根癌农杆菌介导遗传转化稻曲病菌.pdf

根癌农杆菌介导遗传转化稻曲病菌 张震，杜新法’，柴荣耀，毛雪琴，邱海萍，王艳丽，王教瑜，孙国昌 (浙江省农科院植物保护与微生物所，浙江杭州310021： ’通讯联系人，Email：duxf@zaas．org) 【摘要】利用根癌农杆菌介导转化系统，对稻曲病菌分生孢子进行转化，通过潮霉 素抗性筛选转化子。农杆菌经乙酰丁香酮预先诱导，转化效率大约在390-450个转 化子／106个孢予。PCR检钡IJHPH基因表达盒，结果显示被测转化子基因组中均成功整 合了目的片段。同时，在488nm下这些转化子都具有荧光。利用根癌农杆菌成功转 化稻曲病菌，这为进一步研究稻曲病菌致病分子机制的有力工具。 关键词：稻曲病菌；根癌农杆菌：转化 transformationof the Agrobacteriumtumefaciens—mediated of virens phytopathologyUstilaginoidea ZHANG Zhen，DUXin-fa，CItAI Rong-yao，MAOXue-qin，QIUHal—ping，WANG Yah—li，WANG Jiao-yu，SUNGuochang (InstitutePlantProtectionand Academe of of Sciences, Microbiology,ZhejiangAgricultural Hangzhou，310021．China；‘Correspondingauthor，Email：du．xf@zaas．ore) Abstract：Conidiaof virensweretransformedto B Ustilaginoidea hygromycin a resistance transformation a Agrobacterium system by tumefaciens-mediated using vector stable binaryplasmid pBIG2RHPH2一GFP—GUS．AtMTyielded hygromycin B-resistant 200·27010。conidia)，afterstrainC58C1with clones(averageper pre·culture inthetransformantswaschecked AS．The ofthe casket presencefragmentof枷gene PCR．AndalsoalltestedtransformatsshowedEGFPfluorescencein in by mycelium transformationan tumefaciens-mediatedprovidesopportunity 488nm．Agrobacterium for studiesofthemolecularofthe