利用CRISPR/Cas9技术编辑粒长基因GS3改善粳稻花时  

CRISPR/Cas9-mediated Editing of GS3 to Improve Flowering Time in japonica Rice

作　　者：孟帅 徐鹏 张迎信[1] 王宏[1] 曹立勇[1] 程式华[1] 沈希宏[1] MENG Shuai;XU Peng;ZHANG Yingxin;WANG Hong;CAO Liyong;CHENG Shihua;SHEN Xihong(China National Rice Research Institute/State Key Laboratory of Rice Biology/Key Laboratory for Zhejiang Super Rice Research,Hangzhou 311401,China)

机构地区：[1]中国水稻研究所/浙江省超级稻研究重点实验室/水稻生物学国家重点实验室,杭州311401

出　　处：《中国水稻科学》2018年第2期119-127,共9页Chinese Journal of Rice Science

基　　金：国家863计划资助项目(2014AA10A603);国家公益性农业科技研究专项(201403002);中国农业科学院创新工程超级稻育种创新团队和水稻杂种优势研究机理研究创新团队资助项目(CAAS-ASTIP-2013-CNRRI);国家自然科学青年科学基金资助项目(31501290)

摘　　要：【目的】花时不遇是影响杂交水稻制种产量的直接因素之一,已有研究表明水稻粒型差异对花时有影响。本研究采用GS3功能缺失突变来研究粳稻花时差异,以期为粒型影响花时提供佐证。【方法】利用CRISPR/Cas9技术来定向编辑控制粒长基因GS3,获得13对粒型差异的近等基因系粳稻,小区种植,采用目测法来调查花时。【结果】获得转基因T0植株并对其T1植株测序分析,发现长白25、吉粳102、浙粳88、武运粳27和J42均发生单碱基插入移码突变,垦鉴稻6号、空育131、浙粳22、扬粳4227、南粳9108、J5933、J6167和J5938均发生部分碱基缺失突变。对T1植株粒型考查表明,gs3突变体的粒长均显著长于野生型。对稳定的后代花时统计分析发现,粒型变长的突变体均比野生型花时有所提前,且吉粳102、空育131、浙粳88、武运粳27、扬粳4227这5个材料的gs3突变体花时均显著早于野生型,其余材料开花也提早,但不显著。【结论】长粒型粳稻GS3突变体的花时早于短粒粳稻野生型,这一研究可为粳稻粒型育种提供参考,加速长粒粳稻亲本选育,有望推动杂交粳稻发展。【Objective】Flowering time asynchrony is one of the main factors limiting seed production of hybrid rice between male and female parents,and some studies have shown that flowering time is related to grain shape.Loss of function mutant of GS3 was used to study the difference of flowering time in this study so as to elucidate the impact of grain shape on flowering time.【Method】CRISPR/Cas9-mediated editing of GS3 to achieve 13 pairs of near-isogenic line japonica varieties.Then the flowering time was investigated with visual methods.【Result】T1 individuals derived from T0 generation were genotyped.The sequencing results show that some bases were mutated with single base insertion,including Changbai 25,Jijing 102,Zhejing 88,Wuyunjing 27,J42 and others with bases deletion,including Kenjiandao 6,Kongyu 131,Zhejing 22,Yangjing 4227,Nanjing 9108,J5933,J6167,J5938.For the grain shape of T1,the gs3 mutants conferred longer grain length compared to the wild type,and the gs3 mutants of Jijing 102,Kongyu 131,Zhejing 88,Wuyunjing 27 and Yangjing 4227 were significantly earlier in flowering time than those of the wild type.The others were also earlier,but not significant.【Conclusion】Long-grain japonica rice has an earlier flowering time than short-grain japonica rice,which could provide references to study the grain-type breeding of japonica rice,and it would accelerate the breeding process of long-grain japonica parents and promote the development of hybrid japonica rice.

关 键 词：粳稻 花时 CRISPR/Cas9 GS3 

分 类 号：Q785[生物学—分子生物学] S511.032[农业科学—作物学]