水稻条斑病菌avrBs3/pthA家族基因敲除体系的建立  

Establishment of knockout mutagenesis in avrBs3/pthA family genes of Xanthomonas oryzae pv. oryzicola

作　　者：曹燕飞[1] 邹丽芳[2] 赵文祥[1] 纪志远[1] 邹华松[2] 陈功友[1,2] 

机构地区：[1]南京农业大学植物保护学院农业部病虫监测与治理重点开放实验室,江苏南京210095 [2]上海交通大学农业与生物学院,上海200240

出　　处：《浙江大学学报（农业与生命科学版）》2011年第1期40-48,共9页Journal of Zhejiang University:Agriculture and Life Sciences

基　　金：农业部公益行业专项资助项目(NYHYZX07-056);转基因重大专项资助项目(2008ZX08001-002)

摘　　要：水稻细菌性条斑病菌存在至少20个avrBs3/pthA家族成员,但其在水稻上的毒性或无毒性贡献并不清楚.本文依据avrBs3/pthA结构上的保守性,利用同源重组策略,借助自杀性载体pKMS1介导,对水稻条斑病菌中的avrBs3/pthA家族基因进行敲除.结果发现:同源交换可通过基因内和基因间重组实现avrBs3/pthA家族的基因敲除;PCR和Southern杂交结果显示,水稻条斑病菌中有5个avrBs3/pthA家族基因被成功敲除,表明水稻条斑病菌avrBs3/pthA家族基因敲除体系构建成功.这为逐一评价avrBs3/pthA家族基因的毒性或/和无毒性功能奠定了基础.The contribution of at least twenty highly conserved avrBs3/pthA family genes in Xanthomonas oryzae pv.oryzicola（Xoc） to virulence or avirulence in rice is still rudimentary.Based on the conserved structure of the avrBs3/pthA family,a marker-exchange mutagenesis strategy in association with mediation by a suicide vector pKMS1 was used to knock out the avrBs3/pthA family genes of Xoc.The results showed that total five avrBs3/pthA genes were gradually knocked out based on the conserved structure of the family by using marker-exchange mutagenesis mediated by a suicide vector pKMS1.The knock-out mutation occurred either within an avrBs3/pthA gene or between two avrBs3/pthA genes.PCR and Southern hybridization analysis demonstrated that the establishment of knock-out mutagenesis in avrBs3/pthA genes of Xoc was successful,providing a platform to assess virulence or/and avirulence roles contributed by each member of the avrBs3/pthA family in rice.

关 键 词：水稻条斑病菌 avrBs3/pthA基因 基因敲除 遗传重组 

分 类 号：Q78[生物学—分子生物学] S432.1[农业科学—植物病理学]