DinI and RecX modulate RecA-DNA structures in Escherichia coli K-12.

Mol Microbiol. 2007 Jan;63(1):103-15

Authors: Renzette N, Gumlaw N, Sandler SJ

RecA plays a central role in recombination, DNA repair and SOS induction through forming a RecA-DNA helical filament. Biochemical observations show that at low ratios to RecA, DinI and RecX stabilize and destabilize RecA-DNA filaments, respectively, and that the C-terminal 17 residues of RecA are important for RecX function. RecA-DNA filament formation was assayed in vivo using RecA-GFP foci formation in log-phase and UV-irradiated cells. In log-phase cells, dinI mutants have fewer foci than wild type and that recX mutants have more foci than wild type. A recADelta17::gfp mutant had more foci like a recX mutant. dinI recX double mutants have the same number of foci as dinI mutants alone, suggesting that dinI is epistatic to recX. After UV treatment, the dinI, recX and dinI recX mutants differed in their ability to form foci. All three mutants had fewer foci than wild type. The dinI mutant's foci persisted longer than wild-type foci. Roles of DinI and RecX after UV treatment differed from those during log-phase growth and may reflect the different DNA substrates, population of proteins or amounts during the SOS response. These experiments give new insight into the roles of these proteins.

PMID: 17163974 [PubMed - indexed for MEDLINE]

Evolutionary pathways of an ancient gene recX.

Gene. 2007 Jan 31;387(1-2):15-20

Authors: Lin J, Chen ZZ, Tian B, Hua YJ

RecX is a regulator of RecA activity by interacting with RecA protein or RecA filaments. Genes encoding RecX were found in genomes of a wide diversity of bacteria and some plants (e.g., Arabidopsis thaliana and Oryza sativa). Our comparative genome analysis showed that although members of the RecX family are found in many bacterial species, they are not found in archaea and the only gene found in eukaryotes is likely derived from bacteria genomes. It is therefore proposed that RecX is of bacterial origin, and the gene had presented in the common ancestor of bacteria. Moreover, bacterial RecX and plant RecX domain are homologues, and RecX domain in plants may have derived from bacteria via unknown pathways. Plant RecX-like protein was formed by a gene fusion event between a unique N-terminal domain of unknown origin and RecX domain within plant cells. Finally, three possible evolutionary pathways from bacteria to plant were discussed.

PMID: 16996700 [PubMed - indexed for MEDLINE]

Detection of Xanthomonas fragariae and presumptive detection of Xanthomonas arboricola pv. fragariae, from strawberry leaves, by real-time PCR.

J Microbiol Methods. 2007 May 31;

Authors: Weller SA, Beresford-Jones NJ, Hall J, Thwaites R, Parkinson N, Elphinstone JG

Real-time (TaqMan) PCR assays were developed to detect the strawberry angular leaf spot pathogen Xanthomonas fragariae (Xf) and the strawberry bacterial blight pathogen Xanthomonas arboricola pv. fragariae (Xaf). The Xf PCR (Xf gyrB) was designed within regions of the gyraseB gene, unique to Xf, after generating gyraseB DNA sequence data from Xf and other closely related strains. The Xaf PCR (Xaf pep) was designed within regions of the pep prolyl endopeptidase gene that were unique to Xaf, after generating pep DNA sequence data from Xf and Xaf strains. The Xf gyrB PCR detected only Xf strains amongst a panel of 20 Xanthomonas-related spp. and pathovars. The Xaf pep PCR assay detected all Xaf strains tested plus two other (of three tested) X. arboricola pathovars. An existing genomic DNA extraction protocol was modified to facilitate detection of both pathogens to 10(3) cells per strawberry leaf disc.

PMID: 17588695 [PubMed - as supplied by publisher]

Capsicum annuum CCR4-associated factor CaCAF1 is necessary for plant development and defence response.

Plant J. 2007 Jul 21;

Authors: Sarowar S, Oh HW, Cho HS, Baek KH, Seong ES, Joung YH, Choi GJ, Lee S, Choi D

The CCR4-associated factor 1 (CAF1) protein belongs to the CCR4-NOT complex, which is an evolutionary conserved protein complex and plays an important role in the control of transcription and mRNA decay in yeast and mammals. To investigate the function of CAF1 in plants, we performed gain- and loss-of-function studies by overexpression of the pepper CAF1 (CaCAF1) in tomato and virus-induced gene silencing (VIGS) of the gene in pepper plants. Overexpression of CaCAF1 in tomato resulted in significant growth enhancement, with increasing leaf thickness, and enlarged cell size by more than twofold when compared with the control plants. A transmission electron microscopic analysis revealed that the CaCAF1-transgenic tomato plants had thicker cell walls and cuticle layers than the control plants. In addition to developmental changes, overexpression of CaCAF1 in tomato plants resulted in enhanced resistance against the oomycete pathogen Phytophthora infestans. Additionally, microarray, northern and real-time polymerase chain reaction analyses of CaCAF1-transgenic tomato plants revealed that multiple genes were constitutively upregulated, including genes involved in polyamine biosynthesis, defence reactions and cell-wall organogenesis. In contrast, VIGS of CaCAF1 in pepper plants caused significant growth retardation and enhanced susceptibility to the pepper bacterial spot pathogen Xanthomonas axonopodis pv. vesicatoria. Our results suggest roles for plant CAF1 in normal growth and development, as well as in defence against pathogens.

PMID: 17587232 [PubMed - as supplied by publisher]

Transcriptional analysis of the tomato resistance response triggered by recognition of the Xanthomonas type III effector AvrXv3.

Funct Integr Genomics. 2007 Jun 21;

Authors: Balaji V, Gibly A, Debbie P, Sessa G

The type III effector AvrXv3 from Xanthomonas campestris pv. vesicatoria (Xcv) elicits a resistance response in the tomato line Hawaii 7981. To test whether similar genes participate in responses triggered by recognition of different avirulence proteins, we examined the effect of AvrXv3 expression on the plant transcriptome as compared to that of other avirulence proteins. By microarray analysis we monitored expression of approximately 8,600 tomato genes upon inoculation with isogenic Xcv strains differing only by the avrXv3 gene. Changes in transcript levels of 139 genes were observed within 8 h, and a massive shift in expression of 1,294 genes was detected at 12 h. Recognition of AvrXv3 modulated a large number of genes encoding transcription factors and signaling components. In addition, genes involved in defense and stress responses, lipid metabolism, protein degradation, and secondary metabolism were mainly up-regulated. Conversely, genes related to photosynthesis and protein synthesis were generally down-regulated. Many novel genes encoding proteins of unknown function were also identified. A comparison between AvrXv3-modulated genes and those differentially expressed in tomato plants recognizing other bacterial effectors revealed partial overlap and similar distribution in functional classes. The identification of tomato genes modulated by AvrXv3 expression paves the way for dissecting defense networks activated by recognition of this effector in resistant plants.

PMID: 17582538 [PubMed - as supplied by publisher]

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

Mol Microbiol. 2007 Jul;65(1):121-136

Authors: Zhang L, Jia Y, Wang L, Fang R

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.

PMID: 17581124 [PubMed - as supplied by publisher]

A novel pathway for the biodegradation of gamma-hexachlorocyclohexane by a Xanthomonas sp. strain ICH12.

J Appl Microbiol. 2007 Jun;102(6):1468-78

Authors: Manickam N, Misra R, Mayilraj S

AIM: To isolate gamma-hexachlorocyclohexane (HCH)-degrading bacteria from contaminated soil and characterize the metabolites formed and the genes involved in the degradation pathway. METHODS AND RESULTS: A bacterial strain Xanthomonas sp. ICH12, capable of biodegrading gamma- HCH was isolated from HCH-contaminated soil. DNA-colony hybridization method was employed to detect bacterial populations containing specific gene sequences of the gamma-HCH degradation pathway. linA (dehydrodehalogenase), linB (hydrolytic dehalogenase) and linC (dehydrogenase) from a Sphingomonas paucimobilis UT26, reportedly possessing gamma-HCH degradation activity, were used as gene probes against isolated colonies. The isolate was found to grow and utilize gamma-HCH as the sole carbon and energy source. The 16S ribosomal RNA gene sequence of the isolate resulted in its identification as a Xanthomonas species, and we designated it as strain ICH12. During the degradation of gamma-HCH by ICH12, formation of two intermediates, gamma-2,3,4,5,6-pentachlorocyclohexene (gamma-PCCH), and 2,5-dichlorobenzoquinone (2,5-DCBQ), were identified by gas chromatography-mass spectrometric (GC-MS) analysis. While gamma-PCCH was reported previously, 2,5-dichlorohydroquinone was a novel metabolite from HCH degradation. CONCLUSIONS: A Xanthomonas sp. for gamma-HCH degradation from a contaminated soil was isolated. gamma-HCH was utilized as sole source of carbon and energy, and the degradation proceeds by successive dechlorination. Two degradation products gamma-PCCH and 2,5-DCBQ were characterized, and the latter metabolite was not known in contrasts with the previous studies. The present work, for the first time, demonstrates the potential of a Xanthomonas species to degrade a recalcitrant and widespread pollutant like gamma-HCH. SIGNIFICANCE AND IMPACT OF THE STUDY: This study demonstrates the isolation and characterization of a novel HCH-degrading bacterium. Further results provide an insight into the novel degradation pathway which may exist in diverse HCH-degrading bacteria in contaminated soils leading to bioremediation of gamma-HCH.

PMID: 17578411 [PubMed - in process]

Epigenetic Inheritance in Rice Plants.

Ann Bot (Lond). 2007 Jun 18;

Authors: Akimoto K, Katakami H, Kim HJ, Ogawa E, Sano CM, Wada Y, Sano H

Background and Aims Epigenetics is defined as mechanisms that regulate gene expression without base sequence alteration. One molecular basis is considered to be DNA cytosine methylation, which reversibly modifies DNA or chromatin structures. Although its correlation with epigenetic inheritance over generations has been circumstantially shown, evidence at the gene level has been limited. The present study aims to find genes whose methylation status directly correlates with inheritance of phenotypic changes. Methods DNA methylation in vivo was artificially reduced by treating rice (Oryza sativa ssp. japonica) seeds with 5-azadeoxycytidine, and the progeny were cultivated in the field for >10 years. Genomic regions with changed methylation status were screened by the methylation-sensitive amplified polymorphysm (MSAP) method, and cytosine methylation was directly scanned by the bisulfite mapping method. Pathogen infection with Xanthomonas oryzae pv. oryzae, race PR2 was performed by the scissors-dip method on mature leaf blades. Key Results The majority of seedlings were lethal, but some survived to maturity. One line designated as Line-2 showed a clear marker phenotype of dwarfism, which was stably inherited by the progeny over nine generations. MSAP screening identified six fragments, among which two were further characterized by DNA blot hybridization and direct methylation mapping. One clone encoding a retrotransposon gag-pol polyprotein showed a complete erasure of 5-methylcytosines in Line-2, but neither translocation nor expression of this region was detectable. The other clone encoded an Xa21-like protein, Xa21G. In wild-type plants, all cytosines were methylated within the promoter region, whereas in Line-2, corresponding methylation was completely erased throughout generations. Expression of Xa21G was not detectable in wild type but was constitutive in Line-2. When infected with X. oryzae pv. oryzae, against which Xa21 confers resistance in a gene-for-gene manner, the progeny of Line-2 were apparently resistant while the wild type was highly susceptible without Xa21G expression. Conclusions These results indicated that demethylation was selective in Line-2, and that promoter demethylation abolished the constitutive silencing of Xa21G due to hypermethylation, resulting in acquisition of disease resistance. Both hypomethylation and resistant trait were stably inherited. This is a clear example of epigenetic inheritance, and supports the idea of Lamarckian inheritance which suggested acquired traits to be heritable.

PMID: 17576658 [PubMed - as supplied by publisher]

Identification of Xanthomonas campestris pv. vesicatoria Genes Induced in its Interaction with Tomato.

J Bacteriol. 2007 Jun 15;

Authors: Tamir-Ariel D, Navon N, Burdman S

Xanthomonas campestris pv. vesicatoria (Xcv) is the causal agent of bacterial spot disease of tomato and pepper. The disease process is interactive and very intricate, and involves a plethora of genes in the pathogen and in the host. In the pathogen, different genes are activated in response to the changing environment to enable it to survive, adapt, evade host defenses, propagate, and damage the host. To understand the disease process it is imperative to broaden our understanding of the gene machinery that participates in it, and the most reliable way is to identify these genes in vivo. Here, we have adapted a Recombinase-based In Vivo Expression Technology (RIVET) to study the genes activated in Xcv during its interaction with one of its hosts, tomato. This is the first study that demonstrates the feasibility of this approach for identifying in vivo induced genes in a plant pathogen. RIVET revealed 61 unique Xcv genes that delineate a picture of the different processes involved in pathogen-host interaction. To further explore the role of some of these genes, we generated knockout mutants for 13 genes and characterized their ability to grow in planta and to cause disease symptoms. This analysis revealed several genes that may be important for the interaction of the pathogen with its host, including a citH homologue gene, encoding a citrate transporter, which was shown to be required for wild type levels of virulence.

PMID: 17573477 [PubMed - as supplied by publisher]

Qualitative and comparative proteomic analysis of Xanthomonas campestris pv. campestris 17.

Proteomics. 2007 Jun 13;7(12):2047-2058

Authors: Chung WJ, Shu HY, Lu CY, Wu CY, Tseng YH, Tsai SF, Lin CH

The bacterium Xanthomonas campestris pathovar campestris (XCC) 17 is a local isolate that causes crucifer black rot disease in Taiwan. In this study, its proteome was separated using 2-DE and the well-resolved proteins were excised, trypsin digested, and analyzed by MS. Over 400 protein spots were analyzed and 281 proteins were identified by searching the MS or MS/MS spectra against the proteome database of the closely related XCC ATCC 33913. Functional categorization of the identified proteins matched 141 (50%) proteins to 81 metabolic pathways in the Kyoto Encyclopedia of Genes and Genomes (KEGG) database. In addition, we performed a comparative proteome analysis of the pathogenic strain 17 and an avirulent strain 11A to reveal the virulence-related proteins. We detected 22 up-regulated proteins in strain 17 including the degrading enzymes EngXCA, HtrA, and PepA, which had been shown to have a role in pathogenesis in other bacteria, and an anti-host defense protein, Ohr. Thus, further functional studies of these up-regulated proteins with respect to their roles in XCC pathogenicity are suggested.

PMID: 17566974 [PubMed - as supplied by publisher]

The crystallization of apo-form UMP kinase from Xanthomonas campestris is significantly improved in a strong magnetic field.

Acta Crystallograph Sect F Struct Biol Cryst Commun. 2007 May 1;63(Pt 5):438-42

Authors: Tu JL, Chin KH, Wang AH, Chou SH

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 A, a dramatic improvement over the original value of 3.6 A. 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.

PMID: 17565191 [PubMed - in process]

Two type III effector genes of Xanthomonas oryzae pv. oryzae control the induction of the host genes OsTFIIA{gamma}1 and OsTFX1 during bacterial blight of rice.

Proc Natl Acad Sci U S A. 2007 Jun 11;

Authors: Sugio A, Yang B, Zhu T, White FF

Xanthomonas oryzae pv. oryzae strain PXO99(A) induces the expression of the host gene Os8N3, which results in increased host susceptibility to bacterial blight of rice. Here, we show that PXO99(A) affects the expression of two additional genes in a type III secretion system-dependent manner, one encoding a bZIP transcription factor (OsTFX1) and the other the small subunit of the transcription factor IIA located on chromosome 1 (OsTFIIAgamma1). Induction of OsTFX1 and OsTFIIAgamma1 depended on the type III effector genes pthXo6 and pthXo7, respectively, both encoding two previously undescribed members of the transcription activator-like (TAL) effector family. pthXo7 is strain-specific and may reflect adaptation to the resistance mediated by xa5, an allele of OsTFIIAgamma5 encoding a second form of the TFIIA small subunit on chromosome 5 of rice. The loss of pthXo6 resulted in reduced pathogen virulence, and ectopic expression of OsTFX1 abrogated the requirement for pthXo6 for full virulence. X. oryzae pv. oryzae therefore modulates the expression of multiple host genes using multiple TAL effectors from a single strain, and evidence supports the hypothesis that expression of the associated host genes contributes to host susceptibility to disease.

PMID: 17563377 [PubMed - as supplied by publisher]

Regulation of bacterial RecA protein function.

Crit Rev Biochem Mol Biol. 2007 Jan-Feb;42(1):41-63

Authors: Cox MM

The RecA protein is a recombinase functioning in recombinational DNA repair in bacteria. RecA is regulated at many levels. The expression of the recA gene is regulated within the SOS response. The activity of the RecA protein itself is autoregulated by its own C-terminus. RecA is also regulated by the action of other proteins. To date, these include the RecF, RecO, RecR, DinI, RecX, RdgC, PsiB, and UvrD proteins. The SSB protein also indirectly affects RecA function by competing for ssDNA binding sites. The RecO and RecR, and possibly the RecF proteins, all facilitate RecA loading onto SSB-coated ssDNA. The RecX protein blocks RecA filament extension, and may have other effects on RecA activity. The DinI protein stabilizes RecA filaments. The RdgC protein binds to dsDNA and blocks RecA access to dsDNA. The PsiB protein, encoded by F plasmids, is uncharacterized, but may inhibit RecA in some manner. The UvrD helicase removes RecA filaments from RecA. All of these proteins function in a network that determines where and how RecA functions. Additional regulatory proteins may remain to be discovered. The elaborate regulatory pattern is likely to be reprised for RecA homologues in archaeans and eukaryotes.

PMID: 17364684 [PubMed - indexed for MEDLINE]

[Cloning, sequencing and fuctional study of gacA gene from Xanthomonas oryzae pv. oryzicola]

Wei Sheng Wu Xue Bao. 2007 Apr;47(2):208-12

Authors: Yang WF, Chen L, Liu HX, Hu BS, Liu FQ

A gacA homologue, designated gacA(Xooc), was cloned from Xanthomonas oryzae pv. oryzicola (Xooc), a bacterium that causes leaf streak of rice, with degenerated primers by polymerase amplification reaction (PCR). NCBI blast search indicated that GacA(Xooc) had a similar structure to that of other GacA proteins, and had a CheB (Chemotaxis response regulator containing a CheY-like receiver domain)domain. Sequence comparison showed that the gacA(Xooc) was conserved in the Xanthomonas genus. Homology search revealed that the gacA(Xooc) was 99.7% similarities to gacA (AY870457, this lab) of Xanthomonas oryzae pv. oryzae (Xoo). A gacA(Xooc), disruption mutant was successfully generated by a single cross-over event, and confirmed by PCR and Southern blot. But the mutant still had strong pathogenicity,and its virulence was not obviously different from that of wild type strain. The gacA did not globally regulate metabolism in Xooc, which was different from DC3000 of P. syringae pv. tomato, CHAO of P. fluorescens and IC1270 of Serratia plymuthica. Chemotaxis to 0.1% tryptone of the mutants was reduced compared to wild type strain. The results suggest that gacA(X00c) is involved chemotaxis of Xooc. Nevertheless, how gacA to regulate chemotaxis of Xooc, transcription and expression of genes involved in regulation still need to be further studied.

PMID: 17552221 [PubMed - in process]

XC5848, an ORFan protein from Xanthomonas campestris, adopts a novel variant of Sm-like motif.

Proteins. 2007 Jun 1;

Authors: Chin KH, Ruan SK, Wang AH, Chou SH

PMID: 17546661 [PubMed - as supplied by publisher]

Mitogen-activated protein kinase OsMPK6 negatively regulates rice disease resistance to bacterial pathogens.

Planta. 2007 May 31;

Authors: Yuan B, Shen X, Li X, Xu C, Wang S

Mitogen-activated protein kinase (MAPK) cascades play important roles in diverse developmental and physiological processes of plants, including pathogen-induced defense responses. Although at least 17 rice MAPKs have been identified and more than half of these MAPK genes have been shown to be pathogen or elicitor responsive, the exact role of most of the MAPKs in host-pathogen interaction is unknown. Here we report that OsMPK6 is an important regulator in rice disease resistance. Suppressing OsMPK6 or knocking out of OsMPK6 enhanced rice resistance to different races of Xanthomonas oryzae pv. oryzae, causing bacterial blight, one of the most devastating diseases of rice worldwide. The resistant plants showed increased expression of a subset of defense-responsive genes functioning in the NH1 (an Arabidopsis NPR1 orthologue)-involved defense signal transduction pathway. These results suggest that OsMPK6 functions as a repressor to regulate rice defense responses upon bacterial invasion.

PMID: 17541629 [PubMed - as supplied by publisher]

Xanthomonas caspase displays an inherent PARP-like activity.

FEMS Microbiol Lett. 2007 May 22;

Authors: Raju KK, Misra HS, Sharma A

In an earlier study, intracellular accumulation of metabolites such as pyruvate and citrate in Xanthomonas campestris pv. glycines (Xcg) was found to result in a caspase dependent stationary phase rapid cell death (RCD). In the present study, the presence of poly ADP-ribose polymerase (PARP)-like activity associated with caspase-3-like protein of Xcg is reported. This activity was found to be responsible for depletion of cellular NAD(+) levels in RCD-promoting media such as Luria-Bertani medium and starch medium fortified with citrate. Addition of PARP-specific inhibitors such as 3-aminobenzamide to RCD-promoting media restored the intracellular NAD(+) levels and thereby prevented RCD. The inherent association of PARP-like activity with the caspase protein was demonstrated by PARP cellular assay, immuno-precipitation and Western analysis. A truncated polysaccharide deacetylase gene having a caspase-like domain was cloned. The expressed protein though found to be inactive, cross-reacted with human caspase and PARP antibodies. This is the first report demonstrating the presence of a PARP-like activity in a prokaryote and its involvement in cell death.

PMID: 17521358 [PubMed - as supplied by publisher]

Refinement of the Xanthomonas campestris pv. vesicatoria hrpD and hrpE operon structure.

Mol Plant Microbe Interact. 2007 May;20(5):559-67

Authors: Weber E, Berger C, Bonas U, Koebnik R

The plant-pathogenic bacterium Xanthomonas campestris pv. vesicatoria possesses a type III secretion (T3S) system which is encoded in the 23-kb hypersensitive response and pathogenicity (hrp) gene cluster. The T3S system is essential for pathogenicity in susceptible hosts and the induction of the hypersensitive response in resistant plants. In this study, we revisited the operon structure of the right part of the hrp gene cluster. Based on complementation experiments of transposon insertions and reverse-transcription polymerase chain reaction analyses, the hrpD operon contains hrcQ, hrcR, hrcS, and hpaA, whereas hrcD, hrpD6, and hrpE belong to the hrpE operon. We determined the transcriptional start site of the hrpE operon and showed that there is a promoter upstream of hrcD containing a plant-inducible promoter box. Conserved secondary mRNA structures in the intergenic region between hrpD6 and hrpE suggest a posttranscriptional regulated expression of hrpE. Based on comparisons of different hrp gene clusters and the analysis of evolutionary rates, we propose that the hrpE transcriptional unit was integrated into the hrp gene cluster at a later time.

PMID: 17506333 [PubMed - in process]

Molecular and pathotypic characterization of new Xanthomonas oryzae strains from West Africa.

Mol Plant Microbe Interact. 2007 May;20(5):534-46

Authors: Gonzalez C, Szurek B, Manceau C, Mathieu T, Séré Y, Verdier V

DNA polymorphism analysis and pathogenicity assays were used to characterize strains of Xanthomonas oryzae pv. oryzae and Xanthomonas oryzae pv. oryzicola collected from rice leaves in West Africa. Restriction fragment length polymorphism (RFLP), repetitive sequence-based polymerase chain reaction, fluorescent amplified fragment-length polymorphism (FAFLP) analyses were assessed for molecular characterization, while pathogenicity was tested by leaf clipping and leaf infiltration. Dendrograms were generated for the data sets obtained from RFLP analysis and repetitive polymerase chain reaction suggesting that the interrelationships between strains were dependent on the technique used. In all cases, data showed that African strains of X. oryzae pv. oryzae form a group genetically distant from Asian strains. FAFLP analyses separated the X. oryzae strains into three groups with significant bootstrap values. A specific and intriguing feature of African strains of X. oryzae pv. oryzae is a reduction in the number of insertion sequence elements and transcription activator-like (avrBs3/pthA) effector genes, based on the molecular markers employed in the study. In addition, pathogenicity assays conducted with African strains of X. oryzae pv. oryzae on a series of nearly isogenic lines (NILs) identified three new races. Finally, leaf infiltration assays revealed the capacity of African strains of X. oryzae pv. oryzae to induce a nonhost hypersensitive response in Nicotiana benthamiana, in contrast with Asian X. oryzae pv. oryzae and X. oryzae pv. oryzicola strains. Our results reveal substantial differences between genomic characteristics of Asian and African strains of X. oryzae pv. oryzae.

PMID: 17506331 [PubMed - in process]

Identification of the flagellar chaperone FlgN in the phytopathogen Xanthomonas axonopodis pathovar citri by its interaction with hook-associated FlgK.

Arch Microbiol. 2007 May 10;

Authors: Khater L, Alegria MC, Borin PF, Santos TM, Docena C, Tasic L, Farah CS, Ramos CH

Genome annotation of the plant pathogen Xanthomonas axonopodis pv. citri (Xac), identified flagellar genes in a 15.7 kb gene cluster. However, FlgN, a secretion chaperone for hook-associated proteins FlgK and FlgL, was not identified. We performed extensive screening of the X. axonopodis pv. citri genome with the yeast two-hybrid system to identify a protein with the characteristics of the flagellar chaperone FlgN. We found a candidate (XAC1990) encoded by an operon for components of the flagellum apparatus that interacted with FlgK. In order to further support this finding, Xac FlgK and XAC1990 were cloned, expressed, and purified. The recombinant proteins were characterized by spectroscopic methods and their interaction in vitro confirmed by pull-down assays. We, therefore, conclude that XAC1990 and its homologs in other Xanthomonas species are, in fact, FlgN proteins. These observations extend the sequence diversity covered by this family of proteins.

PMID: 17492271 [PubMed - as supplied by publisher]