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当前位置: 首页 > 文章 > 棉花黄萎病菌T-DNA插入突变体库的构建及致病缺陷突变体筛选 河南农业科学 2014 (1) 75-79+89
Position: Home > Articles > Construction of T-DNA Insertional Mutant Library for Verticillium dahliae and Identification of Pathogenicity Defective Mutants Journal of Henan Agricultural Sciences 2014 (1) 75-79+89

棉花黄萎病菌T-DNA插入突变体库的构建及致病缺陷突变体筛选

作  者：
谷素静;汪敏;桑茜;赵静;李洪连
单  位：
河南农业大学植物保护学院
关键词：
棉花黄萎病;大丽轮枝菌;农杆菌介导的遗传转化;T-DNA插入突变;致病性
摘  要：
大丽轮枝菌(Verticillium dahliae)侵染引起的棉花黄萎病是棉花生产上的重要病害之一,而其致病的分子机制尚不清楚,为此,以强致病力落叶型黄萎病菌FGH2为材料,利用农杆菌介导转化技术构建棉花黄萎病菌T-DNA插入突变体库并进行致病缺陷突变体的筛选。当农杆菌与黄萎病菌分生孢子浓度比为250∶1、承载介质为NC膜、乙酰丁香酮浓度400μmol/L、共培养时间48h时,转化效率高达每106个分生孢子产生612.5个转化子。通过采用优化后的转化体系,构建了含1 200个T-DNA插入转化子的棉花黄萎病菌突变体库。利用苗期分生孢子蘸根法测定300个转化子在棉花幼苗上的致病力,获得3个致病力显著降低的突变体181、546、1009,其中546的微菌核形成能力丧失,1009的微菌核形成能力明显下降。黄萎病菌T-DNA插入突变体库的构建,为棉花黄萎病菌致病及微菌核形成机制的研究奠定了基础。
译  名：
Construction of T-DNA Insertional Mutant Library for Verticillium dahliae and Identification of Pathogenicity Defective Mutants
作  者：
GU Su-jing;WANG Min;SANG Xi;ZHAO Jing;LI Hong-lian;College of Plant Protection,Henan Agricultural University;
关键词：
cotton Verticillium wilt;;Verticillium dahliae;;Agrobacterium tumefaciens-mediated transformation;;T-DNA insertional mutagenesis;;pathogenicity
摘  要：
The fungus Verticillium dahliae causes cotton Verticillium wilt,one of the most devastating diseases of cotton,however,the molecular basis of pathogenicity of V.dahliaeis unclear.In this study,Agrobacterium tumefaciens-mediated transformation(ATMT)was applied for insertional mutagenesis of V.dahliae FGH2,a highly virulent and defoliating strain.The collection of 1 200T-DNA random insertion transformants of V.dahliae was generated,using the optimized cocultivating conditions of V.dahliae conidia and Agrobacterium(1∶250ratio)for 48h,with the AS concentration of 400μmol/L and the nitrocellulose filter as the filter type in the co-cultivation,resulting in 612.5transformants per 106 conidia.Three pathogenicity defective V.dahliae mutants were obtained through testing the pathogenicity of 300T-DNA transformants on the cotton variety Yinshan 1seedling by dipping roots with spore suspension,including one T-DNA in-sertion mutant losing the ability for forming microsclerotia and one T-DNA insertion mutant defective of microsclerotia development capability.The T-DNA insertional mutagenesis library of V. dahliae lays a foundation for researching molecular mechanisms of pathogenicity and microsclerotia development of V.dahliae.

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