Open AccessEnumeration of Archaea and Bacteria in seafloor basalt using real‐time quantitative PCR and fluorescence microscopy
Author(s) -
Einen Jørn,
Thorseth Ingunn H.,
Øvreås Lise
Publication year - 2008
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.2008.01119.x
Subject(s) - archaea , bacteria , basalt , 16s ribosomal rna , biology , fluorescence microscope , sybr green i , fluorescence , real time polymerase chain reaction , microbiology and biotechnology , microscopy , chemistry , gene , biochemistry , genetics , paleontology , physics , quantum mechanics , optics
A SYBR Green real‐time quantitative PCR (Q‐PCR) assay for the detection and quantification of Bacteria and Archaea present in the glassy rind of seafloor basalts of different ages and water depths is presented. Two sets of domain‐specific primers were designed and validated for specific detection and quantification of bacterial and archaeal 16S rRNA genes in DNA extracted from basaltic glass. Total cell numbers were also estimated by fluorescence microscopy analysis of SYBR Gold‐stained samples. The results from the two different approaches were concurrent, and Q‐PCR results showed that the total number of cells present in basalts was in the range from 6 × 10 5 to 4 × 10 6 cells g −1 basaltic glass. Further, it was demonstrated that these cells were almost exclusively from the domain Bacteria . When applying the same methods on samples of different ages (22 years–0.1 Ma) and water depths (139–3390 mbsl), no significant differences in cell concentrations or in the relative abundance of Archaea and Bacteria were detected.