Establishment of Protoplast Transformation System in Alternaria tenuissima Using G418 Selection Marker  

以G418为筛选标记的极细链格孢菌(Alternaria tenuissima)原生质体转化体系的建立(英文)

作　　者：万英[1,2] 张大军[2] 黄云[1] 蒋伶活[2] 

机构地区：[1]四川农业大学农学院植物病理系,四川雅安625014 [2]中国农业科学院植物保护研究所植物病虫害生物学国家重点实验室,北京100081

出　　处：《Agricultural Science & Technology》2008年第3期59-62,98,共5页农业科学与技术（英文版）

基　　金：国家重点基础研究项目(2006CB101907)~~

摘　　要：[Objective] The study aimed to establish a protoplast transformation system in Alternaria tenuissima. [Method] The protoplast of A.tenuissima was firstly prepared by enzymolysis method; then the yielded protoplast was transformed by G418 resistant DNA plasmid using PEG/CaCl2 method. [Result] The growth phenotype and PCR detection showed that resistance gene had integrated into A.tenuissima genome. The transformation efficiency of this method reached per μg DNA 3-4 transformants. After subculture thrice under nonselective condition, G418 resistance could still inherit stably. [Conclusion] The transformation system of A.tenuissima was successfully established, which laid basis for studying of the gene function of Alternaria tenuissima.[Objective] The study aimed to establish a protoplast transformation system in Alternaria tenuissima. [Method] The protoplast of A.tenuissima was firstly prepared by enzymolysis method; then the yielded protoplast was transformed by G418 resistant DNA plasmid using PEG/CaCl2 method. [Result] The growth phenotype and PCR detection showed that resistance gene had integrated into A.tenuissima genome. The transformation efficiency of this method reached per μg DNA 3-4 transformants. After subculture thrice under nonselective condition, G418 resistance could still inherit stably. [Conclusion] The transformation system of A.tenuissima was successfully established, which laid basis for studying of the gene function of Alternaria tenuissima.

关 键 词：ALTERNARIA tenuissima G418 RESISTANCE PROTOPLAST TRANSFORMATION 

分 类 号：S432.4[农业科学—植物病理学]