利用CRISPR/Cas9技术编辑水稻温敏不育基因TMS  

CRISPR/Cas9-mediated editing of the thermo-sensitive genic male-sterile gene TMS5 in rice

作　　者：陈日荣 周延彪 王黛君 赵新辉 唐晓丹 许世冲 唐倩莹 符星学 王凯[3] 刘选明 杨远柱 CHEN Ri-Rong;ZHOU Yan-Biao;WANG Dai-Jun;ZHAO Xin-Hui;TANG Xiao-Dan;XU Shi-Chong;TANG Qian-Ying;FU Xing-Xue;WANG Kai;LIU Xuan-Ming;YANG Yuan-Zhu(Hunan Province Key Laboratory of Plant Functional Genomics and Developmental Regulation/College of Biology,Hunan University,Changsha 410082,Hunan,China;Yuan Longping High-Tech Agriculture Co.Ltd./Key Laboratory of Rice Germplasm Enhancement in Southern China,Ministry of Agriculture and Rural Affairs,Changsha 410001,Hunan,China;Yahua Seeds Science Academy of Hunan,Changsha 410119,Hunan,China;College of Plant Science and Technology,Huazhong Agricultural University,Wuhan 430070,Hubei,China;College of Life Sciences,Hunan Normal University,Changsha 410081,Hunan,China)

机构地区：[1]湖南大学生物学院/植物功能基因组学与发育调控湖南省重点实验室,湖南长沙410082 [2]袁隆平农业高科技股份有限公司/农业农村部南方水稻品种创制重点实验室,湖南长沙410001 [3]湖南亚华种业科学研究院,湖南长沙410119 [4]华中农业大学植物科学技术学院,湖北武汉430070 [5]湖南师范大学生命科学学院,湖南长沙410081

出　　处：《作物学报》2020年第8期1157-1165,共9页Acta Agronomica Sinica

基　　金：国家自然科学基金项目(31901516);国家转基因生物新品种培育重大专项(2016ZX08001-004);湖南省自然科学青年基金项目(2019JJ50414)资助。

摘　　要：tms5是生产上应用最广泛的温敏不育基因。为创制新型水稻温敏核不育系,利用CRISPR/Cas9基因编辑技术将6个优异的粳稻和4个优异的籼稻的TMS5基因敲除,获得了温敏核不育系。比较发现,粳稻温敏不育系ZG75S、CYGS、YG0618S、ZG07S、T0361S、7679S的不育起点温度在28~32°C之间,籼稻温敏核不育系2537S、6150S、6379S的不育起点温度在24~28°C之间,而籼稻温敏核不育系1109S的不育起点温度低于23.5°C。说明不同遗传背景材料获得的tms5突变体的不育起点温度不一样,通过TMS5的敲除可能获得不育起点温度较低的温敏核不育系。利用1109S与优质父本8048选配出优质高产杂交稻组合1109S/8048。大田试验表明, 1109S/8048比区试对照丰两优4号增产13.1%。温敏核不育系1109S及高产杂交稻组合1109S/8048的创制为高产育种提供了新的途径。Thermo-sensitive genic male-sterile(TGMS)gene tms5 is most widely used in the two-line hybrid breeding system in China.To develop novel rice thermo-sensitive male sterile lines,we knocked out the TMS5 genes of six elite japonica and four indica rice varieties by using CRISPR/Cas9 gene editing technology.By analyzing the critical sterility-inducing temperature(CIST)of the newly TGMS lines,it was found that the CIST of japonica TGMS lines ZG75 S,CYGS,YG0618 S,ZG07 S,T0361 S,and 7679 S were between 28°C and 32°C,the CIST of indica TGMS lines 2537 S,6150 S and 6379 S were between 24°C and 28°C,and the CIST of indica TGMS line 1109 S was lower than 23.5°C.These results indicated that the CIST of tms5 mutant from different genetic background materials was different.The TGMS lines with lower CIST could be obtained by knocking out the TMS5 from different genetic background materials.A hybrid rice combination 1109 S/8048 had high quality and high yield.The yield of 1109 S/8048 was 13.1%higher than that of Fengliangyou 4.The creation of the TGMS 1109 S and the high-yield cross combination 1109 S/8048 provides a new way for high-yield breeding.

关 键 词：水稻 CRISPR/Cas9 温敏不育 TMS5 杂交组合 

分 类 号：Q943.2[生物学—植物学] S511[农业科学—作物学]