Ferris M J, Muyzer G, Ward D M
Department of Microbiology, Montana State University, Bozeman 59717, USA.
Appl Environ Microbiol. 1996 Feb;62(2):340-6. doi: 10.1128/aem.62.2.340-346.1996.
Denaturing gradient gel electrophoresis (DGGE) of PCR-amplified 16S rRNA gene segments was used to profile microbial populations inhabiting different temperature regions in the microbial mat community of Octopus Spring, Yellowstone National Park. DGGE allowed a rapid evaluation of the distributions of amplifiable sequence types. Profiles were essentially identical within regions of the mat defined by one temperature range but varied between sites with different temperature ranges. Individual DGGE bands were sequenced, and the sequences were compared with those previously obtained from the mat by cloning and from cultivated Octopus Spring isolates. Two known cyanobacterial populations and one known green nonsulfur bacterium-like population were detected by DGGE, as were many new cyanobacterial and green nonsulfur and green sulfur bacterium-like populations and a novel bacterial population of uncertain phylogenetic affiliation. The distributions of several cyanobacterial populations compared favorably with results obtained previously by oligonucleotide probe analyses and suggest that adaptation to temperature has occurred among cyanobacteria which are phylogenetically very similar.
采用变性梯度凝胶电泳(DGGE)对聚合酶链反应(PCR)扩增的16S rRNA基因片段进行分析,以描绘黄石国家公园章鱼泉微生物席群落中不同温度区域的微生物种群分布。DGGE能够快速评估可扩增序列类型的分布情况。在由一个温度范围界定的微生物席区域内,图谱基本相同,但在不同温度范围的位点之间则有所不同。对DGGE的各个条带进行了测序,并将这些序列与之前通过克隆从微生物席以及从章鱼泉培养分离物中获得的序列进行了比较。通过DGGE检测到了两个已知的蓝细菌种群和一个已知的类绿色非硫细菌种群,以及许多新的蓝细菌、类绿色非硫和类绿色硫细菌种群,还有一个系统发育归属不确定的新型细菌种群。几个蓝细菌种群的分布情况与之前通过寡核苷酸探针分析获得的结果相当吻合,这表明在系统发育上非常相似的蓝细菌中已经发生了对温度的适应性变化。