Ravenschlag K, Sahm K, Pernthaler J, Amann R
Molecular Ecology Group, Max-Planck-Institute for Marine Microbiology, D-28359 Bremen, Germany.
Appl Environ Microbiol. 1999 Sep;65(9):3982-9. doi: 10.1128/AEM.65.9.3982-3989.1999.
A 16S ribosomal DNA (rDNA) clone library from permanently cold marine sediments was established. Screening 353 clones by dot blot hybridization with group-specific oligonucleotide probes suggested a predominance of sequences related to bacteria of the sulfur cycle (43.4% potential sulfate reducers). Within this fraction, the major cluster (19.0%) was affiliated with Desulfotalea sp. and other closely related psychrophilic sulfate reducers isolated from the same habitat. The cloned sequences showed between 93 and 100% similarity to these bacteria. Two additional groups were frequently encountered: 13% of the clones were related to Desulfuromonas palmitatis, and a second group was affiliated with Myxobacteria spp. and Bdellovibrio spp. Many clones (18.1%) belonged to the gamma subclass of the class Proteobacteria and were closest to symbiotic or free-living sulfur oxidizers. Probe target groups were further characterized by amplified rDNA restriction analysis to determine diversity within the groups and within the clone library. Rarefaction analysis suggested that the total diversity assessed by 16S rDNA analysis was very high in these permanently cold sediments and was only partially revealed by screening of 353 clones.
构建了一个来自永久寒冷海洋沉积物的16S核糖体DNA(rDNA)克隆文库。用组特异性寡核苷酸探针通过斑点杂交筛选353个克隆,结果表明与硫循环细菌相关的序列占优势(43.4%为潜在硫酸盐还原菌)。在这一部分中,主要类群(19.0%)与脱硫弧菌属及从同一栖息地分离出的其他密切相关的嗜冷硫酸盐还原菌有关。克隆序列与这些细菌的相似性在93%至100%之间。另外还经常遇到两个类群:13%的克隆与棕榈酸脱硫单胞菌有关,第二个类群与黏细菌属和蛭弧菌属有关。许多克隆(18.1%)属于变形菌纲γ亚类,与共生或自由生活的硫氧化菌关系最为密切。通过扩增rDNA限制性分析进一步对探针靶标类群进行表征,以确定类群内部和克隆文库内部的多样性。稀疏分析表明,通过16S rDNA分析评估这些永久寒冷沉积物中的总多样性非常高,而通过筛选353个克隆仅部分揭示了这种多样性。