水稻卷窄叶突变相关基因OsCSLD4的RNAi研究及表达分析  

RNAi AND EXPRESSION ANALYSIS OF A GENE OsCSLD4,WHICH CONTROLS PLANT NARROW AND ROLLED LEAF IN RICE

作　　者：朱丽[1] 胡江[1] 颜美仙[1] 高振宇[1] 刘坚[1] 钱前[1] 郭龙彪[1] 

机构地区：[1]中国水稻研究所/水稻生物学国家重点实验室,浙江杭州310006

出　　处：《核农学报》2010年第5期873-880,共8页Journal of Nuclear Agricultural Sciences

基　　金：中央级公益性科研院所专项资金项目(2009RG002-1);国家转基因专项(2009ZX08001-022B);国家支撑项目(2006BAD13B01)

摘　　要：在图位克隆获得目的基因OsCSLD4的基础上,利用Invitrogen公司的Gateway系统将500bp的OsCSLD4基因cDNA编码序列以反向互补的方式插入pANDA35HK,构建了水稻卷窄叶突变相关基因OsCSLD4的干涉载体pANDAH05。利用农杆菌转化法将干涉载体pANDAH05及对照空载体pANDA35HK分别转化突变体原始亲本日本晴幼胚。转基因阳性植株表型观察结果显示:干涉载体pANDAH05转化日本晴幼胚后,T0代植株出现了类似于突变体1ah137的表型,叶片明显变窄,卷曲程度增加,株高降低;对照空载体pANDA35HK转化日本晴幼胚后,T0代植株仍保持野生型表型;Real-timePCR检测结果显示:日本晴转干涉载体pANDAH05后,目的基因表达量明显降低,干涉强度不同目的基因表达量降低程度不同,研究结果表明目的基因OsCSLD4在水稻叶片形态发育过程中起着非常重要的作用。Based on the cloned gene OsCSLD4,a 500bp OsCSLD4 cDNA fragment was inserted into the binary vector pANDA35HK to generate the transformation vector pANDAH05 for RNA interference by Gateway System.The two vectors,pANDAH05 and pANDA35HK（control）,were introduced into rataria of Nipponbare by Agrobacterium tumefaciens（EHA105） mediated transformation,respectively.RNAi plants designed to repress OsCSLD4 displayed the reduced leaf width,semi-rolling and different extent of dwarfism.Real-time PCR analysis indicated that the expression of target gene from transgenic plant was lower than that of the Nipponbare.These results indicated that OsCSLD4 played an important role in leaf morphogenesis and development in rice.

关 键 词：OsCSLD4基因 RNA干涉 REAL-TIMEPCR 水稻 

分 类 号：S511[农业科学—作物学]