郑58/opaque2近等基因系中opaque2基因突变位点分析与高效分子标记开发  

Analysis of the Mutation Site of opaque2 Gene in Near-isogenic Line Zheng 58/opaque-2 and Development of Functional Molecular Marker

作　　者：韩小花[1] 鲁晓民[1] 周波[1] 魏良明[1] 王延召[1] 张前进[1] 朱卫红[1] 魏昕[1] 郭书磊[1] 刘康 郭金生[1] 王振华[1] 张新[1] HAN Xiao-hua;LU Xiao-min;ZHOU Bo;WEILiang-ming;WANG Yan-zhao;ZHANG Qian-jin;ZHU Wei-hong;WEI Xin;GUO Shu-lei;LIU Kang;GUO Jin-sheng;WANG Zhen-hua;ZHANG Xin(The Cereal Crops Institute,Henan Academy of Agricultural Sciences/Henan Key Labortary of Maize Biology,Zhengzhou 450002,China)

机构地区：[1]河南省农业科学院粮食作物研究所/河南省玉米重点实验室,郑州450002

出　　处：《玉米科学》2019年第2期61-68,共8页Journal of Maize Sciences

基　　金：国家重点研发计划(2016YFD0101205-4);国家重点研发计划项目(2016YFD0300309/03);河南省科技攻关项目(172102110075);河南省基础与前沿技术研究计划项目(162300410178)

摘　　要：opaque2突变体材料是最常用的高赖氨酸玉米供体。对已获得的郑58/o2近等基因系研究表明,胚乳发育不同时期22-kDα-醇溶蛋白的积累均明显低于郑58。荧光定量PCR结果表明,α-醇溶蛋白家族基因Z1A、Z1B、Z1C和Z1D的表达均显著低于郑58。郑58/o2胚乳发育不同时期Opaque2基因均正常表达。测序分析发现,郑58/o2中o2基因ATG后713 bp处缺失10个碱基,ORF预测Opaque2蛋白翻译提前终止。针对突变缺失位点开发基因内分子标记o2-indel-1,利用该标记进行回交转育,结果表明,o2-indel-1与o2突变表型完全连锁,错选率为0。opaque2基因突变位点的解析有助于高效分子标记的开发,有效降低错选率,提高优质蛋白鲜食玉米多基因聚合育种的选择效率。Maize Opaque2(o2)mutant gene enhances lysine content of endosperm as compared with the wild genotype and o2 mutant is the main donor for high-quality protein fresh maize breeding.When compared with Zheng58,Zheng58/o2 had reduction of the accumulation of 22-k Daα-zeins tested by SDS-PAGE.However,q RT-PCR showed that the expression level of the o2 gene in Zheng 58/o2 was normal during the whole endosperm developing period.The expression levels ofα-zein genes(Z1 A,Z1 B,Z1 C and Z1 D)in the o2 mutant were significantly reduced.Sequence analysis of o2 mutant gene in zheng58/o2 showed that a 10-nucleotide deletion mutant occurred at 713 bp downstream of ATG,which resulted in frame shift and early termination of Opaque2 protein translation.Further,a functional molecular marker o2-indel-1 was developed based on the deletion mutation.The marker was completely associated with o2 mutant phenotype when used in marker-assisted backcross breeding.Analysis of o2 mutation site would contribute to the development of functional molecular marker and efficiency increase of high-quality protein fresh maize breeding through marker assisted selection.

关 键 词：玉米 醇溶蛋白 opaque2 基因突变 

分 类 号：S513[农业科学—作物学]