广东金山温泉沉积物中原核与真核微生物多样性初步分析

目的]本研究旨在采用不同的PCR引物对广东省恩平市金山温泉的高温水底沉积物微生物多样性进行初步的分析.方法]采用改进的玻璃珠法抽提温泉沉积物中环境基因组DNA,通过对用4对引物分别扩增得到的原核微生物16S rRNA基因和真核微生物ITS序列的分析,将所得到的数据与国际基因数据库GenBank进行相似性比较并构建系统发育树.结果]研究发现原核类群G的 14个优势克隆中7个都属于蛭弧菌属(Bdellovibrio).与它们最相似的序列是从海洋中分离到的两个菌株 Bacteriovorax sp. NE1 (EF092445)和Bdellovibrio sp. JS5 (AF084859),相似性分别为96%和99%.原核类群X的4个序列主要属于蓝细菌类群,其中JS-X2与在美国黄石公园温泉发现的Uncultured Cyanobacterium (L35331)有95%的相似性,并且与已经全基因组测序的嗜热蓝细菌聚球藻Thermosynechococcus elongatus BP-1 (47118315)有89%的相似性.真核类群Z有三个类群,分别是Penicillium sp.,Lodderomyces sp.和Gloeotinia sp..其中大部分序列与青霉属相似性在88%～ 90%之间.结论]所得到的结果显示金山温泉中的微生物多样性十分丰富.

A molecular view of microbial diversity in sediment of the Jinshan hot spring, Guangdong province

OBJECTIVE: The aim of this study is to analyze the microbial diversity in the sediment of Jingshan hot spring. METHOD: We extracted environment DNA from the sediment and then amplified the 16S rRNA genes of prokarotype and internal transcribed spacers (ITS) DNA of eukarotype by PCR with specific conservative primers. The PCR products were ligated to the T vectors and transformed into Escherichia coli to construct libraries. The libraries were analyzed by amplified ribosomal DNA restriction analysis (ARDRA) approach. The clones of dominant ARDRA patterns were selected to sequence. The sequence similarities were analyzed by using the BLAST programs for searching the GenBank DNA databases. RESULTS: We identified 14 dominant operational taxonomic units (OTUs) from prokarotype G. group. The Phylogenetic tree shows that 7 of them (JS-GU20, JS-GU1, JS-G37, JS-G29, JS-GM2, JS-G42, JS-G41) belong to Bdellovibrio. They had the highest similarity to Bacteriovorax sp. NE1 (EF092445) and Bdellovibrio sp. JS5 (AF084859), 96% and 99% respectively. Four major OTUs of prokarotype X group belong to Cyanobacterium. One of them, named JS-X2, has a similarity of 95% with an uncultured Cyanobacterium (L35331) which was gained from a hot spring of the Yellowstone National Park and has a similarity of 89% with Thermosynechococcus elongatus BP-1 (47118315). Most OTUs of eukaryotic group have 90% sequences similarity to that of Penicillium sp.. CONCLUSION: The results showed that the microbial diversity in the sediment of Jinshan hot spring is rich. And the result of several clones' similarity to Bacteriovorax sp. showed that there might be a new group of thermophilicBacteriovorax sp. in the hot spring sediment.