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[目的]明确hrpZPsg12基因对大豆细菌性斑点病菌致病性的影响。[方法]采用PCR方法从大豆细菌性斑点病菌中克隆hrpZPsg12基因,利用具有自杀特性的敲除质粒pKNOCK-Cm和具有功能互补作用的粘粒pUFR034,构建了hrpZPsg12基因的突变载体pKNOCK477-7和互补载体pU-FR1026-68,并筛选出hrpZPsg12基因的突变体477-1及其功能互补子1026-5。进一步将野生型Psg12、突变体477-1和互补子1026-5等3个菌株同时接种大豆叶片和烟草叶片,进行致病性测定和过敏性反应分析。[结果]所有被接种的大豆和烟草叶片都产生了反应斑。但是,反应斑的大小有差异,Psg12菌株接种的病斑较大,477-1的较小,互补子1026-5的接近野生型菌株Psg12。病斑中细菌繁殖量分析表明,野生型菌株Psg12繁殖量最高,突变体477-1的最低,互补子1026-5的接近野生型菌株Psg12的繁殖量。[结论]hrpZPsg12基因能够增强大豆细菌性斑点病菌对大豆的致病性,并且能够在烟草上产生过敏性反应。
[Objective] The research aimed to clarify the effect of hrpZPsg12 gene on the pathogenicity of soybean bacterial spot pathogens. [Method] The hrpZPsg12 gene was cloned from soybean bacterial spot bacteria by PCR. The mutant pKNOCK477-7 and the complementary of hrpZPsg12 gene were constructed by using the knockout plasmid pKNOCK-Cm with suicide trait and cosmid pUFR034 with function complementarity Vector pU-FR1026-68, and screened the mutant 477-1 of hrpZPsg12 gene and its functional complement 1026-5. Three strains of wild-type Psg12, mutant 477-1 and complementary 1026-5 were further inoculated with soybean leaves and tobacco leaves for pathogenicity determination and anaphylactic reaction analysis. [Result] All the inoculated soybean and tobacco leaves produced reaction spots. However, there was a difference in the size of the reaction spot. The lesion size of the Psg12 strain inoculated was larger, and the 477-1 was smaller. The complement 1026-5 was close to the wild-type strain Psg12. The analysis of bacterial multiplication in the lesion showed that the highest amount of Psg12 was propagated in the wild-type strain, the lowest was in the mutant 477-1, and the number of the complementary clone 1026-5 was close to that of the wild-type strain Psg12. [Conclusion] The hrpZPsg12 gene could enhance the pathogenicity of soybean bacterial spot bacteria to soybean and produce allergic reaction on tobacco.