利用EMS诱变创制抗除草剂粳稻新种质  

New Herbicide-Resistant Japonica Rice Germplasms Created by EMS Mutagenesis

作　　者：王付华[1] 李自超[2] 王亚[1] 付景[1] 杨文博[1] 尹海庆[1] 王生轩[1] 王越涛[1] 白涛[1] 张珍[3] WANG Fuhua;LI Zichao;WANG Ya;FU Jing;YANG Wenbo;YIN Haiqing;WANG Shengxuan;WANG Yuetao;BAI Tao;ZHANG Zhen(Institute of Cereal Crops,Henan Academy of Agricultural Sciences,Zhengzhou 450002,China;Agricultural College,China Agricultural University,Beijing 100193,China;Henan Agricultural Science and Technology Exhibition Hall,Zhengzhou 450000,China)

机构地区：[1]河南省农业科学院粮食作物研究所,河南郑州450002 [2]中国农业大学农学院,北京100193 [3]河南省农业科学技术展览馆,河南郑州450000

出　　处：《河南农业科学》2021年第4期8-16,共9页Journal of Henan Agricultural Sciences

基　　金：国家重点研发计划项目(2017YFD0100400,2016YFD0100101)。

摘　　要：为了创制抗除草剂水稻新种质,用EMS(甲基磺酸乙酯)溶液浸泡郑稻19干种子,在M2幼苗三叶期分别喷施112.5 g/hm2的甲咪唑烟酸和60 g/hm2的烟嘧磺隆,14 d后筛选抗除草剂突变体,并对突变体ALS(Acetolactate synthase)基因序列进行分析,找出ALS基因突变位点。结果表明,喷施甲咪唑烟酸14 d后,抗除草剂植株生长正常、叶色偏绿,敏感植株黄化、生长受抑制;喷施烟嘧磺隆14 d后,抗除草剂植株表现前期受抑制,但能缓慢恢复。用甲咪唑烟酸筛选63000份M2家系,获得抗除草剂突变体6份(HF1—HF6),包括2种ALS氨基酸突变类型:Ser-627-Asn和Gly-628-Glu。用烟嘧磺隆筛选M2家系30000份,获得抗除草剂突变体2份(HF7—HF8),包括2种ALS氨基酸突变类型:Pro-171-Ser和Ala-179-Val。在M2筛选到1个特定ALS碱基突变平均需23250份M2家系。筛选到的抗甲咪唑烟酸突变体HF1(Gly-628-Glu)和HF2(Ser-627-Asn)在1倍大田常规除草剂剂量(112.5 g/hm2)下鲜质量减退率分别为13.09%和20.03%,显著小于郑稻19(P<0.05);在8倍大田常规除草剂剂量(900 g/hm2)下突变体仍能正常生长,表现强的甲咪唑烟酸抗性。对HF1、HF2分别喷施30 g/hm2和60 g/hm2烟嘧磺隆,其鲜质量减退率分别为24.81%、13.13%和34.13%、28.01%,显著小于郑稻19(P<0.05),表现一定的烟嘧磺隆抗性。综上,筛选到8份抗除草剂突变体材料,2份抗烟嘧磺隆,6份抗甲咪唑烟酸,其中2份抗甲咪唑烟酸突变体兼具烟嘧磺隆抗性,创制的抗除草剂材料可用于选育非转基因抗除草剂品种,也可作为创制ALS多位点突变种质的基础材料,进一步增强突变体对除草剂的抗性。In order to create new herbicide-resistant rice germplasms,the dry seeds of Zhengdao 19 were soaked in ethylmethane sulfonate(EMS)solution.At three-leaf stage,M2 seedlings were sprayed with imazapic(112.5 g/ha)and nicosulfuron(60 g/ha).The herbicide-resistant mutants were screened after 14 days.The ALS(acetolactate synthase)gene sequences of mutants were analyzed to find out the mutation sites.The results showed that the herbicide-resistant plants grew normally after spraying imazapic,the leaf kept green,while the leaves of sensitive lines yellowed,and their growth were inhibited;after spraying nicosulfuron,the growth of herbicide-resistant plants were inhibited in the early stage,but could recover slowly.A total of 63000 M2 lines were screened with imazapic,and 6 imazapic-resistant mutants(HF1—HF6)were obtained,including two types of ALS mutations,Ser-627-Asn and Gly-628-Glu.A total of 30000 M2 lines were screened with nicosulfuron,and 2 herbicide-resistant mutants(HF7—HF8)were obtained,including two types of ALS mutations,Pro-171-Ser and Ala-179-Val.To obtain a specific ALS nucleotide mutation,an average of 23250 M2 lines were needed to screen.The fresh weight reduction rates of imazapic-resistant mutants HF1(Gly-628-Glu)and HF2(Ser-627-Asn)were 13.09%and 20.03%respectively after spraying 1 time the conventional herbicide dose(112.5 g/ha),which were significantly less than that of Zhengdao 19(P<0.05);the mutants could still grow normally after spraying 8 times the conventional herbicide dose in the field,showing strong resistance to imazapic herbicides.The fresh weight reduction rates of mutants HF1 and HF2 were 24.81%and 13.13%after spraying 30 g/ha nicosulfuron,and were 34.13%and 28.01%after spraying 60 g/ha nicosulfuron,respectively,which were significantly less than those of Zhengdao 19(P<0.05),showing certain nicosulfuron resistance.In summary,8 herbicide-resistant mutant lines were obtained,2 lines were resistant to nicosulfuron,6 lines were resistant to imazapic,and two of the imazapic-resistant muta

关 键 词：粳稻 抗烟嘧磺隆 抗甲咪唑烟酸 抗除草剂 EMS诱变 种质创建 

分 类 号：S511[农业科学—作物学]