EMS诱变水稻创制抗咪唑啉酮除草剂种质  

Screening of Imidazolinone-Resistant Rice From EMS-Mutated Populations

作　　者：陈天子[1] 余月 凌溪铁 张保龙[1] CHEN Tianzi;YU Yue;LING Xitie;ZHANG Baolong(Provincial Key Laboratory of Agrobiolgy/Jiangsu Academy of Agricultural Sciences,Nanjing,Jiangsu 210014)

机构地区：[1]江苏省农业科学院/江苏省农业生物学重点实验室,江苏南京210014

出　　处：《核农学报》2021年第2期253-261,共9页Journal of Nuclear Agricultural Sciences

基　　金：江苏省重点研发计划(现代农业)项目(BE2018381)。

摘　　要：为了创制水稻非转基因抗除草剂种质,丰富我国水稻轻简化栽培杂草防治所需的配套种质资源,本研究以0.5%(w/v)甲磺酸乙酯(EMS)水溶液诱变水稻种子14 h,然后以咪唑啉酮类除草剂筛选3~4叶期的M2水稻幼苗。结果发现,M2幼苗茎叶喷雾160 mg·L^(-1)甲咪唑烟酸15 d后,非抗性苗黄化、生长被抑制甚至枯死,抗性苗叶色正常、株高增长明显。M3抗性水稻对甲咪唑烟酸的抗性能稳定遗传且其抗性水平不低于1200 mg·L^(-1)甲咪唑烟酸,其抗性水平是野生型对照的100倍以上。分别克隆M3抗除草剂突变体和野生型的乙酰乳酸合成酶基因(ALS),序列比对分析发现M3抗性水稻的ALS基因开放阅读框在第1879、1880 nt由AG碱基突变为CT碱基,导致编码的第627位氨基酸由丝氨酸(AGT)突变为亮氨酸(CTT),或在第1880 nt由G碱基突变为A碱基,导致编码的第627位氨基酸由丝氨酸(AGT)突变为天冬酰胺(AAT)。间接比色法测定M3抗除草剂突变体与野生型的ALS对甲咪唑烟酸的敏感性,结果表明抗性水稻对甲咪唑烟酸的耐受性是野生型的109倍。本研究获得了水稻ALS第627位丝氨酸突变为亮氨酸或天冬酰胺的抗咪唑啉酮新种质材料,获得的抗咪唑啉酮水稻材料的抗性倍数高,能稳定遗传,具有生产应用价值。In order to create non-transgenic herbicide-tolerant rice germplasm which would control the weed easily in rice simplified cultivation in China,rice seeds were treated with 0.5%(w/v)ethyl methanesulfonate for 14 h.The resulted M2 generation at 3-4 leaf stage were sprayed with 160 mg·L^(-1)of imazapic.The non-resistant seedlings turned into yellow,stunted or even died,while the resistant seedlings were normal and the plant height increased evidently.The resistance of M2 to imazapic was stably inherited to their progeny M3,reached up to 1200 mg·L^(-1)imazapic,and 100 times as high as the wild type did.The acetolactate synthase(ALS)genes in the imidazolinone-resistant M3 mutants and the wild-type were cloned by PCR.Sequence analysis revealed that the open reading frame of ALS gene in the imidazolidone-resistant M3 plants carry two points mutations A1879 C and G1880 T,which results in a substitution of serine(AGT)to leucine(CTT)at amino acid 627,or a point mutation G1880 A,which results in a substitution of serine(AGT)to asparagine(AAT)at amino acid 627.A colorimetry method was used to compare the enzyme sensitivity of ALS to imazapic between the herbicide resistant M3 mutant and the wild type.The tolerance of ALS enzyme to imazapic in the imidazolinone resistant rice was 109-fold of that in the wild type.Taken together,imidazolidone-resistant rice mutants with substitution of serine to leucine at amino acid 627 was a novel mutation and was first reported.The imidazolidone-resistant rice obtained in this study showed stable and high resistance to imazapic,make it potential in the application of weed control.

关 键 词：甲基磺酸乙酯(EMS) 水稻 乙酰乳酸合成酶 咪唑啉酮 

分 类 号：S451.21[农业科学—植物保护]