十字花科黑腐病菌中假定糖基水解酶基因与致病力相关研究  

Correlation analysis between putative glycosyl hydrolase gene and pathogenicity in Xanthomonas campestris pv. campestris

作　　者：岑卫健 吴赞[3] 邹海凡[3] 刘娇[3] 姜伯乐[1,3] 姜伟[1,3] 

机构地区：[1]亚热带农业生物资源保护与利用国家重点实验室,南宁530005 [2]广西理工科学实验中心,南宁530005 [3]广西大学生命科学与技术学院,南宁530005

出　　处：《南方农业学报》2015年第5期793-799,共7页Journal of Southern Agriculture

基　　金：广西自然科学基金项目(20130231);亚热带生物资源保护与利用国家重点实验室项目(C190014);广西教育厅高校科学技术研究重点项目(20130300);广西教育厅高校科学技术研究一般项目(KY2015YB012)

摘　　要：【目的】已报道的十字花科黑腐病菌Xcc 8004菌株基因组中XC1077被注释编码一个假定糖基水解酶,初步研究XC1077基因的致病性,为深入研究Xcc 8004中的糖基水解酶在植物病原菌侵染及定殖过程中的作用奠定基础。【方法】通过同源单交换构建XC1077整合突变体NK1077,用带有全长XC1077基因序列的p LAFRJ质粒对NK1077进行功能互补并检测致病性。【结果】PCR及酶切验证表明整合突变体NK1077和互补菌株C1077构建成功。在MMX基本培养基上,NK1077及C1077的生长情况与Xcc 8004一致。致病性试验结果表明,NK1077在满身红萝卜的致病性下降到Xcc 8004的48.3%,过敏反应(HR)检测结果表明NK1077在辣椒ECW-10R上产生的HR反应明显滞后,而互补菌致病力及过敏反应能恢复至野生型水平。【结论】XC1077基因在Xcc 8004中是一个与致病相关的基因。[ Objective ]XC1077 gene was annotated to encode a putative glycosyl hydrolase in the reported genome se- quence of Xanthomonas campestrls pv. campestris strain 8004（Xcc 8004）. The present experiment was concudcted to pre- liminarily investigate pathogenicity of XC1077 gene to lay the foundation for further study on the function of glycosyl hy- drolase protein in the infection and colonization process of plant pathogenic bacteria. [ Method ]The integrate mutant NK1077 of XC1077 gene was constructed by using homologous single exchange method. The pLTRAJ plasmid with whole sequence of XC1077 gene made a functional supplement for NK1077. Then the pathogenicity of eomplementation strain C1077 was detected. [Result]The identification of PCR and enzyme digestion showed that the integrate mutant NK1077 and complementation strain C1077 were constructed successfully. NK1077 and C1077 strains could grow as well as Xcc 8004 on basic medium MMX. The pathogenic experiment demonstrated that the pathogenicity of NK1077 decreased to 48.3% that of Xcc 8004 in Chinese radish （Raphanussativus var. radiculus） cv. Manshenhong. Hypersensitive reaction （HR） results showed that HR of NK1077 was obviously lagging behind in pepper ECW-10R C1077.The pathogenicity and HR of complementation strain C 1077 could returned to the wild-type level. [ Conclusion ]XC1077 gene plays an important role of pathogenicity in Xcc 8004 strain.

关 键 词：十字花科黑腐病菌 XC1077基因 整合突变体 致病力 过敏反应 

分 类 号：Q754[生物学—分子生物学]