木薯细菌性枯萎病菌转座子库的构建及其质量评价  

Construction of Transformants Library in Xanthomonas axonopodis pv. manihotis and It’s Quality Assessment

作　　者：陈江莎[1,2] 李超萍[2] 祝天成[1,2] 时涛[2] 黄贵修[2] 

机构地区：[1]海南大学农学院,海南海口570228 [2]中国热带农业科学院环境与植物保护研究所农业部热带作物有害生物综合治理重点实验室海南省热带农业有害生物监测与控制重点实验室,海南海口571101

出　　处：《热带作物学报》2013年第6期1144-1148,共5页Chinese Journal of Tropical Crops

基　　金：植物基因组学国家重点实验室开放课题(No.2012A0301-08);国家木薯现代产业技术体系建设项目(No.CARS-12-hnhgx);中国热带农业科学院环境与植物保护研究所引进人才科研启动基金项目(No.Hzs1002);海南大学211工程建设项目

摘　　要：为了研究木薯细菌性枯萎病菌的致病分子机理,本研究开展了病原菌的转化子库构建工作。采用电击法,将EZ::TN转座体转化野生型菌株。获得了总数为20 382个转化子并进行了质量评价,采用剪叶法从9 872个转化子中筛选出94个致病性变异的转化子。采用Tail-PCR获得了致病力丧失突变体Xtn62-36的侧翼序列,分析结果表明可能是外源片段插入预测基因AspCXam的编码区而造成致病力丧失。病原菌转化子库的构建及Tail-PCR技术的应用为进一步开展致病相关基因功能的研究提供了基础。In order to study the pathogenic molecular mechanism of Xanthomonos campestris pv. manihotis, a transformant library was conducted in our lab. Using electric shock method, the EZ：：TN〈KAN-2〉Tnp transposome was transformed to the wild strain. A total of 20 382 transformants were obtained and their quality assessments were finished. 94 mutants were screened from 9 872 transformants by the leaf-cutting method. The flanking sequence of mutant Xtn62-36 was confirmed by thermal asymmetric interlaced PCR （TaiL-PCR）. The analysis results showed the insertion of foreign fragments in the coding region of the predicted gene AspCx, possibly resulted in the pathogenicity-defect. The construction of the transformants library and the application of TaiL-PCR would provide a foundation for further study on the function of pathogenicity-related genes.

关 键 词：XANTHOMONAS CAMPESTRIS pv manihotis 转化子库 TAIL-PCR 

分 类 号：S435.33[农业科学—农业昆虫与害虫防治]