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采用 PCR-DGGE 技术对 16S rRNA 基因片段分析盐分层淡水湖中细菌的分布和多样性。

Distribution and diversity of bacteria in a saline meromictic lake as determined by PCR-DGGE of 16S rRNA gene fragments.

机构信息

Dipartimento di Biologia Animale ed Ecologia Marina, Sezione di Ecologia Microbica e Biotecnologie, Università di Messina, V.le F. Stagno d'Alcontres, 31, 98166, Messina, Italy.

出版信息

Curr Microbiol. 2011 Jan;62(1):159-66. doi: 10.1007/s00284-010-9688-0. Epub 2010 Jun 11.

Abstract

The variations in vertical distribution and composition of bacteria in the meromictic Lake Faro (Messina, Italy) were analysed by culture-independent methods in two different mixing conditions. Water samples were collected from a central station from the surface to the bottom (30 m depth) on two different sampling dates--the first characterised by a well-mixed water mass and the second by a marked stratification. A 'red-water' layer, caused by a dense growth of photosynthetic sulphur bacteria, was present at a depth of 25 m in December 2005 and at 15 m in August 2006, defining two different zones in terms of their physicochemical properties. The vertical distribution of bacterioplankton showed that the interface zones were more densely populated than others. In both sampling periods, the highest numbers of live cells were observed within 'red water' layers. The dominant phylotypes of the bacterial community were determined by sequencing the Denaturing Gradient Gel Electrophoresis (DGGE) bands resulting from PCR amplification of 16S rRNA gene fragments. The number of DGGE bands, considered indicative of the total species richness, did not vary predictably across the two different sampling periods. Proteobacteria (α-, γ-, δ- and ε subclass members), Cytophaga-Flavobacterium-Bacteroides, green sulphur bacteria and Cyanobacteria were retrieved from Lake Faro. Most of the bands showed DNA sequences that did not match with other previously described organisms, suggesting the presence of new indigenous bacterial phylotypes.

摘要

利用培养独立方法,分析了混合条件不同的分层湖法罗(意大利墨西拿)中细菌的垂直分布和组成变化。在两个不同的采样日期,从一个中心站从表面到底部(30 米深)采集水样 - 第一个特征是充分混合的水团,第二个特征是明显的分层。2005 年 12 月,在 25 米深处出现了一层“红水”,由密集生长的光合硫细菌引起,在 2006 年 8 月,在 15 米深处出现了一层“红水”,这两层在理化性质上定义了两个不同的区域。浮游细菌的垂直分布表明,界面区域比其他区域更密集。在两个采样期间,活细胞数量最多的是在“红水”层内。通过对 16S rRNA 基因片段 PCR 扩增的 DGGE 条带进行测序,确定了细菌群落的优势 phylotypes。DGGE 条带的数量(被认为是总物种丰富度的指示)在两个不同的采样期间没有可预测的变化。从法罗湖获得了变形菌门(α-、γ-、δ-和ε亚门成员)、噬纤维菌-黄杆菌-拟杆菌门、绿色硫细菌和蓝细菌。大多数条带显示出与其他先前描述的生物体不匹配的 DNA 序列,表明存在新的本土细菌 phylotypes。

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