Probing the Rare Biosphere of the North-West Mediterranean Sea: An Experiment with High Sequencing Effort

PLoS One. 2016 Jul 21;11(7):e0159195. doi: 10.1371/journal.pone.0159195. eCollection 2016.

Abstract

High-throughput sequencing (HTS) techniques have suggested the existence of a wealth of species with very low relative abundance: the rare biosphere. We attempted to exhaustively map this rare biosphere in two water samples by performing an exceptionally deep pyrosequencing analysis (~500,000 final reads per sample). Species data were derived by a 97% identity criterion and various parametric distributions were fitted to the observed counts. Using the best-fitting Sichel distribution we estimate a total species richness of 1,568-1,669 (95% Credible Interval) and 5,027-5,196 for surface and deep water samples respectively, implying that 84-89% of the total richness in those two samples was sequenced, and we predict that a quadrupling of the present sequencing effort would suffice to observe 90% of the total richness in both samples. Comparing the HTS results with a culturing approach we found that most of the cultured taxa were not obtained by HTS, despite the high sequencing effort. Culturing therefore remains a useful tool for uncovering marine bacterial diversity, in addition to its other uses for studying the ecology of marine bacteria.

MeSH terms

  • Bacteria / isolation & purification
  • Base Sequence
  • Biodiversity*
  • Databases, Genetic
  • High-Throughput Nucleotide Sequencing / methods*
  • Mediterranean Sea
  • Species Specificity

Grants and funding

Cruise SUMMER was supported by the Spanish MICINN grant CTM2008-03309/MAR. BGC was supported by a Juan de la Cierva contract from the Spanish “Ministerio de Ciencia e Innovación”. Research was funded by the Spanish “Plan Nacional de Investigación Científica y Técnica” grants Marine Gems (CTM2010-20361) and Blue Genes (CTM2013-48292-C3-1-R).