Authors contributed equally to this work.
A multilocus, temperature stress-related gene expression profile assay in Acropora millepora, a dominant reef-building coral
Version of Record online: 28 SEP 2010
© 2010 Blackwell Publishing Ltd
Molecular Ecology Resources
Volume 11, Issue 2, pages 328–334, March 2011
How to Cite
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: 328–334. doi: 10.1111/j.1755-0998.2010.02923.x
- Issue online: 9 FEB 2011
- Version of Record online: 28 SEP 2010
- Received 2 February 2010; revision received 31 August 2010; accepted 2 September 2010
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.