Diversity of cultivated and metabolically active aerobic anoxygenic phototrophic bacteria along an oligotrophic gradient in the Mediterranean Sea

Abstract

Aerobic anoxygenic phototrophic (AAP) bacteria play significant roles in the bacterioplankton productivity and biogeochemical cycles of the surface ocean. In this study, we applied both cultivation and mRNA-based molecular methods to explore the diversity of AAP bacteria along an oligotrophic gradient in the Mediterranean Sea in early summer 2008. Colony-forming units obtained on three different agar media were screened for the production of bacteriochlorophyll-<i>a</i> (BChl-<i>a</i>), the light-harvesting pigment of AAP bacteria. BChl-<i>a</i>-containing colonies represented a low part of the cultivable fraction. In total, 54 AAP strains were isolated and the phylogenetic analyses based on their 16S rRNA and <i>pufM</i> genes showed that they were all affiliated to the <i>Alphaproteobacteria</i>. The most frequently isolated strains belonged to <i>Citromicrobium bathyomarinum</i>, and <i>Erythrobacter</i> and <i>Roseovarius</i> species. Most other isolates were related to species not reported to produce BChl-<i>a</i> and/or may represent novel taxa. Direct extraction of RNA from seawater samples enabled the analysis of the expression of <i>pufM</i>, the gene coding for the M subunit of the reaction centre complex of aerobic anoxygenic photosynthesis. Clone libraries of <i>pufM</i> gene transcripts revealed that most phylotypes were highly similar to sequences previously recovered from the Mediterranean Sea and a large majority (~94 %) was affiliated to the <i>Gammaproteobacteria</i>. The most abundantly detected phylotypes occurred in the western and eastern Mediterranean basins. However, some were exclusively detected in the eastern basin, reflecting the highest diversity of <i>pufM</i> transcripts observed in this ultra-oligotrophic region. To our knowledge, this is the first study to document extensively the diversity of AAP isolates and to unveil the active AAP community in an oligotrophic marine environment. By pointing out the discrepancies between culture-based and molecular methods, this study highlights the existing gaps in the understanding of the AAP bacteria ecology, especially in the Mediterranean Sea and likely globally.