The 73-kb pIAA plasmid increases competitive fitness of Pseudomonas syringae subspecies savastanoi in oleander

1993 ◽  
Vol 39 (7) ◽  
pp. 659-664 ◽  
Author(s):  
Sara E. Silverstone ◽  
David G. Gilchrist ◽  
Richard M. Bostock ◽  
Tsune Kosuge
Keyword(s):  
Population Density ◽  
Pseudomonas Syringae ◽  
Indoleacetic Acid ◽  
Leaf Tissue ◽  
Insertion Mutant ◽  
Wild Type ◽  
In Planta ◽  
Bacterial Strains ◽  
Growth And Survival ◽  
Iaa Production

Pseudomonas syringae subsp. savastanoi causes tumors on olive and oleander by producing the plant growth regulators indoleacetic acid (IAA) and cytokinins following infection of the plant. The contribution of IAA production to the ability of P. syringae subsp. savastanoi to grow and survive in oleander leaf tissue was studied. Bacterial strains differing only with respect to IAA production were characterized. Growth and survival of wild-type and two mutant strains of P. syringae subsp. savastanoi in oleander leaf tissue were monitored by weekly colony counts and IAA plate assays. Growth rate of the three strains in culture and in planta did not differ significantly. However, the wild-type strain reached a higher population density and maintained its maximum density at least 9 weeks longer than either mutant population. An insertion mutant containing the IAA plasmid (pIAA), but incapable of IAA production, did not maintain a higher population density than a strain cured of the IAA plasmid. The pIAA-cured strain maintained a higher population density when coinoculated with an IAA-producing strain than when inoculated alone. These results suggest that IAA production may contribute to the fitness of P. syringae subsp. savastanoi in oleander tissue and that the iaa operon alone may be responsible for the competitive advantage of cells harboring pIAA.Key words: indoleacetic acid, bacterial ecology.

scholarly journals Virulence of Pseudomonas syringae pv. tomato DC3000 Is Influenced by the Catabolite Repression Control Protein Crc

2017 ◽  
Vol 30 (4) ◽  
pp. 283-294 ◽  
Author(s):  
Suma Chakravarthy ◽  
Bronwyn G. Butcher ◽  
Yingyu Liu ◽  
Katherine D’Amico ◽  
Matthew Coster ◽  
...  
Keyword(s):  
Pseudomonas Syringae ◽  
Catabolite Repression ◽  
Type Strain ◽  
Wild Type Strain ◽  
Rna Binding ◽  
Plant Diseases ◽  
Wild Type ◽  
In Planta ◽  
Growth And Survival ◽  
Control Protein

Pseudomonas syringae infects diverse plant species and is widely used as a model system in the study of effector function and the molecular basis of plant diseases. Although the relationship between bacterial metabolism, nutrient acquisition, and virulence has attracted increasing attention in bacterial pathology, it is largely unexplored in P. syringae. The Crc (catabolite repression control) protein is a putative RNA-binding protein that regulates carbon metabolism as well as a number of other factors in the pseudomonads. Here, we show that deletion of crc increased bacterial swarming motility and biofilm formation. The crc mutant showed reduced growth and symptoms in Arabidopsis and tomato when compared with the wild-type strain. We have evidence that the crc mutant shows delayed hypersensitive response (HR) when infiltrated into Nicotiana benthamiana and tobacco. Interestingly, the crc mutant was more susceptible to hydrogen peroxide, suggesting that, in planta, the mutant may be sensitive to reactive oxygen species generated during pathogen-associated molecular pattern–triggered immunity (PTI). Indeed, HR was further delayed when PTI-induced tissues were challenged with the crc mutant. The crc mutant did not elicit an altered PTI response in plants compared with the wild-type strain. We conclude that Crc plays an important role in growth and survival during infection.

Identification of a DNA region required for growth of Pseudomonas syringae pv. tomato on tomato plants

1992 ◽  
Vol 38 (9) ◽  
pp. 883-890 ◽  
Author(s):  
Dennis P. Jackson ◽  
Douglas A. Gray ◽  
Vincent L. Morris ◽  
Diane A. Cuppels
Keyword(s):  
Organic Acids ◽  
Pseudomonas Syringae ◽  
Acid Metabolism ◽  
Carbon Sources ◽  
Hypersensitive Reaction ◽  
Wild Type ◽  
In Planta ◽  
Tomato Plants ◽  
Tartaric Acids ◽  
Nonpathogenic Strain

The prototrophic Pseudomonas syringae pv. tomato mutant DC3481, which is the result of a single-site Tn5 insertion, cannot grow and cause disease on tomato plants and cannot use the major organic acids of tomato, i.e., citric, malic, succinic, and tartaric acids, as sole carbon sources. Although nonpathogenic, strain DC3481 can still induce a hypersensitive reaction in nonhost plants. We have identified a 30-kb fragment of P. syringae pv. tomato wild-type DNA that can complement this mutant. EcoRI fragments from this region were subcloned and individually subjected to functional complementation analysis. The 3.8-kb fragment, which was the site of the Tn5 insertion, restored pathogenicity and the ability to use all the major organic acids of tomato as carbon sources. It shares sequence homology with several P. syringae pathovars but not other bacterial tomato pathogens. Our results indicate that sequences on the 3.8-kb EcoRI fragment are required for both the ability to grow on tomato leaves (and thus cause disease) and the utilization of carboxylic acids common to tomato. The 3.8-kb fragment may contain a sequence (or sequences) that regulates both traits. Key words: Pseudomonas syringae pv. tomato, phytopathogenicity, Tn5, tricarboxylic acid metabolism, bacterial speck, growth in planta.

scholarly journals Immunogold Labeling of Hrp Pili of Pseudomonas syringae pv. tomato Assembled in Minimal Medium and In Planta

2001 ◽  
Vol 14 (2) ◽  
pp. 234-241 ◽  
Author(s):  
Wenqi Hu ◽  
Jing Yuan ◽  
Qiao-Ling Jin ◽  
Patrick Hart ◽  
Sheng Yang He
Keyword(s):  
Arabidopsis Thaliana ◽  
Pseudomonas Syringae ◽  
Minimal Medium ◽  
Leaf Tissue ◽  
Hypersensitive Reaction ◽  
Immunogold Labeling ◽  
Structural Protein ◽  
In Planta ◽  
Hrp Genes

Hypersensitive reaction and pathogenicity (hrp) genes are required for Pseudomonas syringae pv. tomato (Pst) DC3000 to cause disease in susceptible tomato and Arabidopsis thaliana plants and to elicit the hypersensitive response in resistant plants. The hrp genes encode a type III protein secretion system known as the Hrp system, which in Pst DC3000 secretes HrpA, HrpZ, HrpW, and AvrPto and assembles a surface appendage, named the Hrp pilus, in hrp-gene-inducing minimal medium. HrpA has been suggested to be the Hrp pilus structural protein on the basis of copurification and mutational analyses. In this study, we show that an antibody against HrpA efficiently labeled Hrp pili, whereas antibodies against HrpW and HrpZ did not. Immunogold labeling of bacteria-infected Arabidopsis thaliana leaf tissue with an Hrp pilus antibody revealed a characteristic lineup of gold particles around bacteria and/or at the bacterium-plant contact site. These results confirm that HrpA is the major structural protein of the Hrp pilus and provide evidence that Hrp pili are assembled in vitro and in planta.

scholarly journals Components of the Pseudomonas syringae Type III Secretion System Can Suppress and May Elicit Plant Innate Immunity

2010 ◽  
Vol 23 (6) ◽  
pp. 727-739 ◽  
Author(s):  
Hye-Sook Oh ◽  
Duck Hwan Park ◽  
Alan Collmer
Keyword(s):  
Pseudomonas Syringae ◽  
Type Iii Secretion ◽  
Type Iii Secretion System ◽  
Leaf Tissue ◽  
Secretion System ◽  
Colony Development ◽  
Plant Responses ◽  
In Planta ◽  
Callose Deposition ◽  
Type Iii

The type III secretion system (T3SS) of Pseudomonas syringae translocates into plant cells multiple effectors that suppress pathogen-associated molecular pattern (PAMP)-triggered immunity (PTI). P. syringae pv. tomato DC3000 no longer delivers the T3SS translocation reporter AvrPto-Cya in Nicotiana benthamiana leaf tissue in which PTI was induced by prior inoculation with P. fluorescens(pLN18). Cosmid pLN18 expresses the T3SS system of P. syringae pv. syringae 61 but lacks the hopA1Psy61 effector gene. P. fluorescens(pLN18) expressing HrpHPtoDC3000 or HopP1PtoDC3000, two T3SS-associated putative lytic transglycosylases, suppresses PTI, based on multiple assays involving DC3000 challenge inoculum (AvrPto-Cya translocation, hypersensitive response elicitation, and colony development in planta) or on plant responses (vascular dye uptake or callose deposition). Analysis of additional mutations in pHIR11 derivatives revealed that the pLN18-encoded T3SS elicits a higher level of reactive oxygen species (ROS) than does P. fluorescens without a T3SS, that enhanced ROS production is dependent on the HrpK1 translocator, and that HopA1Psy61 suppresses ROS elicitation attributable to both the P. fluorescens PAMPs and the presence of a functional T3SS.

scholarly journals Identification of a putative DNA-binding protein in Arabidopsis that acts as a susceptibility hub and interacts with multiple Pseudomonas syringae effectors

2020 ◽  
Author(s):  
Karl Schreiber ◽  
Jennifer D Lewis
Keyword(s):  
Pseudomonas Syringae ◽  
Transcriptional Profiling ◽  
Effector Proteins ◽  
Ribosomal Protein Genes ◽  
Loss Of Function ◽  
Wild Type ◽  
In Planta ◽  
Arabidopsis Protein ◽  
Wide Range

Phytopathogens use secreted effector proteins to suppress host immunity and promote pathogen virulence, and there is increasing evidence that the host-pathogen interactome comprises a complex network. In an effort to identify novel interactors of the Pseudomonas syringae effector HopZ1a, we performed a yeast two-hybrid screen that identified a previously uncharacterized Arabidopsis protein that we designate HopZ1a Interactor 1 (ZIN1). Additional analyses in yeast and in planta revealed that ZIN1 also interacts with several other P. syringae effectors. We show that an Arabidopsis loss-of-function zin1 mutant is less susceptible to infection by certain strains of P. syringae, while overexpression of ZIN1 results in enhanced susceptibility. Functionally, ZIN1 exhibits topoisomerase-like activity in vitro. Transcriptional profiling of wild-type and zin1 Arabidopsis plants inoculated with P. syringae indicated that while ZIN1 regulates a wide range of pathogen-responsive biological processes, the list of genes more highly expressed in zin1 versus wild-type plants was particularly enriched for ribosomal protein genes. Altogether, these data illuminate ZIN1 as a potential susceptibility hub that interacts with multiple effectors to influence the outcome of plant-microbe interactions.

scholarly journals Quorum Sensing Regulates Exopolysaccharide Production, Motility, and Virulence in Pseudomonas syringae

2005 ◽  
Vol 18 (7) ◽  
pp. 682-693 ◽  
Author(s):  
Beatriz Quiñones ◽  
Glenn Dulla ◽  
Steven E. Lindow
Keyword(s):  
Quorum Sensing ◽  
Pseudomonas Syringae ◽  
Type Strain ◽  
Double Mutant ◽  
Wild Type Strain ◽  
Homoserine Lactone ◽  
Brown Spot ◽  
Wild Type ◽  
In Planta ◽  
Alginate Production

The N-acyl homoserine lactone (AHL)-mediated quorumsensing system in the phytopathogen Pseudomonas syringae pv. syringae requires the AHL synthase AhlI and the regulator AhlR, and is additionally subject to regulation by AefR. The contribution of quorum sensing to the expression of a variety of traits expected to be involved in epiphytic fitness and virulence of P. syringae were examined. Both an aefR- mutant and an ahlR- double mutant, deficient in AHL production, were significantly impaired in alginate production and had an increased susceptibility to hydrogen peroxide compared with the wild-type strain. These mutants were hypermotile in culture, invaded leaves more rapidly, and caused an increased incidence of brown spot lesions on bean leaves after a 48-h moist incubation. Interestingly, an aefR- mutant was both the most motile and virulent. Like the wild-type strain, the AHL-deficient mutant strains incited water-soaked lesions on bean pods. However, lesions caused by an ahlI- ahlR- double mutant were larger, whereas those incited by an aefR- mutant were smaller. In contrast, tissue maceration of pods, which occurs at a later stage of infection, was completely abolished in the AHL-deficient mutants. Both the incidence of disease and in planta growth of P. syringae pv. tabaci were greatly reduced in transgenic tobacco plants that produced AHL compared with wild-type plants. These results demonstrate that quorum sensing in P. syringae regulates traits that contribute to epiphytic fitness as well as to distinct stages of disease development during plant infection.

Bacteria occurring in onion (Allium cepa L.) foliage in Puerto Rico.

1969 ◽  
pp. 199-219
Author(s):  
Juan Calle-Bellido ◽  
Lydia I. Rivera-Vargas ◽  
Myrna Alameda ◽  
Irma Cabrera
Keyword(s):  
Puerto Rico ◽  
Pseudomonas Syringae ◽  
Allium Cepa ◽  
Leaf Tissue ◽  
Field Conditions ◽  
Bacterial Strains ◽  
Burkholderia Glumae ◽  
Pathogenic Potential ◽  
Allium Cepa L ◽  
Leaf Tissues

Bacteria associated with foliar symptoms of onion (Allium cepa L.) were examined in the southern region of Puerto Rico from January through April 2004. Different symptoms were observed in onion foliage of cultivars 'Mercedes' and 'Excalibur' at Juana Díaz and Santa Isabel, Puerto Rico. Ellipsoidal sunken lesions with soft rot and disruption of tissue were the most common symptoms observed in onion foliage in field conditions. From a total of 39 bacterial strains isolated from diverse symptoms in onion foliage, 38% were isolated from soft rotting lesions. Ninety-two percent of the bacteria isolated from onion foliage was Gram negative. Pantoea spp. with 25%, was the most frequently isolated genus, followed by Pasteurella spp. and Serratia rubidae with 10% each. Fifty- six percent of the strains held plant pathogenic potential; these strains belong to the genera Acidovorax sp., Burkholderia sp., Clavibacter sp., Curtobacterium sp., Enterobacter sp., Pantoea spp., Pseudomonas spp., and Xanthomonas spp. Pathogenicity tests showed that seven out of eight tested bacterial strains evaluated under field conditions caused symptoms in onion foliage for both cultivars. Acidovorax avenae subsp. citrulli, Burkholderia glumae, Pantoea agglomerans, P. dispersa, Pseudomonas sp., Xanthomonas sp., and Xanthomonas-Wke sp. were pathogenic to leaf tissues. Clavibacter michiganensis was not pathogenic to leaf tissues. Other bacteria identified as associated with onion leaf tissue were Curtobacterium flaccumfaciens, Cytophaga sp., Enterobacter cloacae, Flavimonas oryzihabitans, Mannheimia haemolytica, Pantoea stewartii, Pasteurella anatis, P. bettyae, P. langaaensis, Photobacterium damselae, Pseudomonas syringae pv. aptata, Rhizobium radiobacter, Serratia rubidae, Sphingobacterium spiritivorum, Sphingomonas sanguinis, and an unknown strain. This paper is the first survey of bacteria associated with onion foliage in Puerto Rico. The role of non- phytopathogenic bacteria associated with the life cycle of onion under field conditions remains unknown.

scholarly journals The Alternative Sigma Factor RpoN Is Required for hrpActivity in Pseudomonas syringae pv. Maculicola and Acts at the Level of hrpL Transcription

2000 ◽  
Vol 182 (12) ◽  
pp. 3508-3516 ◽  
Author(s):  
Erik L. Hendrickson ◽  
Pablo Guevera ◽  
Frederick M. Ausubel
Keyword(s):  
Pseudomonas Syringae ◽  
Transcriptional Activity ◽  
Constitutive Expression ◽  
Wild Type ◽  
In Planta ◽  
Disease Symptoms ◽  
Nicotiana Tobacum ◽  
Mutant Strains

ABSTRACT β-Glucuronidase (uidA) reporter gene fusions were constructed for the hrpZ, hrpL, andhrpS genes from the phytopathogen Pseudomonas syringae pv. maculicola strain ES4326. These reporters, as well as an avrRpt2-uidA fusion, were used to measure transcriptional activity in ES4326 and a ES4326 rpoNmutant. rpoN was required for the expression ofavrRpt2, hrpZ, and hrpL in vitro in minimal media and in vivo when infiltrated into Arabidopsis thaliana leaves. In contrast, the expression of hrpSwas essentially the same in wild-type and rpoN mutant strains. Constitutive expression of hrpL in anrpoN mutant restored hrpZ transcription to wild-type levels, restored the hypersensitive response when infiltrated into tobacco (Nicotiana tobacum), and partially restored the elicitation of virulence-related symptoms but not growth when infiltrated into Arabidopsis leaves. These data indicate that rpoN-mediated control of hrp gene expression acts at the level of hrpL and that in planta growth of P. syringae is not required for the elicitation of disease symptoms.

scholarly journals Antagonistic Interactions Between Strains of Xanthomonas oryzae pv. oryzae

2003 ◽  
Vol 93 (6) ◽  
pp. 705-711 ◽  
Author(s):  
Christopher Dardick ◽  
Francisco Goes da Silva ◽  
Yuwei Shen ◽  
Pamela Ronald
Keyword(s):  
Xanthomonas Oryzae ◽  
Growth Curve Analysis ◽  
R Gene ◽  
Wild Type ◽  
In Planta ◽  
Bacterial Strains ◽  
Korean Strain ◽  
Rice Bacterial Blight ◽  
Antagonistic Interactions

The ability of some phytopathogenic bacterial strains to inhibit the growth of others in mixed infections has been well documented. Here we report that such antagonistic interactions occur between several wild-type strains of the rice bacterial blight pathogen Xanthomonas oryzae pv. oryzae. In mixed inoculations, a wild-type Philippine strain was found to inhibit the growth of a wild-type Korean strain. Furthermore, a nonpathogenic mutant of the Philippine strain maintained these antagonistic properties. Growth curve analysis indicated that both the wild-type Philippine strain and its nonpathogenic mutant inhibited the growth of the Korean strain 2 days after infection and prior to the onset of disease symptoms. When mixed with the nonpathogenic mutant, 10 out of 18 diverse wild-type X. oryzae pv. oryzae strains did not cause disease. Conversely, three of the strains that were not affected by the nonpathogenic mutant were found to inhibit the growth of both the wild-type and mutant Philippine strains, indicating that antagonism is widespread and strain specific. The observed growth inhibition occurred only in planta and did not correlate with bacteriocin activity in vitro. Antagonistic interactions also were found to affect resistance (R) gene-mediated resistance. The R gene Xa21 was capable of protecting rice plants coinoculated with nonantagonistic virulent and avirulent strains; however, when avirulent strains were coinoculated with virulent antagonistic strains, disease ensued. Taken together, these results indicate that X. oryzae pv. oryzae has evolved strategies to compete with rival strains in a fashion that allows virulent strains to evade R gene-mediated protection even when avirulent strains are present in the inoculum.

scholarly journals The Role of Ethylene Production in Virulence of Pseudomonas syringae pvs. glycinea and phaseolicola

2001 ◽  
Vol 91 (5) ◽  
pp. 511-518 ◽  
Author(s):  
Helge Weingart ◽  
Henriette Ullrich ◽  
Klaus Geider ◽  
Beate Völksch
Keyword(s):  
Pseudomonas Syringae ◽  
Ethylene Production ◽  
Type Strain ◽  
Wild Type Strain ◽  
Wild Type ◽  
In Planta ◽  
Population Sizes ◽  
Soybean Plants ◽  
Type Strains

The importance of ethylene production for virulence of Pseudomonas syringae pvs. glycinea and phaseolicola was assayed by comparing bacterial multiplication and symptom development in bean and soybean plants inoculated with ethylene-negative (efe) mutants and wild-type strains. The efe mutants of Pseudomonas syringae pv. glycinea were significantly reduced in their ability to grow in planta. However, the degree of reduction was strain-dependent. Population sizes of efe mutant 16/83-E1 that did not produce the phototoxin coronatine were 10- and 15-fold lower than those of the wild-type strain on soybean and on bean, and 16/83-E1 produced very weak symptoms compared with the wild-type strain. The coronatine-producing efe mutant 7a/90-E1 reached fourfold and twofold lower population sizes compared with the wild-type strain on soybean and bean, respectively, and caused disease symptoms typical of the wild-type strain. Experiments with ethylene-insensitive soybeans confirmed these results. The virulence of the wild-type strains was reduced to the same extent in ethylene-insensitive soybean plants as the virulence of the efe mutants in ethylene-susceptible soybeans. In contrast, the virulence of Pseudomonas syringae pv. phaseolicola was not affected by disruption of the efe gene.

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