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Quantitative Molecular Analysis of the Microbial Community in Marine Arctic Sediments (Svalbard)

机译:海洋北极沉积物中微生物群落的定量分子分析(斯瓦尔巴特群岛)

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摘要

Fluorescence in situ hybridization (FISH) and rRNA slot blot hybridization with 16S rRNA-targeted oligonucleotide probes were used to investigate the phylogenetic composition of a marine Arctic sediment (Svalbard). FISH resulted in the detection of a large fraction of microbes living in the top 5 cm of the sediment. Up to 65.4% ± 7.5% of total DAPI (4′,6′-diamidino-2-phenylindole) cell counts hybridized to the bacterial probe EUB338, and up to 4.9% ± 1.5% hybridized to the archaeal probe ARCH915. Besides δ-proteobacterial sulfate-reducing bacteria (up to 16% 52) members of the Cytophaga-Flavobacterium cluster were the most abundant group detected in this sediment, accounting for up to 12.8% of total DAPI cell counts and up to 6.1% of prokaryotic rRNA. Furthermore, members of the order Planctomycetales accounted for up to 3.9% of total cell counts. In accordance with previous studies, these findings support the hypothesis that these bacterial groups are not simply settling with organic matter from the pelagic zone but are indigenous to the anoxic zones of marine sediments. Members of the γ-proteobacteria also constituted a significant fraction in this sediment (6.1% ± 2.5% of total cell counts, 14.4% ± 3.6% of prokaryotic rRNA). A new probe (GAM660) specific for sequences affiliated with free-living or endosymbiotic sulfur-oxidizing bacteria was developed. A significant number of cells was detected by this probe (2.1% ± 0.7% of total DAPI cell counts, 13.2% ± 4.6% of prokaryotic rRNA), showing no clear zonation along the vertical profile. Gram-positive bacteria and the β-proteobacteria were near the detection limit in all sediments.
机译:使用荧光原位杂交(FISH)和与16S rRNA靶向的寡核苷酸探针进行rRNA狭点印迹杂交来研究海洋北极沉积物(Svalbard)的系统发育组成。 FISH检测到生活在沉积物顶部5 cm的大部分微生物。与细菌探针EUB338杂交的DAPI(4',6'-diamidino-2-phenylindole)细胞总数中多达65.4%±7.5%,与古细菌探针ARCH915杂交的高达4.9%±1.5%。除了δ减少硫酸盐杆菌的细菌(最多可减少16%52),在此沉积物中检测到的是最富含的细胞噬菌-黄杆菌群,占DAPI细胞总数的12.8%和原核的6.1% rRNA。此外,Planctomycetales的成员占细胞总数的3.9%。根据以前的研究,这些发现支持以下假设:这些细菌群不只是与中上层带的有机物沉降,而是海洋沉积物缺氧带的固有细菌。 γ-变形细菌的成员也占该沉积物中的很大一部分(占总细胞计数的6.1%±2.5%,原核rRNA的14.4%±3.6%)。开发了一种新探针(GAM660),该探针特异于与自由生活或共生硫氧化细菌相关的序列。该探针检测到大量细胞(占DAPI总细胞计数的2.1%±0.7%,原核rRNA的13.2%±4.6%),沿垂直分布没有清晰的分区。在所有沉积物中,革兰氏阳性细菌和β-蛋白细菌均接近检出限。

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