Dynamics of marine bacterial and phytoplankton populations using multiplex liquid bead array technology

Environmental Microbiology
Xavier MayaliRonald S Burton

Abstract

Heterotrophic bacteria and phytoplankton dominate the biomass and play major roles in the biogeochemical cycles of the surface ocean. Here, we designed and tested a fast, high-throughput and multiplexed hybridization-based assay to detect populations of marine heterotrophic bacteria and phytoplankton based on their small subunit ribosomal DNA sequences. The assay is based on established liquid bead array technology, an approach that is gaining acceptance in biomedical research but remains underutilized in ecology. End-labelled PCR products are hybridized to taxon-specific oligonucleotide probes attached to fluorescently coded beads followed by flow cytometric detection. We used ribosomal DNA environmental clone libraries (a total of 450 clones) and cultured isolates to design and test 26 bacterial and 10 eukaryotic probes specific to various ribotypes and genera of heterotrophic bacteria and eukaryotic phytoplankton. Pure environmental clones or cultures were used as controls and demonstrated specificity of the probes to their target taxa. The quantitative nature of the assay was demonstrated by a significant relationship between the number of target molecules and fluorescence signal. Clone library sequencing and bead array flu...Continue Reading

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Citations

Jun 27, 2013·Proceedings of the National Academy of Sciences of the United States of America·Ilnam KangJang-Cheon Cho
Mar 31, 2012·Annual Review of Marine Science·David A CaronAstrid Schnetzer
Jan 19, 2016·Frontiers in Microbiology·Pelin YilmazFrank O Glöckner
Aug 31, 2016·Nature Microbiology·David M Needham, Jed A Fuhrman
Aug 13, 2011·Molecular Ecology Resources·Lani U Gleason, Ronald S Burton
Mar 22, 2018·FEMS Microbiology Ecology·Xavier Mayali, Peter K Weber
Jan 20, 2018·Nature Communications·Antonio M Martin-PlateroMartin F Polz
Feb 2, 2019·Frontiers in Microbiology·Carina BunseJarone Pinhassi

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