Identification of and spatio-temporal differences between microbial assemblages from two neighboring sulfurous lakes:: Comparison by microscopy and denaturing gradient gel electrophoresis
Identification of and spatio-temporal differences between microbial assemblages from two neighboring sulfurous lakes:: Comparison by microscopy and denaturing gradient gel electrophoresis
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DOI：
10.1128/aem.66.2.499-508.2000
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发表时间：
2000-02-01
影响因子：
4.4
通讯作者：
Muyzer, G
中科院分区：
文献类型：
--
作者：
Casamayor, EO;Schäfer, H;Muyzer, G
通过显微镜、pcr -变性梯度凝胶电泳（DGGE）和16S rRNA基因片段序列分析，对西班牙东北部Banyoles的Cis6湖和Vilar湖的微生物群落进行了空间和时间分析。从不同的水深和一年中的两个不同时间（在完全融合的冬季和在浮游植物繁殖的早春）获得的样本进行了分析。虽然湖泊具有相同的气候条件和相同的水源，但湖泊参数不同，显微镜下列举的大多数形态可区分的光合细菌也是如此。通过比较16S rRNA序列分析推断了主要DGGE条带的系统发育关系。从Lake Cis6样品中获得的序列与革兰氏阳性菌和变形菌门的成员有关。从Vilar湖样品中获得的序列与细胞噬菌-黄杆菌-拟杆菌门和蓝藻门的成员有关。因此，我们发现，就像先前报道的形态不同的两湖居民之间的差异一样，形态差异不显著的群落成员之间也存在差异。冬春两季的物种组成变化也很明显。这两个湖泊都含有属于光养绿硫细菌的序列，这与显微镜观察结果一致，但这些序列与以前从湖泊中分离的培养菌株的序列不同。古Euryarchaeal序列（即与甲烷菌和热等离子体相关的序列）也在两个湖泊中存在。这些euryarchaeal群序列在Cis6湖占优势，而在Vilar湖不占优势。在维拉尔湖，一个与绿古菌有关的新的浮游生物种群产生了占优势的古菌带。系统发育分析表明，存在新的细菌和古细菌谱系，这些组合的微生物多样性比以前已知的要大。我们通过比较显微镜和测序结果来评估几种形态型和DGGE条带丰度之间的对应关系。我们的数据提供了证据，证明从DGGE指纹中获得的序列与实际存在于自然系统中浓度较高的微生物相对应。
The microbial assemblages of Lake Cis6 and Lake Vilar (Banyoles, northeast Spain) were analyzed in space and time by microscopy and by performing PCR-denaturing gradient gel electrophoresis (DGGE)and sequence analysis of 16S rRNA gene fragments. Samples obtained from different water depths and at two different times of the year (in the winter during holomixis and in the early spring during a phytoplankton bloom) were analyzed. Although the lakes have the same climatic conditions and the same water source, the limnological parameters were different, as were most of the morphologically distinguishable photosynthetic bacteria enumerated by microscopy. The phylogenetic affiliations of the predominant DGGE bands were inferred by performing a comparative 16S rRNA sequence analysis. Sequences obtained from Lake Cis6 samples were related to gram-positive bacteria and to members of the division Proteobacteria. Sequences obtained from Lake Vilar samples were related to members of the Cytophaga-Flavobacterium-Bacteroides phylum and to cyanobacteria. Thus, we found that like the previously reported differences between morphologically distinct inhabitants of the two lakes, there were also differences among the community members whose morphologies did not differ conspicuously. The changes in the species composition from winter to spring were also marked. The two lakes both contained sequences belonging to phototrophic green sulfur bacteria, which is consistent with microscopic observations, but these sequences were different from the sequences of cultured strains previously isolated from the lakes. Euryarchaeal sequences (i.e., methanogen- and thermoplasma-related sequences) also were present in both lakes. These euryarchaeal group sequences dominated the archaeal sequences in Lake Cis6 but not in Lake Vilar. In Lake Vilar, a new planktonic population related to the crenarchaeota produced the dominant archaeal band. The phylogenetic analysis indicated that new bacterial and archaeal lineages were present and that the microbial diversity of these assemblages was greater than previously known. We evaluated the correspondence between the abundances of several morphotypes and DGGE bands by comparing microscopy and sequencing results. Our data provide evidence that the sequences obtained from the DGGE fingerprints correspond to the microorganisms that are actually present at higher concentrations in the natural system.