Generation and Analysis of Pathogenicity-related Gene Mutants of Colletotrichum gloeosporioides Using a Novel Promoter Trapping System

• 作者: He Chunping Li Rui Liang Yanqiong Liu Yan Wei Xiaohui Wu Weihuai Zheng Fu-cong Zheng Xiaolan
• 来源期刊: Plant Diseases and Pests(植物病虫害研究：英文版)
• 学科: 农学
• 关键词: 致病相关基因 基因突变体 炭疽病菌 T-DNA插入 系统 捕获 侧翼序列 基因组DNA
• 年，卷（期）: 2013,（3）
• 页码范围: 12-15
• 页数: 4页
• 分类号: S432.42

摘要

Agrobacterium tumefaciens-mediated transformation(ATMT) is an efficient tool for insertional mutagenesis and is used in a wide variety of plants. This paper reports a promoter trapping method to generate mutants in the filamentous fungus,Colletotrichum gloeosporioides,by ATMT insertion of a trapping vector(pCAHPH) that carries a promoterless hygromycin phosphotransferase(hph) gene. Transformants were selected on the media containing 200 μg/mL hygromycin B,and screened for pathogenicity-related gene mutants. Their pathogenicity-related mutants T-DNA flanking sequences were then cloned and analyzed. Hph genes were amplified from mutant genomic DNA but not from wild-type DNA,indicating that the phenotypic alternations of these mutants were the results of T-DNA insertion. T-DNA flanking sequences were obtained using modified thermal asymmetric interlaced PCR. Two right-sided flanking sequences were highly homologous to proteins from other species.