EMS诱发小麦京411突变群体构建及Wx-A1基因突变体鉴定

Construction of Mutant Population for Wheat Cultivar Jing411 Induced by EMS and Identification of Wx-A1 Gene

为扩大小麦突变群体类型,提高目的基因点突变挖掘效率,选用北部冬麦区骨干亲本京411为材料,利用甲基磺酸乙酯（ethyl methylsulfonate,EMS）化学诱变剂构建小麦突变群体,以Wx-A1为候选基因,用TILLING技术检测所创建的突变群体。结果表明,0.90%EMS溶液处理8 h和1.50%EMS溶液处理4 h均能获得较好的损伤效果,致死率分别达到38.47%和56.00%;在此基础上创建了包含867个株系的M2突变群体,在该群体中获得Wx-A1基因的7个点突变,突变密度为1/67.0 kb;其中预测有功能变异的4个错义突变系均可以稳定遗传至下一代,其直链淀粉含量降低2.8%~7.4%。本研究所构建的小麦突变群体可以作为其他目的基因的TILLING检测材料,用于小麦目的基因突变挖掘及功能基因组学研究。

In order to increase diversity of mutation population and improve point mutation frequency in wheat,we developed a novel TILLING population using winter wheat cultivar Jing411 by Ethyl methylsulfonate（ EMS） treatment.Mutation frequency was detected by TILLING platform using target gene Wx-A1. The results showed that damage effects of both treatments with 0. 90% EMS solution in 8 h and 1. 50% EMS in 4 h were better than those of other treatments,and lethal rates were 38. 47% and 56. 00%,respectively. A M2 population containing 867 lines was constructed by treatment with 1. 0% and 1. 5% EMS. Seven point mutations of Wx-A1 gene were identified,and its mutation density in average was 1 /67. 0 kb. Four missense mutant lines were stably inherited to next generation,and these mutations might affect its protein function,the amylose contents in seeds was decreased 2. 8% to 7. 4%. As a novel genetic resource,this mutant population could be used for point mutation identification for target genes,and it provides novel alleles for functional genomic studies of wheat.