论文部分内容阅读
【目的】研究新疆红井子盐碱土壤中的放线菌物种多样性。【方法】应用基于16S rRNA基因序列系统发育分析的免培养方法进行放线菌物种多样性分析。利用放线菌特异性引物,以土壤样品总DNA为模板,扩增16S rRNA基因,构建16S rRNA基因克隆文库,并对文库中的插入序列进行RFLP分析。【结果】随机挑选的246个阳性克隆通过酶切筛选出61个不同图谱的重组克隆并测序。分析结果显示这61个克隆序列分属于42个OTUs,分布于放线菌纲(Actinobacteria)的放线菌亚纲(Actinobacteridae)、酸微菌亚纲(Acidimicrobidae)和红色杆菌亚纲(Rubrobacteridae);该环境中有71.4%的序列与已有效发表菌株的序列相似性小于97%,代表着放线菌新类群,其中部分序列形成了几个独立的进化分支,可能代表更高级的新分类单元。【结论】红井子土壤中的放线菌组成具有丰富的多样性,并有新放线菌分类单位的潜在资源,值得进一步进行开发研究。
【Objective】 To study the species diversity of actinomycetes in Hongjingzi saline-alkali soil in Xinjiang. 【Method】 Species diversity of actinomycetes was analyzed using culture-free culture based on 16S rRNA gene sequence phylogenetic analysis. 16S rRNA gene was amplified by using actinomycete specific primer and total DNA of soil sample as template. The 16S rRNA gene clone library was constructed and the inserted sequence in the library was analyzed by RFLP. 【Result】 The results showed that 246 positive clones were screened by enzyme digestion and sequenced 61 different clones. The results showed that the 61 cloned sequences belong to 42 OTUs distributed in Actinobacteria Actinobacteridae, Acidimicrobidae and Rubrobacteridae. In this environment, 71.4% of the sequences shared less than 97% sequence identity with those of the published strains, representing a new class of actinomycetes. Some of these sequences formed several independent evolutionary branches and may represent more advanced new taxonomic units. 【Conclusion】 The composition of actinomycetes in the soil of Hongjingzi is rich in diversity and has the potential resources of the new actinomycete taxonomic unit, so it is worth further research and development.