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Use of real-time qPCR to quantify members of the unculturable heterotrophic bacterial community in a deep sea marine sponge, Vetulina sp

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Title: Use of real-time qPCR to quantify members of the unculturable heterotrophic bacterial community in a deep sea marine sponge, Vetulina sp.
Name(s): Cassler, M., creator
Peterson, C. L., creator
Ledger, Angela, creator
Pomponi, Shirley A., creator
Wright, Amy E., creator
Winegar, R., creator
McCarthy, Peter J., creator
Lopez, Jose V., creator
Type of Resource: text
Genre: Article
Issuance: single unit
Date Issued: 2008
Publisher: ©2007 Springer Science+Business Media, LLC.
Extent: 12 p.
Physical Description: pdf
Language(s): English
Identifier: 2708323 (digitool), FADT2708323 (IID), fau:5503 (fedora), 10.1007/s00248-007-9283-5 (doi)
Note(s): In this report, real-time quantitative PCR (TaqMan® qPCR) of the small subunit (SSU) 16S-like rRNA molecule, a universal phylogenetic marker, was used to quantify the relative abundance of individual bacterial members of a diverse, yet mostly unculturable, microbial community from a marine sponge. Molecular phylogenetic analyses of bacterial communities derived from Caribbean Lithistid sponges have shown a wide diversity of microbes that included at least six major subdivisions; however, very little overlap was observed between the culturable and unculturable microbial communities. Based on sequence data of three culture-independent Lithistid-derived representative bacteria, we designed probe/primer sets for TaqMan® qPCR to quantitatively characterize selected microbial residents in a Lithistid sponge, Vetulina, metagenome. TaqMan® assays included specificity testing, DNA limit of detection analysis, and quantification of specific microbial rRNA sequences such as Nitrospira-like microbes and Actinobacteria up to 172 million copies per microgram per Lithistid sponge metagenome. By contrast, qPCR amplification with probes designed for common previously cultured sponge-associated bacteria in the genera Rheinheimera and Marinomonas and a representative of the CFB group resulted in only minimal detection of the Rheiheimera in total DNA extracted from the sponge. These data verify that a large portion of the microbial community within Lithistid sponges may consist of currently unculturable microorganisms.
This manuscript is a version of an article with the final publication found online at and may be cited as: Cassler, M., C. L. Peterson, A. Ledger, S. A. Pomponi, A. E. Wright, R. Winegar, P. J. McCarthy and J. V. Lopez (2008) Use of Real-Time qPCR to Quantify Members of the Unculturable Heterotrophic Bacterial Community in a Deep Sea Marine Sponge, Vetulina sp, Microbial Ecology 55:384–394 DOI: 10.1007/s00248-007-9283-5
Florida Atlantic University. Harbor Branch Oceanographic Institute contribution #1650.
Subject(s): Microbiology --Research
Sponges --Microbiology
Marine biology --Caribbean Area
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Restrictions on Access: ©2007 Springer Science+Business Media, LLC.
Host Institution: FAU