抗病基因hrpZ_(Psta)转大豆‘吉农18’后代对疫霉根腐病的抗性分析  

Resistance Analysis of Resistant hrpZ_(Psta) Gene Transgenic Soybean'Jinong18'Progenies to Phytophthora Infestans Root Rot

作　　者：郭轩麟 刘添祎 仉祺 张靓 冀采凤 周靖萱 邵瑞超 王丕武[1] Guo Xuanlin;Liu Tianyi;Zhang Qi;Zhang Liang;Ji Caifeng;Zhou Jingxuan;Shao Ruichao;Wang Piwu(Centre for Plant Biotechnology,College of Agronomy,Jilin Agricultural University,Changchun,130118)

机构地区：[1]吉林农业大学农学院,植物生物技术中心,长春130118

出　　处：《分子植物育种》2021年第5期1578-1583,共6页Molecular Plant Breeding

基　　金：国家转基因生物新品种培育重大专项(2016ZX08004-004)资助。

摘　　要：本试验以T7、T8代转基因株系JN18-hrpZ_(Psta)-45为试验材料,运用常规PCR技术、Southern杂交及qRT-PCR技术对T7、T8代大豆株系进行分子检测和稳定性鉴定,随即以下胚轴侵染法为理论依据完成疫霉根腐病抗性鉴定,同时分析其抗病性。结果表明:终止子Nos、启动子35S、筛选标记Bar的目的条带和转化的目的基因在T7、T8代转基因株系中被全部测获;Southern印迹杂交证实:目的基因在大豆基因组中以单拷贝方式完成整合;qRT-PCR检测证实:植株籽粒、叶、茎、根全部有目的基因表达,T7代株系的根、茎、叶、籽粒相对表达量均值分别为2.45、1.73、2.52、1.91,T8代株系的根、茎、叶、籽粒相对表达量均值分别为3.22、2.04、3.46、2.75,证实茎中的表达量最低、叶中最高,以抗病性标准为依据展开对照可以获得如下抗病性鉴定结论:转、非转基因二者的抗病级别分别为高抗、中抗。T7/T8 generation of JN18-hrpZ_(Psta)-45 transgenic plants were used as the source of plant materials.Use PCR,Southern blot and qRT-PCR to test the stability of hrpZ_(Psta) expression,and identification of resistance to Phytophthora infestans root rot by Hypocotyl infection and analysis of resistance to Phytophthora infestans.The results show that target gene,Bar gene,CaMV35S promoter and Nos poly-A terminator,were expressed in transformed soybean.The Southern blot assay revealed that single-copy of the fragment was inserted in the transformed soybean.The qRT-PCR results indicate the target gene was expressed in all organs(Leaves,shoots,roots and seeds)of the transgenic soybeans,with the highest expression level in leaves and lowest in roots,the hrpZ_(Psta) expression of roots,shoots,leaves and seeds in T7 generation are 2.45,1.73,2.52,1.91,the hrpZ_(Psta) expression of roots,shoots,leaves and seeds in T8 generation are 3.22,2.04,3.46,2.75.According to the standard of disease resistance,the results showed that the resistance grade of non-transgenic control was medium resistance,and that of transgenic lines was high resistance.

关 键 词：大豆 hrpZ_(Psta)基因 疫霉根腐病 抗性 

分 类 号：S435.651[农业科学—农业昆虫与害虫防治]