Molecular identification of picoplankton populations in contrasting waters of the Arabian Sea

The composition of picoplankton in the southern oligotrophic, northern mesotrophic waters and deep oxygen minimum zone (OMZ) of the Arabian Sea was determined by 16S ribosomal RNA gene cloning and fluorescence in situ hybridisation (FISH). It was hypothesised that the composition of the heterotrophic picoplankton would be different in these contrasting waters. To reduce the total diversity, cells were sorted by flow cytometry according to their scatter and DNA content before PCR amplification. The 16S rRNA clone libraries resulting from flow-sorted populations were different and often dominated by a small number of clades. Libraries from the Prochlorococcus-dominated southerly waters were dominated by sequences related to uncultured clusters of SAR11, SAR86 and Actinobacteria (HGC 1). From surface waters of the Synechococcus-dominated northern part of the Arabian Sea, mostly sequences related to the uncultured gammaproteobacterial group 'Svalbard' and HGC I were retrieved. The clone libraries from the OMZ were also dominated by sequences falling in the clades SARI 1 and SAR406, but included sequences related to those of sulfate-reducing (Desulfosarcina, Desulfofrigus) and sulfide-oxidising bacteria (endosymbionts of Riftia and Calyptogena). With a recently developed more sensitive FISH protocol approximately 60% of all DAPI stained cells could be identified by general probes as Bacteria, Cren- or Euryarchaeota in both provinces of the Arabian Sea; 40% remained undetected. On this level and on that of the major phylogenetic groups like Alpha- and Gammaproteobacteria only minor differences were detected by FISH. However, the composition of heterotrophic picoplankton clearly differed for the proteobacterial subgroups SAR86, SAR11 and SAR116. These were more abundant in the oligotrophic waters throughout the water column than in the mesotrophic surface waters and the OMZ. This supports our original hypothesis that the contrasting waters in the Arabian Sea harbor different heterotrophic picoplankton communities. In the future, FISH with a larger set of probes for more narrow phylogenetic groups will enable us to quantify these differences in more detail.