National Academy of Agricultural Science, Rural Development Administration, Suwon, Korea.
FEMS Microbiol Lett. 2009 Dec;301(2):149-55. doi: 10.1111/j.1574-6968.2009.01793.x.
Xanthomonas oryzae pathovar oryzae (Xoo) causes bacterial blight disease in rice (Oryza sativa L.). For a study of function, we constructed a random insertion mutant library of Xoo using a Tn5 transposon and isolated the mutant strain (M11; aroK::Tn5) that had extremely low pigment production. In addition, M11 had decreased virulence against the susceptible rice cultivar IR24. Thermal asymmetric interlaced-PCR and sequence analysis of M11 revealed that the transposon was inserted into the aroK gene (which encodes a shikimate kinase). To investigate the expression patterns of the pigment- and virulence-deficient mutant, DNA microarray analysis was performed. In addition, reverse transcriptase-PCR was performed to confirm the expression levels of several genes, including the aro genes of the aroK mutant. Our findings reveal that several crucial genes for virulence, including cellulase and hypersensitive response and pathogenicity (hrp) genes, were regulated by mutations in the aroK gene.
稻黄单胞菌稻致病变种（Xoo）引起水稻（Oryza sativa L.）细菌性条斑病。为了研究功能，我们使用 Tn5 转座子构建了稻黄单胞菌的随机插入突变体文库，并分离出突变菌株（M11；aroK::Tn5），其色素产量极低。此外，M11 对易感水稻品种 IR24 的毒力降低。M11 的热不对称交错-PCR 和序列分析表明，转座子插入到 aroK 基因（编码莽草酸激酶）中。为了研究色素和毒力缺陷突变体的表达模式，进行了 DNA 微阵列分析。此外，还进行了逆转录 PCR 以确认包括 aroK 突变体的 aro 基因在内的几个基因的表达水平。我们的研究结果表明，几个关键的毒力基因，包括纤维素酶和过敏反应和致病性（hrp）基因，受到 aroK 基因突变的调控。
