Community analysis of the bacterial assemblages in the winter cover and pelagic layers of a high mountain lake by in situ hybridization
Community analysis of the bacterial assemblages in the winter cover and pelagic layers of a high mountain lake by in situ hybridization
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DOI：
10.1128/aem.62.6.2138-2144.1996
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发表时间：
1996-06-01
影响因子：
4.4
通讯作者：
Psenner, R
中科院分区：
文献类型：
--
作者：
Alfreider, A;Pernthaler, J;Psenner, R
采用荧光标记rrna靶向寡核苷酸探针原位杂交技术，对某高山湖泊冬盖区和远洋区细菌群落结构进行了分析。将固定在膜过滤器上的细胞与特定于细菌结构域的探针以及Proteobacteria类和Cytophaga-Flavobacterium组的α、β和γ亚类探针杂交。与细菌探针EUB杂交后可检测到的细菌比例为40 ~ 81%的4′，6-二氨基-2-苯基吲哚（DAPI）计数。细菌组合在不同生境（雪、泥和湖水）之间和内部差异很大，但在大多数情况下以β亚类成员为主（探针EUB可检测的细菌占6.5%至116%），与群体特异性探针杂交的细菌总数通常低于探针EUB可检测的比例。虽然测量到的二次生产参数和2-（对碘苯基）-3-（对硝基苯基）-5-苯基氯化四氮唑还原菌的比例在不同的泥层和远洋层中变化超过一个数量级，但荧光探针EUB的可探测性一直很高。在单细胞核糖体含量的背景下，讨论了细菌在营养限制和低温下的生理策略。本研究证实了荧光标记rrna靶向探针对细菌种群结构表征的适用性，即使在低营养栖息地也是如此。
The bacterial community structure in the winter cover and pelagic zone of a high mountain lake was analyzed by in situ hybridization with fluorescently labeled rRNA-targeted oligonucleotide probes. Cells fixed on membrane filters were hybridized with a probe specific for the domain Bacteria as well as with probes for the alpha, beta, and gamma subclasses of the class Proteobacteria and the Cytophaga-Flavobacterium group. The fraction of bacteria detectable after hybridization with the bacterial probe EUB ranged from 40 to 81% of 4',6-diamidino-2-phenylindole (DAPI) counts. The bacterial assemblage varied considerably between and within different habitats (snow, slush, and lake water) but was in most cases dominated by members of the beta subclass (6.5 to 116% of bacteria detectable with probe EUB), The sum of bacteria hybridizing with group-specific probes was usually lower than the fraction detectable with probe EUB. Image analysis was used to characterize morphology and the size-specific biomass distribution of bacterial assemblages, which clearly separated the three habitats, Although the measured secondary production parameters and the fraction of 2-(p-iodophenyl)-3-(p-nitrophenyl)-5-phenyltetrazolium chloride-reducing bacteria varied by more than an order of magnitude in the different slush and pelagic layers, detectability with the fluorescent probe EUB was constantly high, Physiological strategies of bacteria under nutrient limitation and at low temperatures are discussed in the context of the ribosome content of single cells. This study confirms the suitability of fluorescently labeled rRNA-targeted probes for the characterization of bacterial population structures even in oligotrophic habitats.