Laboratoire Génome et Développement des Plantes, Institut de Recherche pour le Développement, UMR 5096 IRD-CNRS-Université de Perpignan, Montpellier, France.
FEMS Microbiol Lett. 2010 Jul 1;308(1):16-23. doi: 10.1111/j.1574-6968.2010.01985.x. Epub 2010 Apr 13.
Xanthomonas oryzae pathovar oryzae (Xoo) and X. oryzae pv. oryzicola (Xoc) cause bacterial diseases in rice: leaf blight and leaf streak, respectively. Although both the Asian and the African strains of Xoo induce similar symptoms, they are genetically different, with the African Xoo strains being more closely related to the Asian Xoc. To identify the sequences responsible for differences between African and Asian Xoo strains and their relatedness to Xoc strains, a suppression-subtractive hybridization (SSH) procedure was performed, using the African Xoo MAI1 strain as a tester and the Philippine Xoo PXO86 strain and Xoc BLS256 strain as drivers. A nonredundant set of 134 sequences from MAI1 was generated. Several DNA fragments isolated by SSH were similar to genes of unknown function, hypothetical proteins, genes related to the type III secretion system, and other pathogenicity-related genes. The specificity of various fragments was validated by Southern blot analysis. SSH sequences were compared with several xanthomonad genomes. In silico analysis revealed SSH sequences as specific to strain MAI1, revealing their potential as specific markers for further epidemiological and diagnostic studies. SSH proved to be a useful method for rapidly identifying specific genes among closely related X. oryzae strains.
稻黄单胞菌亚洲种（Xoo）和稻黄单胞菌非洲种（Xoc）分别引起水稻的细菌性病害：叶枯病和条斑病。尽管亚洲和非洲的 Xoo 菌株都引起类似的症状，但它们在遗传上是不同的，非洲的 Xoo 菌株与亚洲的 Xoc 菌株更为接近。为了鉴定导致非洲和亚洲 Xoo 菌株之间差异以及它们与 Xoc 菌株之间亲缘关系的序列，采用抑制性消减杂交（SSH）程序，以非洲 Xoo MAI1 菌株作为测试菌株，以菲律宾 Xoo PXO86 菌株和 Xoc BLS256 菌株作为驱动菌株。从 MAI1 生成了一组非冗余的 134 个序列。通过 SSH 分离的几个 DNA 片段与功能未知的基因、假设蛋白、与 III 型分泌系统相关的基因和其他致病性相关基因相似。通过 Southern blot 分析验证了各种片段的特异性。SSH 序列与几种黄单胞菌基因组进行了比较。计算机分析显示 SSH 序列特异性地针对菌株 MAI1，这揭示了它们作为进一步进行流行病学和诊断研究的特异性标记的潜力。SSH 被证明是一种在密切相关的稻黄单胞菌菌株中快速鉴定特定基因的有用方法。
