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Bromodeoxyuridine labelling and fluorescence‐activated cell sorting of polyamine‐transforming bacterioplankton in coastal seawater
Author(s) -
Mou Xiaozhen,
Jacob Jisha,
Lu Xinxin,
VilaCosta Maria,
Chan LeongKeat,
Sharma Shalabh,
Zhang Yuqin
Publication year - 2015
Publication title -
environmental microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.954
H-Index - 188
eISSN - 1462-2920
pISSN - 1462-2912
DOI - 10.1111/1462-2920.12550
Subject(s) - bacterioplankton , biology , roseobacter , microcosm , bacteria , spermidine , bacteroidetes , cell sorting , microbiology and biotechnology , polyamine , marine bacteriophage , proteobacteria , biochemistry , gene , 16s ribosomal rna , cell , ecology , genetics , nutrient , phylogenetics , enzyme , phytoplankton , clade
Summary Polyamines ( PAs ) are a group of nitrogen‐rich dissolved organic nitrogen ( DON ) compounds that are ubiquitously distributed in marine environments. To identify bacteria that are involved in PA transformations, coastal bacterioplankton microcosms were amended with a single PA model compound, i.e. putrescine ( PUT ) or spermidine ( SPD ), or with no addition as controls ( CTRs ). Bromodeoxyuridine ( BrdU ) was added to all the microcosms to label newly synthesized DNAs . Fluorescence‐activated cell sorting ( FACS ) analysis indicated significant increases in numbers of total cells and cells with both high and low levels of BrdU incorporation in the PUT and SPD microcosms, but not in the CTRs . 16 S rDNA pyrotag sequencing of FACS ‐sorted cells indicated that PUT ‐ and SPD ‐transforming bacteria were composed similarly of a diverse group of taxa affiliated with A ctinobacteria , B acteroidetes , F irmicutes and P roteobacteria (especially R oseobacter of its alpha lineage). Broad taxonomic distribution of PA ‐transforming bacteria was also indicated by the abundance and distribution of PA transporter gene homologues in a survey of sequenced marine bacterial genomes. Our results suggest that PAs may be common DON substrates for marine bacterioplankton, in line with the hypothesis that bacterially mediated PA transformation accounts for an important proportion of marine DON flux.