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


 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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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)

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 http://www.springerlink.com 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/s00248-007-9283-5Florida Atlantic University. Harbor Branch Oceanographic Institute contribution #1650.

Subject(s): Microbiology --ResearchSponges --MicrobiologyMarine biology --Caribbean Area

Persistent Link to This Record: http://purl.flvc.org/FCLA/DT/2708323

Links: http://dx.doi.org/10.1007/s00248-007-9283-5

Restrictions on Access: ©2007 Springer Science+Business Media, LLC.

Owner Institution: FAU



Autor: Cassler, M., creator Peterson, C.L., creator Ledger, A., creator Pomponi, S.A., creator Wright, A. E., creator Winegar, R., creat

Fuente: http://fau.digital.flvc.org/islandora/object/fau%3A5503



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