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禾谷镰刀菌随机插入突变体文库的构建

发表时间:2016-02-15  浏览量:1843  下载量:636
全部作者: 侯毅平,周明国
作者单位: 南京农业大学植物保护学院
摘 要: 以禾谷镰刀菌Y2021A为出发菌株,建立潮霉素磷酸转移酶基因(hph)随机插入突变体文库。制备禾谷镰刀菌Y2021A原生质体,将hph转化到菌株Y2021A中,共获得hph随机插入突变体10 163个,其中10 125个能在100 mg/L 潮霉素B的马铃薯蔗糖琼脂培养基(potato sucrose medium,PSA)平板上正常生长,被认定为hph随机插入阳性突变体;随机选取5个突变体进行Southern blotting杂交,结果表明hph基因在突变体中均为单拷贝插入,并且插入到不同的位置;在部分突变体中筛选到5个致病性显著下降的突变体,为研究禾谷镰刀菌致病相关基因提供了宝贵材料。该突变体文库为研究禾谷镰刀菌致病机理及其他生物学表型提供了宝贵的资源。
关 键 词: 植物病理学;禾谷镰刀菌;突变体文库;致病性
Title: Constuction of random insertion mutant library of Fusarium graminearum
Author: HOU Yiping, ZHOU Mingguo
Organization: College of Plant Protection, Nanjing Agricultural University
Abstract: In this paper, the mutation library of Fusarium graminearum Y2021A inserted randomly by hygromycin phosphotransferase (hph) was constructed. The protoplast of strain Y2021A was prepared and the DNA fragments of hph was transformed to Y2021A to obtain the randomly inserted mutations. 10 163 mutations inserted by hph were totally obtained, 10 125 out of which considered as positive mutations could grow normally on PSA plate containing 100 mg/L hygromycin B. Southern blotting assay showed that the hph gene inserted different sites in the genome by one copy in five rondomly selected mutants. Five mutants whose pathogenicity significantly reduced were selected and they were the important materials for studying pathogenicity related genes. The mutant library is the precious resouce for studying pathogenicity mechanism and others of F. graminearum.
Key words: phytopathology; Fusarium graminearum; mutant library; pathogenicity
发表期数: 2016年2月第3期
引用格式: 侯毅平,周明国. 禾谷镰刀菌随机插入突变体文库的构建[J]. 中国科技论文在线精品论文,2016,9(3):253-257.
 
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