【摘要】 [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pUFR1026-68,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]hrpZ-Psg12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。更多还原

【Abstract】 [Objective] The paper was to confirm the effect of hrpZPsg12 gene on the pathogenicity of Pseudomonas syringae pv.glycinea.[Method] hrpZPsg12 gene was cloned from P.syringae pv.glycinea using PCR method.The knockout plasmid pKNOCK-Cm with suicide characteristics and clay pUFR034 with complementarity were used to construct the mutation vector pKNOCK477-7 and complementary vector pUFR1026-68 of hrpZPsg12 gene,the mutant 477-1 and the functional complementation unit 1026-5 of the gene was also screened out.Three strains including wild-type Psg12,mutant 477-1 and complementary unit 1026-5 were simultaneously inoculated into soybean leaves and tobacco leaves,then pathogenicity determination and hypersensitive response analysis were carried out.[Result] All the inoculated leaves of soybean and tobacco produced reaction lesion.However,the sizes of reaction lesion were different.The lesion inoculated with Psg12 was relatively large,while the lesion inoculated with 477-1 was relatively small;the lesion of complementary unit 1026-5 was similar to wild-type Psg12.The reproduction amount of bacteria in lesions showed that the reproduction amount was the highest in wild-type Psg12,which was the lowest in mutant 477-1.The reproduction amount of complementary unit 1026-5 was similar to that of wild-type Psg12.[Conclusion] hrpZPsg12 Gene could enhance the pathogenicity of P.syringae pv.glycinea on Soybean and produce hypersensitive response in tobacco.更多还原