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Application of Functional Genomics to the Study of Solanacea's Defense Mechanisms against Xanthomonas sp. Pathogenicity

ABSTRACT: Xanthomonas campestris pv. campestris (Xcc) is a major bacterial pathogen of cruciferous plants worldwide. The pathogen produces polysaccharides including extracellular cyclic glucan, xanthan, and extracellular enzymes that are key virulence factors. Different Xcc mutants (8397-defective in xanthan and 8523- defective in extracellular glucans) have been obtained and characterized in previously, wich shown to be less infective than the wild type strain when inoculated in N.benthamiana, wich has shown to be an excellent model for the study of Xcc-plant interaction. The objective of this work is evaluate this compounds functions in the plant-pathogen interaction, in particular in the plant transcriptoma modulation to confer susceptibility or resistance to the infection. Plant gene expression profiles would be obtained from independently inoculated leaves of N.benthamiana with the following strains of xanthomonas: Xcc. 8004 (wild type), Xcc. 8397 (xanthan minus), Xcc. 8523 (glucan minus), ant water (control). Leaves discs will be collected at 24 hs post infection and immediately submerged in liquid N2. Total RNA will be extracted with plant RNA specific kits (RNA easy QIAGen), treated with DNase, purified and quantified. Keywords: Reference design

ORGANISM(S): Nicotiana benthamiana Solanum tuberosum

PROVIDER: GSE8276 | GEO | 2007/09/01

SECONDARY ACCESSION(S): PRJNA101231

REPOSITORIES: GEO

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Project description:Xanthomonas campestris pathovar campestris (Xcc), the causal agent of black rot disease of cruciferous plants worldwide, is composed of phenotypically heterogeneous groups of strains. The knowledge about the genome diversity and phylogenetic relationships between Xcc strains with different origins are of great interest as they provide insight into the mechanisms of pathogenicity, host preferences and evolution of this pathogen. In our present work, eighteen Xcc strains collected from different geographical area of China mainland were investigated concerning of the genome composition by comparative genomic hybridization (CGH) using microarray slides spotted with PCR-based intragenic DNA fragments of 4273 open reading frames (ORFs) representing the non-redundant genome content of Xcc strain 8004. The common genome backbone of Chinese strains was estimated to contain about 3404 ORFs, which was considered to maintain the basic characteristics of Xcc, i.e. the yellow mucoid colony on nutrient solid medium as well as the pathogenicity to induce black rot disease on host plants. A flexible gene pool of 729 ORFs in Xcc was characterized, of which 402 ORFs were clustered in twenty-seven highly variable genomic regions in Xcc 8004. Of these highly variable genomic regions, five are absolutely absent from Chinese strains, which constitutes the main genomic differences between the Xcc 8004 and Chinese strains. Transcriptome analysis of Xcc 8004 grown in the rich medium NYG and the defined medium XVM2 indicated that the expression of some certain genes in highly variable genomic regions are significantly activated in XVM2, which included the predicted pathogencity and avirulence genes. Candidate genes for cultivar-specificity of Xcc were identified in the variable genomic regions: the avrXccC and avrXccE1 were demonstrated to confer the avirulence on the host plants Mustard cultivar (cv.) Guangtou and Chinese cabbage cv. Zhongbai 83, respectively; and the avrBs1 showed to correlate with the hypersensitive reaction (HR) on the non-host plant pepper ECW10R. This study revealed the common genome backbone of Xcc maintained the basic function in essential metabolisms and basic pathogencity, and the variable genomic determinants contributed to the cultivar-specificity of the pathogen, suggesting that the Xcc genome, with a compact function core carrying essential genes for survival, reproduction, and invasion, is constantly diversifying by acquiring and losing DNA segments, or by DNA degeneration, to improve the genetic novelty for the adaptation during the evolution. Keywords: CGH analysis and transcriptome analysis

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Application of Functional Genomics to the Study of Solanacea's Defense Mechanisms against Xanthomonas sp. Pathogenicity

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