Marine methylotrophs revealed by stable-isotope probing, multiple displacement amplification and metagenomics

JD Neufeld, Y Chen, MG Dumont, JC Murrell

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79 Citations (Scopus)

Abstract

The concentrations of one-carbon substrates that fuel methylotrophic microbial communities in the ocean are limited and the specialized guilds of bacteria that use these molecules may exist at low relative abundance. As a result, these organisms are difficult to identify and are often missed with existing cultivation and gene retrieval methods. Here, we demonstrate a novel proof of concept: using environmentally-relevant substrate concentrations in stable-isotope probing (SIP) incubations to yield sufficient DNA for large-insert metagenomic analysis through multiple displacement amplification (MDA). A marine surface-water sample was labelled sufficiently by incubation with near in situ concentrations of methanol. Picogram quantities of labelled (13)C-DNA were purified from caesium chloride gradients, amplified with MDA to produce microgram amounts of high-molecular-weight DNA (
Original languageEnglish
Pages (from-to)1526-1535
Number of pages10
JournalEnvironmental Microbiology
Volume10
Issue number6
DOIs
Publication statusPublished - Jun 2008

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