新疆艾比湖湿地博乐河入口处土壤细菌多样性分析

Bacterial diversity in Bole river entrance soil of Ebinur Lakewetland,Xinjiang by 16S rRNA gene sequence analysis

【目的】了解新疆艾比湖湿地国家级自然保护区非培养土壤细菌群落组成及多样性。【方法】采用非培养法直接从湿地土壤提取总DNA进行16S r RNA基因扩增,构建细菌16S r RNA基因克隆文库。使用MspⅠ和AfaⅠ限制性内切酶对阳性克隆进行16S r RNA基因扩增片段的限制性酶切分析(Amplified r DNA restriction analysis,ARDRA),挑取具有不同双酶切图谱的克隆进行测序,序列比对并构建16S r RNA基因系统发育树。【结果】从土壤细菌的16S r RNA基因文库中随机挑取75个不同谱型的克隆子,共得到58个OTUs,系统发育归类为8个细菌类群:绿弯菌门(Chloroflexi)、蓝藻门(Cyanobacteria)、变形菌门(Proteobacteria)、厚壁菌门(Firmicutes)、放线菌门(Actinobacteria)、拟杆菌门(Bacteroidetes)、疣微菌门(Verrucomicrob)和芽单胞菌门(Gemmatimonadetes)。其中,变形菌门为第一优势菌群,拟杆菌门为第二优势菌群,两者约占总克隆的65%。【结论】艾比湖湿地博乐河入口处土壤细菌多样性丰富,且存在一定数量的潜在微生物新种。

[Objective] In order to investigate bacteria diversity and community composition in Bole river entrance soil of the Ebinur Lake wetland National Nature Reserve,Xinjiang.[Methods] Total DNA was directly extracted from the soil of the Ebinur Lake wetland Bole river entrance using the culture independent method.16 S r RNA gene was amplified using bacterial primer set Eubac27 F and Eubac1492 R.16 S r RNA gene clone library was constructed.Positive clones were identified by amplified r DNA restriction analysis（ARDRA） using Msp I and Afa I,and unique r DNA pattern clones were sequenced,analysed,and then the phylogenetic tree was constructed.[Results] Seventy-five different clones of Macrorestriction Map was classified into 58 operational taxonomic units（OTUs）,which were associated with 8 phyla by phylogenetic analysis,including Chloroflexi,Cyanobacteria,Proteobacteria,Firmicutes,Actinobacteria,Bacteroidetes,Verrucomicrob and Gemmatimonadetes.Among them,the most abundant phyla was Proteobacteria,followed by Bacteroidetes,accounting for about 65% of the total clones.[Conclusion] The bacterial diversity is abundant in the Ebinur Lake wetland Bole river entrance soil,and exist a certain amount of new unknown taxon in this environment.