大豆R_(SC4)抗病候选基因Glyma.14G204700的克隆及其生物信息学分析  

Cloning and bioinformatics analysis of R_(SC4) resistance candidate gene Glyma.14G204700 in soybean

作　　者：王大刚[1] 陈圣男 杨勇[1] 李杰坤[1] 吴倩[1] 胡国玉[1] 黄志平[1] Wang Dagang;Chen Shengnan;Yang Yong;Li Jiekun;Wu Qian;Hu Guoyu;Huang Zhiping(Key Laboratory of Crop Quality Improvement of Anhui Province,Crop Research Institute,Anhui Academy of Agricultural Sciences,Hefei 230031,Anhui Province,China;College of Resources and Environment,Straits Joint Research Institute,Fujian Agriculture and Forestry University,Fuzhou 350002,Fujian Province,China)

机构地区：[1]安徽省农业科学院作物研究所,安徽省农作物品质改良重点实验室,合肥230031 [2]福建农林大学海峡联合研究院,福建农林大学资源与环境学院,福州350002

出　　处：《植物保护学报》2022年第4期1013-1021,共9页Journal of Plant Protection

基　　金：安徽省重点研发计划(202004a06020034);安徽省农业科学院人才项目(QNYC-201909);财政部和农业农村部国家现代农业产业技术体系建设专项(CARS-04-PS07);安徽省农业科技成果转化专项(2021ZH003)。

摘　　要：为明确大豆抗大豆花叶病毒(soybean mosaic virus,SMV)SC4株系的基因位点R_(SC4)(resistance to SMV strain SC4,R_(SC4))的候选基因Glyma.14G204700在大豆不同抗性品种中的序列结构特征和保守结构域,以齐黄1号、科丰1号、大白麻和南农1138-2共4个大豆品种为材料,通过基因克隆获得大豆Glyma.14G204700基因的cDNA全长序列,并采用生物信息学方法分析其序列特征和编码蛋白的理化特性及结构特征。结果表明,大豆Glyma.14G204700基因在4个大豆品种中的cDNA全长为4719~4776 bp,编码1295~1307个氨基酸,预测蛋白的分子量为148.38~149.33 kD,等电点为5.53~5.61,均为具较强亲水性的非分泌蛋白。Glyma.14G204700蛋白含有植物抗病基因家族蛋白的保守功能域——核苷酸结合域和富含亮氨酸重复结构域。该蛋白的二级结构主要由α-螺旋、无规则卷曲、延伸链和β-折叠组成,所占比例分别为59.45%~61.08%、28.65%~30.15%、8.11%~8.88%和1.61%~2.16%。启动子序列分析发现该基因含有脱落酸、低温及干旱等多种逆境胁迫响应元件。表明大豆R_(SC4)抗病候选基因Glyma.14G204700在大豆不同抗性品种中具有不同的等位变异,优异等位变异的鉴定和开发可为抗SMV大豆育种提供基础材料。To characterize Glyma.14G204700,a candidate gene underlying R_(SC4),a locus for resistance to soybean mosaic virus(SMV)strain SC4 in soybean Glycine max,the full-length cDNA sequences were cloned from four soybean varieties Qihuang No.1,Kefeng No.1,Dabaima and Nannong 1138-2 with contrasting SMV resistances,and sequence features,physicochemical and structural characteristics of the deduced proteins were then characterized using bioinformatics methods.The results showed that Glyma.14G204700 from the four soybean varieties varied from 4719 bp to 4776 bp in length,and encoded non-secretory structural proteins ranging from 1295 to 1307 amino acids with molecular weights ranging from 148.38 kD to 149.33 kD,isoelectric points of 5.53-5.61.These proteins all showed a strong hydrophilicity,and contained conserved functional domains that were typical in plant resistance family proteins,including the nucleotide binding domain and leucine-rich repeat.Analysis of the secondary structure demonstrated that these proteins mainly consisted of an α-helix(59.45%-61.08%),random coil(28.65%-30.15%),an extended chain region(8.11%-8.88%),and a β-sheet(1.61%-2.16%).Promoter sequence analysis indicated that it contained many stress response elements which were responsive to abscisic acid,drought,and low temperature stress etc.The results indicated that the R_(SC4) resistance candidate gene,Glyma.14G204700,contained allelic variation in soybean.The identification and utilization of superior allelic variation could facilitate the breeding for soybean varieties with better SMV resistance.

关 键 词：大豆 抗病基因 克隆 生物信息学分析 

分 类 号：S435.651[农业科学—农业昆虫与害虫防治]