西瓜枯萎病菌T-DNA插入转化子库的构建及其质量评价  

Generation of a Transformant Library of Fusarium oxysporum.f. sp. niveum by ATMT and It's Quality Assessment

作　　者：裴月令[1] 曾凡云[1] 彭军[1] 龙海波[1] 郭建荣[1] 

机构地区：[1]中国热带农业科学院环境与植物保护研究所农业部热带作物有害生物综合治理重点实验室海南省热带农业有害生物监测与控制重点实验室,海南海口571101

出　　处：《热带作物学报》2015年第1期76-82,共7页Chinese Journal of Tropical Crops

基　　金：公益性行业(农业)科研专项经费项目"南繁区生物安全监测预警及控制关键技术研究与示范"(No.201403075);国家自然科学基金(No.31401685);中国热带农业科学院环境与植物保护研究所引进人才科研启动基金项目(No.Hzs1201);人事部留学人员科技活动择优项目

摘　　要：为研究西瓜枯萎病菌的致病分子机理,开展了病原菌的转化子库构建工作。采用携带潮霉素抗性p TFCM双元载体的农杆菌AGL-1介导的ATMT转化方法 ,获得菌株FON-01的转化子,通过表型观察筛选突变体。结果表明:当乙酰丁香酮(AS)浓度为200μmol/L、农杆菌OD600为0.4、分生孢子浓度为1×106个/m L时,于28℃共培养48 h转化效率高达(663.33±24.95)个/106个孢子。构建了总数为3 832个的转化子库并对其进行了质量评价,从2 201个转化子中筛选出菌落形态、生长速度及产孢量等方面有所变异的突变体73个。病原菌转化子库的构建为进一步开展致病分子机理及相关基因克隆等方面研究奠定基础。In order to study the pathogenic molecular mechanism of Fusarium oxysporum, f. sp. Niveum, the transformant library was conducted. Agrobacterium strainAGL-1 containing the binary vector pTFCM was used for the transformation of strain FON-01, and the mutants were screened by penotype observation. Transformation efficiency was（663.33±24.95）, when the acetosyringone concentration was 200 μmol/L, OD600 of A GL-1 was 0.4, conidia concehntration was 106/mL, co-cuhure time was 48 h and co-culture temperature was 28℃. 3 832 transformants were obtained and quality assessment to shape, growth speed and sporulation ability were transformants library would provide the foundation for genes＇ eloning. the library was finished. 73 mutants with variation of colony screened from 2 201 transformants. The construction of further study of pathogenic molecular mechanism and related genes＇ cloning.

关 键 词：西瓜枯萎病菌 ATMT 转化子库 

分 类 号：S432.1[农业科学—植物病理学]