Flourishing deep-sea AAP bacteria detected by flow cytometric sorting and molecular analysis

PLoS One. 2019 Jun 19;14(6):e0218753. doi: 10.1371/journal.pone.0218753. eCollection 2019.

Abstract

Pigmented bacteria cells, including aerobic anoxygenic phototrophic (AAP) bacteria, contribute significantly to secondary production and aquatic carbon cycling but their distribution in the deep sea is still not well understood, especially in the South China Sea. In this study, microscopic, flow cytometric, and molecular analyses were carried out to investigate the abundance and diversity of AAP bacteria at seven stations in the South China Sea. The results revealed the existence of bacteriochlorophyll-containing bacteria below 500 m from two of seven stations. Flow cytometric analysis detected red and infra-red fluorescence under blue (488 nm) light excitation from fluorescent cells. Blue light-excited red fluorescence of these cells from the 1000 m depth at station E403 were verified using epifluorescence microscopy. Based on fluorescence and side scatter features, fluorescent cells were sorted and subjected to molecular analysis. DNA was extracted from these sorted cells from both stations for PCR amplification using 16S rDNA primers. Sequencing of the PCR products showed that the sorted cells from the 1000 m depth at station E403 belonged to the genus Porphyrobacter. The cell population sorted from 500 m at station E703 contained Sphingomonas and a Methylobacterium-like taxon. All these three taxa belong to aerobic anoxygenic phototrophic alpha-proteobacteria. Using flow cytometric analysis, we found that the abundance of Porphyrobacter sp. at 1000 m was 2.71-2.95×104 cells mL-1 whereas cell counts of Sphingomonas sp. and Methylobacterium at 500 m were about 3.75-4.12×105 cells mL-1. These results indicate that albeit not ubiquitous in deep water, bacteriochlorophyll-containing bacteria can be abundant in the deep-sea aphotic zone.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacteria / classification*
  • Bacteria / genetics
  • Bacteria / metabolism
  • Bacteriochlorophylls / metabolism
  • China
  • DNA, Bacterial / genetics
  • DNA, Ribosomal / genetics
  • Flow Cytometry
  • Phototrophic Processes
  • Phylogeny
  • RNA, Ribosomal, 16S / genetics*
  • Seawater / microbiology*
  • Sequence Analysis, DNA / methods*

Substances

  • Bacteriochlorophylls
  • DNA, Bacterial
  • DNA, Ribosomal
  • RNA, Ribosomal, 16S

Grants and funding

This research work was supported by the program of the Natural Science Foundation of China (Grant 411776154 to DQ, 41129001 to SL, 41130855 to LH) and Science Technology Program Guangzhou of China (Grant 201607010289 to DQ).