利用CRISPR-Cas9系统高效创制水稻品系“17318”的OsGW2、OsGN1a基因定向突变体  

Creation Mutants of OsGW2 and OsGN1a in Rice Cultivar “17318” via CRISPR/Cas9

作　　者：周建平[1] 吉琳艺 何瑶 郑雪莲[1] 罗樊 蔡光泽[2] 张勇[1] ZHOU Jian-ping1, JI Lin-yi1, HE Yao1, ZHENG Xue-lian1, LUO Fan2, CAI Guang-ze2, ZHANG Yong1(1. Center for Informational Biology, School of Life Science and Technology, University of Electronic Science and Technology of China, Chengdu 610054, China;2. Xichang University, Xichang, Sichuan 615013, China)

机构地区：[1]电子科技大学生命科学与技术学院信息生物学中心,成都610054 [2]西昌学院,四川西昌615013

出　　处：《西昌学院学报（自然科学版）》2018年第3期1-5,共5页Journal of Xichang University(Natural Science Edition)

基　　金：四川省国际科技合作与交流研发项目(2018HH0112);四川省青年科学基金项目(2017JQ0005);中央高校基本科研基金项目(ZYGX2016J119;ZYGX2016J122)

摘　　要：CRISPR-Cas9系统在水稻功能基因的功能研究、性状改良与分子设计育种中起着重要的作用。本研究成功构建了水稻高效定向敲除OsGW2和OsGN1a的CRISPR/Cas9载体pZJP049。并将该载体通过农杆菌介导转化水稻"17318"愈伤,获得水稻再生苗。PCR扩增的方法进行转基因植株阳性检测,结果显示阳性率为100%。PCR-RFLP结果表明OsGN1a基因位点的突变效率为77.7%。PCR-SSCP检测结果显示OsGW2基因位点突变率为100%。双基因聚合突变体(gw2gw2gn1agn1a)的种子在长度和宽度均有明显增加,与预期一致。本研究通过CRISPR-Cas9系统快速聚合Gn1a, GW2基因,实现了目标性状的精确改良。CRISPR-Cas9 has significant influence on gene function research, agronomic traits improvement andmolecular design breeding. This research successful constructed highly efficient vector pZJP049 that act as OsGW2and OsGN1a double gene knockout. And we transformed pZJP049 into“17318”through agrobacterium mediation,obtained transformation. PCR verified positive rate in seedling is 100% . PCR-RFLP showed mutant efficiency atOsGN1a is 77.7%. PCR-SSCP showed mutant efficiency at OsGW2 is 100%. The grain size （the grain length andwidth） in the double mutant （gw2gw2gn1agn1a） is larger than WT. This research achieved the integration of GN1aand GW2 and precise gene editing by CRISPR-Cas9.

关 键 词：CRISPR-Cas9 水稻 OsGW2 OsGN1a 

分 类 号：Q943.2[生物学—植物学] S511.033[农业科学—作物学]