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中国农学通报

中国农学通报 ›› 2012, Vol. 28 ›› Issue (15): 135-141.doi: 10.11924/j.issn.1000-6850.2011-3383

所属专题： 生物技术

• 生物技术科学 • 上一篇    下一篇

含羞草、山蚂蟥等几种豆科植物根瘤菌的数值分类及16S rDNA PCR-RFLP分析

刘晓云 魏爽 刘庆辉 王立江 郭振国 郅慧梅   

  • 收稿日期:2011-11-15 修回日期:2012-01-13 出版日期:2012-05-25 发布日期:2012-05-25
  • 基金资助:

    国家自然科学基金

Numerical Taxonomy and 16S rDNA PCR-RFLP of Rhizobia of Some Legumes Isolated from Mimosa spp. and Desmodium spp.

  • Received:2011-11-15 Revised:2012-01-13 Online:2012-05-25 Published:2012-05-25

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摘要：

为了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HinfⅠ、MspⅠ、RsaⅠ和HaeⅢ 4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: 白肋烟, 白肋烟, 马里兰烟, 叶绿素酶, 实时定量PCR

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 Ⅰ and 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 Ⅰ belonging to the genera of Burkholderia, include 1 strain; branch Ⅱ belonging to the genera of Cupriavidus, include 5 strains; branch Ⅳ belonging to the genera of Rhizobium, include 29 strains; branch Ⅲ belonging to the genera of Stenotrophomonas with no reference strains. The analysis of 28 phenotypic characteristic of these strains showed that most of these strains had a high level resistance to saline, some strains could grow under severe environment such as high content ration of antibiotics (300 μg/mL) and high temperature (60℃). All the results demonstrated a high genetic and phenotypic diversity of rhizobium nodulating legumes of Yunnan Province.

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