A multilocus, temperature stress-related gene expression profile assay in Acropora millepora, a dominant reef-building coral.

Abstract:

: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.

journal_name

Mol Ecol Resour

authors

Souter P,Bay LK,Andreakis N,Császár N,Seneca FO,van Oppen MJ

doi

10.1111/j.1755-0998.2010.02923.x

subject

Has Abstract

pub_date

2011-03-01 00:00:00

pages

328-34

issue

2

eissn

1755-098X

issn

1755-0998

journal_volume

11

pub_type

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