高建明,胡彩平,郑金龙,张世清,陈河龙,刘巧莲,习金根,易克贤.柱花草炭疽病菌T-DNA 插入突变体库的构建与分析[J].广东农业科学,2014,41(19):142-145 |
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柱花草炭疽病菌T-DNA 插入突变体库的构建与分析 |
Construction and analysis of a T-DNA insertional mutant library for Colletotrichum gloeosporioides |
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DOI: |
中文关键词: 胶孢炭疽菌 柱花草 ATMT 突变体库 致病机理 |
英文关键词: Colletotrichum gloeosporioides曰Stylosanthes guianensis曰ATMT曰mutant library曰pathogenesis |
基金项目:国家自然科学基金(31072076) |
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中文摘要: |
胶孢炭疽菌(Colletotrichum gloeosporioides)引起的柱花草炭疽病是热带牧草柱花草的主要病害,利用根癌农杆菌介导转化(Agrobacterium tumefaciens mediated transformation,ATMT)的T-DNA 插入突变技术,可使炭疽病菌基因组上被插入部位的基因丧失功能,是研究柱花草炭疽病菌致病机理的重要方法。在先前优化根癌农杆菌介导柱花草炭疽病菌遗传转化体系的基础上,构建了4616个转化子的突变体库,从中随机选取部分突变体,对其TDNA插入拷贝数、遗传稳定性、生长速度、产孢量、分生孢子萌发等进行分析。结果表明,所得转化子多为单拷贝,能稳定遗传。一些转化子的生长速度、产孢量、分生孢子萌发率与野生型相比有明显差异。 |
英文摘要: |
The anthracnose caused by Colletotrichum gloeosporioides is a serious threat to Stylosanthes guianensis, an
important tropical forage. Agrobacterium tumefaciens -mediated transformation as a tool for insertional mutagenesis, is the important way to study pathogenesis of C. gloeosporioides. Based on the previously optimized Agrobacterium -mediated
genetic transformation system, a mutant library containing 4 616 putative mutants was constructed, from which some
mutants were randomly selected for analysis of the copy number of T-DNA insertion, genetic stability, growth rate, spore
amount, spore germination and appressorium formation. The results showed that most of the mutants had single T-DNA
insertion and could be stably inherited. The growth rate, spore amount, spore germination rate, appressorium formation rate of some mutants were significantly different from the wild type. |
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