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

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.Key words: auxotroph, plant pathogen, resistance.

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rpfF Mutants of Xanthomonas oryzae pv. oryzae Are Deficient for Virulence and Growth Under Low Iron Conditions

Molecular Plant-Microbe Interactions ◽

10.1094/mpmi.2002.15.5.463 ◽

2002 ◽

Vol 15 (5) ◽

pp. 463-471 ◽

Cited By ~ 85

Author(s):

Subhadeep Chatterjee ◽

Ramesh V. Sonti

Keyword(s):

Xanthomonas Campestris ◽

Iron Supplementation ◽

Extracellular Enzymes ◽

Phenotypic Expression ◽

Tetracycline Resistance ◽

Extracellular Enzyme ◽

Extracellular Polysaccharide ◽

Xanthomonas Oryzae ◽

Uptake System ◽

Serious Disease

Xanthomonas oryzae pv. oryzae causes bacterial leaf blight, a serious disease of rice. In the related bacterium Xanthomonas campestris pv. campestris, the rpfF gene is involved in production of a diffusible extracellular factor (DSF) that positively regulates synthesis of virulence-associated functions like extracellular polysaccharide (EPS) and extracellular enzymes. Transposon insertions in the rpfF homolog of X. oryzae pv. oryzae are deficient for virulence and production of a DSF but are proficient for EPS and extracellular enzyme production. The rpfF X. oryzae pv. oryzae mutants exhibit an unusual tetracycline susceptibility phenotype in which exogenous iron supplementation is required for phenotypic expression of a tetracycline resistance determinant that is encoded on an introduced plasmid. The rpfF X. oryzae pv. oryzae mutants also overproduce one or more siderophores and exhibit a growth deficiency under low iron conditions as well as in the presence of reducing agents that are expected to promote the conversion of Fe+3 to Fe+2. Exogenous iron supplementation promotes migration of rpfF X. oryzae pv. oryzae mutants in rice leaves. The results suggest that rpfF may be involved in controlling an iron-uptake system of X. oryzae pv. oryzae and that an inability to cope with the conditions of low iron availability in the host may be the reason for the virulence deficiency of the rpfF X. oryzae pv. oryzae mutants.

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Role of the FeoB Protein and Siderophore in Promoting Virulence of Xanthomonas oryzae pv. oryzae on Rice

Journal of Bacteriology ◽

10.1128/jb.01558-09 ◽

2010 ◽

Vol 192 (12) ◽

pp. 3187-3203 ◽

Cited By ~ 47

Author(s):

Alok Pandey ◽

Ramesh V. Sonti

Keyword(s):

Gene Cluster ◽

Functional Characterization ◽

Biosynthetic Gene Cluster ◽

Xanthomonas Oryzae ◽

In Planta ◽

Growth Deficiency ◽

Constitutive Production ◽

Serious Disease

ABSTRACT Xanthomonas oryzae pv. oryzae causes bacterial blight, a serious disease of rice. Our analysis revealed that the X. oryzae pv. oryzae genome encodes genes responsible for iron uptake through FeoB (homolog of the major bacterial ferrous iron transporter) and a siderophore. A mutation in the X. oryzae pv. oryzae feoB gene causes severe virulence deficiency, growth deficiency in iron-limiting medium, and constitutive production of a siderophore. We identified an iron regulated xss gene cluster, in which xssABCDE ( X anthomonas siderophore synthesis) and xsuA ( X anthomonas siderophore utilization) genes encode proteins involved in biosynthesis and utilization of X. oryzae pv. oryzae siderophore. Mutations in the xssA, xssB, and xssE genes cause siderophore deficiency and growth restriction under iron-limiting conditions but are virulence proficient. An xsuA mutant displayed impairment in utilization of native siderophore, suggesting that XsuA acts as a specific receptor for a ferric-siderophore complex. Histochemical and fluorimetric assays with gusA fusions indicate that, during in planta growth, the feoB gene is expressed and that the xss operon is not expressed. This study represents the first report describing a role for feoB in virulence of any plant-pathogenic bacterium and the first functional characterization of a siderophore-biosynthetic gene cluster in any xanthomonad.

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Growth Deficiency of a Xanthomonas oryzae pv. oryzae fur Mutant in Rice Leaves Is Rescued by Ascorbic Acid Supplementation

Molecular Plant-Microbe Interactions ◽

10.1094/mpmi-18-0644 ◽

2005 ◽

Vol 18 (7) ◽

pp. 644-651 ◽

Cited By ~ 35

Author(s):

Sujatha Subramoni ◽

Ramesh V. Sonti

Keyword(s):

Ascorbic Acid ◽

Slow Growth ◽

Xanthomonas Oryzae ◽

Ferric Uptake Regulator ◽

Growth Deficiency ◽

Impaired Ability ◽

Rice Leaves ◽

Growth Phenotype ◽

Serious Disease ◽

Ascorbic Acid Supplementation

Xanthomonas oryzae pv. oryzae causes bacterial leaf blight, a serious disease of rice. A mutation was isolated in the ferric uptake regulator (fur) gene of X. oryzae pv. oryzae and it was shown to result in the production of siderophores in a constitutive manner. The fur mutant is hypersensitive to the metallo-antibiotic streptonigrin, a phenotype that is indicative of intracellular free-iron overload, and also exhibits a slow growth phenotype on rich medium. The fur mutant is virulence deficient, hypersensitive to hydrogen peroxide, and exhibits reduced catalase activity. Exogenous supplementation with ascorbic acid (an antioxidant) rescues the growth deficiency of the fur mutant in rice leaves. The virulence deficiency of the X. oryzae pv. oryzae fur mutant is proposed to be due, at least in part, to an impaired ability to cope with the oxidative stress conditions that are encountered during infection.

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PhyA, a Secreted Protein of Xanthomonas oryzae pv. oryzae, Is Required for Optimum Virulence and Growth on Phytic Acid as a Sole Phosphate Source

Molecular Plant-Microbe Interactions ◽

10.1094/mpmi.2003.16.11.973 ◽

2003 ◽

Vol 16 (11) ◽

pp. 973-982 ◽

Cited By ~ 28

Author(s):

Subhadeep Chatterjee ◽

Rajan Sankaranarayanan ◽

Ramesh V. Sonti

Keyword(s):

Phytic Acid ◽

Xanthomonas Campestris ◽

Xanthomonas Oryzae ◽

Bacillus Species ◽

Active Site Residues ◽

Significant Similarity ◽

Reading Frame ◽

Predicted Signal Peptide ◽

Serious Disease ◽

Phosphate Source

Xanthomonas oryzae pv. oryzae causes bacterial leaf blight, a serious disease of rice. We have identified a novel virulence deficient mutant (BXO1691) of X. oryzae pv. oryzae that has a Tn5 insertion in an open reading frame (phyA; putative phytase A) encoding a 373-amino acid (aa) protein containing a 28-aa predicted signal peptide. Extracellular protein profiles revealed that a 38-kDa band is absent in phyA mutants as compared with phyA+ strains. A BLAST search with phyA and its deduced polypeptide sequence indicated significant similarity with conserved hypothetical proteins in Xanthomonas axonopodis pv. citri and Xanthomonas campestris pv. campestris and limited homology to secreted phytases of Bacillus species. Homology modeling with a Bacillus phytase as the template suggests that the PhyA protein has a similar six-bladed β-propeller architecture and exhibits conservation of certain critical active site residues. Phytases are enzymes that are involved in degradation of phytic acid (inositol hexaphosphate), a stored form of phosphate in plants. The phyA mutants exhibit a growth deficiency in media containing phytic acid as a sole phosphate source. Exogenous phosphate supplementation promotes migration of phyA X. oryzae pv. oryzae mutants in rice leaves. These results suggest that the virulence deficiency of phyA mutants is, at least in part, due to inability to use host phytic acid as a source of phosphate. phyA-like genes have not been previously reported to be involved in the virulence of any plant pathogenic bacterium.

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Mutants of Xanthomonas oryzae pv. oryzae Deficient in General Secretory Pathway Are Virulence Deficient and Unable to Secrete Xylanase

Molecular Plant-Microbe Interactions ◽

10.1094/mpmi.2000.13.4.394 ◽

2000 ◽

Vol 13 (4) ◽

pp. 394-401 ◽

Cited By ~ 106

Author(s):

Suvendra K. Ray ◽

R. Rajeshwari ◽

Ramesh V. Sonti

Keyword(s):

Xanthomonas Campestris ◽

Secretory Pathway ◽

Genomic Library ◽

Xanthomonas Oryzae ◽

Enzyme Assays ◽

Specific Primers ◽

Xylanase Production ◽

Protein Secretion System ◽

Serious Disease ◽

Xanthomonas Campestris Pv Campestris

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. Trans-poson mutagenesis and marker exchange studies revealed genes on the cloned DNA that were required for xylanase production and virulence. Sequence analysis with trans-poson-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.

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In VitroActivity of Glucosinolates and Their Degradation Products against Brassica-Pathogenic Bacteria and Fungi

Applied and Environmental Microbiology ◽

10.1128/aem.03142-14 ◽

2014 ◽

Vol 81 (1) ◽

pp. 432-440 ◽

Cited By ~ 39

Author(s):

T. Sotelo ◽

M. Lema ◽

P. Soengas ◽

M. E. Cartea ◽

P. Velasco

Keyword(s):

Pseudomonas Syringae ◽

Xanthomonas Campestris ◽

Pathogenic Bacteria ◽

Degradation Products ◽

Allyl Isothiocyanate ◽

Leaf Extracts ◽

Soil Pathogens ◽

Content Type ◽

Positive Effects

ABSTRACTGlucosinolates (GSLs) are secondary metabolites found inBrassicavegetables that confer on them resistance against pests and diseases. Both GSLs and glucosinolate hydrolysis products (GHPs) have shown positive effects in reducing soil pathogens. Information about theirin vitrobiocide effects is scarce, but previous studies have shown sinigrin GSLs and their associated allyl isothiocyanate (AITC) to be soil biocides. The objective of this work was to evaluate the biocide effects of 17 GSLs and GHPs and of leaf methanolic extracts of different GSL-enrichedBrassicacrops on suppressingin vitrogrowth of two bacterial (Xanthomonas campestrispv. campestris andPseudomonas syringaepv. maculicola) and two fungal (AlternariabrassicaeandSclerotiniascletoriorum)Brassicapathogens. GSLs, GHPs, and methanolic leaf extracts inhibited the development of the pathogens tested compared to the control, and the effect was dose dependent. Furthermore, the biocide effects of the different compounds studied were dependent on the species and race of the pathogen. These results indicate that GSLs and their GHPs, as well as extracts of differentBrassicaspecies, have potential to inhibit pathogen growth and offer new opportunities to study the use ofBrassicacrops in biofumigation for the control of multiple diseases.

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The Contribution of the Predicted Sorting Platform Component HrcQ to Type III Secretion in Xanthomonas campestris pv. vesicatoria Depends on an Internal Translation Start Site

Frontiers in Microbiology ◽

10.3389/fmicb.2021.752733 ◽

2021 ◽

Vol 12 ◽

Author(s):

Christian Otten ◽

Tanja Seifert ◽

Jens Hausner ◽

Daniela Büttner

Keyword(s):

Type Iii Secretion ◽

Structural Component ◽

Xanthomonas Campestris ◽

Pathogenic Bacteria ◽

Start Site ◽

Translation Start Site ◽

Terminal Protein ◽

Type Iii ◽

Translation Start ◽

Membrane Spanning

Pathogenicity of the Gram-negative bacterium Xanthomonas campestris pv. vesicatoria depends on a type III secretion (T3S) system which translocates effector proteins into plant cells. T3S systems are conserved in plant- and animal-pathogenic bacteria and consist of at least nine structural core components, which are designated Sct (secretion and cellular translocation) in animal-pathogenic bacteria. Sct proteins are involved in the assembly of the membrane-spanning secretion apparatus which is associated with an extracellular needle structure and a cytoplasmic sorting platform. Components of the sorting platform include the ATPase SctN, its regulator SctL, and pod-like structures at the periphery of the sorting platform consisting of SctQ proteins. Members of the SctQ family form a complex with the C-terminal protein domain, SctQC, which is translated as separate protein and likely acts either as a structural component of the sorting platform or as a chaperone for SctQ. The sorting platform has been intensively studied in animal-pathogenic bacteria but has not yet been visualized in plant pathogens. We previously showed that the SctQ homolog HrcQ from X. campestris pv. vesicatoria assembles into complexes which associate with the T3S system and interact with components of the ATPase complex. Here, we report the presence of an internal alternative translation start site in hrcQ leading to the separate synthesis of the C-terminal protein region (HrcQC). The analysis of genomic hrcQ mutants showed that HrcQC is essential for pathogenicity and T3S. Increased expression levels of hrcQ or the T3S genes, however, compensated the lack of HrcQC. Interaction studies and protein analyses suggest that HrcQC forms a complex with HrcQ and promotes HrcQ stability. Furthermore, HrcQC colocalizes with HrcQ as was shown by fluorescence microscopy, suggesting that it is part of the predicted cytoplasmic sorting platform. In agreement with this finding, HrcQC interacts with the inner membrane ring protein HrcD and the SctK-like linker protein HrpB4 which contributes to the docking of the HrcQ complex to the membrane-spanning T3S apparatus. Taken together, our data suggest that HrcQC acts as a chaperone for HrcQ and as a structural component of the predicted sorting platform.

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Apis andreniformis associated Actinomycetes show antimicrobial activity against black rot pathogen (Xanthomonas campestris pv. campestris)

PeerJ ◽

10.7717/peerj.12097 ◽

2021 ◽

Vol 9 ◽

pp. e12097

Author(s):

Yaowanoot Promnuan ◽

Saran Promsai ◽

Wasu Pathom-aree ◽

Sujinan Meelai

Keyword(s):

Antimicrobial Activity ◽

16S Rrna ◽

16S Rrna Gene ◽

Honey Bee ◽

Pseudomonas Syringae ◽

Xanthomonas Campestris ◽

Pathogenic Bacteria ◽

Rrna Gene ◽

Apis Andreniformis

This study aimed to investigate cultivable actinomycetes associated with rare honey bee species in Thailand and their antagonistic activity against plant pathogenic bacteria. Actinomycetes were selectively isolated from the black dwarf honey bee (Apis andreniformis). A total of 64 actinomycete isolates were obtained with Streptomyces as the predominant genus (84.4%) followed by Micromonospora (7.8%), Nonomuraea (4.7%) and Actinomadura (3.1%). All isolates were screened for antimicrobial activity against Xanthomonas campestris pv. campestris, Pectobacterium carotovorum and Pseudomonas syringae pv. sesame. Three isolates inhibited the growth of X. campestris pv. campestris during in vitro screening. The crude extracts of two isolates (ASC3-2 and ASC5-7P) had a minimum inhibitory concentration (MIC) of 128 mg L−1against X. campestris pv. campestris. For isolate ACZ2-27, its crude extract showed stronger inhibitory effect with a lower MIC value of 64 mg L−1 against X. campestris pv. campestris. These three active isolates were identified as members of the genus Streptomyces based on their 16S rRNA gene sequences. Phylogenetic analysis based on the maximum likelihood algorithm showed that isolate ACZ2-27, ASC3-2 and ASC5-7P were closely related to Streptomyces misionensis NBRC 13063T (99.71%), Streptomyces cacaoi subsp. cacaoi NBRC 12748T (100%) and Streptomyces puniceus NBRC 12811T (100%), respectively. In addition, representative isolates from non-Streptomyces groups were identified by 16S rRNA gene sequence analysis. High similarities were found with members of the genera Actinomadura, Micromonospora and Nonomuraea. Our study provides evidence of actinomycetes associated with the black dwarf honey bee including members of rare genera. Antimicrobial potential of these insect associated Streptomyces was also demonstrated especially the antibacterial activity against phytopathogenic bacteria.

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Identification of Novel Type III Secretion Effectors in Xanthomonas oryzae pv. oryzae

Molecular Plant-Microbe Interactions ◽

10.1094/mpmi-22-1-0096 ◽

2009 ◽

Vol 22 (1) ◽

pp. 96-106 ◽

Cited By ~ 71

Author(s):

Ayako Furutani ◽

Minako Takaoka ◽

Harumi Sanada ◽

Yukari Noguchi ◽

Takashi Oku ◽

...

Keyword(s):

Pseudomonas Syringae ◽

Type Iii Secretion ◽

Pathogenic Bacteria ◽

Regulatory Protein ◽

Effector Proteins ◽

Xanthomonas Oryzae ◽

Dependent Manner ◽

Plant Pathogenic Bacteria ◽

Type Iii ◽

Genes Encoding

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.

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