Evaluation of reference genes for RT-QPCR studies in the seagrass Zostera muelleri exposed to light limitation

Schliep, M., Pernice, M., Sinutok, S., Bryant, C.V., York, P.H., Rasheed, M.A., and Ralph, P.J. (2015) Evaluation of reference genes for RT-QPCR studies in the seagrass Zostera muelleri exposed to light limitation. Scientific Reports, 5. 17051. pp. 1-11.

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Seagrass meadows are threatened by coastal development and global change. In the face of these pressures, molecular techniques such as reverse transcription quantitative real-time PCR (RT-qPCR) have great potential to improve management of these ecosystems by allowing early detection of chronic stress. In RT-qPCR, the expression levels of target genes are estimated on the basis of reference genes, in order to control for RNA variations. Although determination of suitable reference genes is critical for RT-qPCR studies, reports on the evaluation of reference genes are still absent for the major Australian species Zostera muelleri subsp. capricorni (Z. muelleri). Here, we used three different software (geNorm, NormFinder and Bestkeeper) to evaluate ten widely used reference genes according to their expression stability in Z. muelleri exposed to light limitation. We then combined results from different software and used a consensus rank of four best reference genes to validate regulation in Photosystem I reaction center subunit IV B and Heat Stress Transcription factor A- gene expression in Z. muelleri under light limitation. This study provides the first comprehensive list of reference genes in Z. muelleri and demonstrates RT-qPCR as an effective tool to identify early responses to light limitation in seagrass.

Item ID: 41100
Item Type: Article (Research - C1)
ISSN: 2045-2322
Keywords: dredging, reference genes, light limitation, RT-qPCR, seagrass, Zostera muelleri
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Funders: Australian Research Council (ARC), Gladstone Ports Corporation (GPC)
Projects and Grants: ARC Linkage Grant LP11020045
Date Deposited: 24 Nov 2015 03:52
FoR Codes: 05 ENVIRONMENTAL SCIENCES > 0501 Ecological Applications > 050102 Ecosystem Function @ 30%
06 BIOLOGICAL SCIENCES > 0604 Genetics > 060405 Gene Expression (incl Microarray and other genome-wide approaches) @ 50%
06 BIOLOGICAL SCIENCES > 0602 Ecology > 060203 Ecological Physiology @ 20%
SEO Codes: 96 ENVIRONMENT > 9605 Ecosystem Assessment and Management > 960503 Ecosystem Assessment and Management of Coastal and Estuarine Environments @ 60%
96 ENVIRONMENT > 9606 Environmental and Natural Resource Evaluation > 960604 Environmental Management Systems @ 40%
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