水稻转录因子JAMYB和ERF3的克隆及遗传转化  

Cloning and Genetic Transformation of Transcription Factor JAMYB and ERF3 in Rice

作　　者：祝传书[1,2] 武莹[1] 李威[1] 冯俊涛[1,2] 张兴[1,2] 

机构地区：[1]西北农林科技大学植物保护学院,西北农林科技大学无公害农药研究服务中心,陕西杨陵712100 [2]陕西省生物农药工程技术研究中心,陕西杨陵712100

出　　处：《西北植物学报》2012年第10期1935-1941,共7页Acta Botanica Boreali-Occidentalia Sinica

基　　金：国家重点基础研究发展计划(2006CB102005);国家公益性行业(农业)科研专项(200903052)

摘　　要：以水稻‘秀水11’为材料,通过RT-PCR的方法,从二化螟取食后的水稻中分离出2个特异诱导表达的转录因子基因MYB(MYB transcription factor gene)和ERF(Ethylene Response Factor gene),Blastn比对分析确定所克隆的基因是水稻OsJAMYB(AY026332)和OsERF3(AB036883),基因长度分别为1 026bp和805bp。克隆转化与载体构建,获得了含有过量表达和反义抑制OsJAMYB与OsERF3的4个特异插入片段的表达载体;以农杆菌介导法侵染水稻‘秀水11’愈伤,经共培养、筛选、分化等步骤,获得了含有OsJAMYB和OsERF3正义和反义表达的转基因植株。PCR检测T0代水稻植株结果表明,OsJAMYB和OsERF3已整合到水稻基因组中。该结果为进一步阐明OsJAMYB和OsERF3基因在水稻诱导抗虫反应中的作用和抗虫反应的分子机理奠定了基础。Through RT-PCR,two transcription factor genes,OsJAMYB （MYB transcription factor gene,Genbank no.AY026332） and OsERF3 （Ethylene Response Factor gene,Genbank no.AB036883） were isolated from rice ‘Xiushui 11’ after striped rice borer ［Chilo supperssalis （Walker）］ feeding and were confirmed after Blastn analysis.The length of OsJAMYB and OsERF3 fragments were 1 026 bp and 805 bp,respectively.By using molecular cloning techniques,specific expression vectors were gotten with specific directions and specific insert sequence including OsJAMYB and OsERF3 for transformation by Agrobacterium tumefaciens.After incubating,screening and differentiation with ‘Xiushui 11’ calli and A.tumefaciens,the sense and antisense transgenic plants including OsJAMYB and OsERF3 were gained.The PCR was used to amplify the genes in T0 transformed rice genomics DNA to confirm the inserts of OsJAMYB and OsERF3.The results will provide the knowledge of induced resistance and molecular mechanism of OsJAMYB and OsERF3 in rice.

关 键 词：水稻 转录因子 OsJAMYB OsERF3 遗传转化 

分 类 号：Q785[生物学—分子生物学]