含羞草山蚂蟥等几种豆科植物根瘤菌的数值分类及-中国农学通报.pdfVIP

中国农学通报 2012,28(15): 135-141 Chinese Agricultural Science Bulletin 含羞草、山蚂蟥等几种豆科植物根瘤菌的数值分类 及16SrDNAPCR-RFLP分析 刘晓云，魏 爽，刘庆辉，王立江，郭振国，郅慧梅 （河北大学生命科学学院，河北保定071002） 摘 要：为揭示云南省豆科植物根瘤菌的资源多样性，采用数值分类、16S rDNA PCR-RFLP 和16S rDNA 全序列分析技术对云南省8 个地区含羞草、山蚂蝗等几种不同豆科植物分离的48 株根瘤菌菌株进行了 遗传和表型多样性研究。16S rDNA PCR 产物经Hinf Ⅰ、Msp Ⅰ、Rsa Ⅰ和Hae Ⅲ4 种内切酶消化后产生 22 种遗传图谱类型；数据经MVSP3.0 软件聚类后，所有供试菌株在70%的相似性水平上分为4 个分支， 经16S rDNA 全序列分析，确定分支Ⅰ为伯克霍尔德属(Burkholderia)类群，含1株菌株；分支Ⅱ为贪铜菌 属(Cupriavidus)类群，含5 株菌株；分支Ⅳ为根瘤菌属(Rhizobium)类群，包括29 株菌株；而分支Ⅲ没有和 参比菌株聚在一起，经测序为狭长平胞属(Stenotrophomonas)类群。通过对菌株的抗生素抗性、耐盐性、 生长温度范围、BTB 产酸产碱反应以及对3-酮基乳糖利用情况共计28 个表型性状测定，结果显示所有 菌株耐盐性较强，多数菌株可耐受60℃的高温。实验表明，分离自云南省的这些菌株具有丰富的遗传和 表型多样性。 关键词：含羞草；山蚂蝗；根瘤菌；数值分类；16S rDNA PCR-RFLP 分析 中图分类号：Q939.1 文献标志码：A 论文编号：2011-3383 NumericalTaxonomyand16SrDNAPCR-RFLPofRhizobiaofSomeLegumesIsolated fromMimosaspp.andDesmodiumspp. Liu Xiaoyun, Wei Shuang, Liu Qinghui, Wang Lijiang, Guo Zhenguo, Zhi Huimei (CollegeofLifeSciences,HebeiUniversity, Baoding Hebei 071002) Abstract: In order to characterize the diversity and phylogeny of rhizobium in Yunnan of China, 48 strains isolated from root nodules of Mimosa spp. and Desmodium spp. in 8 regions of Yunnan Province were studied with numerical taxonomy, 16S rDNA PCR-RFLP and 16S rDNA gene sequencing analysis. 23 genotype profiles were obtained from the fingerprinting of the 16S rDNA PCR-RFLP by four restriction endonucleases Hinf Ⅰ, Msp Ⅰ, Rsa Ⅰand Hae Ⅲ. These strains were clustered into 4 branches at the similarity level of 70% by MVSP3.0 software. 16S rDNA sequencing revealed that branch Ⅰbelonging to the genera of Burkholderia, include 1 strain; branch Ⅱbelonging to the