转hrpZ_(Psta)基因大豆株系的分子鉴定及其抗疫霉根腐病分析  

Molecular Identification of Transgenic hrpZ_(Psta) Gene Soybean Lines and Analysis of Resistance to Phytophthora Root Rot

作　　者：邵瑞超 冀采凤 姜南 周靖萱 刘添祎 王丕武[1] SHAO Ruichao;JI Caifeng;JIANG Nan;ZHOU Jingxuan;LIU Tianyi;WANG Piwu(College of Agronomy,Jilin Agricultural University,Changchun 130118,China)

机构地区：[1]吉林农业大学农学院,长春130118

出　　处：《吉林农业大学学报》2024年第4期538-544,共7页Journal of Jilin Agricultural University

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

摘　　要：以T_(7)、T_(8)代转hrpZ_(Psta)基因大豆株系JN29-705-21为供试材料,通过分子鉴定和室内人工接种疫酶菌进行抗病性分析。常规PCR结果表明:在T_(7)、T_(8)代转基因株系中均检测到转化的目的基因hrpZ_(Psta)、筛选标记Badh,启动子35S和终止子Nos,表明hrpZ_(Psta)基因在高世代转基因株系中能够稳定遗传;Southern杂交结果显示:hrpZPsta基因以单拷贝的形式整合到大豆基因组中,且整合位点不同;qRT-PCR检测结果表明:hrpZ_(Psta)基因在转化株系的根、茎、叶中均有表达,其中在叶中表达量最高,在茎中表达量最低,T_(7)代株系的根、茎、叶平均相对表达量分别为2.800,1.772,8.367,T_(8)代株系的根、茎、叶平均相对表达量分别为3.028,1.650,9.036;利用下胚轴侵染法对转化株系进行抗病鉴定分析,结果显示,转基因株系后代对疫霉根腐病的抗病级别为抗病,而非转基因受体株系抗性为中抗,表明转基因株系的抗病能力得到了提高。T_(7) and T_(8) generation transgenic hrpZ_(Psta) gene soybean line JN29-705-21 was used as the test material,and disease resistance analysis was performed by molecular identification and indoor artificial inoculation of phytophthora infestans.The results of routine PCR showed that the target gene hrpZ_(Psta),screening marker Badh,promoter 35S and terminator Nos were detected in T_(7) and T_(8) transgenic lines,which indicated that hrpZ_(Psta) gene could be inherited stably in high generation transgenic lines.Southern hybridization results showed that the hrpZ_(Psta) gene was integrated into the soybean genome as a single copy with different integration sites;the inspection results of qRT-PCR showed that hrpZ_(Psta) gene was expressed in root,stem,and leaf of the transformed lines,among which the highest expression was in leaf and the lowest in stem.The average relative expression of root,stem,leaf in the T_(7) generation lines was 2.800,1.772 and 8.367,respectively;and the average rela⁃tive expression of root,stem,and leaf in the T_(8) generation lines was 3.028,1.650 and 9.036,respectively.The hypocotyl infection method was used to analyze the disease resistance of the transformed lines.The results showed that the resistance level of the offspring of the transgenic lines to phytoph⁃thora root rot was positive,but the resistance of the non-transgenic recipient lines was medium,indi⁃cating that the disease resistance of transgenic lines have improved.

关 键 词：大豆 疫霉根腐病 hrpZ_(Psta)基因 抗性分析 

分 类 号：S565.1[农业科学—作物学]