木薯细菌性枯萎病菌转座子库的构建及其质量评价

摘要

In order to study the pathogenic molecular mechanism of Xanthomonas 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 AspCxam 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.%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.