm formation corroborate these findings. These results demonstrate that, in X. oryzae pv. oryzae, DSF-mediated cell-to-cell signaling coordinates transition from solitary to biofilm lifestyle by promoting expression of attachment functions and negatively regulating expression of motility functions. This is in contrast to X. campestris pv. campestris, a pathogen of crucifers, wherein the DSF system positively regulates motility functions and negatively regulates biofilm formation. These results indicate that virulence-associated functions can be regulated in a completely contrasting fashion by the same signaling system in very closely related bacteria.

  3. Expression and Functional Roles of the Pepper Pathogen-Induced bZIP Transcription Factor CabZIP2 in Enhanced Disease Resistance to Bacterial Pathogen Infection.

    PubMed

    Lim, Chae Woo; Baek, Woonhee; Lim, Sohee; Han, Sang-Wook; Lee, Sung Chul

    2015-07-01

    A pepper bZIP transcription factor gene, CabZIP2, was isolated from pepper leaves infected with a virulent strain of Xanthomonas campestris pv. vesicatoria. Transient expression analysis of the CabZIP2-GFP fusion protein in Nicotiana benthamiana revealed that the CabZIP2 protein is localized in the cytoplasm as well as the nucleus. The acidic domain in the N-terminal region of CabZIP2 that is fused to the GAL4 DNA-binding domain is required to activate the transcription of reporter genes in yeast. Transcription of CabZIP2 is induced in pepper plants inoculated with virulent or avirulent strains of X. campestris pv. vesicatoria. The CabZIP2 gene is also induced by defense-related hormones such as salicylic acid, methyl jasmonate, and ethylene. To elucidate the in vivo function of the CabZIP2 gene in plant defense, virus-induced gene silencing in pepper and overexpression in Arabidopsis were used. CabZIP2-silenced pepper plants were susceptible to infection by the virulent strain of X. campestris pv. vesicatoria, which was accompanied by reduced expression of defense-related genes such as CaBPR1 and CaAMP1. CabZIP2 overexpression in transgenic Arabidopsis plants conferred enhanced resistance to Pseudomonas syringae pv. tomato DC3000. Together, these results suggest that CabZIP2 is involved in bacterial disease resistance.

  4. The unique glutathione reductase from Xanthomonas campestris: Gene expression and enzyme characterization

    SciTech Connect

    Loprasert, Suvit . E-mail: suvit@cri.or.th; Whangsuk, Wirongrong; Sallabhan, Ratiboot; Mongkolsuk, Skorn

    2005-06-17

    The glutathione reductase gene, gor, was cloned from the plant pathogen Xanthomonas campestris pv. phaseoli. Its gene expression and enzyme characteristics were found to be different from those of previously studied homologues. Northern blot hybridization, promoter-lacZ fusion, and enzyme assay experiments revealed that its expression, unlike in Escherichia coli, is OxyR-independent and constitutive upon oxidative stress conditions. The deduced amino acid sequence shows a unique NADPH binding motif where the most highly conserved arginine residue, which is critical for NADPH binding, is replaced by glutamine. Interestingly, a search of the available Gor amino acid sequences from various sources, including other Xanthomonas species, revealed that this replacement is specific to the genus Xanthomonas. Recombinant Gor enzyme was purified and characterized, and was found to have a novel ability to use both, NADPH and NADH, as electron donor. A gor knockout mutant was constructed and shown to have increased expression of the organic peroxide-inducible regulator gene, ohrR.

  5. Environmental control in tea fields to reduce infection by Pseudomonas syringae pv. theae.

    PubMed

    Tomihama, T; Nonaka, T; Nishi, Y; Arai, K

    2009-02-01

    Bacterial shoot blight (BSB) disease, caused by Pseudomonas syringae pv. theae, is a major bacterial disease of tea plants in Japan. BSB mainly occurs in the low-temperature season, and lesion formation by P. syringae pv. theae is enhanced by both low temperature and the presence of ice nucleation-active Xanthomonas campestris (INAX), which catalyzes ice formation at -2 to -4 degrees C and is frequently co-isolated with P. syringae pv. theae from tea plants. Low temperature is thus the most important environmental factor influencing the incidence of BSB; however, the effects of low temperature on infection of the host by P. syringae pv. theae and of environmental controls in fields on the occurrence of the disease are poorly understood. In this study, we show that ice formation on tea leaves by INAX enhanced P. syringae pv. theae invasion into leaf tissue. The natural incidence of BSB in the field was closely related to early autumn frost. Frost protection in late autumn, which prevented ice formation on tea plants, significantly decreased the incidence of BSB, and frost protection combined with bactericide application held the incidence under the economic threshold level. Our data indicate that environmental control in the field based on microbial interactions in the host offers a new strategy for plant disease control.

  6. Open PV Project: Unlocking PV Installation Data (Brochure)

    SciTech Connect

    Not Available

    2012-04-01

    This brochure summarizes the Open PV Project, a collaborative effort of government, industry, and the public to compile a comprehensive database of PV installations in the United States. The brochure outlines the purpose and history of the project as well as the main capabilities and benefits of the online Open PV tool. The brochure also introduces how features of the tool are used, and it describes the sources and characteristics of Open PV's data and data collection processes.

  7. PV System Performance and Standards

    SciTech Connect

    Osterwald, C. R.

    2005-11-01

    This paper presents a brief overview of the status and accomplishments during fiscal year (FY) 2005 of the Photovoltaic (PV) System Performance and Standards Subtask, which is part of the PV Systems Engineering Project (a joint NREL-Sandia project).

  8. The oligopeptide permease (Opp) of the plant pathogen Xanthomonas axonopodis pv. citri.

    PubMed

    Moutran, Alexandre; Quaggio, Ronaldo Bento; Balan, Andrea; Ferreira, Luis Carlos de Souza; Ferreira, Rita de Cássia Café

    2004-05-01

    The oligopeptide permease (Opp), a protein-dependent ABC transporter, has been found in the genome of Xanthomonas axonopodis pv. citri ( Xac), but not in Xanthomonas campestris pv. campestris ( Xcc). Sequence analysis indicated that 4 opp genes ( oppA, oppB, oppC, oppD/F), located in a 33.8-kbp DNA fragment present only in the Xac genome, are arranged in an operon-like structure and share highest sequence similarities with Streptomyces roseofulvus orthologs. Nonetheless, analyses of the GC content, codon usage, and transposon positioning suggested that the Xac opp operon does not have an exogenous origin. The presence of a stop codon at one of the ATP-binding domains of OppD/F would render the uptake system nonfunctional, but detection of a single polycistronic mRNA and periplasmic OppA in actively growing bacteria suggests that the Opp permease is active and could contribute to the distinct nutritional requirements and host specificities of the two Xanthomonas species.

  9. Testing for PV Reliability (Presentation)

    SciTech Connect

    Kurtz, S.; Bansal, S.

    2014-09-01

    The DOE SUNSHOT workshop is seeking input from the community about PV reliability and how the DOE might address gaps in understanding. This presentation describes the types of testing that are needed for PV reliability and introduces a discussion to identify gaps in our understanding of PV reliability testing.

  10. Evolution of Enzymatic Activities in the Enolase Superfamily: L-Fuconate Dehydratase from Xanthomonas campestris

    SciTech Connect

    Yew,W.; Fedorov, A.; Fedorov, E.; Rakus, J.; Pierce, R.; Almo, S.; Gerlt, J.

    2006-01-01

    Many members of the mechanistically diverse enolase superfamily have unknown functions. In this report the authors use both genome (operon) context and screening of a library of acid sugars to assign the L-fuconate dehydratase (FucD) function to a member of the mandelate racemase (MR) subgroup of the superfamily encoded by the Xanthomonas campestris pv. campestris str. ATCC 33913 genome (GI: 21233491). Orthologues of FucD are found in both bacteria and eukaryotes, the latter including the rTS beta protein in Homo sapiens that has been implicated in regulating thymidylate synthase activity. As suggested by sequence alignments and confirmed by high-resolution structures in the presence of active site ligands, FucD and MR share the same active site motif of functional groups: three carboxylate ligands for the essential Mg2+ located at the ends of th third, fourth, and fifth-strands in the (/)7-barrel domain (Asp 248, Glu 274, and Glu 301, respectively), a Lys-x-Lys motif at the end of the second-strand (Lys 218 and Lys 220), a His-Asp dyad at the end of the seventh and sixth-strands (His 351 and Asp 324, respectively), and a Glue at the end of the eighth-strand (Glu 382). The mechanism of the FucD reaction involves initial abstraction of the 2-proton by Lys 220, acid catalysis of the vinylogous-elimination of the 3-OH group by His 351, and stereospecific ketonization of the resulting 2-keto-3-deoxy-L-fuconate product. Screening of the library of acid sugars revealed substrate and functional promiscuity: In addition to L-fuconate, FucD also catalyzes the dehydration of L-galactonate, D-arabinonate, D-altronate, L-talonate, and D-ribonate. The dehydrations of L-fuconate, L-galactonate, and D-arabinonate are initiated by abstraction of the 2-protons by Lys 220. The dehydrations of L-talonate and D-ribonate are initiated by abstraction of the 2-protons by His 351; however, protonation of the enediolate intermediates by the conjugate acid of Lys 220 yields L

  11. Xanthan production by Xanthomonas campestris using whey permeate medium.

    PubMed

    Savvides, A L; Katsifas, E A; Hatzinikolaou, D G; Karagouni, A D

    2012-08-01

    Xanthan gum is a polysaccharide that is widely used as stabilizer and thickener with many industrial applications in food industry. Our aim was to estimate the ability of Xanthomonas campestris ATCC 13951 for the production of xanthan gum by using whey as a growth medium, a by-product of dairy industry. X. campestris ATCC 13951 has been studied in batch cultures using a complex medium for the determination of the optimal concentration of glucose, galactose and lactose. In addition, whey was used under various treatment procedures (de-proteinated, partially hydrolyzed by β-lactamase and partially hydrolyzed and de-proteinated) as culture medium, to study the production of xanthan in a 2 l bioreactor with constant stirring and aeration. A production of 28 g/l was obtained when partially hydrolysed β-lactamase was used, which proved to be one of the highest xanthan gum production reported so far. At the same time, an effort has been made for the control and selection of the most appropriate procedure for the preservation of the strain and its use as inoculant in batch cultures, without loss of its viability and its capability of xanthan gum production. The pre-treatment of whey (whey permeate medium hydrolyzed, WPH) was very important for the production of xanthan by the strain X. campestris ATCC 13951 during batch culture conditions in a 2 l bioreactor. Preservation methods such as lyophilization, cryopreservation at various glycerol solution and temperatures have been examined. The results indicated that the best preservation method for the producing strain X. campestris ATCC 13951 was the lyophilization. Taking into account that whey permeate is a low cost by-product of the dairy industry, the production of xanthan achieved under the studied conditions was considered very promising for industrial application.

  12. [Production of xanthan gum in immobilized cultures of Xanthomonas campestris].

    PubMed

    Anselmo, R J; Viora, S; Carletti, S

    1992-01-01

    The efficiency of xanthan production through surface processes was evaluated. The best porous material was selected first. Thereafter, a comparative study was performed using submerged agitated process vs other without agitation but containing the selected porous material. The culture medium used was white potatoes infusion, buffered with K2HPO4 and supplemented with glucose in diverse concentrations. Besides, to evaluate a different type of surface process, three vegetables were valued: Ipomaea batatus, Solanum tuberosum and Daucus carota, with an without glucose supplement. Larger xanthan production was achieved with immobilization of X. campestris vs the conventional method, when the liquid culture medium was used. The highest yield was obtained when the white potatoes infusion was supplemented with glucose 2.5%, yielding a conversion of this saccharide to xanthan up to 58%. When X. campestris was cultured on fragmented vegetables, the highest xanthan gum yield (5.6g) was obtained with Solanum tuberosum supplemented with glucose. This yield indicators that X. campestris used the glucose added as well as the constitutive polysaccharide of this vegetable.

  13. A gene encoding stellacyanin is induced in Capsicum annuum by pathogens, methyl jasmonate, abscisic acid, wounding, drought and salt stress.

    PubMed

    Kong, Hye Young; Jung, Ho Won; Lee, Sung Chul; Choi, Doil; Hwang, Byung Kook

    2002-08-01

    A stellacyanin cDNA clone (CASLP1) was isolated from a pepper cDNA library from hypersensitive response (HR) lesions of leaves infected with an avirulent strain of Xanthomonas campestris pv. vesicatoria. The deduced amino acid sequences of CASLP1 are homologous to those of stellacyanins from cucumber, maize, pea and Arabidopsis. The CASLP1 mRNA was not constitutively expressed in all organs of pepper, but strongly induced and accumulated in pepper tissues infected with X. campestris pv. vesicatoria, Colletotrichum coccodes, Phytophthora capsici or C. gloeosporioides. In situ hybridization results revealed that CASLP1 transcripts were strongly localized in the phloem areas of vascular bundles in infected tissues of pepper stems and fruits. CASLP1 mRNA accumulation was found in lower pepper leaves infected by either virulent or avirulent strains of X. campestris pv. vesicatoria and non-pathogenic Pseudomonas fluorescens, but not in uninoculated upper leaves. Induction of this CASLP1 gene occurred in pepper leaves applied with methyl jasmonate (MeJA), but not with ethylene, salicylic acid, dl-beta-amino-n-butyric acid and benzothiadiazole. Accumulation of CASLP1 transcripts was locally or systemically induced in pepper leaves upon mechanical wounding and was activated in a MeJA-dependent manner. The CASLP1 transcript was also strongly induced in leaf and stem tissues after exposure of pepper plants to abscisic acid, salt and drought.

  14. Xanthomonas axonopodis pv. citri uses a plant natriuretic peptide-like protein to modify host homeostasis.

    PubMed

    Gottig, Natalia; Garavaglia, Betiana S; Daurelio, Lucas D; Valentine, Alex; Gehring, Chris; Orellano, Elena G; Ottado, Jorgelina

    2008-11-25

    Plant natriuretic peptides (PNPs) are a class of extracellular, systemically mobile molecules that elicit a number of plant responses important in homeostasis and growth. The bacterial citrus pathogen, Xanthomonas axonopodis pv. citri, also contains a gene encoding a PNP-like protein, XacPNP, that shares significant sequence similarity and identical domain organization with plant PNPs but has no homologues in other bacteria. We have expressed and purified XacPNP and demonstrated that the bacterial protein alters physiological responses including stomatal opening in plants. Although XacPNP is not expressed under standard nutrient rich culture conditions, it is strongly induced under conditions that mimic the nutrient poor intercellular apoplastic environment of leaves, as well as in infected tissue, suggesting that XacPNP transcription can respond to the host environment. To characterize the role of XacPNP during bacterial infection, we constructed a XacPNP deletion mutant. The lesions caused by this mutant were more necrotic than those observed with the wild-type, and bacterial cell death occurred earlier in the mutant. Moreover, when we expressed XacPNP in Xanthomonas axonopodis pv. vesicatoria, the transgenic bacteria caused less necrotic lesions in the host than the wild-type. In conclusion, we present evidence that a plant-like bacterial PNP can enable a plant pathogen to modify host responses to create conditions favorable to its own survival.

  15. Quantifying PV power Output Variability

    SciTech Connect

    Hoff, Thomas E.; Perez, Richard

    2010-10-15

    This paper presents a novel approach to rigorously quantify power Output Variability from a fleet of photovoltaic (PV) systems, ranging from a single central station to a set of distributed PV systems. The approach demonstrates that the relative power Output Variability for a fleet of identical PV systems (same size, orientation, and spacing) can be quantified by identifying the number of PV systems and their Dispersion Factor. The Dispersion Factor is a new variable that captures the relationship between PV Fleet configuration, Cloud Transit Speed, and the Time Interval over which variability is evaluated. Results indicate that Relative Output Variability: (1) equals the inverse of the square root of the number of systems for fully dispersed PV systems; and (2) could be further minimized for optimally-spaced PV systems. (author)

  16. PV Hourly Simulation Tool

    SciTech Connect

    Dean, Jesse; Metzger, Ian

    2010-12-31

    This software requires inputs of simple general building characteristics and usage information to calculate the energy and cost benefits of solar PV. This tool conducts and complex hourly simulation of solar PV based primarily on the area available on the rooftop. It uses a simplified efficiency calculation method and real panel characteristics. It includes a detailed rate structure to account for time-of-use rates, on-peak and off-peak pricing, and multiple rate seasons. This tool includes the option for advanced system design inputs if they are known. This tool calculates energy savings, demand reduction, cost savings, incentives and building life cycle costs including: simple payback, discounted payback, net-present value, and savings to investment ratio. In addition this tool also displays the environmental benefits of a project.

  17. XCC2731, a GGDEF domain protein in Xanthomonas campestris, is involved in bacterial attachment and is positively regulated by Clp.

    PubMed

    Hsiao, Yi-Min; Liu, Yu-Fan; Fang, Mei-Chiung; Song, Wan-Ling

    2011-10-20

    In Xanthomonas campestris pv. campestris (Xcc), which is the causative agent of black rot in crucifers, the virulence factor level is substantially decreased in the mutant deficient in RpfG, a phosphodiesterase that degrades the second messenger cyclic di-GMP. The rpfG mutant also grew in an aggregated state. It is indicated that expression of Pseudomonas GGDEF domain protein WspR (a diguanylate cyclase that synthesizes cyclic di-GMP) in wild-type Xcc can produce a phenocopy of the rpfG mutant. In this study, we showed that over-expression of GGDEF domain protein XCC2731 in wild-type Xcc caused (i) aggregation of cells, (ii) reduction in motility, and (iii) decrease in production of virulence factor extracellular enzymes and exopolysaccharides. Site-directed mutagenesis of the conserved G, G, and E residues of the GGDEF domain in XCC2731 abolished its function. The XCC2731 mutant has attenuated virulence. Furthermore, XCC2731 mutant was affected in surface attachment. Using the 5' RACE method, the XCC2731 transcription initiation site was mapped at nucleotide G, 15nt upstream of the XCC2731 start codon. Transcriptional fusion assay and gel retardation analysis indicated that Clp (cAMP receptor protein-like protein) positively regulates XCC2731 transcription in a direct manner. Reporter analysis also revealed that XCC2731 transcription is subject to catabolite repression, and reduced under conditions of oxygen limitation and high osmolarity. Our findings not only extend previous work on Clp regulation to show that it influences the expression of XCC2731 in Xcc, but also are the first to characterize the GGDEF domain protein gene expression in this phytopathogen. Copyright © 2010 Elsevier GmbH. All rights reserved.

  18. Expression, purification, and characterization of an aminopeptidase (Xac2987) with broad specificity from Xanthomonas axonopodis pv. citri.

    PubMed

    Santos, Kelly; Medrano, Francisco J

    2007-03-01

    We report here, the cloning, expression, and purification of a broad specificity aminopeptidase from Xanthomonas campestris pv. citri in fusion with a hexa-histidine tag at the N-terminal portion of the protein to facilitate purification. The protein was expressed in the soluble fraction and could be purified in one step by IMAC, yielding approximately 50mg pure protein per liter of cells. We show that the protein is folded and presents aminopeptidase activity against synthetic substrates. Also, we present the characterization of its specificity, showing that the protein was, indeed, able to catalyze the removal of N-terminal residues from synthetic substrates.

  19. Effect of different fertilizers on nitrogen isotope composition and nitrate content of Brassica campestris.

    PubMed

    Yuan, Yuwei; Zhao, Ming; Zhang, Zhiheng; Chen, Tianjin; Yang, Guiling; Wang, Qiang

    2012-02-15

    The effect of different fertilizers on the δ(15)N value, nitrate concentration, and nitrate reductase activity of Brassica campestris and the δ(15)N value of soil has been investigated through a pot experiment. The δ(15)N mean value of B. campestris at the seedling stage observed in the composted chicken treatment (+8.65‰) was higher than that of chemical fertilizer treatment (+5.73‰), compost-chemical fertilizer (+7.53‰), and control check treatment (+7.86‰). There were significantly different δ(15)N values (p < 0.05) between B. campestris cultivated with composted chicken manure treatment and with chemical fertilizer treatment. The similar results were also found at the middle stage and the terminal stage. The variation of δ(15)N value in soil for different treatments was smaller than that of B. campestris, which was +6.71-+8.12‰, +6.83-+8.24‰, and +6.85-8.4‰, respectively, at seedling stage, middle stage, and terminal stage. With the growth of B. campestris, the nitrate content decreased in all treatments, and the nitrate reductase activity in B. campestris increased except for the CK. Results suggested that the δ(15)N values of B. campestris and soil were more effected by the fertilizer than by the dose level, and the δ(15)N value analysis could be used as a tool to discriminate the B. campestris cultivated with composted manure or chemical fertilizer.

  20. Draft genome of the xanthan producer Xanthomonas campestris NRRL B-1459 (ATCC 13951).

    PubMed

    Wibberg, Daniel; Alkhateeb, Rabeaa S; Winkler, Anika; Albersmeier, Andreas; Schatschneider, Sarah; Albaum, Stefan; Niehaus, Karsten; Hublik, Gerd; Pühler, Alfred; Vorhölter, Frank-Jörg

    2015-06-20

    Xanthomonas campestris NRRL B-1459 was used in pioneering studies related to the biotechnological production of xanthan, the commercially most important polysaccharide of bacterial origin. The analysis of its genome revealed a 5.1Mb chromosome plus the first complete plasmid of an X. campestris strain applied in biotechnology.

  1. The RpfB-Dependent Quorum Sensing Signal Turnover System Is Required for Adaptation and Virulence in Rice Bacterial Blight Pathogen Xanthomonas oryzae pv. oryzae.

    PubMed

    Wang, Xing-Yu; Zhou, Lian; Yang, Jun; Ji, Guang-Hai; He, Ya-Wen

    2016-03-01

    Xanthomonas oryzae pv. oryzae, the bacterial blight pathogen of rice, produces diffusible signal factor (DSF) family quorum sensing signals to regulate virulence. The biosynthesis and perception of DSF family signals require components of the rpf (regulation of pathogenicity factors) cluster. In this study, we report that RpfB plays an essential role in DSF family signal turnover in X. oryzae pv. oryzae PXO99A. The production of DSF family signals was boosted by deletion of the rpfB gene and was abolished by its overexpression. The RpfC/RpfG-mediated DSF signaling system negatively regulates rpfB expression via the global transcription regulator Clp, whose activity is reversible in the presence of cyclic diguanylate monophosphate. These findings indicate that the DSF family signal turnover system in PXO99A is generally consistent with that in Xanthomonas campestris pv. campestris. Moreover, this study has revealed several specific roles of RpfB in PXO99A. First, the rpfB deletion mutant produced high levels of DSF family signals but reduced extracellular polysaccharide production, extracellular amylase activity, and attenuated pathogenicity. Second, the rpfB/rpfC double-deletion mutant was partially deficient in xanthomonadin production. Taken together, the RpfB-dependent DSF family signal turnover system is a conserved and naturally presenting signal turnover system in Xanthomonas spp., which plays unique roles in X. oryzae pv. oryzae adaptation and pathogenesis.

  2. Xanthomonas campestris FabH is required for branched-chain fatty acid and DSF-family quorum sensing signal biosynthesis

    PubMed Central

    Yu, Yong-Hong; Hu, Zhe; Dong, Hui-Juan; Ma, Jin-Cheng; Wang, Hai-Hong

    2016-01-01

    Xanthomonas campestris pv. campestris (Xcc), a Gram-negative phytopathogenic bacterium, causes black rot disease of cruciferous vegetables. Although Xcc has a complex fatty acid profile comprised of straight-chain fatty acids and branched-chain fatty acids (BCFAs), and encodes a complete set of genes required for fatty acid synthesis, there is still little known about the mechanism of BCFA synthesis. We reported that expression of Xcc fabH restores the growth of Ralstonia solanacearum fabH mutant, and this allows the R. solanacearum fabH mutant to produce BCFAs. Using in vitro assays, we demonstrated that Xcc FabH is able to condense branched-chain acyl-CoAs with malonyl-ACP to initiate BCFA synthesis. Moreover, although the fabH gene is essential for growth of Xcc, it can be replaced with Escherichia coli fabH, and Xcc mutants failed to produce BCFAs. These results suggest that Xcc does not have an obligatory requirement for BCFAs. Furthermore, Xcc mutants lost the ability to produce cis-11-methyl-2-dodecenoic acid, a diffusible signal factor (DSF) required for quorum sensing of Xcc, which confirms that the fatty acid synthetic pathway supplies the intermediates for DSF signal biosynthesis. Our study also showed that replacing Xcc fabH with E. coli fabH affected Xcc pathogenesis in host plants. PMID:27595587

  3. Xanthomonas campestris FabH is required for branched-chain fatty acid and DSF-family quorum sensing signal biosynthesis.

    PubMed

    Yu, Yong-Hong; Hu, Zhe; Dong, Hui-Juan; Ma, Jin-Cheng; Wang, Hai-Hong

    2016-09-06

    Xanthomonas campestris pv. campestris (Xcc), a Gram-negative phytopathogenic bacterium, causes black rot disease of cruciferous vegetables. Although Xcc has a complex fatty acid profile comprised of straight-chain fatty acids and branched-chain fatty acids (BCFAs), and encodes a complete set of genes required for fatty acid synthesis, there is still little known about the mechanism of BCFA synthesis. We reported that expression of Xcc fabH restores the growth of Ralstonia solanacearum fabH mutant, and this allows the R. solanacearum fabH mutant to produce BCFAs. Using in vitro assays, we demonstrated that Xcc FabH is able to condense branched-chain acyl-CoAs with malonyl-ACP to initiate BCFA synthesis. Moreover, although the fabH gene is essential for growth of Xcc, it can be replaced with Escherichia coli fabH, and Xcc mutants failed to produce BCFAs. These results suggest that Xcc does not have an obligatory requirement for BCFAs. Furthermore, Xcc mutants lost the ability to produce cis-11-methyl-2-dodecenoic acid, a diffusible signal factor (DSF) required for quorum sensing of Xcc, which confirms that the fatty acid synthetic pathway supplies the intermediates for DSF signal biosynthesis. Our study also showed that replacing Xcc fabH with E. coli fabH affected Xcc pathogenesis in host plants.

  4. Genetics of xanthan production in Xanthomonas campestris: the xanA and xanB genes are involved in UDP-glucose and GDP-mannose biosynthesis.

    PubMed Central

    Köplin, R; Arnold, W; Hötte, B; Simon, R; Wang, G; Pühler, A

    1992-01-01

    The nucleotide sequence of a 3.4-kb EcoRI-PstI DNA fragment of Xanthomonas campestris pv. campestris revealed two open reading frames, which were designated xanA and xanB. The genes xanA and xanB encode proteins of 448 amino acids (molecular weight of 48,919) and 466 amino acids (molecular weight of 50,873), respectively. These genes were identified by analyzing insertion mutants which were known to be involved in xanthan production. Specific tests for the activities of enzymes involved in the biosynthesis of UDP-glucose and GDP-mannose indicated that the xanA gene product was involved in the biosynthesis of both glucose 1-phosphate and mannose 1-phosphate. The deduced amino acid sequence of xanB showed a significant degree of homology (59%) to the phosphomannose isomerase of Pseudomonas aeruginosa, a key enzyme in the biosynthesis of alginate. Moreover, biochemical analysis and complementation experiments with the Escherichia coli manA fragment revealed that xanB encoded a bifunctional enzyme, phosphomannose isomerase-GDP-mannose pyrophosphorylase. Images PMID:1370280

  5. The PAS domain-containing histidine kinase RpfS is a second sensor for the diffusible signal factor of Xanthomonas campestris.

    PubMed

    An, Shi-Qi; Allan, John H; McCarthy, Yvonne; Febrer, Melanie; Dow, J Maxwell; Ryan, Robert P

    2014-05-01

    A cell-cell signalling system mediated by the fatty acid signal DSF controls the virulence of Xanthomonas campestris pv. campestris (Xcc) to plants. The synthesis and recognition of the DSF signal depends upon different Rpf proteins. DSF signal generation requires RpfF whereas signal perception and transduction depends upon the sensor RpfC and regulator RpfG. Detailed analyses of the regulatory roles of different Rpf proteins have suggested the occurrence of further sensors for DSF. Here we have used a mutagenesis approach coupled with high-resolution transcriptional analysis to identify XC_2579 (RpfS) as a second sensor for DSF in Xcc. RpfS is a complex sensor kinase predicted to have multiple Per/Arnt/Sim (PAS) domains, a histidine kinase domain and a C-terminal receiver (REC) domain. Isothermal calorimetry showed that DSF bound to the isolated N-terminal PAS domain with a Kd of 1.4 μM. RpfS controlled expression of a sub-set of genes distinct from those controlled by RpfC to include genes involved in type IV secretion and chemotaxis. Mutation of XC_2579 was associated with a reduction in virulence of Xcc to Chinese Radish when assayed by leaf spraying but not by leaf inoculation, suggesting a role for RpfS-controlled factors in the epiphytic phase of the disease cycle.

  6. The use of stable and unstable green fluorescent proteins for studies in two bacterial models: Agrobacterium tumefaciens and Xanthomonas campestris pv. campestris.

    PubMed

    Sabuquillo, Pilar; Gea, Adela; Matas, Isabel M; Ramos, Cayo; Cubero, Jaime

    2017-05-01

    Fluorescent proteins have been used to track plant pathogens to understand their host interactions. To be useful, the transgenic pathogens must present similar behaviour than the wild-type isolates. Herein, a GFP marker was used to transform two plant pathogenic bacteria, Agrobacterium and Xanthomonas, to localize and track the bacteria during infection. The transgenic bacteria were evaluated to determine whether they showed the same fitness than the wild-type strains or whether the expression of the GFP protein interfered in the bacterial activity. In Agrobacterium, the plasmid used for transformation was stable in the bacteria and the strain kept the virulence, while Xanthomonas was not able to conserve the plasmid and transformed strains showed virulence variations compared to wild-type strains. Although marking bacteria with GFP to track infection in plants is a common issue, works to validate the transgenic strains and corroborate their fitness are not usual. Results, presented here, confirm the importance of proper fitness tests on the marked strains before performing localization assays, to avoid underestimation of the microbe population or possible artificial effects in its interaction with the plant.

  7. First Report of Severe Outbreaks of Bacterial Leaf Spot of Leafy Brassica Greens Caused by Xanthomonas Campestris pv. Campestris in South Carolina

    USDA-ARS?s Scientific Manuscript database

    Severe outbreaks of leaf spot disease of leafy vegetable Brassica have occurred from early spring to late fall for at least the past five years in Lexington County, the major growing region for leafy greens in South Carolina. Significant economic losses to this disease have been reported by both la...

  8. PV water pumping: NEOS Corporation recent PV water pumping activities

    SciTech Connect

    Lane, C.

    1995-11-01

    NEOS Corporation has been very active in PV-powered water pumping, particularly with respect to electric utilities. Most of the recent activity has been through the Photovoltaic Services Network (PSN). The PSN is an independent, not-for-profit organization comprised of all types of electric utilities: rural electric coops, public power districts, investor-owned utilities, and power marketing agencies. The PSN`s mission is to work pro-actively to promote utility involvement in PV through education and training. PV information is distributed by the PSN in three primary forms: (1) consultation with PSN technical service representatives: (2) literature generated by the PSN; and (3) literature published by other organizations. The PSN can also provide assistance to members in developing PV customer service programs. The PSN`s product support activities include consolidation of information on existing packaged PV systems and facilitation of the development of new PV product packages that meet utility-defined specifications for cost performance, and reliability. The PSN`s initial product support efforts will be focused on commercially available packaged PV systems for a variety of off-grid applications. In parallel with this effort, if no products exist that meet the PSN`s functional specifications, the PSN will initiate the second phase of product development support process by encouraging the development of new packaged systems. Through these services and product support activities, the PSN anticipates engaging all segments for the PV industry, thus providing benefits to PV systems suppliers as well as local PV service contractors.This paper describes field testing of pv power systems for water pumping.

  9. Outdoor PV Degradation Comparison

    SciTech Connect

    Jordan, D. C.; Smith, R. M.; Osterwald, C. R.; Gelak, E.; Kurtz, S. R.

    2011-02-01

    As photovoltaic (PV) penetration of the power grid increases, it becomes vital to know how decreased power output; may affect cost over time. In order to predict power delivery, the decline or degradation rates must be determined; accurately. At the Performance and Energy Rating Testbed (PERT) at the Outdoor Test Facility (OTF) at the; National Renewable Energy Laboratory (NREL) more than 40 modules from more than 10 different manufacturers; were compared for their long-term outdoor stability. Because it can accommodate a large variety of modules in a; limited footprint the PERT system is ideally suited to compare modules side-by-side under the same conditions.

  10. Composition and functional properties of Lupinus campestris protein isolates.

    PubMed

    Rodríguez-Ambriz, S L; Martínez-Ayala, A L; Millán, F; Dávila-Ortíz, G

    2005-09-01

    Protein isolates from L. campestris and soybean seeds were prepared using isoelectric precipitation (PI) and micellization (MI) procedures. The amount of protein recovered was considerably higher with the isoelectric precipitation than with the micellization procedure (60% and 30%, respectively). Protein contents were higher than 90% in protein isolates. Antinutritional factors content (alkaloids, lectins, and tannins) were reduced to innocuous levels after protein isolate preparation. Minimum protein solubility for the precipitated lupin protein isolate (LPI) was at pH 4.0, and between pH 4 and 6 for the micellized lupin protein isolate (LMI), increasing at both extremes of the pH scale. Water absorption for the LMI was 1.3 ml/g of protein and its oil absorption 2.2 ml/g of protein. The LPI had 1.7 ml/g of protein in both water and oil absorption. Foaming capacity and stability was pH-dependent. Foaming capacity was higher at pH 2 and lower near the protein isoelectric points. Minimum protein concentration for gelation in LMI was 8% w/v at pH 4, while for LPI was 6% at pH 4 and 6. Amino acid composition in L. campestris flour and protein isolates was high in lysine and low in methionine. Most of the essential amino acids in lupin protein isolates were at acceptable levels compared to a reference pattern for infants and adults. The electrophoretic pattern of both protein isolates showed three bands with different mobilities, suggesting that the protein fractions belong to alpha-conglutin (11S-like protein), beta-conglutin (7S-like protein) and gamma-conglutin. It is proven that some of the functional properties of L. campestris protein isolates are similar to those soybean protein isolates recovered under equal conditions.

  11. Xanthomonas campestris Overcomes Arabidopsis Stomatal Innate Immunity through a DSF Cell-to-Cell Signal-Regulated Virulence Factor1[OA

    PubMed Central

    Gudesblat, Gustavo E.; Torres, Pablo S.; Vojnov, Adrián A.

    2009-01-01

    Pathogen-induced stomatal closure is part of the plant innate immune response. Phytopathogens using stomata as a way of entry into the leaf must avoid the stomatal response of the host. In this article, we describe a factor secreted by the bacterial phytopathogen Xanthomonas campestris pv campestris (Xcc) capable of interfering with stomatal closure induced by bacteria or abscisic acid (ABA). We found that living Xcc, as well as ethyl acetate extracts from Xcc culture supernatants, are capable of reverting stomatal closure induced by bacteria, lipopolysaccharide, or ABA. Xcc ethyl acetate extracts also complemented the infectivity of Pseudomonas syringae pv tomato (Pst) mutants deficient in the production of the coronatine toxin, which is required to overcome stomatal defense. By contrast, the rpfF and rpfC mutant strains of Xcc, which are unable to respectively synthesize or perceive a diffusible molecule involved in bacterial cell-to-cell signaling, were incapable of reverting stomatal closure, indicating that suppression of stomatal response by Xcc requires an intact rpf/diffusible signal factor system. In addition, we found that guard cell-specific Arabidopsis (Arabidopsis thaliana) Mitogen-Activated Protein Kinase3 (MPK3) antisense mutants were unresponsive to bacteria or lipopolysaccharide in promotion of stomatal closure, and also more sensitive to Pst coronatine-deficient mutants, showing that MPK3 is required for stomatal immune response. Additionally, we found that, unlike in wild-type Arabidopsis, ABA-induced stomatal closure in MPK3 antisense mutants is not affected by Xcc or by extracts from Xcc culture supernatants, suggesting that the Xcc factor might target some signaling component in the same pathway as MPK3. PMID:19091877

  12. Interband cascade (IC) photovoltaic (PV) architecture for PV devices

    DOEpatents

    Yang, Rui Q.; Tian, Zhaobing; Mishima, Tetsuya D.; Santos, Michael B.; Johnson, Matthew B.; Klem, John F.

    2015-10-20

    A photovoltaic (PV) device, comprising a PV interband cascade (IC) stage, wherein the IC PV stage comprises an absorption region with a band gap, the absorption region configured to absorb photons, an intraband transport region configured to act as a hole barrier, and an interband tunneling region configured to act as an electron barrier. An IC PV architecture for a photovoltaic device, the IC PV architecture comprising an absorption region, an intraband transport region coupled to the absorption region, and an interband tunneling region coupled to the intraband transport region and to the adjacent absorption region, wherein the absorption region, the intraband transport region, and the interband tunneling region are positioned such that electrons will flow from the absorption region to the intraband transport region to the interband tunneling region.

  13. Helping enhances productivity in campo flicker ( Colaptes campestris) cooperative groups

    NASA Astrophysics Data System (ADS)

    Dias, Raphael Igor; Webster, Michael S.; Macedo, Regina H.

    2015-06-01

    Reproductive adults in many bird species are assisted by non-breeding auxiliary helpers at the nest, yet the impact of auxiliaries on reproduction is variable and not always obvious. In this study, we tested Hamilton's rule and evaluated the effect of auxiliaries on productivity in the facultative cooperative breeder campo flicker ( Colaptes campestris campestris). Campo flickers have a variable mating system, with some groups having auxiliaries and others lacking them (i.e., unassisted pairs). Most auxiliaries are closely related to the breeding pair (primary auxiliaries), but some auxiliaries (secondary auxiliaries) are unrelated females that joined established groups. We found no effect of breeder quality (body condition) or territory quality (food availability) on group productivity, but the presence of auxiliaries increased the number of fledglings produced relative to unassisted pairs. Nonetheless, the indirect benefit of helping was small and did not outweigh the costs of delayed breeding and so seemed insufficient to explain the evolution of cooperative breeding in campo flickers. We concluded that some ecological constraints must limit dispersal or independent breeding, making staying in the group a "best-of-a-bad-job" situation for auxiliaries.

  14. A Proteomic Analysis of Seed Development in Brassica campestri L

    PubMed Central

    Li, Wenlan; Gao, Yi; Xu, Hong; Zhang, Yu; Wang, Jianbo

    2012-01-01

    To gain insights into the protein dynamics during seed development, a proteomic study on the developing Brassica campestri L. seeds with embryos in different embryogenesis stages was carried out. The seed proteins at 10, 16, 20, 25 and 35 DAP (days after pollination), respectively, were separated using two-dimensional gel electrophoresis and identities of 209 spots with altered abundance were determined by matrix-assisted laser desorption ionization time-of-flight/time-of-flight mass spectrometry (MALDI-TOF/TOF MS). These proteins were classified into 16 groups according to their functions. The most abundant proteins were related to primary metabolism, indicating the heavy demand of materials for rapid embryo growth. Besides, the high amount of proteins involved in protein processing and destination indicated importance of protein renewal during seed development. The remaining were those participated in oxidation/detoxification, energy, defense, transcription, protein synthesis, transporter, cell structure, signal transduction, secondary metabolism, transposition, DNA repair, storage and so on. Protein abundance profiles of each functional class were generated and hierarchical cluster analysis established 8 groups of dynamic patterns. Our results revealed novel characters of protein dynamics in seed development in Brassica campestri L. and provided valuable information about the complex process of seed development in plants. PMID:23189193

  15. Helping enhances productivity in campo flicker (Colaptes campestris) cooperative groups.

    PubMed

    Dias, Raphael Igor; Webster, Michael S; Macedo, Regina H

    2015-06-01

    Reproductive adults in many bird species are assisted by non-breeding auxiliary helpers at the nest, yet the impact of auxiliaries on reproduction is variable and not always obvious. In this study, we tested Hamilton's rule and evaluated the effect of auxiliaries on productivity in the facultative cooperative breeder campo flicker (Colaptes campestris campestris). Campo flickers have a variable mating system, with some groups having auxiliaries and others lacking them (i.e., unassisted pairs). Most auxiliaries are closely related to the breeding pair (primary auxiliaries), but some auxiliaries (secondary auxiliaries) are unrelated females that joined established groups. We found no effect of breeder quality (body condition) or territory quality (food availability) on group productivity, but the presence of auxiliaries increased the number of fledglings produced relative to unassisted pairs. Nonetheless, the indirect benefit of helping was small and did not outweigh the costs of delayed breeding and so seemed insufficient to explain the evolution of cooperative breeding in campo flickers. We concluded that some ecological constraints must limit dispersal or independent breeding, making staying in the group a "best-of-a-bad-job" situation for auxiliaries.

  16. Antioxidant and antifungal activities of Smilax campestris Griseb. (Smilacaceae).

    PubMed

    Morais, Marcela Isis; Pinto, Maria Eduarda Amaral; Araújo, Sthéfane Guimarães; Castro, Ana Hortência Fonsêca; Duarte-Almeida, Joaquim Mauricio; Rosa, Luiz Henrique; Rosa, Carlos Augusto; Johann, Susana; Lima, Luciana Alves Rodrigues dos Santos

    2014-01-01

    Ethanol extract and fractions obtained from aerial parts of Smilax campestris were examined in order to determine their phenolic composition, antioxidant capacity and antifungal activities. High-performance liquid chromatography coupled with DAD analysis indicated that quercetin and rutin were the main phenolic compounds present in butanol fraction and ethanol extract, respectively. The antioxidant activity assessed by the scavenging ability on 1,1-diphenyl-2-picrylhydrazyl radical was significantly more pronounced for the ethanol extract and butanol fraction than that of the commercial antioxidant 2,6-di-tert-butyl-4-methylphenol. The antifungal activity of extract and fractions was investigated by using microdilution method against five Candida and two Cryptococcus yeast strains. Ethanol extract and fractions exhibited antifungal activities against Candida albicans, Candida glabrata, Candida krusei, Candida parapsilosis, Candida tropicalis and Cryptococcus gattii. This work provides the knowledge of profile and content of flavonoids and their antioxidant and antifungal activities in the extract and fractions of aerial parts of S. campestris.

  17. Identification of plant-induced genes of the bacterial pathogen Xanthomonas campestris pathovar campestris using a promoter-probe plasmid.

    PubMed

    Osbourn, A E; Barber, C E; Daniels, M J

    1987-01-01

    A promoter-probe plasmid suitable for use in Xanthomonas campestris pathovar campestris (causal agent of crucifer black rot) was constructed by ligating a broad host range IncQ replicon into the promoter-probe plasmid pKK232-8, which contains a promoterless chloramphenicol acetyltransferase gene. Xanthomonas chromosomal DNA fragments were 'shotgun' cloned into a restriction site in front of this gene, and the resulting library was transferred en masse into Xanthomonas. Individual transconjugants possessing DNA insertions with promoter activity in plants were identified by virtue of their ability to infect chloramphenicol-treated turnip seedlings. Of 19 transconjugants identified in this way five were chloramphenicol resistant both in turnip seedlings and on agar plates. However the remaining 14 were only chloramphenicol resistant in planta, and thus apparently contained plant-inducible promoter fragments. Resistance to chloramphenicol was correlated with increased chloramphenicol acetyltransferase activity for the transconjugants assayed. The promoter fragments were used to isolate genomic clones from a library, and the role of the genes contained in these clones in pathogenicity is being investigated.

  18. Patterns of mitochondrial DNA instability in Brassica campestris cultured cells.

    PubMed

    Shirzadegan, M; Palmer, J D; Christey, M; Earle, E D

    1991-01-01

    We previously showed that the mitochondrial DNA (mtDNA) of a Brassica campestris callus culture had undergone extensive rearrangements (i.e. large inversions and a duplication) relative to DNA of the control plant [54]. In this study we observed that after continued growth, the mtDNA of this culture continues to change, with rearranged forms amplifying and diminishing to varying proportions. Strikingly similar changes were detected in the mtDNA profiles of a variety of other long- and short-term callus and cell suspension lines. However, the proportions of parental ('unrearranged') and novel ('rearranged') forms varied in different cultured cell mtDNAs. To address the source of this heterogeneity, we compared the mtDNA organization of 28 individual plants from the parental seed stock. With the exception of one plant containing high levels of a novel plasmid-like mtDNA molecule, no significant variation was detected among individual plants and therefore source plant variation is unlikely to have contributed to the diversity of mitochondrial genomes observed in cultured cells. The source of this culture-induced heterogeneity was also investigated in 16 clones derived from single protoplasts. A mixed population of unrearranged and rearranged mtDNA molecules was apparent in each protoclone, suggesting that the observed heterogeneity in various cultures might reflect the genomic composition of each individual cell; however, the induction of an intercellular heterogeneity subsequent to the protoplast isolation was not tested and therefore cannot be ruled out. The results of this study support our earlier model that the rapid structural alteration of B. campestris mtDNA in vitro results from preferential amplification and reassortment of minor pre-existing forms of the genome rather than de novo rearrangement. Infrequent recombination between short dispersed repeated elements is proposed as the underlying mechanism for the formation of these minor mtDNA molecules.

  19. Pressure-equalizing PV assembly and method

    DOEpatents

    Dinwoodie, Thomas L.

    2004-10-26

    Each PV assembly of an array of PV assemblies comprises a base, a PV module and a support assembly securing the PV module to a position overlying the upper surface of the base. Vents are formed through the base. A pressure equalization path extends from the outer surface of the PV module, past the PV module, to and through at least one of the vents, and to the lower surface of the base to help reduce wind uplift forces on the PV assembly. The PV assemblies may be interengaged, such as by interengaging the bases of adjacent PV assemblies. The base may include a main portion and a cover and the bases of adjacent PV assemblies may be interengaged by securing the covers of adjacent bases together.

  20. PV module mounting method and mounting assembly

    DOEpatents

    Lenox, Carl J.S.; Johnson, Kurt M.

    2013-04-23

    A method for mounting PV modules to a deck includes selecting PV module layout pattern so that adjacent PV module edges are spaced apart. PV mounting and support assemblies are secured to the deck according to the layout pattern using fasteners extending into the deck. The PV modules are placed on the PV mounting and support assemblies. Retaining elements are located over and secured against the upper peripheral edge surfaces of the PV modules so to secure them to the deck with the peripheral edges of the PV modules spaced apart from the deck. In some examples a PV module mounting assembly, for use on a shingled deck, comprises flashing, a base mountable on the flashing, a deck-penetrating fastener engageable with the base and securable to the deck so to secure the flashing and the base to the shingled deck, and PV module mounting hardware securable to the base.

  1. PSCAD Modules Representing PV Generator

    SciTech Connect

    Muljadi, E.; Singh, M.; Gevorgian, V.

    2013-08-01

    Photovoltaic power plants (PVPs) have been growing in size, and the installation time is very short. With the cost of photovoltaic (PV) panels dropping in recent years, it can be predicted that in the next 10 years the contribution of PVPs to the total number of renewable energy power plants will grow significantly. In this project, the National Renewable Energy Laboratory (NREL) developed a dynamic modeling of the modules to be used as building blocks to develop simulation models of single PV arrays, expanded to include Maximum Power Point Tracker (MPPT), expanded to include PV inverter, or expanded to cover an entire PVP. The focus of the investigation and complexity of the simulation determines the components that must be included in the simulation. The development of the PV inverter was covered in detail, including the control diagrams. Both the current-regulated voltage source inverter and the current-regulated current source inverter were developed in PSCAD. Various operations of the PV inverters were simulated under normal and abnormal conditions. Symmetrical and unsymmetrical faults were simulated, presented, and discussed. Both the three-phase analysis and the symmetrical component analysis were included to clarify the understanding of unsymmetrical faults. The dynamic model validation was based on the testing data provided by SCE. Testing was conducted at SCE with the focus on the grid interface behavior of the PV inverter under different faults and disturbances. The dynamic model validation covers both the symmetrical and unsymmetrical faults.

  2. Locational Sensitivity Investigation on PV Hosting Capacity and Fast Track PV Screening

    SciTech Connect

    Ding, Fei; Mather, Barry; Ainsworth, Nathan; Gotseff, Peter; Baker, Kyri

    2016-05-05

    A 15% PV penetration threshold is commonly used by utilities to define photovoltaic (PV) screening methods where PV penetration is defined as the ratio of total solar PV capacity on a line section to peak load. However, this method doesn't take into account PV locational impact or feeder characteristics that could strongly change the feeder's capability to host PVs. This paper investigates the impact of PV location and phase connection type on PV hosting capacity, and then proposes a fast-track PV screening approach that leverages various PV hosting capacity metric responding to different PV locations and types. The proposed study could help utilities to evaluate PV interconnection requests and also help increase the PV hosting capacity of distribution feeders without adverse impacts on system voltages.

  3. Complete Genome Sequences of Six Copper-Resistant Xanthomonas Strains Causing Bacterial Spot of Solaneous Plants, Belonging to X. gardneri, X. euvesicatoria, and X. vesicatoria, Using Long-Read Technology

    PubMed Central

    Richard, Damien; Boyer, Claudine; Lefeuvre, Pierre; Canteros, Blanca I.; Beni-Madhu, Shyam; Portier, Perrine

    2017-01-01

    ABSTRACT Xanthomonas vesicatoria, Xanthomonas euvesicatoria, and Xanthomonas gardneri cause bacterial spot disease. Copper has been applied since the 1920s as part of integrated management programs. The first copper-resistant strains were reported some decades later. Here, we fully sequenced six Xanthomonas strains pathogenic to tomato and/or pepper and having a copper-resistant phenotype. PMID:28232425

  4. Low concentrator PV optics optimization

    NASA Astrophysics Data System (ADS)

    Sharp, Leonard; Chang, Ben

    2008-08-01

    Purpose: Cost reduction is a major focus of the solar industry. Thin film technologies and concentration systems are viable ways to reducing cost, with unique strengths and weakness for both. Most of the concentrating PV work focuses on high concentration systems for reducing energy cost. Meanwhile, many believe that low concentrators provide significant cost reduction potential while addressing the mainstream PV market with a product that acts as a flat panel replacement. This paper analyzes the relative benefit of asymmetric vs. symmetric optics for low-concentrators in light of specific PV applications. Approach: Symmetric and asymmetric concentrating PV module performance is evaluated using computer simulation to determine potential value across various geographic locations and applications. The selected optic design is modeled against standard cSi flat panels and thin film to determine application fit, system level energy density and economic value. Results: While symmetric designs may seem ideal, asymmetric designs have an advantage in energy density. Both designs are assessed for aperture, optimum concentration ratio, and ideal system array configuration. Analysis of performance across climate specific effects (diffuse, direct and circumsolar) and location specific effects (sunpath) are also presented. The energy density and energy production of low concentrators provide a compelling value proposition. More significantly, the choice of optics for a low concentrating design can affect real world performance. With the goal of maximizing energy density and return on investment, this paper presents the advantages of asymmetric optic concentration and illustrates the value of this design within specific PV applications.

  5. PV system testing and standards

    NASA Astrophysics Data System (ADS)

    DeBlasio, Richard

    1999-03-01

    The U.S. Department of Energy (DOE) PV Program System Performance and Engineering Project is being conducted by The National Renewable Energy Laboratory (NREL), Sandia National Laboratories (SNL), Southwest Technology Development Institute (SWTDI), and Florida Solar Energy Center (FSEC). It provides PV system, subsystem, and component-level technology-performance characterization testing; test-method development and validation; national and international consensus standards and codes development, test-facility product certification, and laboratory-accreditation program implementation; and information exchange and technical assistance to the PV community. Emphasis is placed on reducing technical and infrastructural barriers to system acceptance, reducing life-cycle cost of systems, providing systems-engineering best practices and guidelines, and leading the national effort in performance and reliability testing, and consensus standards, codes, and certification program development and implementation—thereby ensuring that PV systems meet customers' needs and expectations. A summary of project activities, accomplishments, and future plans is provided and highlighted by an overview of PV system test-procedure and standards development.

  6. Cell-cell signal-dependent dynamic interactions between HD-GYP and GGDEF domain proteins mediate virulence in Xanthomonas campestris.

    PubMed

    Ryan, Robert P; McCarthy, Yvonne; Andrade, Maxuel; Farah, Chuck S; Armitage, Judith P; Dow, J Maxwell

    2010-03-30

    RpfG is a paradigm for a class of widespread bacterial two-component regulators with a CheY-like receiver domain attached to a histidine-aspartic acid-glycine-tyrosine-proline (HD-GYP) cyclic di-GMP phosphodiesterase domain. In the plant pathogen Xanthomonas campestris pv. campestris (Xcc), a two-component system comprising RpfG and the complex sensor kinase RpfC is implicated in sensing and responding to the diffusible signaling factor (DSF), which is essential for cell-cell signaling. RpfF is involved in synthesizing DSF, and mutations of rpfF, rpfG, or rpfC lead to a coordinate reduction in the synthesis of virulence factors such as extracellular enzymes, biofilm structure, and motility. Using yeast two-hybrid analysis and fluorescence resonance energy transfer experiments in Xcc, we show that the physical interaction of RpfG with two proteins with diguanylate cyclase (GGDEF) domains controls a subset of RpfG-regulated virulence functions. RpfG interactions were abolished by alanine substitutions of the three residues of the conserved GYP motif in the HD-GYP domain. Changing the GYP motif or deletion of the two GGDEF-domain proteins reduced Xcc motility but not the synthesis of extracellular enzymes or biofilm formation. RpfG-GGDEF interactions are dynamic and depend on DSF signaling, being reduced in the rpfF mutant but restored by DSF addition. The results are consistent with a model in which DSF signal transduction controlling motility depends on a highly regulated, dynamic interaction of proteins that influence the localized expression of cyclic di-GMP.

  7. Structure of the Full-Length Bacteriophytochrome from the Plant Pathogen Xanthomonas campestris Provides Clues to its Long-Range Signaling Mechanism.

    PubMed

    Otero, Lisandro Horacio; Klinke, Sebastián; Rinaldi, Jimena; Velázquez-Escobar, Francisco; Mroginski, María Andrea; Fernández López, María; Malamud, Florencia; Vojnov, Adrián Alberto; Hildebrandt, Peter; Goldbaum, Fernando Alberto; Bonomi, Hernán Ruy

    2016-09-25

    Phytochromes constitute a major superfamily of light-sensing proteins that are reversibly photoconverted between a red-absorbing (Pr) and a far-red-absorbing (Pfr) state. Bacteriophytochromes (BphPs) are found among photosynthetic and non-photosynthetic bacteria, including pathogens. To date, several BphPs have been biophysically characterized. However, it is still not fully understood how structural changes are propagated from the photosensory module to the output module during the signal transduction event. Most phytochromes share a common architecture consisting of an N-terminal photosensor that includes the PAS2-GAF-PHY domain triad and a C-terminal variable output module. Here we present the crystal structure of the full-length BphP from the plant pathogen Xanthomonas campestris pv. campestris (XccBphP) bearing its photosensor and its complete output module, a PAS9 domain. In the crystals, the protein was found to be in the Pr state, whereas diffraction data together with resonance Raman spectroscopic and theoretical results indicate a ZZZssa and a ZZEssa chromophore configuration corresponding to a mixture of Pr and Meta-R state, the precursor of Pfr. The XccBphP quaternary assembly reveals a head-to-head dimer in which the output module contributes to the helical dimer interface. The photosensor, which is shown to be a bathy-like BphP, is influenced in its dark reactions by the output module. Our structural analyses suggest that the photoconversion between the Pr and Pfr states in the full-length XccBphP may involve changes in the relative positioning of the output module. This work contributes to understand the light-induced structural changes propagated from the photosensor to the output modules in phytochrome signaling.

  8. Transcriptional regulation and molecular characterization of the manA gene encoding the biofilm dispersing enzyme mannan endo-1,4-beta-mannosidase in Xanthomonas campestris.

    PubMed

    Hsiao, Yi-Min; Liu, Yu-Fan; Fang, Mei-Chiung; Tseng, Yi-Hsiung

    2010-02-10

    Exopolysaccharide and several extracellular enzymes of Xanthomonas campestris pv. campestris (Xcc), the causative agent of black rot in crucifers, are important virulence determinants. It is known that Clp (cAMP receptor protein-like protein) and RpfF (an enoyl-CoA hydratase homologue required for the synthesis of diffusible signal factor, DSF) regulate the production of these determinants. Addition of DSF or Xcc extracellular protein containing partially purified mannanase (EC 3.2.1.78, encoded by manA) can disperse the cell aggregates formed by rpfF mutant. In this study, nucleotide G 64 nt upstream of the manA translation start codon was determined as the transcription initiation site by the 5' RACE technique. Transcriptional fusion assays showed that manA transcription is positively regulated by Clp and RpfF and induced by locust bean gum. The manA coding region was cloned and expressed in E. coli as recombinant ManA (rManA). The rManA was purified by affinity chromatography, and its biochemical properties were characterized. The rManA had a pH optimum at 7.0 (0.1 M Hepes) and a temperature optimum at about 37 degrees C. Sequence and mutational analyses demonstrated that Xcc manA encodes the major mannanase, a member of family 5 of glycosyl hydrolases. This study not only extends previous work on Clp and RpfF regulation by showing that they both influence the expression of manA in Xcc, but it also for the first time characterizes Xanthomonas mannanase at the protein level.

  9. [Effects of exogenous dimethylarsinic acid on Brassica campestris growth and soil arsenic bioavailability].

    PubMed

    Bai, Ling-Yu; Zeng, Xi-Bai; Hu, Liu-Jie; Li, Lian-Fang; He, Qiu-Hong

    2011-02-01

    A pot experiment was conducted to study the effects of exogenous dimethylarsinic acid (DMA) on the growth of Brassica campestris and the bioavailability of soil arsenic (As). With the increasing concentration of applied DMA, the emergence rate and biomass of B. campestris increased at low concentration DMA, but decreased at high concentration DMA. When the DMA concentration reached 90 mg x kg(-1), the emergence rate and biomass of B. campestris in the second cropping decreased by 9.5% and 57.0%, respectively, compared with those in the control, indicating that exogenous DMA had longer-term effects on the growth of B. campestris. The soil available As and the As uptake by B. campestris all increased with increasing concentration of exogenous DMA, and there existed significant correlations among them. After applied into soil, the exogenous DMA demethylated, with As(V) as the main product and lesser amount of As (III), and the concentrations of soil As(V) and As(III) increased with increasing application rate of exogenous DMA.

  10. Flexible packaging for PV modules

    NASA Astrophysics Data System (ADS)

    Dhere, Neelkanth G.

    2008-08-01

    Economic, flexible packages that provide needed level of protection to organic and some other PV cells over >25-years have not yet been developed. However, flexible packaging is essential in niche large-scale applications. Typical configuration used in flexible photovoltaic (PV) module packaging is transparent frontsheet/encapsulant/PV cells/flexible substrate. Besides flexibility of various components, the solder bonds should also be flexible and resistant to fatigue due to cyclic loading. Flexible front sheets should provide optical transparency, mechanical protection, scratch resistance, dielectric isolation, water resistance, UV stability and adhesion to encapsulant. Examples are Tefzel, Tedlar and Silicone. Dirt can get embedded in soft layers such as silicone and obscure light. Water vapor transmittance rate (WVTR) of polymer films used in the food packaging industry as moisture barriers are ~0.05 g/(m2.day) under ambient conditions. In comparison, light emitting diodes employ packaging components that have WVTR of ~10-6 g/(m2.day). WVTR of polymer sheets can be improved by coating them with dense inorganic/organic multilayers. Ethylene vinyl acetate, an amorphous copolymer used predominantly by the PV industry has very high O2 and H2O diffusivity. Quaternary carbon chains (such as acetate) in a polymer lead to cleavage and loss of adhesional strength at relatively low exposures. Reactivity of PV module components increases in presence of O2 and H2O. Adhesional strength degrades due to the breakdown of structure of polymer by reactive, free radicals formed by high-energy radiation. Free radical formation in polymers is reduced when the aromatic rings are attached at regular intervals. This paper will review flexible packaging for PV modules.

  11. Comparison of Pyranometers vs. PV Reference Cells for Evaluation of PV Array Performance

    SciTech Connect

    Dunn, L.; Gostein, M.; Emery, K.

    2012-09-01

    As the photovoltaics (PV) industry has grown, the need for accurately monitoring the solar resource of PV power plants has increased. Historically, the PV industry has relied on thermopile pyranometers for irradiance measurements, and a large body of historical irradiance data taken with pyranometers exists. However, interest in PV reference devices is increasing. In this paper, we discuss why PV reference devices are better suited for PV applications, and estimate the typical uncertainties in irradiance measurements made with both pyranometers and PV reference devices. We assert that the quantity of interest in monitoring a PV power plant is the equivalent irradiance under the IEC 60904-3 reference solar spectrum that would produce the same electrical response in the PV array as the incident solar radiation. For PV-plant monitoring applications, we find the uncertainties in irradiance measurements of this type to be on the order of +/-5% for thermopile pyranometers and +/-2.4% for PV reference devices.

  12. Bioconcentration factors (BCF) of silver in wild Agaricus campestris

    SciTech Connect

    Falandysz, J.; Danisiewicz, D.

    1995-07-01

    Silver is an element naturally occurring in small concentrations in different environmental sites. However, many anthropogenic sources of silver led to contamination of this element in soil surfaces, pastures, and coastal marine areas in different parts of the world. Estimates are that 40% of the 1.15x10{sup 4}t of silver produced annually worldwide, will escape into the environment. Due to municipal waste discharge and/or industrial effluents with high silver concentrations, 100 x above the background level have been reported in invertebrate species from polluted marine areas. The meta-stabile radioisotope, {sup 110m}Ag, is a main component of the liquid effluents from nuclear facilities under normal operating conditions. The presence of {sup 111}Ag and {sup 110m}Ag also has been widely found throughout Europe in the 1986 Chernobyl fallout. Silver ions are environmentally harmful. High toxic effects have been observed at low concentrations, especially in aquatic species. Species of lower fungi as well as the mushroom Agaricus bisporus are know to bioaccumulate high concentrations of silver when grown on an artificially enriched substrate. This study looks at the relationship between the silver content of soil and bioconcentration potential of wild Agaricus campestris from sites under different use and with different concentrations of heavy metals. 28 refs., 2 figs., 2 tabs.

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

    PubMed

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

    2014-01-01

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

  14. Progress & Frontiers in PV Performance

    SciTech Connect

    Deline, Chris; DiOrio, Nick; Jordan, Dirk; Toor, Fatima

    2016-09-12

    PowerPoint slides for a presentation given at Solar Power International 2016. Presentation includes System Advisor Model (SAM) introduction and battery modeling, bifacial PV modules and modeling, shade modeling and module level power electronics (MLPE), degradation rates, and PVWatts updates and validation.

  15. In silico model of DSF synthase RpfF protein from Xanthomonas oryzae pv. Oryzae: a novel target for bacterial blight of rice disease.

    PubMed

    Reddy, Vidavaluru Sravani; Kumar, Yellapu Nanda; Raghavendra, Aminedi; Sowjenya, Gopal; Kumar, Suman; Ramyasree, Guggulla; Reddy, Gottiprolu Rajarami

    2012-01-01

    Rice plant diseases play a major role as biological constraints on production. One of such rice disease is bacterial leaf blight, caused by Xanthomonas oryzae pv. Oryzae (Xoo). The diffusible signal factor (DSF) synthesized by Xoo has a major role in virulence to rice plant. The DSF synthase RpfF protein, which is related to crotonase superfamily is responsible for the maintaining concentration of DSF. DSF-dependent quorum sensing (QS) system adopts protein- protein interaction mechanism to auto regulates the production of DSF. The antibacterial activity of pesticides against Xoo has not yet been completely understood. Three dimensional structure of RpfF protein was predicted using homology modeling method by MODELLER 9V9 software, SWISS MODEL and GENO3D online tools and structures were validated by Ramachandran plot, TM-Score and RMSD. 3D structure of RpfF (accession number AAL06345) was predicted using DSF synthase of Xanthomonas campestris pv. campestris (Xcc) (PDB ID: 3M6M) as a template. The stereo chemical check reveals the structure developed from the modeller was the best one and the potential ligand binding sites were identified by CASTp Server. The predicted RpfF model provides insight into its structure, active sites and aid in the development of novel inhibitors to control bacterial leaf blight in rice plant. DSF synthase RpfF protein could be used as a novel target to control infection.

  16. Genome sequencing and comparative analysis of the carrot bacterial blight pathogen, Xanthomonas hortorum pv. carotae M081, for insights into pathogenicity and applications in molecular diagnostics.

    PubMed

    Kimbrel, Jeffrey A; Givan, Scott A; Temple, Todd N; Johnson, Kenneth B; Chang, Jeff H

    2011-08-01

    Xanthomonas hortorum pv. carotae (Xhc) is an economically important pathogen of carrots. Its ability to epiphytically colonize foliar surfaces and infect seeds can result in bacterial blight of carrots when grown in warm and humid regions. We used high-throughput sequencing to determine the genome sequence of isolate M081 of Xhc. The short reads were de novo assembled and the resulting contigs were ordered using a syntenic reference genome sequence from X. campestris pv. campestris ATCC 33913. The improved, high-quality draft genome sequence of Xhc M081 is the first for its species. Despite its distance from other sequenced xanthomonads, Xhc M081 still shared a large inventory of orthologous genes, including many clusters of virulence genes common to other foliar pathogenic species of Xanthomonas. We also mined the genome sequence and identified at least 21 candidate type III effector genes. Two were members of the avrBs2 and xopQ families that demonstrably elicit effector-triggered immunity. We showed that expression in planta of these two type III effectors from Xhc M081 was sufficient to elicit resistance gene-mediated hypersensitive responses in heterologous plants, indicating a possibility for resistance gene-mediated control of Xhc. Finally, we identified regions unique to the Xhc M081 genome sequence, and demonstrated their potential in the design of molecular diagnostics for this pathogen.

  17. A multiplex real-time PCR assay for detection of Xanthomonas campestris from brassicas.

    PubMed

    Berg, T; Tesoriero, L; Hailstones, D L

    2006-06-01

    To develop a sensitive real-time PCR-based protocol for the detection of Xanthomonas campestris pathovars from Brassica seed. A 5' nuclease real-time PCR assay was developed to screen Brassica spp. seed for the presence of X. campestris pathovars that cause black rot. The assay amplifies a 78-bp segment of the X. campestris hrpF gene and a 100-bp segment of the Brassica spp. 18S-25S internal transcribed spacer region. The Brassica spp. target provides an internal control for the amplification process to prevent false negatives that may arise from inhibitors that are often present in extracts from plant material. Whilst the primers were compatible with SYBR Green I assays, the use of fluorescently labelled probes in a 5' nuclease assay afforded greatest sensitivity and specificity. Seed batches carrying one artificially infected seed among 10,000 were readily detected using the assay. The multiplex real-time PCR assay permitted the rapid detection of pathogenic strains of X. campestris from bacterial colonies, Brassica seed and plants. Strains of X. campestris pathogenic to brassicas were readily detected from seed via a multiplex 5' nuclease real-time PCR assay. The real-time assay offers an improvement in sensitivity and a reduced turn-around time over the conventional multiplex PCR. Real-time PCR can be used to rapidly screen Brassica spp. seed batches for the presence of X. campestris pathovars. This assay provides a means for growers and the seed industry to be aware of the black rot status of their planting material, so that they may more effectively employ disease control measures or seed disinfection.

  18. Grid Integrated Distributed PV (GridPV) Version 2.

    SciTech Connect

    Reno, Matthew J.; Coogan, Kyle

    2014-12-01

    This manual provides the documentation of the MATLAB toolbox of functions for using OpenDSS to simulate the impact of solar energy on the distribution system. The majority of the functio ns are useful for interfacing OpenDSS and MATLAB, and they are of generic use for commanding OpenDSS from MATLAB and retrieving information from simulations. A set of functions is also included for modeling PV plant output and setting up the PV plant in th e OpenDSS simulation. The toolbox contains functions for modeling the OpenDSS distribution feeder on satellite images with GPS coordinates. Finally, example simulations functions are included to show potential uses of the toolbox functions. Each function i n the toolbox is documented with the function use syntax, full description, function input list, function output list, example use, and example output.

  19. Lightweight IMM PV Flexible Blanket Assembly

    NASA Technical Reports Server (NTRS)

    Spence, Brian

    2015-01-01

    Deployable Space Systems (DSS) has developed an inverted metamorphic multijunction (IMM) photovoltaic (PV) integrated modular blanket assembly (IMBA) that can be rolled or z-folded. This IMM PV IMBA technology enables a revolutionary flexible PV blanket assembly that provides high specific power, exceptional stowed packaging efficiency, and high-voltage operation capability. DSS's technology also accommodates standard third-generation triple junction (ZTJ) PV device technologies to provide significantly improved performance over the current state of the art. This SBIR project demonstrated prototype, flight-like IMM PV IMBA panel assemblies specifically developed, designed, and optimized for NASA's high-voltage solar array missions.

  20. Minimal phenotypic test for simple differentiation of Xanthomonas campestris from other yellow-pigmented bacteria isolated from soil

    PubMed Central

    Soudi, MR; Alimadadi, N; Ghadam, P

    2011-01-01

    Background and Objectives Isolation of Xanthomonas campestris from soil has a wide range of applications from monitoring of phytopathogenic populations in soil to screening of improved xanthan-producing strains. Identification of Xanthomonas campestris and its pathovars requires pathogenicity tests in addition to phenotypic and molecular characterization. Materials and Methods Thirty phenotypic tests were carried out on 57 yellow-pigmented bacterial isolates obtained from soil of cabbage farms after screening on Selective Xanthomonas (SX) agar and transferring on Yeast Malt agar. Absorption spectra of pigments and capability of biopolymer production were determined for the isolates. Some characteristics of the biopolymer produced and presence of a X. campestris-specific gene marker were investigated for nine putative X. campestris isolates. Results The present study introduces a set of simple phenotypic tests including urease, acid production from sucrose, mucoid growth on 5% sucrose, starch hydrolysis, growth in 4% NaCl, motility and utilization of asparagine as sole carbon and nitrogen source for quick and inexpensive tentative identification of Xanthomonas campestris. Validation of these tests was confirmed in 100% of the cases by characterization of bacterial exopolysaccharide as xanthan and production of genus-specific xanthomonadin pigment. Moreover, tracking of hrc gene among putative X. campestris isolates gave positive results in 80% of cases. Conclusion The Minimal simple phenotypic tests facilitate the screening and differentiation of putative X. campestris isolates from other false bacterial strains isolated from soil on semiselective SX agar. PMID:22347588

  1. Minimal phenotypic test for simple differentiation of Xanthomonas campestris from other yellow-pigmented bacteria isolated from soil.

    PubMed

    Soudi, Mr; Alimadadi, N; Ghadam, P

    2011-06-01

    Isolation of Xanthomonas campestris from soil has a wide range of applications from monitoring of phytopathogenic populations in soil to screening of improved xanthan-producing strains. Identification of Xanthomonas campestris and its pathovars requires pathogenicity tests in addition to phenotypic and molecular characterization. Thirty phenotypic tests were carried out on 57 yellow-pigmented bacterial isolates obtained from soil of cabbage farms after screening on Selective Xanthomonas (SX) agar and transferring on Yeast Malt agar. Absorption spectra of pigments and capability of biopolymer production were determined for the isolates. Some characteristics of the biopolymer produced and presence of a X. campestris-specific gene marker were investigated for nine putative X. campestris isolates. The present study introduces a set of simple phenotypic tests including urease, acid production from sucrose, mucoid growth on 5% sucrose, starch hydrolysis, growth in 4% NaCl, motility and utilization of asparagine as sole carbon and nitrogen source for quick and inexpensive tentative identification of Xanthomonas campestris. Validation of these tests was confirmed in 100% of the cases by characterization of bacterial exopolysaccharide as xanthan and production of genus-specific xanthomonadin pigment. Moreover, tracking of hrc gene among putative X. campestris isolates gave positive results in 80% of cases. The Minimal simple phenotypic tests facilitate the screening and differentiation of putative X. campestris isolates from other false bacterial strains isolated from soil on semiselective SX agar.

  2. A multilocus sequence analysis of Xanthomonas campestris reveals a complex structure within crucifer-attacking pathovars of this species.

    PubMed

    Fargier, E; Fischer-Le Saux, M; Manceau, C

    2011-04-01

    Previous classification of Xanthomonas campestris has defined six pathovars (aberrans, armoraciae, barbareae, campestris, incanae, and raphani) that cause diseases on cruciferous plants. However, pathogenicity assays with a range of strains and different hosts identifies only three types of symptom: black rot, leaf spot and bacterial blight. These findings raise the question of the genetic relatedness between strains assigned to different pathovars or symptom phenotypes. Here we have addressed this issue by multilocus sequence analysis of 42 strains. The X. campestris species was polymorphic at the 8 loci analysed and had a high genetic diversity; 23 sequence types were identified of which 16 were unique. All strains that induce black rot (pathovars aberrans and campestris) were genetically close but split in two groups. Only three clonal complexes were found, all within pathovar campestris. The assignment of the genome-sequenced strain 756C to pathovar raphani suggested from disease symptoms was confirmed, although this group of strains was particularly polymorphic. Strains belonging to pathovars barbareae and incanae were closely related, but distinct from pathovar campestris. There is evidence of genetic exchanges of housekeeping genes within this species as deduced from a clear incongruence between individual gene phylogenies and from network structures from SplitsTree analysis. Overall this study showed that the high genetic diversity derived equally from recombination and point mutation accumulation. However, X. campestris remains a species with a clonal evolution driven by a differential adaptation to cruciferous hosts.

  3. Heritage Park Facilities PV Project

    SciTech Connect

    Hobaica, Mark

    2013-09-26

    Project Objective: To procure a photovoltaic array (PV) system which will generate approximately 256kW of power to be used for the operations of the Aquatic Complex and the adjacent Senior Facility at the Heritage Park. This project complies with the EERE’s work and objectives by promoting the development and deployment of an energy system that will provide current and future generations with clean, efficient, affordable, and reliable energy.

  4. Predation of Indianmeal moth larvae by Lyctocoris campestris(F.) (Hemiptera: Anthocoridae) in different stored commodities

    USDA-ARS?s Scientific Manuscript database

    Predation rates for the anthocorid predator Lyctocoris campestris (F.) against varying densities of late-instar Plodia interpunctello (Hubner) were compared in whole corn, whole wheat, or folled oat stored commodities. More prey were attacked in corn and wheat than in oats, and female predators gene...

  5. Draft Genome Sequence of the Xanthan Producer Xanthomonas campestris LMG 8031

    PubMed Central

    Huptas, Christopher; Wenning, Mareike

    2016-01-01

    Here, we report the draft genome sequence of Xanthomonas campestris LMG 8031, for which nearly no genetic information is available, despite its good xanthan-producing properties. We performed an Illumina-based sequencing approach of LMG 8031. The genome revealed a 5.0-Mb chromosome having 4,434 coding sequences and a G+C content of 65%. PMID:27789631

  6. Over-expression of miR158 causes pollen abortion in Brassica campestris ssp. chinensis.

    PubMed

    Ma, Zhiming; Jiang, Jianxia; Hu, Ziwei; Lyu, Tianqi; Yang, Yang; Jiang, Jingjing; Cao, Jiashu

    2017-02-01

    We identified and cloned the two precursors of miR158 and its target gene in Brassica campestris ssp. chinensis, which both had high relative expression in the inflorescences. Further study revealed that over-expression of miR158 caused reduced pollen varbility, which was caused by the degradation of pollen contents from the binucleate microspore stage. These results first suggest the role of miR158 in pollen development of Brassica campestris ssp. chinensis. MicroRNAs (miRNAs) play crucial roles in many important growth and development processes both in plants and animals by regulating the expression of their target genes via mRNA cleavage or translational repression. In this study, miR158, a Brassicaceae specific miRNA, was functionally characterized with regard to its role in pollen development of non-heading Chinese cabbage (Brassica campestris ssp. chinensis). Two family members of miR158 in B. campestris, namely bra-miR158a1 and bra-miR158a2, and their target gene bra027656, which encodes a pentatricopeptide repeat (PPR) containing protein, were identified. Then, qRT-PCR analysis and GUS-reporter system revealed that both bra-miR158 and its target gene had relatively high expression levels in the inflorescences. Further study revealed that over-expression of miR158 caused reduced pollen varbility and pollen germination ratio, and the degradation of pollen contents from the binucleate microspore stage was also found in those deformed pollen grains, which led to pollen shrinking and collapse in later pollen development stage. These results first shed light on the importance of miR158 in pollen development of Brassica campestris ssp. chinensis.

  7. Evaluation of the roles that alkyl hydroperoxide reductase and Ohr play in organic peroxide-induced gene expression and protection against organic peroxides in Xanthomonas campestris.

    PubMed

    Vattanaviboon, Paiboon; Whangsuk, Wirongrong; Panmanee, Warunya; Klomsiri, Chananat; Dharmsthiti, Saovanee; Mongkolsuk, Skorn

    2002-11-29

    Alkyl hydroperoxide reductase (ahpC) and organic hydroperoxide resistance (ohr) are distinct genes, structurally and regulatory, but have similar physiological functions. In Xanthomonas campestris pv. phaseoli inactivation of either gene results in increased sensitivity to killing with organic peroxides. An ahpC1-ohr double mutant was highly sensitive to both growth inhibition and killing treatment with organic peroxides. High level expression of ahpC or ohr only partially complemented the phenotype of the double mutant, suggesting that these genes function synergistically, but through different pathways, to protect Xanthomonas from organic peroxide toxicity. Functional analyses of Ohr and AhpC abilities to degrade organic hydroperoxides revealed that both Ohr and AhpC could degrade tert-butyl hydroperoxide (tBOOH) while the former was more efficient at degrading cumene hydroperoxide (CuOOH). Expression analysis of these genes in the mutants showed no compensatory alterations in the levels of AhpC or Ohr. However, CuOOH induced expression of these genes in the mutants was affected. CuOOH induced ahpC expression was higher in the ohr mutant than in the parental strain; in contrast, the ahpC mutation has no effect on the level of induced ohr expression. These analyses reveal complex physiological roles and expression patterns of seemingly functionally similar genes.

  8. PV output smoothing with energy storage.

    SciTech Connect

    Ellis, Abraham; Schoenwald, David Alan

    2012-03-01

    This report describes an algorithm, implemented in Matlab/Simulink, designed to reduce the variability of photovoltaic (PV) power output by using a battery. The purpose of the battery is to add power to the PV output (or subtract) to smooth out the high frequency components of the PV power that that occur during periods with transient cloud shadows on the PV array. The control system is challenged with the task of reducing short-term PV output variability while avoiding overworking the battery both in terms of capacity and ramp capability. The algorithm proposed by Sandia is purposely very simple to facilitate implementation in a real-time controller. The control structure has two additional inputs to which the battery can respond. For example, the battery could respond to PV variability, load variability or area control error (ACE) or a combination of the three.

  9. Ensuring Quality of PV Modules: Preprint

    SciTech Connect

    Kurtz, S.; Wohlgemuth, J.; Hacke, P.; Kempe, M.; Sample, T.; Yamamichi, M.; Kondo, M.; Doi, T.; Otani, K.; Amano, J.

    2011-07-01

    Photovoltaic (PV) customers need to have confidence in the PV modules they purchase. Currently, no test can quantify a module's lifetime with confidence, but stress tests are routinely used to differentiate PV product designs. We suggest that the industry would be strengthened by using the wisdom of the community to develop a single set of tests that will help customers quantify confidence in PV products. This paper evaluates the need for quality assurance (QA) standards and suggests a path for creating these. Two types of standards are needed: 1) QA of the module design and 2) QA of the manufacturing process.

  10. 46 CFR 153.355 - PV venting systems.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 46 Shipping 5 2011-10-01 2011-10-01 false PV venting systems. 153.355 Section 153.355 Shipping... Systems § 153.355 PV venting systems. When Table 1 requires a PV venting system, the cargo tank must have a PV valve in its vent line. The PV valve must be located between the tank and any connection to...

  11. 46 CFR 153.355 - PV venting systems.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 46 Shipping 5 2010-10-01 2010-10-01 false PV venting systems. 153.355 Section 153.355 Shipping... Systems § 153.355 PV venting systems. When Table 1 requires a PV venting system, the cargo tank must have a PV valve in its vent line. The PV valve must be located between the tank and any connection to...

  12. Distributed PV Adoption - Sensitivity to Market Factors

    SciTech Connect

    Gagnon, Pieter; Sigrin, Ben

    2016-02-01

    NREL staff used the dSolar (distributed solar) model to forecast the adoption of distributed, behind-the-meter PV through the year 2050 for 9 different scenarios. The scenarios varied in their assumptions about a carbon tax, the cost of PV systems in the future, and what credit would be given for excess generation once current net metering policies expire.

  13. PV reference cells: Calibration and stability problems

    SciTech Connect

    Koltun, M.M.

    1995-09-01

    The PV reference cells described in this paper were made and calibrated in Russia. Their parameters have also been measured in Italy, Germany and Israel, in recent years. Different calibration methods are discussed, as well as stability data of PV reference cell parameters, as measured from 1962 to date.

  14. PV integration into a CSP plant

    NASA Astrophysics Data System (ADS)

    Carvajal, Javier López; Barea, Jose M.; Barragan, Jose; Ortega, Carlos

    2017-06-01

    This paper describes a preliminary techno-economic analysis of the integration of a PV plant into an optimized Parabolic Trough Plant in order to reduce the online consumptions and thus, increase the net electricity injected into the grid. The idea is to assess the feasibility of such project and see what configuration would be the optimal. An extra effort has been made in terms of modelling as the analysis has to be done to the integrated CSP + PV plant instead of analyzing them independently. Two different technologies have been considered for the PV plant, fix and one-axis tracking. Additionally three different scenarios have been considered for the CSP plant auxiliary consumptions as they are essential for determining the optimal PV plant (the higher the auxiliary consumption the higher the optimal PV plant). As could be expected, the results for all cases with PV show an improvement in terms of electricity generation and also in terms of LCOE with respect to the CSP plant. Such improvement is slightly higher with tracking technology for this specific study. Although this exercise has been done to an already designed CSP plant (so only the PV plant had to be optimized), the methodology could be applied for the optimization of an integrated CSP + PV plant during the design phase.

  15. Requirement of the cytosolic interaction between PATHOGENESIS-RELATED PROTEIN10 and LEUCINE-RICH REPEAT PROTEIN1 for cell death and defense signaling in pepper.

    PubMed

    Choi, Du Seok; Hwang, In Sun; Hwang, Byung Kook

    2012-04-01

    Plants recruit innate immune receptors such as leucine-rich repeat (LRR) proteins to recognize pathogen attack and activate defense genes. Here, we identified the pepper (Capsicum annuum) pathogenesis-related protein10 (PR10) as a leucine-rich repeat protein1 (LRR1)-interacting partner. Bimolecular fluorescence complementation and coimmunoprecipitation assays confirmed the specific interaction between LRR1 and PR10 in planta. Avirulent Xanthomonas campestris pv vesicatoria infection induces PR10 expression associated with the hypersensitive cell death response. Transient expression of PR10 triggers hypersensitive cell death in pepper and Nicotiana benthamiana leaves, which is amplified by LRR1 coexpression as a positive regulator. LRR1 promotes the ribonuclease activity and phosphorylation of PR10, leading to enhanced cell death signaling. The LRR1-PR10 complex is formed in the cytoplasm, resulting in its secretion into the apoplastic space. Engineered nuclear confinement of both proteins revealed that the cytoplasmic localization of the PR10-LRR1 complex is essential for cell death-mediated defense signaling. PR10/LRR1 silencing in pepper compromises resistance to avirulent X. campestris pv vesicatoria infection. By contrast, PR10/LRR1 overexpression in Arabidopsis thaliana confers enhanced resistance to Pseudomonas syringae pv tomato and Hyaloperonospora arabidopsidis. Together, these results suggest that the cytosolic LRR-PR10 complex is responsible for cell death-mediated defense signaling.

  16. Requirement of the Cytosolic Interaction between PATHOGENESIS-RELATED PROTEIN10 and LEUCINE-RICH REPEAT PROTEIN1 for Cell Death and Defense Signaling in Pepper[W

    PubMed Central

    Choi, Du Seok; Hwang, In Sun; Hwang, Byung Kook

    2012-01-01

    Plants recruit innate immune receptors such as leucine-rich repeat (LRR) proteins to recognize pathogen attack and activate defense genes. Here, we identified the pepper (Capsicum annuum) pathogenesis-related protein10 (PR10) as a leucine-rich repeat protein1 (LRR1)–interacting partner. Bimolecular fluorescence complementation and coimmunoprecipitation assays confirmed the specific interaction between LRR1 and PR10 in planta. Avirulent Xanthomonas campestris pv vesicatoria infection induces PR10 expression associated with the hypersensitive cell death response. Transient expression of PR10 triggers hypersensitive cell death in pepper and Nicotiana benthamiana leaves, which is amplified by LRR1 coexpression as a positive regulator. LRR1 promotes the ribonuclease activity and phosphorylation of PR10, leading to enhanced cell death signaling. The LRR1-PR10 complex is formed in the cytoplasm, resulting in its secretion into the apoplastic space. Engineered nuclear confinement of both proteins revealed that the cytoplasmic localization of the PR10-LRR1 complex is essential for cell death–mediated defense signaling. PR10/LRR1 silencing in pepper compromises resistance to avirulent X. campestris pv vesicatoria infection. By contrast, PR10/LRR1 overexpression in Arabidopsis thaliana confers enhanced resistance to Pseudomonas syringae pv tomato and Hyaloperonospora arabidopsidis. Together, these results suggest that the cytosolic LRR-PR10 complex is responsible for cell death–mediated defense signaling. PMID:22492811

  17. The pepper E3 ubiquitin ligase RING1 gene, CaRING1, is required for cell death and the salicylic acid-dependent defense response.

    PubMed

    Lee, Dong Hyuk; Choi, Hyong Woo; Hwang, Byung Kook

    2011-08-01

    Ubiquitination is essential for ubiquitin/proteasome-mediated protein degradation in plant development and defense. Here, we identified a novel E3 ubiquitin ligase RING1 gene, CaRING1, from pepper (Capsicum annuum). In pepper, CaRING1 expression is induced by avirulent Xanthomonas campestris pv vesicatoria infection. CaRING1 contains an amino-terminal transmembrane domain and a carboxyl-terminal RING domain. In addition, it displays in vitro E3 ubiquitin ligase activity, and the RING domain is essential for E3 ubiquitin ligase activity in CaRING1. CaRING1 also localizes to the plasma membrane. In pepper plants, virus-induced gene silencing of CaRING1 confers enhanced susceptibility to avirulent X. campestris pv vesicatoria infection, which is accompanied by compromised hypersensitive cell death, reduced expression of PATHOGENESIS-RELATED1, and lowered salicylic acid levels in leaves. Transient expression of CaRING1 in pepper leaves induces cell death and the defense response that requires the E3 ubiquitin ligase activity of CaRING1. By contrast, overexpression of CaRING1 in Arabidopsis (Arabidopsis thaliana) confers enhanced resistance to hemibiotrophic Pseudomonas syringae pv tomato and biotrophic Hyaloperonospora arabidopsidis infections. Taken together, these results suggest that CaRING1 is involved in the induction of cell death and the regulation of ubiquitination during the defense response to microbial pathogens.

  18. The Pepper E3 Ubiquitin Ligase RING1 Gene, CaRING1, Is Required for Cell Death and the Salicylic Acid-Dependent Defense Response1[C][W][OA

    PubMed Central

    Lee, Dong Hyuk; Choi, Hyong Woo; Hwang, Byung Kook

    2011-01-01

    Ubiquitination is essential for ubiquitin/proteasome-mediated protein degradation in plant development and defense. Here, we identified a novel E3 ubiquitin ligase RING1 gene, CaRING1, from pepper (Capsicum annuum). In pepper, CaRING1 expression is induced by avirulent Xanthomonas campestris pv vesicatoria infection. CaRING1 contains an amino-terminal transmembrane domain and a carboxyl-terminal RING domain. In addition, it displays in vitro E3 ubiquitin ligase activity, and the RING domain is essential for E3 ubiquitin ligase activity in CaRING1. CaRING1 also localizes to the plasma membrane. In pepper plants, virus-induced gene silencing of CaRING1 confers enhanced susceptibility to avirulent X. campestris pv vesicatoria infection, which is accompanied by compromised hypersensitive cell death, reduced expression of PATHOGENESIS-RELATED1, and lowered salicylic acid levels in leaves. Transient expression of CaRING1 in pepper leaves induces cell death and the defense response that requires the E3 ubiquitin ligase activity of CaRING1. By contrast, overexpression of CaRING1 in Arabidopsis (Arabidopsis thaliana) confers enhanced resistance to hemibiotrophic Pseudomonas syringae pv tomato and biotrophic Hyaloperonospora arabidopsidis infections. Taken together, these results suggest that CaRING1 is involved in the induction of cell death and the regulation of ubiquitination during the defense response to microbial pathogens. PMID:21628629

  19. [Diffusible signal factor production and virulence expression in deltarpfFxoo, deltarpfCxoo and deltarpfGxoo, the gene deletion mutants of DSF/Rpf signaling proteins of Xanthomonas oryzae pv. oryzae].

    PubMed

    Sun, Lei; Wu, Maosen; Chen, Huamin; He, Chenyang

    2010-06-01

    To better elucidate the functions of RpfFxoo, RpfCxoo and RpfGxoo, 3 proteins of diffusible signal factor (DSF)-dependent cell-cell signaling system in regulation of virulence expression of Xanthomonas oryzae pv. oryzae (Xoo). Deltarpfxoo, the gene deletion mutants were generated from PXO99(A), the wild-type strain of Xoo via marker-exchanging and DSF biosynthesis and extracellular polysaccharide production and virulence to rice of the mutants were assayed. rpfFxoo,rpfCxoo and rpfGxoo were cloned from the genomic DNA of PXO99(A) and the relative single or double mutants were constructed. Compared to PXO99(A), DSF production was deficient in deltarpfFxoo, deltarpfFCxoo and deltarpfFGxoo, while DSF was overproduced in deltarpfCxoo and reduced in deltarpfGxoo. DSF production of deltarpfFxcc, deltarpfCxcc and deltarpfGxcc, the mutants of X. campestris pv. campestris can be restored as XC1, the wild-type strain by in trans complementation of rpfFxoo, rpfCxoo and rpfGxoo. All the mutants except deltarpfFxoo were remarkably deficient in production All the mutants significantly exhibited the reduced bacterial virulence to rice. of extracellular polysaccharide. DSF signaling proteins RpfFxoo, RpfCxoo and RpfGxoo might function in regulation of DSF biogenesis and EPS production and bacterial virulence.

  20. Variation suggestive of horizontal gene transfer at a lipopolysaccharide (lps) biosynthetic locus in Xanthomonas oryzae pv. oryzae, the bacterial leaf blight pathogen of rice

    PubMed Central

    Patil, Prabhu B; Sonti, Ramesh V

    2004-01-01

    Background In animal pathogenic bacteria, horizontal gene transfer events (HGT) have been frequently observed in genomic regions that encode functions involved in biosynthesis of the outer membrane located lipopolysaccharide (LPS). As a result, different strains of the same pathogen can have substantially different lps biosynthetic gene clusters. Since LPS is highly antigenic, the variation at lps loci is attributed to be of advantage in evading the host immune system. Although LPS has been suggested as a potentiator of plant defense responses, interstrain variation at lps biosynthetic gene clusters has not been reported for any plant pathogenic bacterium. Results We report here the complete sequence of a 12.2 kb virulence locus of Xanthomonas oryzae pv. oryzae (Xoo) encoding six genes whose products are homologous to functions involved in LPS biosynthesis and transport. All six open reading frames (ORFs) have atypical G+C content and altered codon usage, which are the hallmarks of genomic islands that are acquired by horizontal gene transfer. The lps locus is flanked by highly conserved genes, metB and etfA, respectively encoding cystathionine gamma lyase and electron transport flavoprotein. Interestingly, two different sets of lps genes are present at this locus in the plant pathogens, Xanthomonas campestris pv. campestris (Xcc) and Xanthomonas axonopodis pv. citri (Xac). The genomic island is present in a number of Xoo strains from India and other Asian countries but is not present in two strains, one from India (BXO8) and another from Nepal (Nepal624) as well as the closely related rice pathogen, Xanthomonas oryzae pv. oryzicola (Xoor). TAIL-PCR analysis indicates that sequences related to Xac are present at the lps locus in both BXO8 and Nepal624. The Xoor strain has a hybrid lps gene cluster, with sequences at the metB and etfA ends, being most closely related to sequences from Xac and the tomato pathogen, Pseudomonas syringae pv. tomato respectively

  1. Small scale production and characterization of xanthan gum synthesized by local isolates of Xanthomonas campestris.

    PubMed

    Barua, Rajesh; Alam, Md Jahangir; Salim, Mohammad; Ashrafee, Tamzida Shamim

    2016-02-01

    Xanthan gum is a commercially important microbial exopolysaccharide (EPS) produced by Xanthomonas campestris. X. campestris is a plant pathogen causing various plant diseases such as black rot of crucifers, bacterial leaf blight and citrus canker disease resulting in crop damage. In this study, we isolated efficient local bacterial isolates which are capable to produce xanthan gum utilizing different sources of carbon (maltose, sucrose and glucose). Bacterial isolates from different plant leaves and fruits were identified as Xanthomonas campestris based on their morphological and biochemical characteristics. Among the 23 isolates, 70% were capable of producing gum. Taro plant, considered as new bacterial host, also have the capability to produce xanthan gum. Production conditions of xanthan gum and their relative viscosity by these bacterial isolates were optimized using basal medium containing commercial carbon and nitrogen sources and various temperature and rotation. Highest level of xanthan gum (18.286 g/l) with relative viscosity (7.2) was produced (Host, Citrus macroptera) at 28 degrees C, pH 7.0, 150 rpm using sucrose as a carbon source at orbital shaker. Whereas, in lab fermenter, same conditions gave best result (19.587 g/l gum) with 7.8 relative viscosity. Chilled alcohol (96%) was used to recover the xanthan gum. FTIR studies also carried out for further confirmation of compatibility by detecting the chemical groups.

  2. Clustering of mutations blocking synthesis of xanthan gum by Xanthomonas campestris.

    PubMed Central

    Thorne, L; Tansey, L; Pollock, T J

    1987-01-01

    Mutations that block the synthesis of xanthan gum by Xanthomonas campestris B1459S-4L-II were isolated as nonmucoid colonies after treatment with ethyl methanesulfonate. Complete libraries of DNA fragments from wild-type X. campestris were cloned into Escherichia coli by using a broad-host-range cosmid vector and then transferred into each mutant strain by conjugal mating. Cloned fragments that restored xanthan gum synthesis (Xgs+; mucoidy) were compared according to restriction pattern, DNA sequence homology, and complementation of a subset of Xgs- mutations. Groups of clones that contained overlapping homologous DNA were found to complement specific Xgs- mutations. The results suggest clustering of the genetic loci involved in xanthan synthesis. The clustering occurred within three unlinked regions. Two forms of complementation were observed. In most instances, independently isolated cosmid clones that complemented a single mutation were found to be partially homologous. Less frequent was the second form of complementation, in which two cosmid clones that lacked any homologous sequences restored the mucoid phenotype to a single mutant. Finally, xanthan production was measured for wild-type X. campestris carrying multiple plasmid copies of the cloned xanthan genes. Images PMID:3038845

  3. Location and cloning of the ketal pyruvate transferase gene of Xanthomonas campestris.

    PubMed Central

    Marzocca, M P; Harding, N E; Petroni, E A; Cleary, J M; Ielpi, L

    1991-01-01

    Genes required for xanthan polysaccharide synthesis (xps) are clustered in a DNA region of 13.5 kb in the chromosome of Xanthomonas campestris. Plasmid pCHC3 containing a 12.4-kb insert of xps genes has been suggested to include a gene involved in the pyruvylation of xanthan gum (N.E. Harding, J.M. Cleary, D.K. Cabañas, I. G. Rosen, and K. S. Kang, J. Bacteriol. 169:2854-2861, 1987). An essential step toward understanding the biosynthesis of xanthan gum and to enable genetic manipulation of xanthan structure is the determination of the biochemical function encoded by the xps genes. On the basis of biochemical characterization of an X. campestris mutant which produces pyruvate-free xanthan gum, complementation studies, and heterologous expression, we have identified the gene coding for the ketal pyruvate transferase (kpt) enzyme. This gene was located on a 1.4-kb BamHI fragment of pCHC3 and cloned in the broad-host-range cloning vector pRK404. An X. campestris kpt mutant was constructed by mini-Mu(Tetr) mutagenesis of the cloned gene and then by recombination of the mutation into the chromosome of the wild-type strain. PMID:1657892

  4. Low-Pressure Plasma Application for the Inactivation of the Seed-borne Pathogen Xanthomonas campestris.

    PubMed

    Nishioka, Terumi; Takai, Yuichiro; Mishima, Tomoko; Kawaradani, Mitsuo; Tanimoto, Hideo; Okada, Kiyotsugu; Misawa, Tatsuya; Kusakari, Shinichi

    2016-01-01

    The aim of this study was to investigate the effect of low-pressure plasma treatment on seed disinfection and the possible mechanisms underlying this effect. Seed-borne disease refers to plant diseases that are transmitted by seeds; seed disinfection is an important technique for prevention of such diseases. In this study, the effectiveness of low-pressure plasma treatment in the inactivation of the seed-borne plant pathogenic bacterium, Xanthomonas campestris, inoculated on cruciferous seeds, was evaluated. The highest inactivation effect was observed when the treatment voltage and argon gas flow rate were 5.5 kV and 0.5 L/min, respectively. The viable cell number of X. campestris was 6.6 log cfu/seed before plasma treatment, and decreased by 3.9 log after 5 min of treatment and by 6.6 log after 40 min. Ethidium monoazide treatment and quantitative real-time PCR results indicated that both the cell membrane and target DNA region were damaged following 5 min of plasma treatment. Although both heat and ozone were generated during the plasma treatment, the contribution of both factors to the inactivation of X. campestris was small by itself in our low-pressure plasma system. Overall, we have shown that our low-pressure plasma system has great applicability to controlling plant pathogenic bacterium contamination of seeds.

  5. GsmR, a response regulator with an HD-related output domain in Xanthomonas campestris, is positively controlled by Clp and is involved in the expression of genes responsible for flagellum synthesis.

    PubMed

    Liu, Yu-Fan; Liao, Chao-Tsai; Song, Wan-Ling; Hsu, Pei-Chi; Du, Shin-Chiao; Lo, Hsueh-Hsia; Hsiao, Yi-Min

    2013-01-01

    In prokaryotes, two-component signal transduction systems, consisting of a histidine kinase and a response regulator, play a critical role in regulating a range of cellular functions. A recent study suggests that XCC3315, a response regulator with a CheY-like receiver domain attached to an uncharacterized HD-related output domain (HDOD domain), plays a role in the general stress response of the Gram-negative bacterium Xanthomonas campestris pv. campestris (Xcc), the causal agent of black rot in cruciferous plants. Here, we demonstrated genetically that XCC3315, designated as gsmR (general stress and motility regulator), is involved in the expression of genes responsible for flagellum synthesis, including rpoN2, flhF, flhB, and fliC. Site-directed mutagenesis revealed that Glu9 and Arg100 in the receiver domain and Gly205, Asp263, His287, Trp298 and His311 in the HDOD are critical amino acids for GsmR function in cell motility regulation. The gsmR transcription initiation site was mapped. Promoter analysis and gel retardation assay revealed that the expression of gsmR is positively controlled by the global transcriptional regulator Clp in a direct manner, and is subject to catabolite repression. Our findings not only extend the previous work on Clp regulation to show that it influences the expression of gsmR in Xcc, but are also the first to characterize the expression of this response regulator gene in this phytopathogen. Furthermore, GsmR is the first HDOD-containing protein of bacteria in which key amino acids have been experimentally identified and characterized.

  6. Terawatt Challenge for Thin-Film PV

    SciTech Connect

    Zweibel, K.

    2005-08-01

    The evolution of PV into one of the world's largest industries is not going to happen without major unforeseen problems. However, this study attempts to address the obvious ones, so that we can put aside the mythology of PV (for example, that it is only ''boutique power'' or that one must pave the world with it to be useful) and get on with changing the world's energy infrastructure. With the years of rapid market growth now under way in PV, the author is sure this will not be the last effort to understand the real potential and pitfalls of meeting the Challenge.

  7. A new flexible a-Si PV module and its application to rooftop PV systems

    SciTech Connect

    Ichikawa, Yukimi; Ihara, Takuro; Hama, Toshio

    1994-12-31

    A novel photovoltaic (PV) module for roof top systems, solar roofing, was proposed. Solar roofing is a flexible amorphous silicon (a-Si) PV sheet having the function of roofing. Tempered glass is used as roof covering material. Technical items for roof top systems using solar roofing were discussed. Preliminary studies on module component materials showed feasibility of solar roofing. The authors designed a construction technology for tempered glass covered roofs. Effects of shadows on PV module upon its performance were also analyzed.

  8. Rating PV Power and Energy: Cell, Module, and System Measurements

    SciTech Connect

    Emery, Keith

    2016-06-02

    A summary of key points related to research-level measurements of current vs. voltage measurement theory including basic PV operation, equivalent circuit, and concept of spectral error; PV power performance including PV irradiance sensors, simulators and commercial and generic I-V systems; PV measurement artifacts, intercomparisons, and alternative rating methods.

  9. PV Module Reliability Research (Fact Sheet)

    SciTech Connect

    Not Available

    2013-06-01

    This National Center for Photovoltaics sheet describes the capabilities of its PV module reliability research. The scope and core competencies and capabilities are discussed and recent publications are listed.

  10. Distributed PV Adoption in Maine Through 2021

    SciTech Connect

    Gagnon, Pieter; Sigrin, Ben

    2015-11-06

    NREL has used its dSolar (distributed solar) model to generate low-medium-high estimates of distributed PV adoption in Maine through 2021. This presentation gives a high-level overview of the model and modeling results.

  11. PV System Energy Evaluation Method (Presentation)

    SciTech Connect

    Kurtz, S.

    2014-01-01

    This presentation describes a comparison of the "predicted" energy (based on historical weather data) with the "expected" energy (based on the measured weather data) to determine whether a PV system is performing as modeled in order to verify the accuracy of a model. A key factor in defining this energy test is determining the test boundary so that weather variations are not inadvertently included in what is considered to be PV system performance.

  12. Updating Technical Screens for PV Interconnection: Preprint

    SciTech Connect

    Coddington, M.; Ellis, A.; Lynn, K.; Razon, A.; Key, T.; Kroposki, B.; Mather, B.; Hill, R.; Nicole, K.; Smith, J.

    2012-08-01

    Solar photovoltaics (PV) is the dominant type of distributed generation (DG) technology interconnected to electric distribution systems in the United States, and deployment of PV systems continues to increase rapidly. Considering the rapid growth and widespread deployment of PV systems in United States electric distribution grids, it is important that interconnection procedures be as streamlined as possible to avoid unnecessary interconnection studies, costs, and delays. Because many PV interconnection applications involve high penetration scenarios, the process needs to allow for a sufficiently rigorous technical evaluation to identify and address possible system impacts. Existing interconnection procedures are designed to balance the need for efficiency and technical rigor for all DG. However, there is an implicit expectation that those procedures will be updated over time in order to remain relevant with respect to evolving standards, technology, and practical experience. Modifications to interconnection screens and procedures must focus on maintaining or improving safety and reliability, as well as accurately allocating costs and improving expediency of the interconnection process. This paper evaluates the origins and usefulness of the capacity penetration screen, offers potential short-term solutions which could effectively allow fast-track interconnection to many PV system applications, and considers longer-term solutions for increasing PV deployment levels in a safe and reliable manner while reducing or eliminating the emphasis on the penetration screen.

  13. Antiulcerogenic Activity of the Hydroalcoholic Extract of Leaves of Croton campestris A. St.-Hill in Rodents

    PubMed Central

    Júnior, Francisco E. B.; de Oliveira, Dayanne R.; Bento, Elizângela B.; Leite, Laura H. I.; Souza, Daniele O.; Siebra, Ana Luiza A.; Sampaio, Renata S.; Martins, Anita O. P. B.; Ramos, Andreza G. B.; Tintino, Saulo R.; Lacerda-Neto, Luiz J.; Figueiredo, Patricia R. L.; Oliveira, Larissa R.; Rodrigues, Cristina K. S.; Sales, Valterlúcio S.; Figueiredo, Francisco R. S. D. N.; Nascimento, Emmily P.; Monteiro, Alefe B.; Amaro, Érika N.; Costa, José G. M.; Douglas Melo Coutinho, Henrique; de Menezes, Irwin R. A.; Kerntopf, Marta R.

    2013-01-01

    Croton campestris A. St.-Hill., popularly known as “velame do campo,” is a species native to the savannah area of Northeast Brazil, which is used by traditional communities in folk medicine for variety of health problems, especially detoxification, inflammation, and gastritis. The hydroalcoholic extract of C. campestris leaves (HELCC) was assessed for its antiulcerogenic effect in gastric lesion models and effect on intestinal motility in mice, and possible mechanisms of action were examined. HELCC showed significant gastroprotective action in all models of gastric ulcer evaluated; the results suggest that this action probably involves the nitric oxide pathway. HELCC did not show alteration of intestinal motility in mice. It was also found that C. campestris represents a promising natural source with important biological potential, justifying some of its uses in folk medicine. PMID:23864894

  14. [Role of the surface and extracellular substances of the phytopathogenic bacterium Xanthomonas campestris in its interactions with cabbage plants].

    PubMed

    Stadnik, G I; Kalashnikova, E E; Konnova, S A; Ignatov, V V

    2001-01-01

    Changes in some physiological and biochemical characteristics of cabbage (cv. Slava) seedling roots in response to inoculation with the phytopathogen Xanthomonas campestris and its surface and extracellular substances were evaluated. Seven days after the inoculation, the growth of the roots was slightly suppressed and they contained increased amounts of peroxidase. The effect of the lipopolysaccharides stripped from the cell surface or isolated from the culture liquid of X. campestris was similar to that of the whole cells of the phytopathogen. The bacterial lectin isolated from the cell surface material did not induce any defense response in cabbage plants but, presumably, could play a role in the contact interactions between bacteria and plants.

  15. Significant alterations in anisotropic ice growth rate induced by the ice nucleation-active bacteria Xanthomonas campestris

    NASA Astrophysics Data System (ADS)

    Nada, Hiroki; Zepeda, Salvador; Miura, Hitoshi; Furukawa, Yoshinori

    2010-09-01

    In the present study, we found that the ice nucleation-active bacteria Xanthomonas campestris significantly altered anisotropic ice growth rate. Results of ice growth experiments in the presence of X. campestris showed that this bacterium decreased the ice crystal growth rate in the c-axis, whereas it increased growth rates in directions normal to the c-axis. These results indicate that these alterations in anisotropic growth rate are the result of selective binding of bacterial ice-nucleating proteins along the {0 0 0 1} basal plane.

  16. PilR enhances the sensitivity of Xanthomonas axonopodis pv. citri to the infection of filamentous bacteriophage Cf.

    PubMed

    Yang, Yen-Chun; Chou, Chun-Ping; Kuo, Tsong-Teh; Lin, Shan-Hwa; Yang, Mei-Kwei

    2004-04-01

    The pilA gene, which encodes the major structure of pili, is required for infection of Xanthomonas axonopodis pv. citri ( X. a. pv. citri) by the filamentous bacteriophage Cf. Two open reading frames (ORFs) located downstream of pilA were cloned and characterized. One 1392-bp ORF encodes a protein of 464 amino acids which shares substantial similarity with pilR of other bacterial species; the second ORF ( orf618), of 1854-bp, shares sequence similarity with pilS. The existence of the pilR-like and pilS-like genes in various X. campestris pathovars indicated that these two genes are well conserved in Xanthomonas. pilR and pilS mutants were constructed by gene replacement. We found that a pilR mutant, resistant to the infection of phage Cf, was unable to synthesize PilA protein; however, the abundance of the PilA protein and of the pilA transcript was markedly increased by the introduction of a plasmid containing the cloned pilR gene. The restoration of the normal pilus-specific sensitivity of this transformed clone to Cf indicated that the pilR gene functions as a transcriptional regulator of pilA. The pilS mutant, however, was susceptible to Cf infection, and the level of pilA expression in this mutant was similar to that of wild-type cells. Promoter analysis of luciferase reporter gene constructs containing the 5' untranslated regions of pilR or pilS genes revealed that, although the pilR and pilS are contiguous in X. a. pv. citri, the two genes are expressed independently, and the strong pilR promoter leads to the accumulation of PilR in X. a. pv. citri, which positively regulates the biosynthesis of PilA. These results revealed the enhanced sensitivity of X. a. pv. citri to phage Cf in the presence of PilR and indicated that the filamentous phage Cf utilize bacterial pili as a receptor site for its infection.

  17. Winter warming facilitates range expansion: cold tolerance of the butterfly Atalopedes campestris.

    PubMed

    Crozier, Lisa

    2003-05-01

    Our ability to predict ecological and evolutionary responses to climate change requires an understanding of the mechanistic links between climate and range limits. The warming trend over the past half-century has generated numerous opportunities to develop much-needed case studies of these links. Species that are only limited by climatic factors are likely to shift range quickly during periods of warming. Such species directly impact recipient communities and indicate trends that will become more widespread. Because minimum temperature (T (min)) is rising at twice the rate of maximum temperature, species with this range-limiting factor may be especially responsive to global warming. In this study, I test the hypothesis that rising T (min) has directly affected the range of a skipper butterfly. Atalopedes campestris has moved northward rapidly this century, recently colonizing eastern Washington where January T (min) has risen 3 degrees C in 50 years. The results show that: 1. A. campestris' range lies completely within the -4 degrees C January average minimum isotherm, and that recently colonized areas were below this threshold earlier this century. 2. In acute cold stress experiments, -4 to -7 degrees C proved to be a critical thermal limit: median supercooling point was -6.3 degrees C, and minimum lethal temperature (LT50 with 12-h exposure) was -5.7 degrees C. 3. In chronic cold stress experiments, survivorship declined sharply in diurnally fluctuating thermal regimes typical of the current range edge. High mortality occurred under constant 0 degrees C conditions as well as in fluctuating regimes, implying that thermal insulation from snow would not protect A. campestris. 4. There was no evidence of evolution in cold tolerance at the range margin, despite strong selection. Thus, winter warming was apparently a prerequisite for the range expansion. Characteristics of this species that seem to be associated with its rapid response are that it is an opportunistic

  18. Induction of extracellular matrix glycoproteins in Brassica petioles by wounding and in response to Xanthomonas campestris.

    PubMed

    Davies, H A; Daniels, M J; Dow, J M

    1997-09-01

    A panel of monoclonal antibodies that recognize plant extracellular matrix glycoproteins previously implicated in plant-microbe interactions was used to study the effects of pathogen inoculation and wounding on glycoproteins in petioles of Brassica campestris. The panel of monoclonals comprised two sets: JIM11, JIM12, and JIM20 recognize epitopes carried on hydroxyproline-rich glycoproteins (HRGPs) (M. Smallwood, A. Beven, N. Donovan, S. J. Neitl, J. Peart, K. Roberts, and J. P. Knox, Plant J. 5:237-246, 1994); MAC204 and MAC265 recognize glycoproteins of the Rhizobium infection thread (K. A. VandenBosch, D. J. Bradley, S. Perotto, G. W. Butcher, and N. J. Brewin, EMBO J. 8:335-342, 1989). Wounding or inoculation of petioles with avirulent strains of pathovars of Xanthomonas campestris induced the synthesis of two new groups of antigens: gp160 ran as a smear on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) with apparent molecular mass from 120 to 200 kDa and was recognized by JIM20 and MAC204; gpS remained in the stacking gel on SDS-PAGE and was recognized by JIM11, JIM20, and MAC204. The response to virulent strains of pathovars of X. campestris was either less pronounced or absent. gpS comprised several components that were resolved by cation-exchange chromatography. Some of these components were characterized as extensin-like HRGPs. The level of induction of the gpS group of antigens by virulent strains was not altered by mutation of a number of genes required for basic pathogenicity or by heat-killing the bacteria.

  19. Antihypertensive and vasorelaxant effects of aqueous extract of Artemisia campestris L. from Eastern Morocco.

    PubMed

    Dib, Ikram; Tits, Monique; Angenot, Luc; Wauters, Jean Noel; Assaidi, Asmae; Mekhfi, Hassane; Aziz, Mohammed; Bnouham, Mohammed; Legssyer, Abdelkhaleq; Frederich, Michel; Ziyyat, Abderrahim

    2017-07-12

    Artemisia campestris L. (Asteraceae) has many traditional uses, among which treatment of diabetes and hypertension. This study was conducted in order to confirm the antihypertensive and hypotensive effects of A. campestris L. aqueous extract (AcAE) and to explore the underlying mechanism of action of its vasorelaxant effect, besides the acute toxicity. Also, the chemical composition of AcAE was investigated. the chemical content of AcAE was determined by using HPLC and NMR techniques. The antihypertensive effect was assessed indirectly by tail-cuff method on L-NAME induced hypertensive rats, while the hypotensive action was monitored intravenously by invasive method on normotensive rats. The vasorelaxant effect and vascular mechanism of action were studied in the presence of antagonists and blockers on aorta isolated from normotensive rats. On the other side, the acute toxicity was studied by oral feeding of extract to the mice. The global phytochemical profile of AcAE reveals the presence of several polyphenols as main components. A. campestris L. infusion was characterized by mono- and di-cinnamoyl compounds, with 3,5-dicaffeoylquinic (isochlorogenic A) acid being the main compound, followed by 5-caffeoylquinic (chlorogenic) acid. Vicenin-2 (apigenin 6,8-di-C-glucoside) appeared to be the most abundant compound among flavonoids. The daily treatment with AcAE at 150mg/kg/day prevented the installation of hypertension on L-NAME hypertensive rats, and reduced SBP from 172mmHg up to 144mmHg. At the dose 40mg/kg, AcAE provoked reduction of systolic (SBP), diastolic (DBP) and mean arterial pressure (MAP), without affecting the heart rate. Also, AcAE (10(-2)-2mg/ml) relaxed the precontracted aorta by 95.8±1.3%. The denudation and preincubation of aorta with atropine, calmidazolium, L-NAME, hydroxycobalamin, ODQ, 8-RP-Br-PET-cGMP, thapsigargin and verapamil attenuated the vasorelaxant response, while the pre-treatment with 4-AP, TEA, glibenclamide and BaCl2 did not

  20. Maintenance procedures for the curtailment of genetic instability: Xanthomonas campestris NRRL B-1459.

    PubMed Central

    Kidby, D; Sandford, P; Herman, A; Cadmus, M

    1977-01-01

    Characteristics are described of small-colony variants of Xanthomonas campestris NRRL B-1459 which are frequently encountered when routine culture maintenance procedures are employed. In contrast to the parental type, smallcolony variants were shown to be resistant to a number of antibiotics, to acridine orange, and to phage which are virulent for the parent colony type. Sensitivity to ultraviolet radiation was similar in both colony types. A simple method for preservation of viable cells is described. The suitability of the method for providing reproducible inocula free from variant cell types is examined. PMID:326188

  1. Production of xanthan gum by Sphingomonas bacteria carrying genes from Xanthomonas campestris.

    PubMed

    Pollock, T J; Mikolajczak, M; Yamazaki, M; Thorne, L; Armentrout, R W

    1997-08-01

    Twelve genes coding for assembly, acetylation, pyruvylation, polymerization, and secretion of the polysaccharide xanthan gum are clustered together on the chromosome of the bacterium Xanthomonas campestris. These genes (gumBCDEFGHIJKLM) are sufficient for synthesis of xanthan gum when placed in bacteria from a different genus, Sphingomonas. The polysaccharide from the recombinant microorganism is largely indistinguishable, structurally and functionally, from native xanthan gum. These results demonstrate that a complex pathway for biosynthesis of a specific polysaccharide can be acquired by a single inter-generic transfer of genes between bacteria. This suggests the biological and commercial feasibility of synthesizing xanthan gum or other polysaccharides in non-native hosts.

  2. Pepper gene encoding a basic class II chitinase is inducible by pathogen and ethephon.

    PubMed

    Hong; Jung; Kim; Hwang

    2000-10-16

    A chitinase cDNA clone (designated CAChi2) was isolated from the cDNA library of pepper leaves infected with Xanthomonas campestris pv. vesicatoria. The 1004-bp full-length CAChi2 cDNA encodes a basic chitinase with an N-terminal 24 amino acid signal peptide followed by a catalytic region. An analysis of its sequence indicates that CAChi2 is a class II chitinase, because it does not have chitin-binding domain and C-terminal extension sequences. The deduced amino acid sequence of CAChi2 has a high level of identity with class II chitinases from potato, tomato, tobacco and petunia. Southern analysis demonstrated that the CAChi2 chitinase is encoded by a single or two copy genes in the pepper genome. Following X. campestris pv. vesicatoria or Phytophthora capsici infection, the CAChi2 chitinase mRNA was more highly expressed in the incompatible interaction, compared to expression in the compatible interaction. Treatment with ethylene-releasing ethephon resulted in a strong accumulation of the transcripts in the leaves. In contrast, DL-beta-amino-n-butyric acid, salicylic acid and methyl jasmonate were not effective in inducing CAChi2 transcripts in pepper leaves.

  3. Identification of a novel pathogen-induced gene encoding a leucine-rich repeat protein expressed in phloem cells of Capsicum annuum.

    PubMed

    Jung, Eui Hwan; Jung, Ho Won; Lee, Sung Chul; Han, Sang Wook; Heu, Sunggi; Hwang, Byung Kook

    2004-02-20

    The CALRR1 gene, expressed in pepper leaves following infection by Xanthomonas campestris pv. vesicatoria, encodes a secreted leucine-rich repeat (LRR) with five tandem repeats of a 24-amino-acid LRR motif. Northern blot analyses revealed that CALRR1 is not constitutively expressed in pepper plants, but is strongly induced upon the infection by X. campestris pv. vesicatoria, Phytophthora capsici, Colletotrichum coccodes and Colletotrichum gloeosporioides on leaves. CALRR1 was not systemically induced in upper leaves by bacterial infection. The inoculation of bacterial live cells, and treatment with dead cells and culture filtrates of pathogenic or nonpathogenic bacteria triggered the accumulation of CALRR1 transcripts. Treatment with signaling molecules, including salicylic acid (SA), ethylene (ET), methyl jasmonate (MeJA), dl-beta-amino-n-butyric acid (BABA) and benzothiadiazole (BTH), did not activate the transcription of the CALRR1 gene, indicating that CALRR1 expression is not regulated by defense signaling pathways activated by these molecules. CALRR1 was induced by treatment with high salinity, abscisic acid (ABA) and wounding, but not by drought and cold stress. An in situ hybridization study showed that CALRR1 mRNA was localized in phloem tissues of leaves, stems, and green fruits of pepper plants during the pathogen infection and ABA exposure. The location characteristics and the spatio-temporal expression pattern of CALRR1 suggest that it may play a role in protecting phloem cells against biotic and abiotic stresses affecting phloem function.

  4. Defense response of a pepper cultivar cv. Sy-2 is induced at temperatures below 24°C.

    PubMed

    Koeda, Sota; Hosokawa, Munetaka; Kang, Byoung-Cheorl; Tanaka, Chihiro; Choi, Doil; Sano, Satoshi; Shiina, Takashi; Doi, Motoaki; Yazawa, Susumu

    2012-01-01

    Temperature is one of the most important environmental factors that influence plant growth and development. Recent studies imply that plants show various responses to non-extreme ambient temperatures. Previously, we have found that a pepper cultivar cv. Sy-2 (Capsicum chinense) shows developmental defects at temperatures below 24°C. In this study, to gain new insights into the temperature sensitivity of cv. Sy-2, temperature-sensitive genes were screened using microarray techniques. At restrictive temperature of 20°C, almost one-fourth of the 411 up-regulated genes were defense related or predicted to be defense related. Further expression analyses of several defense-related genes showed that defense-related genes in cv. Sy-2 were constitutively expressed at temperatures below 24°C. Moreover, accumulation of high level of salicylic acid (SA) in cv. Sy-2 grown at 20°C suggests that the defense response is activated in the absence of pathogens. To confirm that the defense response is induced in cv. Sy-2 below 24°C, we evaluated the resistance to biotrophic bacterial pathogen Xanthomonas campestris pv. vesicatoria and necrotrophic fungal pathogen Cercospora capsici. Cv. Sy-2 showed enhanced resistance to X. campestris pv. vesicatoria, but not to C. capsici.

  5. The Pepper Extracellular Xyloglucan-Specific Endo-β-1,4-Glucanase Inhibitor Protein Gene, CaXEGIP1, Is Required for Plant Cell Death and Defense Responses1[C][W][OA

    PubMed Central

    Choi, Hyong Woo; Kim, Nak Hyun; Lee, Yeon Kyeong; Hwang, Byung Kook

    2013-01-01

    Plants produce various proteinaceous inhibitors to protect themselves against microbial pathogen attack. A xyloglucan-specific endo-β-1,4-glucanase inhibitor1 gene, CaXEGIP1, was isolated and functionally characterized in pepper (Capsicum annuum) plants. CaXEGIP1 was rapidly and strongly induced in pepper leaves infected with avirulent Xanthomonas campestris pv vesicatoria, and purified CaXEGIP1 protein significantly inhibited the hydrolytic activity of the glycoside hydrolase74 family xyloglucan-specific endo-β-1,4-glucanase from Clostridium thermocellum. Soluble-modified green fluorescent protein-tagged CaXEGIP1 proteins were mainly localized to the apoplast of onion (Allium cepa) epidermal cells. Agrobacterium tumefaciens-mediated overexpression of CaXEGIP1 triggered pathogen-independent, spontaneous cell death in pepper and Nicotiana benthamiana leaves. CaXEGIP1 silencing in pepper conferred enhanced susceptibility to virulent and avirulent X. campestris pv vesicatoria, accompanied by a compromised hypersensitive response and lowered expression of defense-related genes. Overexpression of dexamethasone:CaXEGIP1 in Arabidopsis (Arabidopsis thaliana) enhanced resistance to Hyaloperonospora arabidopsidis infection. Comparative histochemical and proteomic analyses revealed that CaXEGIP1 overexpression induced a spontaneous cell death response and also increased the expression of some defense-related proteins in transgenic Arabidopsis leaves. This response was also accompanied by cell wall thickening and darkening. Together, these results suggest that pathogen-inducible CaXEGIP1 positively regulates cell death-mediated defense responses in plants. PMID:23093361

  6. PV performance modeling workshop summary report.

    SciTech Connect

    Stein, Joshua S.; Tasca, Coryne Adelle; Cameron, Christopher P.

    2011-05-01

    During the development of a solar photovoltaic (PV) energy project, predicting expected energy production from a system is a key part of understanding system value. System energy production is a function of the system design and location, the mounting configuration, the power conversion system, and the module technology, as well as the solar resource. Even if all other variables are held constant, annual energy yield (kWh/kWp) will vary among module technologies because of differences in response to low-light levels and temperature. A number of PV system performance models have been developed and are in use, but little has been published on validation of these models or the accuracy and uncertainty of their output. With support from the U.S. Department of Energy's Solar Energy Technologies Program, Sandia National Laboratories organized a PV Performance Modeling Workshop in Albuquerque, New Mexico, September 22-23, 2010. The workshop was intended to address the current state of PV system models, develop a path forward for establishing best practices on PV system performance modeling, and set the stage for standardization of testing and validation procedures for models and input parameters. This report summarizes discussions and presentations from the workshop, as well as examines opportunities for collaborative efforts to develop objective comparisons between models and across sites and applications.

  7. DOE High Performance Concentrator PV Project

    SciTech Connect

    McConnell, R.; Symko-Davies, M.

    2005-08-01

    Much in demand are next-generation photovoltaic (PV) technologies that can be used economically to make a large-scale impact on world electricity production. The U.S. Department of Energy (DOE) initiated the High-Performance Photovoltaic (HiPerf PV) Project to substantially increase the viability of PV for cost-competitive applications so that PV can contribute significantly to both our energy supply and environment. To accomplish such results, the National Center for Photovoltaics (NCPV) directs in-house and subcontracted research in high-performance polycrystalline thin-film and multijunction concentrator devices with the goal of enabling progress of high-efficiency technologies toward commercial-prototype products. We will describe the details of the subcontractor and in-house progress in exploring and accelerating pathways of III-V multijunction concentrator solar cells and systems toward their long-term goals. By 2020, we anticipate that this project will have demonstrated 33% system efficiency and a system price of $1.00/Wp for concentrator PV systems using III-V multijunction solar cells with efficiencies over 41%.

  8. PV system field experience and reliability

    NASA Astrophysics Data System (ADS)

    Durand, Steven; Rosenthal, Andrew; Thomas, Mike

    1997-02-01

    Hybrid power systems consisting of battery inverters coupled with diesel, propane, or gasoline engine-driven electrical generators, and photovoltaic arrays are being used in many remote locations. The potential cost advantages of hybrid systems over simple engine-driven generator systems are causing hybrid systems to be considered for numerous applications including single-family residential, communications, and village power. This paper discusses the various design constraints of such systems and presents one technique for reducing hybrid system losses. The Southwest Technology Development Institute under contract to the National Renewable Energy Laboratory and Sandia National Laboratories has been installing data acquisition systems (DAS) on a number of small and large hybrid PV systems. These systems range from small residential systems (1 kW PV - 7 kW generator), to medium sized systems (10 kW PV - 20 kW generator), to larger systems (100 kW PV - 200 kW generator). Even larger systems are being installed with hundreds of kilowatts of PV modules, multiple wind machines, and larger diesel generators.

  9. Investigation of indirect benefits of PV rooftop in Thailand

    NASA Astrophysics Data System (ADS)

    Khumkrong, T.; Chuangchote, S.; Chenvidhya, D.; Kirtikara, K.

    2017-05-01

    Other than electricity generation, which is the direct benefit of PV rooftop, cooling load reduction due to PV shading is a benefit impact in the uses of PV rooftop. This report is a study of those indirect benefits of PV rooftop. Relation of shading of PV modules and reduction of cooling load was studied in a real testing cite at the office building of CES Solar Cell Testing Center (CSSC). Several data, i.e. solar radiation, rooftop temperatures before/after PV-panel installation, and electricity consumed by equipment, were monitored and collected. This data could be further estimated for cooling load via transient heat conduction approach.

  10. PV Systems Reliability Final Technical Report.

    SciTech Connect

    Lavrova, Olga; Flicker, Jack David; Johnson, Jay; Armijo, Kenneth Miguel; Gonzalez, Sigifredo; Schindelholz, Eric John; Sorensen, Neil R.; Yang, Benjamin Bing-Yeh

    2015-12-01

    The continued exponential growth of photovoltaic technologies paves a path to a solar-powered world, but requires continued progress toward low-cost, high-reliability, high-performance photovoltaic (PV) systems. High reliability is an essential element in achieving low-cost solar electricity by reducing operation and maintenance (O&M) costs and extending system lifetime and availability, but these attributes are difficult to verify at the time of installation. Utilities, financiers, homeowners, and planners are demanding this information in order to evaluate their financial risk as a prerequisite to large investments. Reliability research and development (R&D) is needed to build market confidence by improving product reliability and by improving predictions of system availability, O&M cost, and lifetime. This project is focused on understanding, predicting, and improving the reliability of PV systems. The two areas being pursued include PV arc-fault and ground fault issues, and inverter reliability.

  11. Evaluation of Encapsulant Materials for PV Applications

    SciTech Connect

    Kempe, M.

    2010-01-01

    Encapsulant materials used in PV modules serve multiple purposes. They physically hold components in place, provide electrical insulation, optically couple superstrate materials (e.g., glass) to PV cells, protect components from mechanical stress by mechanically de-coupling components via strain relief, and protect materials from corrosion. To do this, encapsulants must adhere well to all surfaces, remain compliant, and transmit light after exposure to temperature, humidity, and UV radiation histories. Encapsulant materials by themselves do not completely prevent water vapour ingress [1-3], but if they are well adhered, they will prevent the accumulation of liquid water providing protection against corrosion as well as electrical shock. Here, a brief review of some of the polymeric materials under consideration for PV applications is provided, with an explanation of some of their advantages and disadvantages.

  12. Development of non-destructive quality measurement technique for cabbage seed (Brassica campestris L) using hyperspectral reflectance imaging

    USDA-ARS?s Scientific Manuscript database

    Cabbage (Brassica campestris L) is an important crop for Asian countries especially in Korea, Japan and China. In order to achieve uniform and high-yield rate of cabbage product, the seed lot quality needs to be controlled. Non-destructive evaluation of seed viability is an important technique for i...

  13. Asteraceae Artemisia campestris and Artemisia herba-alba Essential Oils Trigger Apoptosis and Cell Cycle Arrest in Leishmania infantum Promastigotes

    PubMed Central

    Messaoud, Chokri; Haoues, Meriam; Neffati, Noura; Bassoumi Jamoussi, Imen; Essafi-Benkhadir, Khadija; Boussaid, Mohamed; Karoui, Habib

    2016-01-01

    We report the chemical composition and anti-Leishmania and antioxidant activity of Artemisia campestris L. and Artemisia herba-alba Asso. essential oils (EOs). Our results showed that these extracts exhibit different antioxidant activities according to the used assay. The radical scavenging effects determined by DPPH assay were of IC50 = 3.3 mg/mL and IC50 = 9.1 mg/mL for Artemisia campestris and Artemisia herba-alba essential oils, respectively. However, antioxidant effects of both essential oils, determined by ferric-reducing antioxidant power (FRAP) assay, were in the same range (2.3 and 2.97 mg eq EDTA/g EO, resp.), while the Artemisia herba-alba essential oil showed highest chelating activity of Fe2+ ions (27.48 mM Fe2+). Interestingly, we showed that both EOs possess dose-dependent activity against Leishmania infantum promastigotes with IC50 values of 68 μg/mL and 44 μg/mL for A. herba-alba and A. campestris, respectively. We reported, for the first time, that antileishmanial activity of both EOs was mediated by cell apoptosis induction and cell cycle arrest at the sub-G0/G1 phase. All our results showed that EOs from A. herba-alba and A. campestris plants are promising candidates as anti-Leishmania medicinal products. PMID:27807464

  14. Glyphosate inhibits the translocation of green fluorescent protein and sucrose from a transgenic tobacco host to Cuscuta campestris Yunk.

    PubMed

    Nadler-Hassar, Talia; Goldshmidt, Alexander; Rubin, Baruch; Wolf, Shmuel

    2004-09-01

    The parasitic plant Cuscuta campestris is dependent on its host for water, assimilates and amino acids. It can be controlled by the herbicide glyphosate, which inhibits 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS), resulting in shikimate accumulation. In this study, C. campestris was parasitic on transgenic tobacco plants expressing green fluorescent protein (GFP) in the phloem. Changes in [14C]sucrose and GFP accumulation in the parasite were used as indicators of the herbicide's effect on translocation between the host and parasite. Host plants were treated with glyphosate 22 days after sowing. Shikimate accumulation in the parasite 1 day after glyphosate treatment (DAGT) confirmed EPSPS inhibition in C. campestris. No damage was visible in the host plants for the first 3 DAGT, while during that same time, a significant reduction in [14C]sucrose and GFP accumulation was observed in the parasite. Thus, we propose that the parallel reduction in GFP and sucrose accumulation in C. campestris is a result of a glyphosate effect on the parasite's ability to withdraw assimilates from the host.

  15. The crystallization of apo-form UMP kinase from Xanthomonas campestris is significantly improved in a strong magnetic field

    SciTech Connect

    Tu, Jhe-Le; Chin, Ko-Hsin; Wang, Andrew H.-J.; Chou, Shan-Ho

    2007-05-01

    A bacterial UMP kinase from the plant pathogen X. campestris pathovar campestris has been overexpressed in E. coli, purified and crystallized in a strong magnetic field. The crystals diffracted to 2.35 Å. Bacterial UMP kinases (UMPKs) are crucial enzymes that are responsible for microbial UTP biosynthesis. Interestingly, eukaryotic and prokaryotic cells use different enzymes for UMP-phosphorylation reactions. Prokaryotic UMPKs are thus believed to be potential targets for antimicrobial drug development. Here, the cloning, expression and crystallization of SeMet-substituted XC1936, a bacterial UMPK from Xanthomonas campestris pathovar campestris, are reported. The crystallization of the apo-form UMPK was found to be significantly improved in a strong magnetic field; the crystals diffracted to a resolution of 2.35 Å, a dramatic improvement over the original value of 3.6 Å. Preliminary structural analyses of apo-form XC1936 using crystals grown in a strong magnetic field clearly reveal well defined loop regions involved in substrate-analogue binding that were previously not visible. Crystallization in a strong magnetic field thus was found to be indispensable in determining the flexible region of the XC1936 UMPK structure.

  16. Crystallization and preliminary X-ray analysis of XC1015, a histidine triad-like protein from Xanthomonas campestris

    SciTech Connect

    Lo, Wen-Ting; Chin, Ko-Hsin; Shr, Hui-Lin; Gao, Fei Philip; Lyu, Ping-Chiang; Wang, Andrew H.-J.; Chou, Shan-Ho

    2006-12-01

    A HIT-like protein from the plant pathogen X. campestris pathovar campestris has been overexpressed in E. coli, purified and crystallized. The crystals diffract to 1.3 Å. Histidine-triad (HIT) proteins are a superfamily of nucleotide hydrolases and transferases that contain a conserved HϕHϕHϕϕ motif (where ϕ is a hydrophobic amino acid) and are found in a variety of organisms. In addition to binding to a variety of nucleotides, other biological functions of the HIT superfamily proteins have been discovered and HIT malfunction has been implicated in several human diseases. Structural studies of HIT superfamily proteins are thus of particular interest. In this manuscript, the cloning, expression, crystallization and preliminary X-ray analysis of XC1015, a HIT protein present in the plant pathogen Xanthomonas campestris pathovar campestris, are reported. The XC1015 crystals diffracted to a resolution of 1.3 Å. They are tetragonal and belong to space group P4{sub 3}2{sub 1}2, with unit-cell parameters a = 40.52, b = 40.52, c = 126.89 Å.

  17. Lead (Pb)-inhibited radicle emergence in Brassica campestris involves alterations in starch-metabolizing enzymes.

    PubMed

    Singh, Harminder Pal; Kaur, Gurpreet; Batish, Daizy R; Kohli, Ravinder K

    2011-12-01

    Lead (Pb) is a toxic heavy metal released into the natural environment and known to cause oxidative damage and alter antioxidant mechanism in plants. However, not much is known about the interference of Pb with the biochemical processes and carbohydrate metabolism during seed germination. We, therefore, investigated the effect of Pb (50-500 μM) upon biochemical alterations in germinating seeds (at 24-h stage) of Brassica campestris L. Pb treatment significantly enhanced protein and carbohydrate contents that increased by ~43% and 200%, respectively, at 500-μM Pb over control. In contrast, the activities of starch/carbohydrate-metabolizing enzymes--α-amylases, β-amylases, acid invertases, and acid phosphatases--decreased by ~54%, 60%, 74%, and 52%, respectively, over control. Activities of peroxidases and polyphenol oxidases, involved in stress acclimation, however, increased by ~1.2- to 3.9-folds and 0.4- to 1.4-folds upon 50-500-μM Pb treatment. Pb enhanced oxidizing ability by 10 to 16.7 times over control suggesting interference with emerging root's oxidizing capacity. The study concludes that Pb exposure inhibits radicle emergence from B. campestris by interfering with the biochemical processes linked to protein and starch metabolism.

  18. PECTATE LYASE-LIKE10 is associated with pollen wall development in Brassica campestris.

    PubMed

    Jiang, Jingjing; Yao, Lina; Yu, Youjian; Lv, Meiling; Miao, Ying; Cao, Jiashu

    2014-11-01

    PECTATE LYASE-LIKE10 (PLL10) was previously identified as one of the differentially expressed genes both in microspores during the late pollen developmental stages and in pistils during the fertilization process in Chinese cabbage (Brassica campestris ssp. chinensis). Here, antisense-RNA was used to study the functions of BcPLL10 in Chinese cabbage. Abnormal pollen was identified in the transgenic lines (bcpll10-4, -5, and -6). In fertilization experiments, fewer seeds were harvested when the antisense-RNA lines were used as pollen donor. In vivo and in vitro pollen germination assays less germinated pollen tubes were observed in bcpll10 lines. Scanning electron microscopy observation verified that the tryphine materials were over accumulated around the pollen surface and sticked them together in bcpll10. Moreover, transmission electron microscopy observation revealed that the internal endintine was overdeveloped and predominantly occupied the intine, and disturbed the normal proportional distribution of the two layers in the non-germinal furrow region; and no obvious demarcation existed between them in the germinal furrow region in the bcpll10 pollen. Collectively, this study presented a novel PLL gene that played an important role during the pollen wall development in B. campestris, which may also possess potential importance for male sterility usage in agriculture.

  19. Increase of xanthan production by cloning xps genes into wild-type Xanthomonas campestris.

    PubMed

    Tseng, Y H; Ting, W Y; Chou, H C; Yang, B Y; Chen, C C

    1992-02-01

    Previously, genomic banks of Xanthomonas campestris were constructed in Escherichia coli, using mobilizable broad-host-range cosmids as the vectors. Following conjugal transfer, genes involved in the biosynthesis of xanthan polysaccharide (XPS) were cloned by the ability to restore the mucoid phenotype to the non-mucoid mutants. In this study, all clones were transferred into the wild-type strain Xc17 to evaluate the effects of the cloned genes on XPS production. Most clones showed no significant effect; however, two plasmids, pP2401 and pP2201, caused 10 and 15% yield increases, respectively, compared with that of controls. While it was not clear how pP2201 caused the yield increase, the effect of pP2401 seemed to result from elevated phosphomannose isomerase activity. Since XPS synthesis in X. campestris is a very efficient process, only relatively small increases are to be expected; an enhancement of productivity by 10-15% is important to the commercial production of xanthan.

  20. [Reclamation of the plant regeneration efficiency of Brassica campestris subsp. chinensis var.Parachinensis].

    PubMed

    Yu, X L; Cao, J S; Xu, S Y

    2001-06-01

    This investigation has developed an efficient and fast method for plant regeneration from petiole of cotyledon explants of Brassica campestris L. subsp. chinensis Makino var. parachinensis Tsen et Lee. A medium was designed for B. campestris subsp. chinensis var. parachinensis to obtain the high frequency of shoot regeneration, which contained BAP 2 mg/L, NAA 0.75-1.0 mgL and 7.5 mg/L AgNO3 solution to the half of NH4+ concentration's MS basic medium. 60 mL/L coconut milk were added to all of media. In this method, frequency of shoot regeneration of "youqing caixin" reached as high as 91.2% and the number of shoots per explant reached as high as 4.7 plants. The result showed that there was a positive correlation between frequency of shoot regeneration and number of shoots per explant. The little shoots could be observed five days after inoculation and were formed directly. The inducing rate of roots of the shoots reached as high as 100% and the rate of viability of transferred mature plant reached higher than 95%. The regeneration period from petiole with cotyledon to a seedling was shorten to about 49 days. Factors influencing in vitro explant regeneration were studied.

  1. Ozone affects gas exchange, growth and reproductive development in Brassica campestris (Wisconsin fast plants).

    PubMed

    Black, V J; Stewart, C A; Roberts, J A; Black, C R

    2007-01-01

    Exposure to ozone (O(3)) may affect vegetative and reproductive development, although the consequences for yield depend on the effectiveness of the compensatory processes induced. This study examined the impact on reproductive development of exposing Brassica campestris (Wisconsin Fast Plants) to ozone during vegetative growth. Plants were exposed to 70 ppb ozone for 2 d during late vegetative growth or 10 d spanning most of the vegetative phase. Effects on gas exchange, vegetative growth, reproductive development and seed yield were determined. Impacts on gas exchange and foliar injury were related to pre-exposure stomatal conductance. Exposure for 2 d had no effect on growth or reproductive characteristics, whereas 10-d exposure reduced vegetative growth and reproductive site number on the terminal raceme. Mature seed number and weight per pod and per plant were unaffected because seed abortion was reduced. The observation that mature seed yield per plant was unaffected by exposure during the vegetative phase, despite adverse effects on physiological, vegetative and reproductive processes, shows that indeterminate species such as B. campestris possess sufficient compensatory flexibility to avoid reductions in seed production.

  2. Production of xanthan gum by free and immobilized cells of Xanthomonas campestris and Xanthomonas pelargonii.

    PubMed

    Niknezhad, Seyyed Vahid; Asadollahi, Mohammad Ali; Zamani, Akram; Biria, Davoud

    2016-01-01

    Production of xanthan gum using immobilized cells of Xanthomonas campestris and Xanthomonas pelargonii grown on glucose or hydrolyzed starch as carbon sources was investigated. Calcium alginate (CA) and calcium alginate-polyvinyl alcohol-boric acid (CA-PVA) beads were used for the immobilization of cells. Xanthan titers of 8.2 and 9.2g/L were obtained for X. campestris cells immobilized in CA-PVA beads using glucose and hydrolyzed starch, respectively, whereas those for X. pelargonii were 8 and 7.9 g/L, respectively. Immobilized cells in CA-PVA beads were successfully employed in three consecutive cycles for xanthan production without any noticeable degradation of the beads whereas the CA beads were broken after the first cycle. The results of this study suggested that immobilized cells are advantageous over the free cells for xanthan production. Also it was shown that the cells immobilized in CA-PVA beads are more efficient than cells immobilized in CA beads for xanthan production.

  3. The polygalacturonase gene BcMF2 from Brassica campestris is associated with intine development

    PubMed Central

    Huang, Li; Cao, Jiashu; Zhang, Aihong; Ye, Yiqun; Zhang, Yuchao; Liu, Tingting

    2009-01-01

    Brassica campestris Male Fertility 2 (BcMF2) is a putative polygalacturonase (PG) gene previously isolated from the flower bud of Chinese cabbage (Brassica campestris L. ssp. chinensis Makino, syn. B. rapa ssp. chinensis). This gene was found to be expressed specifically in tapetum and pollen after the tetrad stage of anther development. Antisense RNA technology was used to study the function of BcMF2 in Chinese cabbage. Scanning and transmission electron microscopy revealed that there were deformities in the transgenic mature pollen grains such as abnormal location of germinal furrows. In addition, the homogeneous pectic exintine layer facing the exterior seemed to be overdeveloped and predominantly occupied the intine, thus reversing the normal proportional distribution of the internal endintine layer and the external exintine layer. Since it is a continuation of the intine layer, the pollen tube wall could not grow normally. This resulted in the formation of a balloon-like swelling structure in the pollen tube tip in nearly 80% of the transgenic pollen grains. Premature degradation of tapetum was also found in these transgenic plants, which displayed decreased expression of the BcMF2 gene. BcMF2 might therefore encode a new PG with an important role in pollen wall development, possibly via regulation of pectin's dynamic metabolism. PMID:19039102

  4. Developing standards for PV packaging materials

    NASA Astrophysics Data System (ADS)

    Wohlgemuth, John; Kempe, Michael; Miller, David; Kurtz, Sarah

    2011-09-01

    The initial qualification standards for photovoltaic modules were designed to help develop a product that is safe, and able to survive reasonably long time periods when deployed in the field. To accomplish this, TC-82 of the International Electro-Technical Commission (IEC), developed and published module qualification standards (IEC 61215 for crystalline Si, IEC 61646 for thin films and IEC 62108 for concentrating modules) and a module safety standard (IEC 61730 -1 and 2). As PV has developed and the technology has become better understood, the properties of materials used in the module package play an increasingly important part in achieving long-term durability and safety. Certain basic properties are required of the materials in order for the modules to be safe and to be able to survive in the field for 25 years or more. Therefore Working Group 2 (Modules) of TC-82 began work to develop new material-level standards for PV that will utilize existing standards, whenever available, but tailored for characterizing the properties that are important for PV modules and modified to take into account the environmental conditions specific to PV applications. The goal is to provide a uniform approach to characterizing candidate materials, providing the necessary information to designers selecting materials for use in their PV products as well as to certification bodies assessing the quality and safety of the products made from these materials. This paper will describe the details of the effort underway to determine what PV material standards are necessary and the progress on developing those standards.

  5. Real time PV manufacturing diagnostic system

    SciTech Connect

    Kochergin, Vladimir; Crawford, Michael A.

    2015-09-01

    The main obstacle Photovoltaic (PV) industry is facing at present is the higher cost of PV energy compared to that of fossil energy. While solar cell efficiencies continue to make incremental gains these improvements are so far insufficient to drive PV costs down to match that of fossil energy. Improved in-line diagnostics however, has the potential to significantly increase the productivity and reduce cost by improving the yield of the process. On this Phase I/Phase II SBIR project MicroXact developed and demonstrated at CIGS pilot manufacturing line a high-throughput in-line PV manufacturing diagnostic system, which was verified to provide fast and accurate data on the spatial uniformity of thickness, an composition of the thin films comprising the solar cell as the solar cell is processed reel-to-reel. In Phase II project MicroXact developed a stand-alone system prototype and demonstrated the following technical characteristics: 1) ability of real time defect/composition inconsistency detection over 60cm wide web at web speeds up to 3m/minute; 2) Better than 1mm spatial resolution on 60cm wide web; 3) an average better than 20nm spectral resolution resulting in more than sufficient sensitivity to composition imperfections (copper-rich and copper-poor regions were detected). The system was verified to be high vacuum compatible. Phase II results completely validated both technical and economic feasibility of the proposed concept. MicroXact’s solution is an enabling technique for in-line PV manufacturing diagnostics to increase the productivity of PV manufacturing lines and reduce the cost of solar energy, thus reducing the US dependency on foreign oil while simultaneously reducing emission of greenhouse gasses.

  6. Radiometric instrumentation for PV system performance monitoring

    SciTech Connect

    Stoffel, T.

    1995-09-01

    This paper provides an overview of existing instrumentation options and solicits user needs for improving outdoor solar radiation measurements for determining PV system performance. The following topics are discussed: (1) historical overview and terminology; (2) radiometer calibration and characterization methods used by NREL; (3) sample calibration results for various commercial instruments; (4) current research topics; (5) user needs for improved PV performance monitoring. Emphasis is placed on the need for the user to understand the measurement capabilities of commercially available radiometers and interpret the measurement results accordingly. A list of radiometer manufacturers is also provided.

  7. Large-Scale PV Integration Study

    SciTech Connect

    Lu, Shuai; Etingov, Pavel V.; Diao, Ruisheng; Ma, Jian; Samaan, Nader A.; Makarov, Yuri V.; Guo, Xinxin; Hafen, Ryan P.; Jin, Chunlian; Kirkham, Harold; Shlatz, Eugene; Frantzis, Lisa; McClive, Timothy; Karlson, Gregory; Acharya, Dhruv; Ellis, Abraham; Stein, Joshua; Hansen, Clifford; Chadliev, Vladimir; Smart, Michael; Salgo, Richard; Sorensen, Rahn; Allen, Barbara; Idelchik, Boris

    2011-07-29

    This research effort evaluates the impact of large-scale photovoltaic (PV) and distributed generation (DG) output on NV Energy’s electric grid system in southern Nevada. It analyzes the ability of NV Energy’s generation to accommodate increasing amounts of utility-scale PV and DG, and the resulting cost of integrating variable renewable resources. The study was jointly funded by the United States Department of Energy and NV Energy, and conducted by a project team comprised of industry experts and research scientists from Navigant Consulting Inc., Sandia National Laboratories, Pacific Northwest National Laboratory and NV Energy.

  8. Updating Interconnection Screens for PV System Integration

    SciTech Connect

    Coddington, M.; Mather, B.; Kroposki, B.; Lynn, K.; Razon, A.; Ellis, A.; Hill, R.; Key, T.; Nicole, K.; Smith, J.

    2012-02-01

    This white paper evaluates the origins and usefulness of the capacity penetration screen, offer short-term solutions which could effectively allow fast-track interconnection to many PV system applications, and considers longer-term solutions for increasing PV deployment levels in a safe and reliable manner while reducing or eliminating the emphasis on the penetration screen. Short-term and longer-term alternatives approaches are offered as examples; however, specific modifications to screening procedures should be discussed with stakeholders and must ultimately be adopted by state and federal regulatory bodies.

  9. International PV QA Task Force's Proposed Comparative Rating System for PV Modules: Preprint

    SciTech Connect

    Wohlgemuth, J.; Kurtz, S.

    2014-10-01

    The International PV Quality Assurance Task Force is developing a rating system that provides comparative information about the relative durability of PV modules. Development of accelerated stress tests that can provide such comparative information is seen as a major step toward being able to predict PV module service life. This paper will provide details of the ongoing effort to determine the format of such an overall module rating system. The latest proposal is based on using three distinct climate zones as defined in IEC 60721-2-1 for two different mounting systems. Specific stresses beyond those used in the qualification tests are being developed for each of the selected climate zones.

  10. PV Cephei: Young Star Caught Speeding?

    NASA Astrophysics Data System (ADS)

    Goodman, Alyssa A.; Arce, Héctor G.

    2004-06-01

    Three independent lines of evidence imply that the young star PV Cep is moving at roughly 20 km s-1 through the interstellar medium. The first and strongest suggestion of motion comes from the geometry of the Herbig-Haro (HH) knots in the ``giant'' HH flow associated with PV Cep. Bisectors of lines drawn between pairs of knots at nearly equal distances from PV Cep imply an east-west motion of the source, and a plasmon model fitted to the knot positions gives a good fit of 22 km s-1 motion for the star. The second bit of damning evidence comes from a redshifted trail of molecular gas pointing in the same east-west direction implied by the HH knot geometry. The third exhibit we offer in accusing PV Cep of speeding involves the apparent tilt in the high-velocity molecular jet now emanating from the star. This tilt is best explained if the true, current jet direction is north-south, as it is in Hubble Space Telescope WFPC2 images, and the star is moving, again at roughly 20 km s-1. Tracing the motion of PV Cep backward in time to the nearest cluster from which it might have been ejected, we find that it is very likely to have been thrown out of the massive star-forming cluster NGC 7023, more than 10 pc away. PV Cep and NGC 7023 are at similar distances, and the backward trace of PV Cep's motion is astonishingly well aligned with a dark, previously unexplained rift in NGC 7023. We propose that PV Cep was ejected, at a speed large enough to escape NGC 7023, at least 100,000 yr ago but that it did not enter the molecular cloud in which it now finds itself until more like 35,000 yr ago. Our calculations show that the currently observable molecular outflow associated with PV Cep is about 10,000 yr old, so the flow has had plenty of time to form while in its current molecular cloud. However, the question of what PV Cep was doing and what gas/disk it took along with it in the time it was traveling through the low-density region between NGC 7023 and its current home is an open

  11. PV Ceph: Young Star Caught Speeding?

    NASA Astrophysics Data System (ADS)

    Goodman, A. A.; Arce, H. G.

    2003-12-01

    Three independent lines of evidence imply that the young star PV Ceph is moving at roughly 20 km s-1 through the interstellar medium. The first, and strongest, suggestion of motion comes from the geometry of the HH knots in the ``giant" Herbig-Haro flow associated with PV Ceph. Bisectors of lines drawn between pairs of knots at nearly equal distances from PV Ceph imply an E-W motion of the source, and a plasmon model fit to the knot positions gives a good fit of 22 km s-1 motion for the star. The second bit of damning evidence comes from a redshifted ``trail" of molecular gas, pointing in the same E-W direction implied by the HH knot geometry. The third exhibit we offer in accusing PV Ceph of speeding involves the tilt apparent in the high-velocity molecular jet now emanating from the star. This tilt is best explained if the true, current, jet direction is N-S, as it is in HST WFPC images, and the star is moving--again at roughly 20 km s-1. Tracing the motion of PV Ceph backward in time, to the nearest cluster from which it might have been ejected, we find that it is very likely to have been thrown out of the massive star-forming cluster NGC7023--more than 10 pc away. PV Ceph and NGC7023 are at similar distances, and the backward-trace of PV Ceph's motion is astonishingly well-aligned with a dark, previously unexplained, rift in NGC7023. We propose that PV Ceph was ejected, at a speed large enough to escape NGC7023, at least 100,000 years ago, but that it did not enter the molecular cloud in which it now finds itself until more like 10,000 years ago. Our calculations show that currently-observable molecular outflow associated with PV Ceph is about 10,000 years old, so that the flow has had plenty of time to form while in its current molecular cloud. But, the question of what PV Ceph was doing, and what gas/disk it took along with it in the time it was traveling through the low-density region between NGC7023 and its current home is open to question. Recent numerical

  12. Spectral radiometric instrumentation and modelling for PV applications

    SciTech Connect

    Cannon, T.W.

    1995-09-01

    This paper describes issues dealing with instrumentation and atmospheric transmission modelling for determining solar spectral irradiance for outdoor photovoltaic (PV) applications. The relevance of these determinations to PV testing and evaluation is discussed with examples.

  13. Transcription Factors PvERF15 and PvMTF-1 Form a Cadmium Stress Transcriptional Pathway1[OPEN

    PubMed Central

    Lin, Tingting; Yang, Wanning; Lu, Wen; Wang, Ying

    2017-01-01

    In plants, cadmium (Cd)-responsive transcription factors are key downstream effectors of Cd stress transcriptional pathways, which are capable of converging Cd stress signals through triggering the expression of Cd detoxification genes. However, the upstream transcriptional regulatory pathways that modulate their responses to Cd are less clear. Previously, we identified the bean (Phaseolus vulgaris) METAL RESPONSE ELEMENT-BINDING TRANSCRIPTION FACTOR1 (PvMTF-1) that responds to Cd and confers Cd tolerance in planta. Here, we demonstrate an upstream transcriptional regulation of the PvMTF-1 response to Cd. Using a yeast one-hybrid system, we cloned the bean ETHYLENE RESPONSE FACTOR15 (PvERF15) that binds to the PvMTF-1 promoter. PvERF15 was strongly induced by Cd stress, and its overexpression resulted in the up-regulation of PvMTF-1. DNA-protein interaction assays further revealed that PvERF15 binds directly to a 19-bp AC-rich element in the PvMTF-1 promoter. The AC-rich element serves as a positive element bound by PvERF15 to activate gene expression. More importantly, knockdown of PvERF15 by RNA interference resulted in reduced Cd-induced expression of PvMTF-1. PvERF15 seems to be involved in Cd tolerance, since knockdown of PvERF15 by RNA interference in bean leaf discs decreased Cd tolerance in a transient assay. Since PvERF15 is a component of the Cd stress transcriptional pathway in beans and PvMTF-1 is one of its downstream targets, our findings provide a PvERF15/PvMTF-1 transcriptional pathway and thereby contribute to the understanding of Cd stress transcriptional regulatory pathways in plants. PMID:28073984

  14. National solar technology roadmap: Film-silicon PV

    SciTech Connect

    Keyes, Brian

    2007-06-01

    Silicon photovoltaic (PV) technologies are addressed in two different technology roadmaps: Film-Silicon PV and Wafer-Silicon PV. This Film-Silicon PV roadmap applies to all silicon-film technologies that rely on a supporting substrate such as glass, polymer, aluminum, stainless steel, or metallurgical-grade silicon. Such devices typically use amorphous, nanocrystalline, fine-grained polycrystalline, or epitaxial silicon layers that are 1–20 μm thick.

  15. TRNSYS HYBRID wind diesel PV simulator

    SciTech Connect

    Quinlan, P.J.A.; Mitchell, J.W.; Klein, S.A.; Beckman, W.A.; Blair, N.J.

    1996-12-31

    The Solar Energy Laboratory (SEL) has developed a wind diesel PV hybrid systems simulator, UW-HYBRID 1.0, an application of the TRNSYS 14.2 time-series simulation environment. An AC/DC bus links up to five diesels and wind turbine models, along with PV modules, a battery bank, and an AC/DC converter. Multiple units can be selected. PV system simulations include solar angle and peak power tracking options. Weather data are Typical Meteorological Year data, parametrically generated synthesized data, or external data files. PV performance simulations rely on long-standing SEL-developed algorithms. Loads data are read as scalable time series. Diesel simulations include estimated fuel-use and waste heat output, and are dispatched using a least-cost of fuel strategy. Wind system simulations include varying air density, wind shear and wake effects. Time step duration is user-selectable. UW-HYBRID 1.0 runs in Windows{reg_sign}, with TRNSED providing a customizable user interface. 12 refs., 6 figs.

  16. Final Technical Report: PV Fault Detection Tool.

    SciTech Connect

    King, Bruce Hardison; Jones, Christian Birk

    2015-12-01

    The PV Fault Detection Tool project plans to demonstrate that the FDT can (a) detect catastrophic and degradation faults and (b) identify the type of fault. This will be accomplished by collecting fault signatures using different instruments and integrating this information to establish a logical controller for detecting, diagnosing and classifying each fault.

  17. PV Theories For Planetary Atmospheric Circulations

    NASA Technical Reports Server (NTRS)

    Allison, Michael; Travis, Larry (Technical Monitor)

    2000-01-01

    Potential vorticity (or PV) has become an important tool for the diagnosis and modeling of the Earth's atmospheric and oceanic circulations. More recently, the application of PV thinking to numerical simulations and spacecraft observations of other atmospheres, including those of Mars, Venus, and Titan, has encouraged the hope for a unified understanding of planetary circulation regimes encompassing a wide range of rotation, stratification, and forcing parameters. Specifically, the accumulated evidence suggests that zonal-mean winds and temperatures at upper tropospheric levels approximate a state of zero potential vorticity within the bounding latitudes of the westerly jets, while the poleward regions of cyclonic shear conform to a PV state that is well mixed with respect to its polar limit. This review of the prospects for conceptual planetary circulation models will explore the possibility that the zonal-mean state of an atmosphere can be calculated in terms of a simple link between the latitudinal PV and potential temperature variation at an upper baroclinic steering level.

  18. PV Theories For Planetary Atmospheric Circulations

    NASA Technical Reports Server (NTRS)

    Allison, Michael; Travis, Larry (Technical Monitor)

    2000-01-01

    Potential vorticity (or PV) has become an important tool for the diagnosis and modeling of the Earth's atmospheric and oceanic circulations. More recently, the application of PV thinking to numerical simulations and spacecraft observations of other atmospheres, including those of Mars, Venus, and Titan, has encouraged the hope for a unified understanding of planetary circulation regimes encompassing a wide range of rotation, stratification, and forcing parameters. Specifically, the accumulated evidence suggests that zonal-mean winds and temperatures at upper tropospheric levels approximate a state of zero potential vorticity within the bounding latitudes of the westerly jets, while the poleward regions of cyclonic shear conform to a PV state that is well mixed with respect to its polar limit. This review of the prospects for conceptual planetary circulation models will explore the possibility that the zonal-mean state of an atmosphere can be calculated in terms of a simple link between the latitudinal PV and potential temperature variation at an upper baroclinic steering level.

  19. Role of Polycrystalline Thin-Film PV Technologies in Competitive PV Module Markets: Preprint

    SciTech Connect

    von Roedern, B.; Ullal, H. S.

    2008-05-01

    This paper discusses the developments in thin-film PV technologies and provides an outlook on future commercial module efficiencies achievable based on today's knowledge about champion cell performance.

  20. Commercialization of PV-powered pumping systems for use in utility PV service programs. Final report

    SciTech Connect

    1997-03-01

    The project described in this report was a commercialization effort focused on cost-effective remote water pumping systems for use in utility-based photovoltaic (PV) service programs. The project combined a commercialization strategy tailored specifically for electric utilities with the development of a PV-powered pumping system that operates conventional ac pumps rather than relying on the more expensive and less reliable PV pumps on the market. By combining these two attributes, a project goal was established of creating sustained utility purchases of 250 PV-powered water pumping systems per year. The results of each of these tasks are presented in two parts contained in this Final Summary Report. The first part summarizes the results of the Photovoltaic Services Network (PSN) as a new business venture, while the second part summarizes the results of the Golden Photon system installations. Specifically, results and photographs from each of the system installations are presented in this latter part.

  1. Water Extract from Spent Mushroom Substrate of Hericium erinaceus Suppresses Bacterial Wilt Disease of Tomato

    PubMed Central

    Kwak, A Min; Min, Kyeong Jin; Lee, Sang Yeop

    2015-01-01

    Culture filtrates of six different edible mushroom species were screened for antimicrobial activity against tomato wilt bacteria Ralstonia solanacearum B3. Hericium erinaceus, Lentinula edodes (Sanjo 701), Grifola frondosa, and Hypsizygus marmoreus showed antibacterial activity against the bacteria. Water, n-butanol, and ethyl acetate extracts of spent mushroom substrate (SMS) of H. erinaceus exhibited high antibacterial activity against different phytopathogenic bacteria: Pectobacterium carotovorum subsp. carotovorum, Agrobacterium tumefaciens, R. solanacearum, Xanthomonas oryzae pv. oryzae, X. campestris pv. campestris, X. axonopodis pv. vesicatoria, X. axonopodis pv. citiri, and X. axonopodis pv. glycine. Quantitative real-time PCR revealed that water extracts of SMS (WESMS) of H. erinaceus induced expressions of plant defense genes encoding β-1,3-glucanase (GluA) and pathogenesis-related protein-1a (PR-1a), associated with systemic acquired resistance. Furthermore, WESMS also suppressed tomato wilt disease caused by R. solanacearum by 85% in seedlings and promoted growth (height, leaf number, and fresh weight of the root and shoot) of tomato plants. These findings suggest the WESMS of H. erinaceus has the potential to suppress bacterial wilt disease of tomato through multiple effects including antibacterial activity, plant growth promotion, and defense gene induction. PMID:26539048

  2. High Resolution PV Power Modeling for Distribution Circuit Analysis

    SciTech Connect

    Norris, B. L.; Dise, J. H.

    2013-09-01

    NREL has contracted with Clean Power Research to provide 1-minute simulation datasets of PV systems located at three high penetration distribution feeders in the service territory of Southern California Edison (SCE): Porterville, Palmdale, and Fontana, California. The resulting PV simulations will be used to separately model the electrical circuits to determine the impacts of PV on circuit operations.

  3. Beacons In Brief. P/PV In Brief. Issue 2

    ERIC Educational Resources Information Center

    Blank, Susan; Farley, Chelsea

    2004-01-01

    This second issue in P/PV's "In Brief" series focuses on the San Francisco Beacon Initiative and P/PV's recently released evaluation results. The Beacon Initiative established after-school programs in eight public schools in low-income San Francisco neighborhoods. P/PV's 36-month evaluation examined key developmental and academic outcomes.…

  4. Beacons In Brief. P/PV In Brief. Issue 2

    ERIC Educational Resources Information Center

    Blank, Susan; Farley, Chelsea

    2004-01-01

    This second issue in P/PV's "In Brief" series focuses on the San Francisco Beacon Initiative and P/PV's recently released evaluation results. The Beacon Initiative established after-school programs in eight public schools in low-income San Francisco neighborhoods. P/PV's 36-month evaluation examined key developmental and academic outcomes.…

  5. Determination of Parameters of PV Concentrating System With Heliostat

    NASA Astrophysics Data System (ADS)

    Vardanyan, R.; Norsoyan, A.; Dallakyan, V.

    2010-10-01

    The structure of PV concentrating system with heliostat is analyzed. The mathematical model of system consisting of PV concentrating module and heliostat is developed. With the use of developed mathematical model the optimal parameters of the system are determined. The results of this work can be used during the design of PV concentrating systems with heliostats.

  6. The possibility of developing hybrid PV/T solar system

    NASA Astrophysics Data System (ADS)

    Dobrnjac, M.; Zivkovic, P.; Babic, V.

    2017-05-01

    An alternative and cost-effective solution to developing integrated PV system is to use hybrid photovoltaic/thermal (PV/T) solar system. The temperature of PV modules increases due to the absorbed solar radiation that is not converted into electricity, causing a decrease in their efficiency. In hybrid PV/T solar systems the reduction of PV module temperature can be combined with a useful fluid heating. In this paper we present the possibility of developing a new hybrid PV/T solar system. Hybrid PV/T system can provide electrical and thermal energy, thus achieving a higher energy conversion rate of the absorbed solar radiation. We developed PV/T prototype consisted of commercial PV module and thermal panel with our original solution of aluminium absorber with special geometric shapes. The main advantages of our combined PV/T system are: removing of heat from the PV panel; extending the lifetime of photovoltaic cells; excess of the removing heat from PV part is used to heat the fluid in the thermal part of the panel; the possibility of using on the roof and facade constructions because less weight.

  7. Technologies to Increase PV Hosting Capacity in Distribution Feeders

    SciTech Connect

    Ding, Fei; Mather, Barry; Gotseff, Peter

    2016-11-14

    This paper studies the distributed photovoltaic (PV) hosting capacity in distribution feeders by using the stochastic analysis approach. Multiple scenario simulations are conducted to analyze several factors that affect PV hosting capacity, including the existence of voltage regulator, PV location, the power factor of PV inverter and Volt/VAR control. Based on the conclusions obtained from simulation results, three approaches are then proposed to increase distributed PV hosting capacity, which can be formulated as the optimization problem to obtain the optimal solution. All technologies investigated in this paper utilize only existing assets in the feeder and therefore are implementable for a low cost. Additionally, the tool developed for these studies is described.

  8. Technologies to Increase PV Hosting Capacity in Distribution Feeders: Preprint

    SciTech Connect

    Ding, Fei; Mather, Barry; Gotseff, Peter

    2016-08-01

    This paper studies the distributed photovoltaic (PV) hosting capacity in distribution feeders by using the stochastic analysis approach. Multiple scenario simulations are conducted to analyze several factors that affect PV hosting capacity, including the existence of voltage regulator, PV location, the power factor of PV inverter and Volt/VAR control. Based on the conclusions obtained from simulation results, three approaches are then proposed to increase distributed PV hosting capacity, which can be formulated as the optimization problem to obtain the optimal solution. All technologies investigated in this paper utilize only existing assets in the feeder and therefore are implementable for a low cost. Additionally, the tool developed for these studies is described.

  9. PV_LIB Toolbox v. 1.3

    SciTech Connect

    2015-12-09

    PV_LIB comprises a library of Matlab? code for modeling photovoltaic (PV) systems. Included are functions to compute solar position and to estimate irradiance in the PV system’s plane of array, cell temperature, PV module electrical output, and conversion from DC to AC power. Also included are functions that aid in determining parameters for module performance models from module characterization testing. PV_LIB is open source code primarily intended for research and academic purposes. All algorithms are documented in openly available literature with the appropriate references included in comments within the code.

  10. Characterization of xanthan gum produced from glycerol by a mutant strain Xanthomonas campestris CCTCC M2015714.

    PubMed

    Wang, Zichao; Wu, Jianrong; Zhu, Li; Zhan, Xiaobei

    2017-02-10

    Xanthan gum was produced by a mutant strain X. campestris CCTCC M2015714 with glycerol as the sole carbon source. The monosaccharide composition and molar ratio of xanthan gum produced from glycerol are glucose: mannose: glucuronic acid=2.0:1.65:1.0. Meanwhile, chemical structure of xanthan gum produced from glycerol is similar to that of the commercial xanthan through FT-IR and NMR. Remarkably, the molecular weight of xanthan gum produced using our method (3.0±0.14×10(6)Da) is about half that of the commercial one (5.8±0.25×10(6)Da), and the consistency index (K) of which is less than 1/10 that of the commercial xanthan. This work paves the way for xanthan production from glycerol and is useful for studying the structure/application of xanthan gum.

  11. Ticks (Acari: Ixodidae) of small antelopes: steenbok, Raphicerus campestris and suni, Neotragus moschatus.

    PubMed

    Golezardy, H; Horak, I G

    2006-09-01

    During surveys on the tick burdens of various wildlife species in South Africa, nine small antelopes became available for study. Six of these were steenbok, Raphicerus campestris and three sunis, Neotragus moschatus, and their tick burdens are recorded here. The steenbok were examined in three nature reserves and harboured nine tick species. The sunis were examined in a fourth reserve and were infested with eight species. The steenbok and sunis were generally infested with the immature stages of the same tick species that infest larger animals in the same geographic regions. In addition the sunis harboured Haemaphysalis parmata, which in South Africa is present only in the eastern and north-eastern coastal and adjacent areas of KwaZulu-Natal Province. They were also infested with Rhipicephalus kochi, which in South Africa occurs only in the far north-east of the KwaZulu-Natal and Limpopo Provinces.

  12. Soxhlet-assisted matrix solid phase dispersion to extract flavonoids from rape (Brassica campestris) bee pollen.

    PubMed

    Ma, Shuangqin; Tu, Xijuan; Dong, Jiangtao; Long, Peng; Yang, Wenchao; Miao, Xiaoqing; Chen, Wenbin; Wu, Zhenhong

    2015-11-15

    Soxhlet-assisted matrix solid phase dispersion (SA-MSPD) method was developed to extract flavonoids from rape (Brassica campestris) bee pollen. Extraction parameters including the extraction solvent, the extraction time, and the solid support conditions were investigated and optimized. The best extraction yields were obtained using ethanol as the extraction solvent, silica gel as the solid support with 1:2 samples to solid support ratio, and the extraction time of one hour. Comparing with the conventional solvent extraction and Soxhlet method, our results show that SA-MSPD method is a more effective technique with clean-up ability. In the test of six different samples of rape bee pollen, the extracted content of flavonoids was close to 10mg/g. The present work provided a simple and effective method for extracting flavonoids from rape bee pollen, and it could be applied in the studies of other kinds of bee pollen.

  13. Insulin-releasing and insulin-like activity of Agaricus campestris (mushroom).

    PubMed

    Gray, A M; Flatt, P R

    1998-05-01

    Agaricus campestris (mushroom) has been documented as a traditional treatment for diabetes. Here the administration of mushroom in the diet (62.5 g/kg) and drinking water (2.5 g/l) countered the hyperglycaemia of streptozotocin-diabetic mice. An aqueous extract of mushroom (1 mg/ml) stimulated 2-deoxyglucose transport (2.0-fold), glucose oxidation (1.5-fold) and incorporation of glucose into glycogen (1.8-fold) in mouse abdominal muscle. In acute 20 min tests, 0.25-1 mg/ml aqueous extract of mushroom evoked a stepwise 3.5- to 4.6-fold stimulation of insulin secretion from the BRIN-BD11 pancreatic B-cell line. This effect was abolished by 0.5 mM diazoxide and prior exposure to extract did not affect subsequent stimulation of insulin secretion by 10 mM L-alanine, thereby negating a detrimental effect on cell viability. The effect of extract was potentiated by 16.7 mM glucose, L-alanine (10 mM) and IBMX (1 mM), and a depolarising concentration of KCl (25 mM) did not augment the insulin-releasing activity of mushroom. Activity of the extract was found to be heat stable, acetone soluble and unaltered by exposure to alkali, but decreased with exposure to acid. Dialysis to remove components with molecular mass < 2000 Da caused a 40% reduction in activity. Sequential extraction with solvents revealed insulin-releasing activity to be greatest in polar fractions. Lack of haemagglutinin activity with extract activity indicated that activity was unlikely to be due to a lectin-mediated event. These results demonstrate the presence of antihyperglycaemic, insulin-releasing and insulin-like activity in A. campestris.

  14. Uptake and transformation of arsenic during the reproductive life stage of Agaricus bisporus and Agaricus campestris.

    PubMed

    Nearing, Michelle M; Koch, Iris; Reimer, Kenneth J

    2016-11-01

    Fruiting bodies from the Agaricus genus have been found to contain non-toxic arsenobetaine (AB) as a major compound. It is unknown whether AB is formed during the vegetative or reproductive life stages of the fungus, or by the surrounding microbial community, but AB's structural similarity to glycine betaine has led to the hypothesis that AB may be adventitiously accumulated as an osmolyte. To investigate the potential formation of AB during the reproductive life stage of Agaricus species, growth substrate and fungi were collected during the commercial growth of Agaricus bisporus and analyzed for arsenic speciation using HPLC-ICP-MS. AB was found to be the major arsenic compound in the fungus at the earliest growth stage of fruiting (the primordium). The growth substrate mainly contained arsenate (As(V)). The distribution of arsenic in an A. bisporus primordium grown on As(V) treated substrate, and in a mature Agaricus campestris fruiting body collected from arsenic contaminated mine tailings, was mapped using two dimensional XAS imaging. The primordium and stalk of the mature fruiting body were both found to be growing around pockets of substrate material containing higher As concentrations, and AB was found exclusively in the fungal tissues. In the mature A. campestris the highest proportion of AB was found in the cap, supporting the AB as an osmolyte hypothesis. The results have allowed us to pinpoint the fungus life stage at which AB formation takes place, namely reproduction, which provides a direction for further research. Copyright © 2016. Published by Elsevier B.V.

  15. Phytotoxic activity in Flourensia campestris and isolation of (--)-hamanasic acid A as its active principle compound.

    PubMed

    Silva, Mariana P; Piazza, Leonardo A; López, Daniela; López Rivilli, Marisa J; Turco, Mauricio D; Cantero, Juan J; Tourn, Mónica G; Scopel, Ana L

    2012-05-01

    An aqueous extract from Flourensia campestris (Asteraceae) dry aerial parts showed strong inhibition on the germination and growth of Lactuca sativa. Based on bio-guided chromatographic fractionation of aq. extracts from dry and fresh leaves and spectroscopic means, (-)-hamanasic acid A (7-carboxy-8-hydroxy-1(2), 12(13)-dien-bisabolene (1)) was isolated as the most inhibitory active principle on germination (ECg(50)=2.9 mM) and on root (ECr(50)=1.5 mM)/shoot (ECs(50)=2.0 mM) growth. As measured by GC, and correlated with a simple designed 2D-TLC, compound 1 was distributed throughout the plant, with a remarkably high concentration (1.6%) in the leaves and the inflorescences. At least a quarter of the amount of 1 was found in aqueous extracts suggesting that leaching would be a key route for its release into the environment. By contrast, leaf essential oils (HD) between 0.5 and 1.5 μl ml(-1) did not show herbicidal effects and 1 was not found in them (TLC) nor among volatiles (HS-SPME). Volatile compositions were assessed by GC-FID and GC-MS and led to the identification of 23 compounds (4 monoterpenes and 19 sesquiterpenes) with a wide seasonal (spring-summer%) variation, represented principally by bicyclo-germacrene (37-6%), spathulenol (4-32%), globulol (20-0%), beta-caryophyllene (15-6%), caryophyllene oxide (1-13%) and bicycloelemene (10-1%), respectively. The high amount of 1 in F. campestris together with its feasibility of being extracted with water suggest that (-)-hamanasic acid A is an allelochemical in this species. Species-specific studies must be carried out to evaluate the potential of 1 as a natural herbicidal compound. Published by Elsevier Ltd.

  16. Materials Testing for PV Module Encapsulation

    SciTech Connect

    Jorgensen, G.; Terwilliger, K.; Glick, S.; Pern, J.; McMahon, T.

    2003-05-01

    Important physical properties of materials used in PV module packaging are presented. High-moisture-barrier, high-resistivity, adhesion-promoting coatings on polyethyl-ene terephthalate (PET) films have been fabricated and characterized for use in PV module application and com-pared to standard polymer backsheet materials. Ethylene vinyl acetate (EVA) and an encapsulant replacement for EVA are studied for their water vapor transmission rate (WVTR) and adhesion properties. WVTR, at test conditions up to 85C/100% relative humidity (RH), and adhesion val-ues are measured before and after filtered xenon arc lamp ultraviolet (UV) exposure and damp heat exposure at 85C/85% RH. Water ingress is quantified by weight gain and embedded humidity sensors.

  17. Cascaded Microinverter PV System for Reduced Cost

    SciTech Connect

    Bellus, Daniel R.; Ely, Jeffrey A.

    2013-04-29

    In this project, a team led by Delphi will develop and demonstrate a novel cascaded photovoltaic (PV) inverter architecture using advanced components. This approach will reduce the cost and improve the performance of medium and large-sized PV systems. The overall project objective is to develop, build, and test a modular 11-level cascaded three-phase inverter building block for photovoltaic applications and to develop and analyze the associated commercialization plan. The system will be designed to utilize photovoltaic panels and will supply power to the electric grid at 208 VAC, 60 Hz 3-phase. With the proposed topology, three inverters, each with an embedded controller, will monitor and control each of the cascade sections, reducing costs associated with extra control boards. This report details the final disposition on this project.

  18. A study on the essential oil of Ferulago campestris: how much does extraction method influence the oil composition?

    PubMed

    Riela, Serena; Bruno, Maurizio; Rosselli, Sergio; Saladino, Maria L; Caponetti, Eugenio; Formisano, Carmen; Senatore, Felice

    2011-02-01

    The essential oil of different parts of Ferulago campestris (Bess.) collected in Sicily has been extracted by microwave-assisted hydrodistillation (MAHD) and by classic hydrodistillation (HD). A comparative qualitative-quantitative study on the composition of the oils was carried out. A total of 100 compounds were identified in the oils obtained by MAHD, whereas 88 compounds characterized the HD oils. The most prominent components were, in all different parts of F. campestris and in both extraction methods, 2,4,5-trimethylbenzaldehyde and 2,4,6-trimethylbenzaldehyde isomers; the latter was not previously found. The attempt to evaluate where the oil components are located in all parts of the plant was carried out by means of a kinetic study. Then, electron microscopy observation on the different parts before and after MAHD and HD was performed.

  19. PV Manufacturing R&D Project -- Trends in the U.S. PV Industry

    SciTech Connect

    Brown, K. E.; Mitchell, R. L.; Bower, W. I.; King, R.

    2005-01-01

    To foster continued growth in the U.S. photovoltaic (PV) industry, the U.S. Department of Energy initiated the PV Manufacturing R&D (PVMR&D) Project--a partnership with U.S. PV industry participants to perform cost-shared manufacturing research and development. Throughout FY 2004, PVMR&D managed fourteen subcontracts across the industry. The impact of PVMR&D is quantified by reductions in direct module manufacturing costs, scale-up of existing PV production capacity, and accrual of cost savings to the public and industry. An analysis of public and industry investment shows that both recaptured funds by mid-1998 based on estimated manufacturing cost savings from PVMR&D participation. Since project inception, total PV manufacturing capacity has increased from 14 MW to 201 MW at the close of 2003, while direct manufacturing costs declined from $5.55/W to $2.49/W. These results demonstrate continued progress toward the overriding goals of the PVMR&D project.

  20. Energy balance of the global photovoltaic (PV) industry--is the PV industry a net electricity producer?

    PubMed

    Dale, Michael; Benson, Sally M

    2013-04-02

    A combination of declining costs and policy measures motivated by greenhouse gas (GHG) emissions reduction and energy security have driven rapid growth in the global installed capacity of solar photovoltaics (PV). This paper develops a number of unique data sets, namely the following: calculation of distribution of global capacity factor for PV deployment; meta-analysis of energy consumption in PV system manufacture and deployment; and documentation of reduction in energetic costs of PV system production. These data are used as input into a new net energy analysis of the global PV industry, as opposed to device level analysis. In addition, the paper introduces a new concept: a model tracking energetic costs of manufacturing and installing PV systems, including balance of system (BOS) components. The model is used to forecast electrical energy requirements to scale up the PV industry and determine the electricity balance of the global PV industry to 2020. Results suggest that the industry was a net consumer of electricity as recently as 2010. However, there is a >50% that in 2012 the PV industry is a net electricity provider and will "pay back" the electrical energy required for its early growth before 2020. Further reducing energetic costs of PV deployment will enable more rapid growth of the PV industry. There is also great potential to increase the capacity factor of PV deployment. These conclusions have a number of implications for R&D and deployment, including the following: monitoring of the energy embodied within PV systems; designing more efficient and durable systems; and deploying PV systems in locations that will achieve high capacity factors.

  1. Population genetics and phylogeography of endangered Oxytropis campestris var. chartacea and relatives: arctic-alpine disjuncts in eastern North America.

    PubMed

    Chung, Melissa; Gelembiuk, Greg; Givnish, Thomas J

    2004-12-01

    Fassett's locoweed (Oxytropis campestris var. chartacea, Fabaceae) is an endangered perennial endemic to Wisconsin. Patterns of genetic variation within and among six remaining populations and their relationship to other members of the O. campestris complex were analysed using AFLPs from 140 accessions across northern North America. Within-population measures of genetic diversity were high (mean expected heterozygosity HE = 0.16; mean nucleotide diversity pi = 0.015) compared with other herbaceous plants. Estimates of among-population differentiation were low (FST = 0.12; PhiST = 0.29), consistent with outcrossing. Genetic and geographical distances between populations were significantly correlated within Fassett's locoweed (r2 = 0.73, P < 0.002 for Mantel test) and O. campestris as a whole (r2 = 0.63, P < 0.0001). Individual and population-based phylogenetic analyses showed that Fassett's locoweed is monophyletic and sister to O. campestris var. johannensis. Morphometric analyses revealed significant differences between Fassett's locoweed and populations of var. johannensis. The first chromosome count for Fassett's locoweed indicates that it is tetraploid (2n = 32), unlike hexaploid var. johannensis. High within-population diversity and relatively low among-population differentiation are consistent with populations of Fassett's locoweed being relicts of a more continuous Pleistocene distribution. Our data support the continued recognition of Fassett's locoweed and protection under federal and state regulations. High levels of genetic diversity within populations suggest that maintain-ing the ecological conditions that favour the life cycle of this plant may be a more pressing concern than the erosion of genetic variation.

  2. Structure of XC6422 from Xanthomonas campestris at 1.6 Å resolution: a small serine α/β-hydrolase

    SciTech Connect

    Yang, Chao-Yu; Chin, Ko-Hsin; Chou, Chia-Cheng; Wang, Andrew H.-J.; Chou, Shan-Ho

    2006-06-01

    The crystal structure of a conserved hypothetical protein from X. campestris has been determined to a resolution of 1.6 Å. The determined X. campestris structure shows that it belongs to the superfamily of serine α/β hydrolase, with an extra strand preceding the first β-strand to lead to extensive subunit interactions in the crystal. XC6422 is a conserved hypothetical protein from Xanthomonas campestris pathovar campestris (Xcc), a Gram-negative yellow-pigmented pathogenic bacterium that causes black rot, one of the major worldwide diseases of cruciferous crops. The protein consists of 220 amino acids and its structure has been determined to 1.6 Å resolution using the multi-wavelength anomalous dispersion (MAD) method. Although it has very low sequence identity to protein sequences in the PDB (less than 20%), the determined structure nevertheless shows that it belongs to the superfamily of serine α/β-hydrolases, with an active site that is fully accessible to solvent owing to the absence of a lid domain. Modelling studies with the serine esterase inhibitor E600 indicate that XC6422 adopts a conserved Ser-His-Asp catalytic triad common to this superfamily and has a preformed oxyanion hole for catalytic activation. These structural features suggest that XC6422 is most likely to be a hydrolase active on a soluble ester or a small lipid. An extra strand preceding the first β-strand in the canonical α/β-hydrolase fold leads to extensive subunit interactions between XC6422 monomers, which may explain why XC6422 crystals of good diffraction quality can grow to dimensions of up to 1.5 mm in a few days.

  3. Integrating Solar PV in Utility System Operations

    SciTech Connect

    Mills, A.; Botterud, A.; Wu, J.; Zhou, Z.; Hodge, B-M.; Heany, M.

    2013-10-31

    This study develops a systematic framework for estimating the increase in operating costs due to uncertainty and variability in renewable resources, uses the framework to quantify the integration costs associated with sub-hourly solar power variability and uncertainty, and shows how changes in system operations may affect these costs. Toward this end, we present a statistical method for estimating the required balancing reserves to maintain system reliability along with a model for commitment and dispatch of the portfolio of thermal and renewable resources at different stages of system operations. We estimate the costs of sub-hourly solar variability, short-term forecast errors, and day-ahead (DA) forecast errors as the difference in production costs between a case with “realistic” PV (i.e., subhourly solar variability and uncertainty are fully included in the modeling) and a case with “well behaved” PV (i.e., PV is assumed to have no sub-hourly variability and can be perfectly forecasted). In addition, we highlight current practices that allow utilities to compensate for the issues encountered at the sub-hourly time frame with increased levels of PV penetration. In this analysis we use the analytical framework to simulate utility operations with increasing deployment of PV in a case study of Arizona Public Service Company (APS), a utility in the southwestern United States. In our analysis, we focus on three processes that are important in understanding the management of PV variability and uncertainty in power system operations. First, we represent the decisions made the day before the operating day through a DA commitment model that relies on imperfect DA forecasts of load and wind as well as PV generation. Second, we represent the decisions made by schedulers in the operating day through hour-ahead (HA) scheduling. Peaking units can be committed or decommitted in the HA schedules and online units can be redispatched using forecasts that are improved

  4. Susceptibility of Anopheles campestris-like and Anopheles barbirostris species complexes to Plasmodium falciparum and Plasmodium vivax in Thailand.

    PubMed

    Thongsahuan, Sorawat; Baimai, Visut; Junkum, Anuluck; Saeung, Atiporn; Min, Gi-Sik; Joshi, Deepak; Park, Mi-Hyun; Somboon, Pradya; Suwonkerd, Wannapa; Tippawangkosol, Pongsri; Jariyapan, Narissara; Choochote, Wej

    2011-02-01

    Nine colonies of five sibling species members of Anopheles barbirostris complexes were experimentally infected with Plasmodium falciparum and Plasmodium vivax. They were then dissected eight and 14 days after feeding for oocyst and sporozoite rates, respectively, and compared with Anopheles cracens. The results revealed that Anopheles campestris-like Forms E (Chiang Mai) and F (Udon Thani) as well as An. barbirostris species A3 and A4 were non-potential vectors for P. falciparum because 0% oocyst rates were obtained, in comparison to the 86.67-100% oocyst rates recovered from An. cracens. Likewise, An. campestris-like Forms E (Sa Kaeo) and F (Ayuttaya), as well as An. barbirostris species A4, were non-potential vectors for P. vivax because 0% sporozoite rates were obtained, in comparison to the 85.71-92.31% sporozoite rates recovered from An. cracens. An. barbirostris species A1, A2 and A3 were low potential vectors for P. vivax because 9.09%, 6.67% and 11.76% sporozoite rates were obtained, respectively, in comparison to the 85.71-92.31% sporozoite rates recovered from An. cracens. An. campestris-like Forms B and E (Chiang Mai) were high-potential vectors for P. vivax because 66.67% and 64.29% sporozoite rates were obtained, respectively, in comparison to 90% sporozoite rates recovered from An. cracens.

  5. Characterization of a putative pollen-specific arabinogalactan protein gene, BcMF8, from Brassica campestris ssp. chinensis.

    PubMed

    Huang, Li; Cao, Jia-Shu; Zhang, Ai-Hong; Ye, Yi-Qun

    2008-12-01

    The BcMF8 (Brassica campestris male fertility 8) gene, possessing the features of 'classical' arabinogalactan protein (AGP) was isolated from Brassica campestris L. ssp. chinensis, Makino syn. B. rapa L. ssp. chinensis. This gene was highly abundant in the fertile flower buds but silenced in the sterile ones of genic male sterile A/B line ('ZUBajh97-01A/B') in B. campestris. Expression patterns analysis suggested BcMF8 was a pollen-specific gene, whose transcript started to be expressed at the uninucleate stage and maintained throughout to the pollen at pollination stage. BcMF8 is highly homologous to the known pollen-specific AGP genes Sta 39-4 and Sta 39-3 from B. napus. Isolation and multiple alignment of the homologs of BcMF8 gene in the family Cruciferae indicated that BcMF8 was highly conserved in this family, which reflect the conservation in biological function and importance of this putative AGP gene in plant development. Similarity analysis also demonstrated Sta 39-4 and Sta 39-3 may originate from different genomes.

  6. Cadmium-Induced Hydrogen Accumulation Is Involved in Cadmium Tolerance in Brassica campestris by Reestablishment of Reduced Glutathione Homeostasis.

    PubMed

    Wu, Qi; Su, Nana; Chen, Qin; Shen, Wenbiao; Shen, Zhenguo; Xia, Yan; Cui, Jin

    2015-01-01

    Hydrogen gas (H2) was recently proposed as a therapeutic antioxidant and signaling molecule in clinical trials. However, the underlying physiological roles of H2 in plants remain unclear. In the present study, hydrogen-rich water (HRW) was used to characterize the physiological roles of H2 in enhancing the tolerance of Brassica campestris against cadmium (Cd). The results showed that both 50 μM CdCl2 and 50%-saturated HRW induced an increase of endogenous H2 in Brassica campestris seedlings, and HRW alleviated Cd toxicity related to growth inhibition and oxidative damage. Seedlings supplied with HRW exhibited increased root length and reduced lipid peroxidation, similar to plants receiving GSH post-treatment. Additionally, seedlings post-treated with HRW accumulated higher levels of reduced glutathione (GSH) and ascorbic acid (AsA) and showed increased GST and GPX activities in roots. Molecular evidence illustrated that the expression of genes such as GS, GR1 and GR2, which were down-regulated following the addition of Cd, GSH or BSO, could be reversed to varying degrees by the addition of HRW. Based on these results, it could be proposed that H2 might be an important regulator for enhancing the tolerance of Brassica campestris seedlings against Cd, mainly by governing reduced glutathione homeostasis.

  7. Cadmium-Induced Hydrogen Accumulation Is Involved in Cadmium Tolerance in Brassica campestris by Reestablishment of Reduced Glutathione Homeostasis

    PubMed Central

    Chen, Qin; Shen, Wenbiao; Shen, Zhenguo; Xia, Yan; Cui, Jin

    2015-01-01

    Hydrogen gas (H2) was recently proposed as a therapeutic antioxidant and signaling molecule in clinical trials. However, the underlying physiological roles of H2 in plants remain unclear. In the present study, hydrogen-rich water (HRW) was used to characterize the physiological roles of H2 in enhancing the tolerance of Brassica campestris against cadmium (Cd). The results showed that both 50 μM CdCl2 and 50%-saturated HRW induced an increase of endogenous H2 in Brassica campestris seedlings, and HRW alleviated Cd toxicity related to growth inhibition and oxidative damage. Seedlings supplied with HRW exhibited increased root length and reduced lipid peroxidation, similar to plants receiving GSH post-treatment. Additionally, seedlings post-treated with HRW accumulated higher levels of reduced glutathione (GSH) and ascorbic acid (AsA) and showed increased GST and GPX activities in roots. Molecular evidence illustrated that the expression of genes such as GS, GR1 and GR2, which were down-regulated following the addition of Cd, GSH or BSO, could be reversed to varying degrees by the addition of HRW. Based on these results, it could be proposed that H2 might be an important regulator for enhancing the tolerance of Brassica campestris seedlings against Cd, mainly by governing reduced glutathione homeostasis. PMID:26445361

  8. PV solar electricity: status and future

    NASA Astrophysics Data System (ADS)

    Hoffmann, Winfried

    2006-04-01

    Within the four main market segments of PV solar electricity there are already three areas competitive today. These are off-grid industrial and rural as well as consumer applications. The overall growth within the past 8 years was almost 40 % p.a. with a "normal" growth of about 18 % p.a. for the first three market segments whereas the grid connected market increased with an astonishing 63 % p.a. The different growth rates catapulted the contribution of grid connected systems in relation to the total market from about one quarter 6 years ago towards more than three quarters today. The reason for this development is basically due to industry-politically induced market support programs in the aforementioned countries. It is quite important to outline under which boundary conditions grid connected systems will be competitive without support programs like the feed in tariff system in Germany, Spain and some more to come in Europe as well as investment subsidies in Japan, US and some other countries. It will be shown that in a more and more liberalized utility market worldwide electricity produced by PV solar electricity systems will be able to compete with their generating cost against peak power prices from utilities. The point of time for this competitiveness is mainly determined by the following facts: 1. Price decrease for PV solar electricity systems leading to an equivalent decrease in the generated cost for PV produced kWh. 2. Development of a truly liberalized electricity market. 3. Degree of irradiation between times of peak power demand and delivery of PV electricity. The first topic is discussed using price experience curves. Some explanations will be given to correlate the qualitative number of 20 % price decrease for doubling cumulative worldwide sales derived from the historic price experience curve with a more quantitative analysis based on our EPIA-Roadmap (productivity increase and ongoing improvements for existing technologies as well as development

  9. Involvement of the pepper antimicrobial protein CaAMP1 gene in broad spectrum disease resistance.

    PubMed

    Lee, Sung Chul; Hwang, In Sun; Choi, Hyong Woo; Hwang, Byung Kook

    2008-10-01

    Pathogen-inducible antimicrobial defense-related proteins have emerged as key antibiotic peptides and enzymes involved in disease resistance in plants. A novel antimicrobial protein gene, CaAMP1 (for Capsicum annuum ANTIMICROBIAL PROTEIN1), was isolated from pepper (C. annuum) leaves infected with Xanthomonas campestris pv vesicatoria. Expression of the CaAMP1 gene was strongly induced in pepper leaves not only during pathogen infection but also after exposure to abiotic elicitors. The purified recombinant CaAMP1 protein possessed broad-spectrum antimicrobial activity against phytopathogenic bacteria and fungi. CaAMP1:smGFP fusion protein was localized mainly in the external and intercellular regions of onion (Allium cepa) epidermal cells. The virus-induced gene silencing technique and gain-of-function transgenic plants were used to determine the CaAMP1 gene function in plant defense. Silencing of CaAMP1 led to enhanced susceptibility to X. campestris pv vesicatoria and Colletotrichum coccodes infection, accompanied by reduced PATHOGENESIS-RELATED (PR) gene expression. In contrast, overexpression of CaAMP1 in Arabidopsis (Arabidopsis thaliana) conferred broad-spectrum resistance to the hemibiotrophic bacterial pathogen Pseudomonas syringae pv tomato, the biotrophic oomycete Hyaloperonospora parasitica, and the fungal necrotrophic pathogens Fusarium oxysporum f. sp. matthiolae and Alternaria brassicicola. CaAMP1 overexpression induced the salicylic acid pathway-dependent genes PR1 and PR5 but not the jasmonic acid-dependent defense gene PDF1.2 during P. syringae pv tomato infection. Together, these results suggest that the antimicrobial CaAMP1 protein is involved in broad-spectrum resistance to bacterial and fungal pathogen infection.

  10. Involvement of the Pepper Antimicrobial Protein CaAMP1 Gene in Broad Spectrum Disease Resistance1[C][OA

    PubMed Central

    Lee, Sung Chul; Hwang, In Sun; Choi, Hyong Woo; Hwang, Byung Kook

    2008-01-01

    Pathogen-inducible antimicrobial defense-related proteins have emerged as key antibiotic peptides and enzymes involved in disease resistance in plants. A novel antimicrobial protein gene, CaAMP1 (for Capsicum annuum ANTIMICROBIAL PROTEIN1), was isolated from pepper (C. annuum) leaves infected with Xanthomonas campestris pv vesicatoria. Expression of the CaAMP1 gene was strongly induced in pepper leaves not only during pathogen infection but also after exposure to abiotic elicitors. The purified recombinant CaAMP1 protein possessed broad-spectrum antimicrobial activity against phytopathogenic bacteria and fungi. CaAMP1:smGFP fusion protein was localized mainly in the external and intercellular regions of onion (Allium cepa) epidermal cells. The virus-induced gene silencing technique and gain-of-function transgenic plants were used to determine the CaAMP1 gene function in plant defense. Silencing of CaAMP1 led to enhanced susceptibility to X. campestris pv vesicatoria and Colletotrichum coccodes infection, accompanied by reduced PATHOGENESIS-RELATED (PR) gene expression. In contrast, overexpression of CaAMP1 in Arabidopsis (Arabidopsis thaliana) conferred broad-spectrum resistance to the hemibiotrophic bacterial pathogen Pseudomonas syringae pv tomato, the biotrophic oomycete Hyaloperonospora parasitica, and the fungal necrotrophic pathogens Fusarium oxysporum f. sp. matthiolae and Alternaria brassicicola. CaAMP1 overexpression induced the salicylic acid pathway-dependent genes PR1 and PR5 but not the jasmonic acid-dependent defense gene PDF1.2 during P. syringae pv tomato infection. Together, these results suggest that the antimicrobial CaAMP1 protein is involved in broad-spectrum resistance to bacterial and fungal pathogen infection. PMID:18676663

  11. The Xanthomonas Type III Effector XopD Targets the Arabidopsis Transcription Factor MYB30 to Suppress Plant Defense[W

    PubMed Central

    Canonne, Joanne; Marino, Daniel; Jauneau, Alain; Pouzet, Cécile; Brière, Christian; Roby, Dominique; Rivas, Susana

    2011-01-01

    Plant and animal pathogens inject type III effectors (T3Es) into host cells to suppress host immunity and promote successful infection. XopD, a T3E from Xanthomonas campestris pv vesicatoria, has been proposed to promote bacterial growth by targeting plant transcription factors and/or regulators. Here, we show that XopD from the B100 strain of X. campestris pv campestris is able to target MYB30, a transcription factor that positively regulates Arabidopsis thaliana defense and associated cell death responses to bacteria through transcriptional activation of genes related to very-long-chain fatty acid (VLCFA) metabolism. XopD specifically interacts with MYB30, resulting in inhibition of the transcriptional activation of MYB30 VLCFA-related target genes and suppression of Arabidopsis defense. The helix-loop-helix domain of XopD is necessary and sufficient to mediate these effects. These results illustrate an original strategy developed by Xanthomonas to subvert plant defense and promote development of disease. PMID:21917550

  12. PV technology and success of solar electricity in Vietnam

    SciTech Connect

    Dung, T.Q.

    1997-12-31

    Since 1990 the PV Technology and the Solar electricity have been strongly developed in Vietnam. The PV experts of Solarlab have studied and set up an appropriate PV Technology responding to local Market needs. It has not only stood well but has been also transferred to Mali Republic and Lao P.D.R. The PV off grid systems of Solarlab demonstrate good efficiency and low prices. Over 60 solar stations and villages have been built to provide solar lighting for about 3000 families along the country in remote, mountainous areas and islands. 400 families are using stand-alone Solar Home Systems. The Solar electricity has been chosen for Rural Electrification and National Telecommunication Network in remote and mountainous regions. Many International projects in cooperation with FONDEM-France, SELF USA and Governmental PV projects have been realized by Solarlab. The experiences of maintenance, management and finance about PV development in Vietnam are also mentioned.

  13. Photovoltaic (PV) Power Systems for Enhancing Energy Security

    DTIC Science & Technology

    2012-05-24

    Energy and Environment Technology Transition – Supporting DoD Readiness, Sustainability, and the Warfighter Photovoltaic (PV) Power Systems for...to 00-00-2012 4. TITLE AND SUBTITLE Photovoltaic (PV) Power Systems for Enhancing Energy Security 5a. CONTRACT NUMBER 5b. GRANT NUMBER 5c...use of 235W solar panels Note [2] System AC Rating based upon typical .77 conversion factor from DC power to AC power ConsiderationNo. PV LAYOUT OPTION

  14. Long-Term Performance of the SERF PV Systems

    SciTech Connect

    Marion, B.; Adelstein, J.

    2003-05-01

    This paper provides the changes in performance ratings of two photovoltaic (PV) systems located on the roof of the Solar Energy Research Facility (SERF) building at the National Renewable Energy Laboratory (NREL) in Golden, Colorado. For the period of May 1994 to April 2002, the performance rating of the two PV systems decreased at the rate of 1% per year. Most of the changes in performance rating are attributed to changes in the performance of the PV arrays. But about a fifth of the observed changes were from the inverter not tracking the peak-power as effectively as the PV arrays aged.

  15. Coherent PV anomalies associated with deep moist convection

    NASA Astrophysics Data System (ADS)

    Weijenborg, C.; Chagnon, J.; Friederichs, P.; Hense, A.

    2016-12-01

    Potential Vorticity (PV) elegantly describes synoptic and planetary scale dynamics, but it has received less attention on smaller scales. On the convective scale PV is characterised by dipoles associated with convective cells. We hypothesise that the PV dipoles are quasi-balanced. If so, one should see a coherent evolution of the PV dipoles. Moreover, the PV dipoles might be described by a reduced order model. This hypothesis is tested by tracking convective cells in the nonhydrostatic COSMO-DE Numerical Weather Prediction (NWP) model during nine severe weather events. The 3135 convective cells used in this study are representative of deep moist convection over western Europe in the COSMO-DE model. Composites of the evolution of convective cells are discussed. Even when averaging over 3135 cells during nine cases, a clear horizontal PV dipole pattern can be seen, with associated flow anomalies. A normal mode analysis is performed, starting with the Chagnon-Gray model. We compare the quasi-stationary PV found using the normal modes analysis with the dominant patterns in the modeled convection. This is compared with the dominant patterns in modeled deep moist convection. We discuss a multivariate linear regression between the steady state wind and potential temperature fields obtained using the normal modes expansion with the dominant structures in the modeled data. The consistency of the PV dipoles implies that PV could be quasi-invertible on the convective weather scale. An application could therefore be data reduction.

  16. PV modules with optimized energy balance

    NASA Astrophysics Data System (ADS)

    Weixlberger, Johann; Bruckner, Richard

    2011-09-01

    The overall energy balance of a solar PV-module across its life time needs a consideration incl. its energy consumption during manufacturing process versus its energy harvesting capabilities during life time. A glass-glass-module based on thin tempered glass on front and backside can dramatically influence this overall balance, since more than 50 % of encapsulation materials manufacturing energy can be saved, followed by a an further impact on frameless mounting of light-weighted modules, reducing mounting costs and enabling simpler BIPV.

  17. Development of a Dispatchable PV Peak Shainv System. PV: Bonus Program - Phase 1 Report. Volume 1

    SciTech Connect

    1995-10-01

    This report summarizes the work performed by Delmarva Power and Light and its subcontractors in Phase 1 of the US Department of Energy's PV:BONUS Program. The purpose of the program is to develop products and systems for buildings which utilize photovoltaic (N) technology. Beginning with a cooperative research effort with the University of Delaware's Center for Energy and Environmental Policy Research Delmarva Power developed and demonstrated the concept of Dispatchable PV Peak Shaving. This concept and the system which resulted horn the development work are unique from other grid-connected PV systems because it combines a PV, battery energy storage, power conversion and control technologies into an integrated package. Phase 1 began in July 1993 with the installation of a test and demonstration system at Delmarva's Northern Division General Office building near Newark, Delaware. Following initial testing throughout the summer and fall of 1993, significant modifications were made under an amendment to the DOE contract. Work on Phase 1 concluded in the early spring of 1995. Significant progress towards the goal of commercializing the system was made during Phase 1, and is summarized. Based on progress in Phase 1, a proposal to continue the work in Phase 2 was submitted to the US DOE in May 1995. A contract amendment and providing funds for the Phase 2 work is expected in July 1995.

  18. A fatal case of poisoning related to new cathinone designer drugs, 4-methoxy PV8, PV9, and 4-methoxy PV9, and a dissociative agent, diphenidine.

    PubMed

    Kudo, Keiko; Usumoto, Yosuke; Kikura-Hanajiri, Ruri; Sameshima, Naomi; Tsuji, Akiko; Ikeda, Noriaki

    2015-09-01

    A woman in her thirties was found dead on a bed. Considerable amounts of "aroma liquid" and "bath salt" products and hypnotic drug tablets were scattered beside the bed. Autopsy showed pulmonary congestion and edema. Gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-mass spectrometry (LC-MS) analyses of "aroma liquid" and "bath salt" products showed the presence of new cathinone designer drugs, 4-methoxy PV8 (4-methoxy PHPP), PV9 (α-POP), and 4-methoxy PV9 (4-methoxy α-POP), and a dissociative agent, diphenidine. Drug screening in stomach contents, blood and hydrolyzed urine of the woman by GC-MS and liquid chromatography-tandem mass spectrometry (LC-MS/MS) revealed the presence of the above 4 types of drugs and 3 types of benzodiazepines, triazolam, flunitrazepam, and nitrazepam, and their metabolites. The above 7 drugs and 3 benzodiazepine metabolites were simultaneously determined by LC-MS/MS after modified QuEChERS (Quick, Easy, Cheap, Effective, Rugged, Safe) extraction using diazepam-d5 as the internal standard. The concentrations of 4-methoxy PV8, PV9, 4-methoxy PV9, and diphenidine in the femoral blood were 2.69, 0.743, 0.261, and 1.38μg/ml, respectively, which were significantly higher than concentrations reported in previous cases. Alcohol concentration in the femoral blood was 1.52mg/ml. Based on the pathological and toxicological findings, the cause of death was determined to be 3 types of cathinone drugs, 4-methoxy PV8, PV9, and 4-methoxy PV9, and diphenidine poisoning under the influence of 3 benzodiazepines and alcohol.

  19. Oxynitride Thin Film Barriers for PV Packaging

    SciTech Connect

    Glick, S. H.; delCueto, J. A.; Terwilliger, K. M.; Jorgensen, G. J.; Pankow, J. W.; Keyes, B. M.; Gedvilas, L. M.; Pern, F. J.

    2005-11-01

    Dielectric thin-film barrier and adhesion-promoting layers consisting of silicon oxynitride materials (SiOxNy, with various stoichiometry) were investigated. For process development, films were applied to glass (TCO, conductive SnO2:F; or soda-lime), polymer (PET, polyethylene terephthalate), aluminized soda-lime glass, or PV cell (a-Si, CIGS) substrates. Design strategy employed de-minimus hazard criteria to facilitate industrial adoption and reduce implementation costs for PV manufacturers or suppliers. A restricted process window was explored using dilute compressed gases (3% silane, 14% nitrous oxide, 23% oxygen) in nitrogen (or former mixtures, and 11.45% oxygen mix in helium and/or 99.999% helium dilution) with a worst-case flammable and non-corrosive hazard classification. Method employed low radio frequency (RF) power, less than or equal to 3 milliwatts per cm2, and low substrate temperatures, less than or equal to 100 deg C, over deposition areas less than or equal to 1000 cm2. Select material properties for barrier film thickness (profilometer), composition (XPS/FTIR), optical (refractive index, %T and %R), mechanical peel strength and WVTR barrier performance are presented.

  20. Selenium alleviates chromium toxicity by preventing oxidative stress in cabbage (Brassica campestris L. ssp. Pekinensis) leaves.

    PubMed

    Qing, Xuejiao; Zhao, Xiaohu; Hu, Chengxiao; Wang, Peng; Zhang, Ying; Zhang, Xuan; Wang, Pengcheng; Shi, Hanzhi; Jia, Fen; Qu, Chanjuan

    2015-04-01

    The beneficial role of selenium (Se) in alleviation of chromium (Cr)-induced oxidative stress is well established. However, little is known about the underlying mechanism. The impacts of exogenous Se (0.1mg/L) on Cr(1mg/L)-induced oxidative stress and antioxidant systems in leaves of cabbage (Brassica campestris L. ssp. Pekinensis) were investigated by using cellular and biochemical approaches. The results showed that supplementation of the medium with Se was effective in reducing Cr-induced increased levels of lipid peroxides and superoxide free radicals (O(-)2(·)), as well as increasing activities of superoxide dismutase (SOD) and peroxidase (POD). Meanwhile, 1mg/L Cr induced loss of plasma membrane integrity, growth inhibition, as well as ultrastructural changes of leaves were significantly reversed due to Se supplementation in the medium. In addition, Se application significantly altered the subcellular distribution of Cr which transported from mitochondria, nucleus and the cell-wall material to the soluble fraction and chloroplasts. However, Se application did no significant alteration of Cr effects on osmotic adjustment accumulating products. The study suggested that Se is able to protect leaves of cabbage against Cr toxicity by alleviation of Cr induced oxidative stress, and re-distribution of Cr in the subcellular of the leaf. Furthermore, free radicals, lipid peroxides, activity of SOD and POD, and subcellular distribution of Cr can be considered the efficient biomarkers to indicate the efficiency of Se to detoxification Cr.

  1. Roseomonas soli sp. nov., isolated from an agricultural soil cultivated with Chinese cabbage (Brassica campestris).

    PubMed

    Kim, Dong-Uk; Ka, Jong-Ok

    2014-03-01

    A bacterial strain, designated 5N26(T), was isolated from an agricultural soil cultivated with Chinese cabbage (Brassica campestris). Cells of this strain were Gram-reaction-negative, strictly aerobic, motile, non-spore-forming rods, and catalase- and urease-negative. The major fatty acids of strain 5N26(T) were C16 : 0 (7.5 %), C18 : 1 2-OH (13.4 %) and summed feature 8 (C18:1ω6c and/or C18:1ω7c; 63.2%). The polar lipid profile contained diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylcholine, phosphatidylmonomethylethanolamine and one unidentified aminolipid. The G+C content of the genomic DNA of strain 5N26(T) was 68.3 mol%. 16S rRNA gene sequence analysis showed that strain 5N26(T) was phylogenetically related to Roseomonas lacus TH-G33(T) and Roseomonas terrae DS-48(T) (97.0 % and 96.6 % sequence similarity, respectively). The results of genotypic and phenotypic data showed that strain 5N26(T) could be distinguished from phylogenetically related species, and that this strain represented a novel species within the genus Roseomonas, for which the name Roseomonas soli sp. nov. (type strain 5N26(T) = KACC 16376(T) = NBRC 109097(T)) is proposed.

  2. Structure of a novel N-acetyl-L-citrulline deacetylase from Xanthomonas campestris.

    PubMed

    Shi, Dashuang; Yu, Xiaolin; Roth, Lauren; Tuchman, Mendel; Allewell, Norma M

    2007-03-01

    The structure of a novel acetylcitrulline deacetylase from the plant pathogen Xanthomonas campestris has been solved by multiple-wavelength anomalous dispersion (MAD) using crystals grown from selenomethionine-substituted protein and refined at 1.75 A resolution. The asymmetric unit of the crystal contains one monomer consisting of two domains, a catalytic domain and a dimerization domain. The catalytic domain is able to bind a single Co(II) ion at the active site with no change in conformation. The dimerization domain forms an interface between two monomers related by a crystallographic two-fold symmetry axis. The interface is maintained by hydrophobic interactions between helices and hydrogen bonding between two beta strands that form a continuous beta sheet across the dimer interface. Because the dimers are also related by two-fold crystallographic axes, they pack together across the crystal via the dimerization domain, suggesting that higher order oligomers may form in solution. The polypeptide fold of the monomer is similar to the fold of Pseudomonas sp. carboxypeptidase G2 and Neisseria meningitidis succinyl diaminopimelate desuccinylase. Structural comparison among these enzymes allowed modeling of substrate binding and suggests a possible catalytic mechanism, in which Glu130 functions as a bifunctional general acid-base catalyst and the metal ion polarizes the carbonyl of the acetyl group.

  3. Photosynthetic carbon fixation characteristics of fruiting structures of Brassica campestris L

    SciTech Connect

    Singal, H.R.; Sheoran, I.S.; Singh, R.

    1987-04-01

    Activities of key enzymes of the Calvin cycle and C/sub 4/ metabolism, rates of CO/sub 2/ fixation, and the initial products of photosynthetic /sup 14/CO/sub 2/ fixation were determined in the podwall, seed coat (fruiting structures), and the subtending leaf (leaf below a receme) of Brassica campestris L. cv Toria. Compared to activities of ribulose-1,5-bisphosphate carboxylase and other Calvin cycle enzymes, e.g. NADP-glyceraldehyde-3-phosphate-dehydrogenase and ribulose-5-phosphate kinase, the activities of phosphoenol pyruvate carboxylase and other enzymes of C/sub 4/ metabolism, viz. NADP-malate dehydrogenase, NADP-malic enzyme, glutamate pyruvate transaminase, and glutamate oxaloacetate transaminase, were generally much higher in seed than in podwall and leaf. Podwall and leaf were comparable to each other. Pulse-chase experiments showed that in seed the major product of /sup 14/CO/sub 2/ assimilation was malate (in short time), whereas in podwall and leaf, the label initially appeared in 3-PGA. With time, the label moved to sucrose. In contrast to legumes, Brassica pods were able to fix net CO/sub 2/ during light. However, respiratory losses were very high during the dark period.

  4. Antioxidant, antibacterial, anti-inflammatory and wound healing effects of Artemisia campestris aqueous extract in rat.

    PubMed

    Ghlissi, Zohra; Sayari, Nadhim; Kallel, Rim; Bougatef, Ali; Sahnoun, Zouheir

    2016-12-01

    This study investigated some biological properties of Artemisia campestris aqueous extract (ACAE) as well its global chemical compositions. Twenty four rats were excised on the posterior neck skin area and divided into 4 groups, treated respectively with: sterile saline, glycerol, CICAFLORA and ACAE. The wound closure rate, histopathology evolution and the superoxide dismutase (SOD), catalase (CAT) and malondialdehyde (MDA) level in skin tissue were evaluated. Anti-inflammatory activity was studied by carrageenan-induced rat paw edema. Animals were divided into 3 groups pre-treated respectively with sterile saline, acetylsalicylic acid (AA) and ACAE. The antibacterial activity was tested against six bacteria and the antioxidant activity was estimated by the 1,1-diphenyl-2-picrylhydrazyl (DPPH), reducing power and β-carotene activities. Our results demonstrated a significant improvement in wound healing progression and in oxidative stress damage in the wounds tissues of ACAE-treated rats, compared to control. ACAE-treated rats revealed also a significant inhibition of carrageenan-induced hind paws edema as confirmed by the histological analysis. In addition to the antioxidant activity, ACAE showed considerable antibacterial activities. ACAE exhibited important wound healing effect probably due to the anti-inflammatory, antibacterial and antioxidant activities of its phytochemical contents. Therefore, this study confirms its popular use and highlights its promise in the development of new drugs. Copyright © 2016 Elsevier Masson SAS. All rights reserved.

  5. Pathology of Sarcocystis campestris infection in Richardson's ground squirrels (Spermophilus richardsoni).

    PubMed Central

    Wobeser, G; Cawthorn, R J; Gajadhar, A A

    1983-01-01

    Richardson's ground squirrels were infected with 1500 or 9000 sporocysts of Sarcocystis campestris from badgers. No lesions were found in animals killed one to three days postinfection (pi). Hepatitis and phlebitis of hepatic veins were present in animals killed between four and eight days pi. No meronts were detected in these animals, but the lesions suggested that a generation of merogony occurred in the hepatic veins. Meronts were found in endothelial cells in many tissues beginning on day 9 pi. They were most numerous on day 10 pi, and less so on day 11, and subsequently. Meronts were most numerous in the lung; none was found in liver or spleen. Four of ten squirrels infected with 1500 sporocysts in one trial died between days 11 and 13 pi. There were petechial hemorrhages in skeletal muscle, lung, serosal membranes, and brain in these animals, with microscopic evidence of pulmonary, myocardial, and brain injury. One animal infected with 9000 sporocysts had petechiae in the liver at six days pi. Foci of inflammation were visible in the myocardium and brain of animals killed to 64 days pi. This species may serve as an experimental model for sarcocystosis in domestic animals. Images Fig. 1. Fig. 2. Fig. 3. Fig. 4. Fig. 5. Fig. 6. PMID:6411306

  6. Alfalfa dodder (Cuscuta campestris) toxicity in horses: clinical, haematological and serum biochemical findings.

    PubMed

    Abutarbush, S M

    2013-07-27

    The objective of this observational study is to describe clinical, haematological and serum biochemical findings of horses affected with alfalfa dodder (Cuscuta campestris) toxicity. Twenty horses naturally exposed to alfalfa dodder toxicity were examined and information was collected on history and clinical signs. Physical examination was done on horses in the premises (n=20), and venous blood samples of 12 horses were submitted for haematology and serum biochemical examination for each horse. Abnormal clinical signs started around 36 hours after horses were fed the contaminated alfalfa. Abnormal signs were seen in 11 horses and those included diarrhoea (n=8), decreased appetite (n=7), neurological signs (n=4) and abdominal pain (n=1). Some horses had multiple clinical signs of the above. The results of complete blood cell count revealed leukocytopenia, neutropenia and thrombocytopenia. Serum biochemical analysis revealed decreased ALP, AST and CPK levels and increased direct bilirubin level. The used alfalfa was stopped immediately and a different alfalfa from a new container that did not contain any weeds was fed. Horses on the premises were observed closely, and the abnormal clinical signs resolved within three days. No treatment was implemented. Knowledge about toxicity of horses by Cuscuta species is scarce in the English veterinary literature and very limited.

  7. [Production and cytogenetics of intergeneric hybrids between Ogura CMS Brassica campestris var. purpuraria and Raphanus sativus].

    PubMed

    Huang, B Q; Chang, L; Ju, C M; Chen, J G

    2001-01-01

    Crosses between Ogura CMS Brassica campestris var. purpuraria (AA, 2n = 20) and Raphanus sativus (RR, 2n = 18) were made and many intergeneric hybrids were produced. The F1 seedlings did not show chlorosis at low temperature. When red Raphanus sativus varieties were used as male parent, the leaf petiole and leaf vein of F1 plants were purple, and when white Raphanus sativus varieties were used as male parent, the leaf petiole and leaf vein of F1 plants were not purple. All the F1 plants had white flowers and normal honey glands. Male gametes of the F1 were highly sterile and female gametes of the F1 were partly fertile. Cytological studies indicated that chromosome number of the F1 was 2n = 19 as expected, the mean chromosome pairing pattern was 15.53 I + 1.34 II + 0.25 III + 0.01 IV. Most chromosomes exsistet as univalents, but there also exsisted some bivalents, trivalents and even tetravalents, suggesting that chromosome set A was partly homologous with chromosome set R.

  8. Crystallization and preliminary X-ray diffraction analysis of Xaa-Pro dipeptidase from Xanthomonas campestris.

    PubMed

    Kumar, Ashwani; Are, Venkata Narayana; Ghosh, Biplab; Agrawal, Utsavi; Jamdar, Sahayog N; Makde, Ravindra D; Sharma, Surinder M

    2014-09-01

    Xaa-Pro dipeptidase (XPD; prolidase; EC 3.4.13.9) specifically hydrolyzes dipeptides with a prolyl residue at the carboxy-terminus. Xanthomonas spp. possess two different isoforms of XPD (48 and 43 kDa) which share ∼24% sequence identity. The XPD of 43 kDa in size (XPD43) from Xanthomonas spp. is unusual as it lacks the strictly conserved tyrosine residue (equivalent to Tyr387 in Escherichia coli aminopeptidase P) that is suggested to be important in the proton-shuttle transfer required for catalysis in the M24B (MEROPS) family. Here, the crystallization and preliminary X-ray analysis of XPD43 from X. campestris (GenBank accession No. NP_637763) are reported. Recombinant XPD43 was crystallized using the microbatch-under-oil technique. Diffraction data were collected on the recently commissioned protein crystallography beamline (PX-BL21) at the Indian synchrotron (Indus-2, 2.5 GeV) to 1.83 Å resolution with 100% completeness. The crystal belonged to space group P212121, with unit-cell parameters a = 84.32, b = 105.51, c = 111.35 Å. Two monomers are expected to be present in the asymmetric unit of the crystal, corresponding to a solvent content of 58%. Structural analysis of XPD43 will provide new insights into the role of the conserved residues in catalysis in the M24B family.

  9. Method for improving specific xanthan productivity during continuous fermentation. [Xanthomonas sp. , Xanthomonas campestris

    SciTech Connect

    Weisrock, W.P.

    1982-01-19

    The heteropolysaccharides produced by the action of Xanthomonas bacteria on carbohydrate media have a potential application as film forming agents and as thickeners for oil field drilling fluids, fracturing liquids, and emulsifying, stabilizing, and sizing agents. Heteropolysaccharides, particularly xanthan gum, have significant potential as mobility control agents in micellar polymer flooding. Xanthan gum has excellent viscosifying properties at low concentration; it is resistant to shear degradation and exhibits only minimal losses in viscosity as a function of temperature, pH, and ionic strength. During continuous culture, the concentration of biomass is set by the concentration of the limiting nutrient being fed with the medium and biomass concentration is varied by raising or lowering the limiting nutrient concentration. By growing a species of the genus Xanthomonas such as Xanthomonas campestris, in continuous culture in a medium containing glucose, mineral salts, and NH/sub 4/Cl and either glutamate or glutamate plus yeast extract, the specific productivity is improved by first operating and then raising the cell concentration. 16 claims.

  10. Structure of a Novel N-acetyl-L-citrulline Deacetylase from Xanthomonas campestris

    SciTech Connect

    Shi,D.; Yu, X.; Roth, L.; Tuchman, M.; Allewell, N.

    2007-01-01

    The structure of a novel acetylcitrulline deacetylase from the plant pathogen Xanthomonas campestris has been solved by multiple-wavelength anomalous dispersion (MAD) using crystals grown from selenomethionine-substituted protein and refined at 1.75 {angstrom} resolution. The asymmetric unit of the crystal contains one monomer consisting of two domains, a catalytic domain and a dimerization domain. The catalytic domain is able to bind a single Co(II) ion at the active site with no change in confirmation. the dimerization domain forms an interface between two monomers related by a crystallographic two-fold symmetry axis. The interface is maintained by hydrophobic interactions between helices and hydrogen bonding between two {beta} strands that form a continuous {beta} sheet across the dimer interface. Because the dimers are also related by two-fold crystallographic axes, they pack together across the crystal via the dimerization domain, suggesting that higher order oligomers may form in solution. The polypeptide fold of the monomer is similar to the fold of Pseudomonas sp. carboxypeptidase G2 and Neisseria meningitidis succinyl diaminopimelate desuccinylase. Structural comparison among these enzymes allowed modeling of substrate binding and suggests a possible catalytic mechanism, in which Glu130 functions as a bifunctional general acid-base catalyst and the metal ion polarizes the carbonyl of the acetyl group.

  11. Method for improving specific Xanthan productivity during continuous fermentation. [Xanthomonas sp. , Xanthomonas campestris

    SciTech Connect

    Weisrock, W.P.

    1982-01-19

    The heteropolysaccharides produced by the action of Xanthomonas bacteria on carbohydrate media have a potential application as film forming agents and as thickeners for oil field drilling fluids, fracturing liquids, and emulsifying, stabilizing, and sizing agents. Heteropolysaccharides, particularly xanthan gum, have significant potential as mobility control agents in micellar polymer flooding. Xanthan gum has excellent viscosifying properties at low concentration; it is resistant to shear degradation and exhibits only minimal losses in viscosity as a function of temperature, pH, and ionic strength. During continuous culture, the concentration of biomass is set by the concentration of the limiting nutrient being fed with the medium and biomass concentration is varied by raising or lowering the limiting nutrient concentration. By growing a species of the genus Xanthomonas such as Xanthomonas campestris, in continuous culture in a medium containing glucose, mineral salts, and NH/sub 4/Cl and either glutamate or glutamate plus yeast extract, the specific productivity is improved by first operating and then raising the cell concentration. 16 claims.

  12. Xanthan production by a native strain of X. campestris and evaluation of application in EOR.

    PubMed

    Nasr, Shaghayegh; Soudi, Mohammad Reza; Haghighi, Manouchehr

    2007-09-01

    In this study, we used a native strain of X. campestris for xanthan production in lab-scale fermentor and the product was recovered with organic solvents and dried. Then we studied the potential usage of our products in different harsh conditions, including heat, pH and salinity treatments. Furthermore, we used 2D-micromodel for microbial oil recovery investigations. According to present experiments, temperature and salt contents did not have a significant influence on rheological behavior of xanthan solutions and these aqueous solutions maintained at least 80% of their primary viscosity. In addition, these solutions were resistant to a broad range of pH variations. Viscosity of the xanthan solution was increased as it was heated over 120 degrees C. Micro-model experiments showed that the most efficient concentration of xanthan for Enhanced Oil Recovery (EOR) is 1000 mg L(-1) and 53% of original oil in place was recovered, which showed remarkable increase comparing to original oil in place that was recovered (31%) from sole water flooding. The same or even better results were obtained from native xanthan, when its properties were compared to those of a commercial sample which was gifted by NIOC.

  13. Step-Stress Accelerated Degradation Testing (SSADT) for Photovoltaic (PV) Devices and Cells (Presentation)

    SciTech Connect

    Lee, J.; Elmore, R.; Suh, C.; Jones, W.

    2010-10-01

    Presentation on step-stress accelerated degradation testing (SSADT) for photovoltaics (PV). Developed are a step-stress degradation test (SSADT) for PV reliability tests and a lifetime prediction model for PV products.

  14. Impact of r