百脉根根瘤菌nod-lacZ转录融合子构建及表达分析  

Construction and expression study of Mesorhizobium loti nod-lacZ fusions

作　　者：裴俊清 段柳剑 张忠明[1] PEI Junqing, DUAN Liujian, ZHANG Zhongming(State Key Laboratory of Agricultural Microbiology, 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1-8,共8页Journal of Huazhong Agricultural University

基　　金：国家自然科学基金项目(31670240)

摘　　要：根据Mesorhizobium loti基因组信息,克隆nodA、nodB、nodD等基因的启动子序列,分别与报告基因构建相应的转录融合子(nod-lacZ),并分别导入中慢生型百脉根根瘤菌MAFF303099和广谱根瘤菌NGR234(Rhizobium sp.)中,利用百脉根根的抽提物作为诱导物,检测结瘤基因的表达。结果表明:在MAFF303099菌株中,百脉根根抽提物不能诱导nodA的表达;能诱导nodB和nodD1的表达;nodD2呈组成型表达;供试的5种不同的类黄酮化合物都不能诱导nodA基因表达;在NGR234菌株中,毛地黄酮(Luteolin)能诱导nodA表达。The promoter sequences of nodA,nodB,nodD were cloned based on the genomic information of Mesorhizobium loti MAFF303099 and fused with the reporter gene lacZ to construct the nodlacZ fusion separately.Then these fusions were separately transformed into M.loti MAFF303099 and Rhizobium sp.NGR234 to detect the expression of these nod genes after induced by Lotus japonicus root extract（RE）.The results showed that there was no expression of nodA when treated with root extracts.RE induced expression of nodBand nodD1,but showed upregulation of just 2-3 times.nodD2 expressed constitutively in M.loti MAFF303099.Neither of these induced the expression of nodA in M.loti MAFF303099 induced by 5 different flavonoid compounds separately,but the expression of nodA was detected in NGR234 strain treated by Luteolin.

关 键 词：百脉根根瘤菌 结瘤基因 根抽提物 nod-lacZ转录融合子 

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