Quantification of bacterial and archaeal symbionts in high and low microbial abundance sponges using real-time PCR

Handle URI:
http://hdl.handle.net/10754/599422
Title:
Quantification of bacterial and archaeal symbionts in high and low microbial abundance sponges using real-time PCR
Authors:
Bayer, Kristina; Kamke, Janine ( 0000-0001-5475-1238 ) ; Hentschel, Ute
Abstract:
In spite of considerable insights into the microbial diversity of marine sponges, quantitative information on microbial abundances and community composition remains scarce. Here, we established qPCR assays for the specific quantification of four bacterial phyla of representative sponge symbionts as well as the kingdoms Eubacteria and Archaea. We could show that the 16S rRNA gene numbers of Archaea, Chloroflexi, and the candidate phylum Poribacteria were 4-6 orders of magnitude higher in high microbial abundance (HMA) than in low microbial abundance (LMA) sponges and that actinobacterial 16S rRNA gene numbers were 1-2 orders higher in HMA over LMA sponges, while those for Cyanobacteria were stable between HMA and LMA sponges. Fluorescence in situ hybridization of Aplysina aerophoba tissue sections confirmed the numerical dominance of Chloroflexi, which was followed by Poribacteria. Archaeal and actinobacterial cells were detected in much lower numbers. By use of fluorescence-activated cell sorting as a primer- and probe-independent approach, the dominance of Chloroflexi, Proteobacteria, and Poribacteria in A. aerophoba was confirmed. Our study provides new quantitative insights into the microbiology of sponges and contributes to a better understanding of the HMA/LMA dichotomy. The authors quantified sponge symbionts in eight sponge species from three different locations by real time PCR targetting 16S rRNA genes. Additionally, FISH was performed and diversity and abundance of singularized microbial symbionts from Aplysina aerophoba was determined for a comprehensive quantification work. © 2014 Federation of European Microbiological Societies.
Citation:
Bayer K, Kamke J, Hentschel U (2014) Quantification of bacterial and archaeal symbionts in high and low microbial abundance sponges using real-time PCR. FEMS Microbiology Ecology 89: 679–690. Available: http://dx.doi.org/10.1111/1574-6941.12369.
Publisher:
Oxford University Press (OUP)
Journal:
FEMS Microbiology Ecology
Issue Date:
9-Jul-2014
DOI:
10.1111/1574-6941.12369
PubMed ID:
24942664
Type:
Article
ISSN:
0168-6496
Sponsors:
We thank Christine Gernert, Anne Müller, Anne Trimbach, Susanne Schmitt and Lucas Moitinho-Silva (all University of Wuerzburg), as well as Sabine Masanetz (Bio-Rad Laboratories GmbH, Munich) for useful contributions on particular aspects of the study. We further thank Martin Pfannkuchen and Renato Batel (Ruder Boscovic Institute, Rovinj/Croatia) for their help in sponge sampling and T. Ravasi (King Abdullah University of Science and Technology, Saudi Arabia) for providing sponge samples from the Red Sea.
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Full metadata record

DC FieldValue Language
dc.contributor.authorBayer, Kristinaen
dc.contributor.authorKamke, Janineen
dc.contributor.authorHentschel, Uteen
dc.date.accessioned2016-02-28T05:50:50Zen
dc.date.available2016-02-28T05:50:50Zen
dc.date.issued2014-07-09en
dc.identifier.citationBayer K, Kamke J, Hentschel U (2014) Quantification of bacterial and archaeal symbionts in high and low microbial abundance sponges using real-time PCR. FEMS Microbiology Ecology 89: 679–690. Available: http://dx.doi.org/10.1111/1574-6941.12369.en
dc.identifier.issn0168-6496en
dc.identifier.pmid24942664en
dc.identifier.doi10.1111/1574-6941.12369en
dc.identifier.urihttp://hdl.handle.net/10754/599422en
dc.description.abstractIn spite of considerable insights into the microbial diversity of marine sponges, quantitative information on microbial abundances and community composition remains scarce. Here, we established qPCR assays for the specific quantification of four bacterial phyla of representative sponge symbionts as well as the kingdoms Eubacteria and Archaea. We could show that the 16S rRNA gene numbers of Archaea, Chloroflexi, and the candidate phylum Poribacteria were 4-6 orders of magnitude higher in high microbial abundance (HMA) than in low microbial abundance (LMA) sponges and that actinobacterial 16S rRNA gene numbers were 1-2 orders higher in HMA over LMA sponges, while those for Cyanobacteria were stable between HMA and LMA sponges. Fluorescence in situ hybridization of Aplysina aerophoba tissue sections confirmed the numerical dominance of Chloroflexi, which was followed by Poribacteria. Archaeal and actinobacterial cells were detected in much lower numbers. By use of fluorescence-activated cell sorting as a primer- and probe-independent approach, the dominance of Chloroflexi, Proteobacteria, and Poribacteria in A. aerophoba was confirmed. Our study provides new quantitative insights into the microbiology of sponges and contributes to a better understanding of the HMA/LMA dichotomy. The authors quantified sponge symbionts in eight sponge species from three different locations by real time PCR targetting 16S rRNA genes. Additionally, FISH was performed and diversity and abundance of singularized microbial symbionts from Aplysina aerophoba was determined for a comprehensive quantification work. © 2014 Federation of European Microbiological Societies.en
dc.description.sponsorshipWe thank Christine Gernert, Anne Müller, Anne Trimbach, Susanne Schmitt and Lucas Moitinho-Silva (all University of Wuerzburg), as well as Sabine Masanetz (Bio-Rad Laboratories GmbH, Munich) for useful contributions on particular aspects of the study. We further thank Martin Pfannkuchen and Renato Batel (Ruder Boscovic Institute, Rovinj/Croatia) for their help in sponge sampling and T. Ravasi (King Abdullah University of Science and Technology, Saudi Arabia) for providing sponge samples from the Red Sea.en
dc.publisherOxford University Press (OUP)en
dc.subjectFluorescence in situ hybridizationen
dc.subjectMarine spongesen
dc.subjectMicrobial symbiontsen
dc.subjectQuantitative real-time PCRen
dc.subjectSingle-cell sortingen
dc.titleQuantification of bacterial and archaeal symbionts in high and low microbial abundance sponges using real-time PCRen
dc.typeArticleen
dc.identifier.journalFEMS Microbiology Ecologyen
dc.contributor.institutionDepartment of Botany II; Julius-von-Sachs Institute for Biological Sciences; University of Wuerzburg; Wuerzburg Germanyen

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