水稻锌指蛋白基因OsWIP6的克隆、表达及RNAi载体转化

Cloning, Expression and RNAi Vector Transformation of a Zinc Finger Protein Gene OsWIP6 from Rice

为了解水稻C2H2型锌指蛋白转录因子基因的表达调控机理及生物学功能,克隆了水稻的一个C2H2型锌指蛋白转录因子基因(OsWIP6),并构建其RNA干涉植物表达载体,通过农杆菌介导转化获得转基因植株.生物信息学分析显示,OsWIP6氨基酸序列与拟南芥两个WIP家族转录因子高度同源.组织差异性表达分析表明,该基因在水稻花发育上具有表达偏向性.与野生型相比,转基因水稻表现为抽穗延迟,结实率及千粒重降低,半定量分析显示转基因植株OsWIP6表达量明显下降.因此,推断OsWIP6基因与水稻育性密切相关.

For understanding expression regulation mechanism and biological functions of C2H2-type zinc-finger transcription factor in rice,in this study,OsWIP6,a C2H2-type zinc-finger transcription factor gene in rice was cloned,and its RNA interference（RNAi） vector was constructed.The vector was introduced into rice cv.Nipponbare callus by means of Agrobacterium-mediated transformation.Bioinformatics analysis indicated that OsWIP6 had a high homology with two WIP family transcription factors in Arabidopsis thaliana.Tissue differential expression analysis showed that OsWIP6 preferred to express in flowers.Compared with wild-type plants,the heading date of transgenic plants was significantly delayed;the seed setting rate and 1 000-grain weight also decreased.Semi-quantitative RT-PCR revealed a significant reduction of endogenous OsWIP6 expression levels in the RNAi transgenic plants.Consequently,OsWIP6 may be closely related with rice fertility.