陕西太白尾矿废弃地豆科植物根瘤菌16S rDNA PCR-RFLP及全序列分析  

16S rDNA PCR-RFLP and Sequencing of Legume Rhizobia Isolated from Taibai Mining Tailing of Shaanxi Province

作　　者：余建福[1] 韦革宏[1] 丁小平[1] 曹鹏[1] 

机构地区：[1]西北农林科技大学生命科学学院陕西省农业分子生物学重点实验室,陕西杨陵712100

出　　处：《西北植物学报》2007年第3期502-508,共7页Acta Botanica Boreali-Occidentalia Sinica

基　　金：高等学校博士学科点专项科研基金(20050712013);霍英东青年教师基金(101029);全国优秀博士学位论文作者基金(200254)

摘　　要：应用16S rDNA-RFLP和16S rDNA全序列测定方法,对分离自陕西太白金矿尾矿废弃地的55株根瘤菌和12株参比菌株进行了遗传多样性和系统发育地位研究。采用平均连锁法(UPMGA)对16S rDNA PCR-RFLP聚类,结果显示所有菌株在72%的水平上聚到一起。根据参比菌株的种属关系,将供试菌株初步分成6个遗传发育群。群Ⅰ为根瘤菌属,群Ⅱ为中华根瘤菌属,群Ⅲ是中慢生根瘤菌属,群Ⅳ为土壤杆菌属,群V为一未知群,群Ⅵ为慢生根瘤菌属。分离自天蓝苜蓿的根瘤菌主要分布在群Ⅱ,截叶胡枝子根瘤菌在各个群内均有分布,表现出丰富的遗传多样性。选取群Ⅰ、Ⅱ的代表菌株TB17-1、TB50-1进行16S rDNA全序列测定分析,结果显示TB17-1与Rhizonbium leguminosarumUSDA2370的同源性高达99.7%,TB50-1与Sinorhizobium melilotiLMG6133的同源性为100%。全序列测定结果与RFLP分析结果基本一致。The genetic diversity and phylogeny of 55 rhizobia isolated from eight genera of legume plant originating from Taibai gold mining tailing in Shaanxi province and 12 recognized reference strains was evaluated by RFLP analysis of 16S rRNA genes and 16S rDNA sequencing. The dendrogram of 16S rDNA PCR-RFLP revealed the isolates were assembled at 72% similarity level. According to reference strains genera position,the tested strains were classified into 6 distinct 16S rRNA gene groups. The group Ito group IV belonged to the genera of Rhizobium,Sinorhizobium,Mesorhizobium and Agrobacterium; group V was an unknown group and group Ⅵ was within Bradyrkizobiurn respectively. Isolates from Medicago lupulina were clustered into group Ⅱ within the genus of Sinorhizobium and isolates from Lespedeza cuneata distributed into each group and characterized versatile genetic diversity. Phylogenetic tree of 16S rDNA sequences constructed from representative strains of group Ⅰ TB17-1 and group Ⅱ TB50-1. Similarity analysis demonstrated TB17-1 was merged into Rhizobium genus,and high to 99.7% similarity level with Rkizonbium legurainosarum USDA2370 ; TB50-1 was 100 ; similar to Sinorhizobium meliloti LMG6133.The results of 16S rDNA PCR-RFLP were in good agreement with sequencing analysis.

关 键 词：根瘤菌 16S rDNA RFLP 系统发育 尾矿废弃地 

分 类 号：Q939.114[生物学—微生物学]