A multilocus, temperature stress-related gene expression profile assay in Acropora millepora, a dominant reef-building coral
Souter, P., Bay, L.K., Andreakis, N., Császár, N., Seneca, F.O., and van Oppen, M.J.H. (2011) A multilocus, temperature stress-related gene expression profile assay in Acropora millepora, a dominant reef-building coral. Molecular Ecology Resources, 11 (2). pp. 328-334.
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We report an accurate multiplex reverse transcription quantitative polymerase chain reaction (RT–qPCR) assay, capable of reproducing gene expression profiles from 16 target genes [12 genes of interest (GOIs) and four reference genes (RGs)] in Acropora millepora, a common reef-building model coral species. The 12 GOIs have known or putative roles in the coral bleaching response, yet the method is not restricted to this particular assay and gene set. The procedure is based on the Beckman Coulter (Fullerton, CA, USA) GenomeLab™ GeXP Genetic Analysis System and bridges the gap between quantitative real-time PCR (qPCR) expression analysis of a single or a small number of genes and microarray gene expression surveys of thousands of genes. Despite large variation among biological replicates, the majority of GOIs were up-regulated (up to 4000%) in most colonies during a laboratory-based thermal stress experiment. Two genes, Nf-kβ2 and MnSod, were consistently up-regulated in all colonies tested, and we therefore propose these as candidate markers useful for population-level evaluations of thermal stress. Our assay provides an important new tool for coral bleaching studies; because of the lower cost, labour and amount of cDNA required compared with singleplex qPCR, population-level studies with large biological replication are feasible.
|Item Type:||Article (Refereed Research - C1)|
|Keywords:||Acropora, bleaching, GeXP, microarray, RT–qPCR, stress|
|FoR Codes:||06 BIOLOGICAL SCIENCES > 0602 Ecology > 060205 Marine and Estuarine Ecology (incl Marine Ichthyology) @ 100%|
|SEO Codes:||97 EXPANDING KNOWLEDGE > 970106 Expanding Knowledge in the Biological Sciences @ 100%|
|Deposited On:||26 Mar 2012 15:57|
|Last Modified:||18 Oct 2013 01:28|
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