拟南芥中根瘤共生基因线路的搭建及其表达分析  

Construction and expression analyses of nodule symbiosis genes route in Arabidopsisthaliana

作　　者：梅江鹏 李豪[1] 张忠明[1] MEI Jiangpeng, LI Hao ,ZHANG Zhongming(State Key Laboratory of Agricultural Microbiology ,Life Sciences and Technology Huazhong Agricultural University ,Wuhan 430070, Chin)

机构地区：[1]华中农业大学生命科学技术学院/农业微生物学国家重点实验室,武汉430070

出　　处：《华中农业大学学报》2018年第2期28-33,共6页Journal of Huazhong Agricultural University

基　　金：国家自然科学基金项目(31670240);教育部高等学校博士学科点专项(20130146130003)

摘　　要：为研究共生信号途径相关基因在非宿主植物中的表达情况,通过生物技术及合成生物学方法,将10个能在拟南芥根中表达的启动子和10个百脉根共生信号传递的关键基因分别构建2个多基因表达载体,即pUNS6(含6个基因)和pDNS4(含4个基因)。结果表明:利用毛根转化互补相应豆科植物的突变体,都能恢复突变体的结瘤功能,说明克隆的目的基因都具有生物学功能;通过根癌农杆菌介导的花序侵染,将pUNS6和pDNS4分别转入拟南芥中,对阳性植株分离和鉴定,在转基因T_0、T_1、T_2代植株中都鉴定到目的基因,表明多个共生基因稳定存在于拟南芥基因组上;利用RT-PCR在阳性转基因植株根中都能检测到mRNA水平的基因表达。Ten key genes in symbiosis signaling fromLotus japonicus were controlled under ten different individual promoters expressed in roots of Arabidopsis thaliana with biotechnology and synthetic biology approaches.Two multigenes vectors,pUNS6 containing 6 genes and pDNS4 containing 4 genes,were constructed respectively to transform Arabidopsisthaliana.The result of complement assay showed that ten genes controlled in CaMV35 Spromoter were able to restore symbiotic function of relative genes mutant in leguminous plants with hairy root transformation.The pUNS6 and pDNS4 were transformed into Arabidopsis with Agrobacterium tumefaciens-mediated inflorescence infection respectively.The result of identifying the transgenic T_0,T_1 and T_2 plants by PCR and RT-PCR showed that those of foreign symbiotic genes were stably existed on genome and were able to express at the RNA level in Arabidopsis thaliana.

关 键 词：百脉根 拟南芥 共生信号基因 花序侵染 

分 类 号：S182[农业科学—农业基础科学] Q78[生物学—分子生物学]