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Environmental Microbiology |
Department of Biological Sciences, University of Warwick, Coventry CV4 7AL, UK1
Max-Planck-Institut für Marine Mikrobiologie, Bremen, Germany2
Unité de Physiologie Microbienne (CNRS URA 2172), Institut Pasteur, Paris, France3
The H. Steinitz Marine Biology Laboratory, InterUniversity Institute of Eilat, Eilat, Israel4
Author for correspondence: David J. Scanlan. Tel: +44 24 76 528363. Fax: +44 24 76 523701. e-mail: dscanlan{at}bio.warwick.ac.uk
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.
Keywords: cyanobacteria, Red Sea, oligonucleotide probes, 16S rRNA, TSA
Abbreviations: DAPI, 4’,6-diamidino-2-phenylindole; HL, high light; HRP, horseradish peroxidase; LL, low light; TETON, 4-(1,4,7,10-tetra-oxadecyl)-1-naphthol; TSA, tyramide signal amplification
The GenBank accession numbers for the sequences reported in this paper are AF311217 (RCC278, EQPAC1), AF311218 (RCC277, NATL1MIT), AF311219 (RCC280, NATL2B), AF311220 (RCC264, TAK9803-2), AF311291 (WH7803), AF311292 (WH8018) and AF311293 (WH8103).
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