West Nyree J, Schönhuber Wilhelm A, Fuller Nicholas J, Amann Rudolf I, Rippka Rosmarie, Post Anton F, Scanlan David J
Department of Biological Sciences, University of Warwick, Coventry CV4 7AL, UK1.
Unité de Physiologie Microbienne (CNRS URA 2172), Institut Pasteur, Paris, France3.
Microbiology (Reading). 2001 Jul;147(Pt 7):1731-1744. doi: 10.1099/00221287-147-7-1731.
An in situ hybridization method was applied to the identification of marine cyanobacteria assignable to the genus Prochlorococcus using horseradish-peroxidase-labelled 16S rRNA-targeted oligonucleotide probes in combination with tyramide signal amplification (TSA). With this method very bright signals were obtained, in contrast to hybridizations with oligonucleotides monolabelled with fluorochromes, which failed to give positive signals. Genotype-specific oligonucleotides for high light (HL)- and low light (LL)-adapted members of this genus were identified by 16S rRNA sequence analyses and their specificities confirmed in whole-cell hybridizations with cultured strains of Prochlorococcus marinus Chisholm et al., 1992, Prochlorococcus sp. and Synechococcus sp. In situ hybridization of these genotype-specific probes to field samples from stratified water bodies collected in the North Atlantic Ocean and the Red Sea allowed a rapid assessment of the abundance and spatial distribution of HL- and LL-adapted Prochlorococcus. In both oceanic regions the LL-adapted Prochlorococcus populations were localized in deeper water whereas the HL-adapted Prochlorococcus populations were not only distinct in each region but also exhibited strikingly different depth distributions, HLI being confined to shallow water in the North Atlantic, in contrast to HLII, which was present throughout the water column in the Red Sea.
应用原位杂交方法,结合酪胺信号放大(TSA)技术,使用辣根过氧化物酶标记的靶向16S rRNA的寡核苷酸探针,鉴定属于原绿球藻属的海洋蓝细菌。与用荧光染料单标记的寡核苷酸杂交相比,该方法获得了非常明亮的信号,而后者未能给出阳性信号。通过16S rRNA序列分析鉴定了该属适应高光(HL)和低光(LL)的基因型特异性寡核苷酸,并在与1992年Chisholm等人的海洋原绿球藻、原绿球藻属和聚球藻属培养菌株的全细胞杂交中证实了它们的特异性。将这些基因型特异性探针与从北大西洋和红海采集的分层水体的野外样本进行原位杂交,可以快速评估适应HL和LL的原绿球藻的丰度和空间分布。在这两个海洋区域中,适应LL的原绿球藻种群位于较深的水体中,而适应HL的原绿球藻种群不仅在每个区域都不同,而且在深度分布上也表现出显著差异,HLI局限于北大西洋的浅水区域,而HLII则存在于红海的整个水柱中。