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通过反向遗传学方法和半定量RT-PCR方法,研究了十字花科黑腐病菌中被注释的应急反应相关基因spoT_(Xcc)与Ⅲ型分泌系统(T3SS)基因的调控关系。基于自杀质粒pK18mob同源整合方法构建了十字花科黑腐病菌spoT_(Xcc)基因(XC_0956)的非极性整合突变体,检测了突变体的多种表型,发现spoT_(Xcc)突变体延迟了在非寄主植物辣椒ECW-10R上引发的过敏反应(HR)。利用GUS(β-葡糖苷酸酶)活性检测、原位组织染色和半定量RT-PCR的方法,发现T3SS相关基因在spoT_(Xcc)基因的非极性整合突变体中表达量降低,表明十字花科黑腐病菌中被注释的应急反应相关基因spoT_(Xcc)显著正调控T3SS相关基因的表达。
The reverse genetics method and semi-quantitative RT-PCR method were used to study the regulation of the annotated response gene spoT_ (Xcc) and the type Ⅲ secretion system (T3SS) in the crucian caring bacteria. Based on the homologous recombination method of suicide plasmid pK18mob, a nonpolar integrated mutant of spoC gene (XC_0956) was constructed and the phenotypes of mutants were examined. The spoT_ (Xcc) mutant was found to be delayed The anaphylactic reaction (HR) elicited on non-host plant peppers ECW-10R. Using GUS assay, in situ staining and semi-quantitative RT-PCR, it was found that the expression of T3SS-related genes decreased in nonpolar integrated mutants of spoT_ (Xcc) gene, indicating that cross The annotated emergency response-related gene spoT_ (Xcc) in C. arborescens significantly positively regulates T3SS-related gene expression.