Appl Environ Microbiol. 1996 Jun;62(6):2138-44. doi: 10.1128/aem.62.6.2138-2144.1996.
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(prm1),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.
采用荧光标记的 rRNA 靶向寡核苷酸探针原位杂交的方法分析了高山湖泊冬季覆盖层和浮游区的细菌群落结构。将固定在膜滤器上的细胞与针对细菌域的探针以及针对 α、β和 γ 变形菌纲和噬纤维菌-黄杆菌组的探针进行杂交。与细菌探针 EUB 杂交后可检测到的细菌分数在 4（prm1），6-二脒基-2-苯基吲哚（DAPI）计数的 40%至 81%之间。细菌组合在不同栖息地（雪、雪泥和湖水）之间和内部差异很大，但在大多数情况下，β亚纲（可通过探针 EUB 检测到的细菌的 6.5%至 116%）成员占主导地位。与特异性探针杂交的细菌总和通常低于可通过探针 EUB 检测到的分数。图像分析用于表征细菌组合的形态和大小特异性生物量分布，这些分布清楚地区分了三个栖息地。尽管不同雪泥和浮游层中的测量次生生产力参数和 2-（对碘苯基）-3-（对硝基苯基）-5-苯基四唑氯化物还原菌的分数变化超过一个数量级，但荧光探针 EUB 的可检测性始终很高。在营养限制和低温下细菌的生理策略在核糖体含量的背景下进行了讨论。这项研究证实了荧光标记的 rRNA 靶向探针用于表征即使在贫营养栖息地中的细菌种群结构的适用性。
