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中文摘要：
本文以青枯菌致病力分化菌株Po82的Ⅲ型分泌系统调控基因hrpB为研究对象,采用同源重组双交换法,构建获得青枯菌Po82菌株的hrpB基因缺失突变株Po82ΔhrpB及其互补菌株Po82ΔhrpB-pML123-hrpB,并对野生型菌株、突变株和互补菌株进行了致病力及生物学功能的验证。致病力测定结果表明,青枯菌hrpB基因缺失突变株Po82ΔhrpB的致病力较Po82野生型菌株显著下降,而互补菌株Po82ΔhrpB-pML123-hrpB能够部分恢复突变菌株的致病力。生长曲线测定结果表明,在营养贫瘠型培养基中,hrpB基因缺失突变株Po82ΔhrpB的生长速率较野生型青枯菌Po82菌株快,但是在营养丰富型培养基中,两者生长速率基本一致。野生型和突变株的运动性测定结果显示,两者的运动性无显著差异。表明hrpB基因在青枯菌致病过程中具有重要影响,并对进一步发掘鉴定Po82菌株中新的Ⅲ型效应子,进而深入解析其致病力分化的分子机理具有重要的作用。
英文摘要：
Ralstonia solanacearum strain Po82,a pathogenic variation strain,possesses pathogenic traits of both NPB and Moko strains.hrpB gene encodes a positive regulator and controls the expression of the type Ⅲ secretion system（T3SS） and pathogenicity effectors transiting through this pathway.The mutant of hrpB gene was constructed by using homologous recombination,and named Po82ΔhrpB.Based on the mutant strain,the complement strain was also constructed.The pathogenicity and the biological function of the wild type,the mutant strain and the complement strain were tested.Pathogenicity test showed that disease index of the mutant Po82ΔhrpB was decreased compared with that of the wild type Po82.Growth curve analysis indicated that Po82ΔhrpB mutant grew as fast as the Po82 strain in rich medium,whereas,in Boucher＇s minimal medium,Po82ΔhrpB mutant grew faster than the Po82.There were no significant differences between the mutants and wild type strain in the ability of mobility.The hrpB gene was a critical factor in pathogenicity of R.solanacearum and played an important role in identifying new type Ⅲ effectors and analyzing the molecular mechanism of pathogenicity variation.
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