根癌农杆菌介导的香蕉枯萎病菌4号生理小种的转化  

Agrobacterium tumefaciens-mediated transformation of Fusarium oxysporum f.sp.cubense race 4

作　　者：李敏慧[1] 张荣[1,2] 姜大刚[3] 习平根[1] 庄楚雄[3] 姜子德[1] 

机构地区：[1]华南农业大学资源与环境学院,广州510642 [2]华南农业大学园艺学院,广州510642 [3]华南农业大学生命科学院,广州510642

出　　处：《植物病理学报》2009年第4期405-412,共8页Acta Phytopathologica Sinica

基　　金：国家自然科学基金(30571207);广东省自然科学基金(E05202480)

摘　　要：本文针对香蕉枯萎病菌4号生理小种这一对我国香蕉生产构成了极大威胁的检疫性有害生物,建立了根癌农杆菌介导的该生理小种的转化体系,确定了影响转化效率主要因子的最佳条件分别是:共培养乙酰丁香酮(AS)浓度为200μmol/L、共培养时间为48 h、培养温度为25℃、诱导培养基pH值为5.5。此条件下,转化效率达到21～24个转化子/104香蕉枯萎病菌孢子。PCR和Southern杂交分析表明外源的T-DNA已经成功随机地整合到该病原菌基因组中,且多为单拷贝。应用该转化体系已获得近25 000个转化子,为研究该生理小种致病机制奠定了基础。Fusarium oxysporum f. sp. cubense （FOC） race 4, the quarantine fungus caused a highly destructive vascular wilt disease of banana, had been transformed by using Agrobacterium tumefaciens-mediated trans- formation. The objective of the study was to optimize the transformation system to obtain T-DNA inserted-mutants for gene functional study. The key conditions included co-cultivation with 200μmol/L acetosyringone for 48 h, induction cultivation at pH 5.5 and 25℃. Under the conditions, transformation efficiency was in the range of 21 -24 transformants per 104 FOC race 4 conidia. PCR and Southern blot analysis of the transformants showed that the T-DNA had been successfully integrated into genome of the fungus with the majority of single copy at random sites. A total of 25 000 transformants had been obtained in the study, which provided a foundation for better understuding molecular mechanism of FOC pathogenicity.

关 键 词：香蕉枯萎病菌4号生理小种 农杆菌介导的转化 影响因子优化 

分 类 号：S432.1[农业科学—植物病理学] Q789[农业科学—农业昆虫与害虫防治]