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• W2277919100 abstract "Sponges, phylum Porifera, are amongst the most ancient metazoans. As prodigious filter feeders, they influence both benthic and pelagic ecosystems and their association with a diverse range of microorganisms raises many questions in the ecology, evolution and biotechnology realms. Despite this recent attention, much remains unknown about the microbial communities associated within marine sponges. The research described within this thesis utilised DNA (16S rRNA-gene) and/or RNA (16S rRNA) next-generation sequencing technologies to address the specificity, stability and community-level ecology of sponge-microbe associations. Numerous 16S rRNA-based studies have identified microbial phylotypes found exclusively in sponge hosts and not in the seawater or surrounding environment, i.e. so-called sponge-specific clusters (SCs) or sponge- and coral-specific clusters (SCCs). With more than 7500 sponge-derived rRNA sequences (from clone, isolate and denaturing gradient gel electrophoresis data) available, the aim was to determine whether the current notion of sponge-specific sequence clusters remains valid. Comprehensive phylogenetic analyses were performed on the 7546 sponge-derived 16S and 18S rRNA sequences. Overall, 27% of all sequences fell into monophyletic, sponge-specific sequence clusters. These results strongly support the existence of sponge-specific microbes, with the ARB database generated during this work providing a valuable resource for further studies of rare and abundant sponge symbionts. A systematic evaluation of sponge tissue preservation (RNAlater, liquid nitrogen, lyophilized, frozen) and nucleic acid extraction (CTAB-based DNA extraction, RNA isolation via TRIzol and two coextraction methods) protocols, for the recovery of DNA and/or RNA, was assessed using the New Zealand sponge Ancorina alata. Although all protocols resulted in sufficiently high DNA and/or RNA quantity and quality for downstream applications, there were significant differences in yield of nucleic acids extracted. Denaturing gradient gel electrophoresis (DGGE) analysis of community 16S rRNA gene- and 16S rRNA-derived fragments revealed no major changes attributable to either preservation or extraction method. All protocols performed favourably and therefore the choice of protocol can be made based on practical considerations, including ease of use, time availability and cost. Based on these assessments, RNAlater and a Qiagen co-extraction kit were used for all subsequent experiments of this thesis. Understanding variability in microbial communities is a fundamental goal when examining any microbe-host association. RNA (16S rRNA)???based amplicon pyrosequencing was used to investigate the temporal stability of sponge-associated bacterial communities. Temporal variation between a host and its symbiotic community can also be tracked by isotopic composition (??15N and ??13C values) of the host tissue. Two New Zealand sponge species (A. alata and Tethya stolonifera) were sampled over a two year period. A total of 4468 operational taxonomic…" @default.
• W2277919100 created "2016-06-24" @default.
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• W2277919100 date "2013-01-01" @default.
• W2277919100 modified "2023-09-27" @default.
• W2277919100 title "The microbial ecology of marine sponge-associated microorganisms" @default.
• W2277919100 hasPublicationYear "2013" @default.
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