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【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】 To understand the composition and diversity of bacterial community in uncultured soil in Aibi Lake Wetland National Nature Reserve of Xinjiang. 【Method】 The 16S rRNA gene was amplified by direct DNA extraction from wetland soils using non-culture method, and a bacterial 16S rRNA gene clone library was constructed. The positive clones were sequenced using Amplified r DNA restriction analysis (ARDRA) using Msp I and Afa I restriction enzymes, and clones with different double digestion maps were sequenced. The sequences The 16S rRNA gene phylogenetic tree was aligned and constructed. 【Result】 Fifty-eight OTUs were randomly selected from 16S rRNA gene library of soil bacteria. The phylogenetic relationships were classified into eight bacterial groups: Chloroflexi, Cyanobacteria, Proteobacteria, Firmicutes, Actinobacteria, Bacteroidetes, Verrucomicrob and Gemmatimonadetes ). Among them, Proteobacteria was the first dominant bacteria, and Bacteroidetes was the second dominant bacteria, accounting for about 65% of the total clones. 【Conclusion】 The bacterial diversity in the soil at the Bole River inlet of the Aibi Lake wetland is abundant, and a certain number of new potential microorganisms exist.