title: Evaluation of Reference Genes for RT-qPCR Studies in the Seagrass Zostera muelleri Exposed to Light Limitation.
abstract: 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.
authors.affiliation: Plant Functional Biology and Climate Change Cluster (C3), University of Technology Sydney, 15 Broadway, Ultimo, 2007, NSW, Australia.
authors.forename: M
authors.initials: M
authors.lastname: Schliep
authors.affiliation: Plant Functional Biology and Climate Change Cluster (C3), University of Technology Sydney, 15 Broadway, Ultimo, 2007, NSW, Australia.
authors.forename: M
authors.initials: M
authors.lastname: Pernice
authors.affiliation: Plant Functional Biology and Climate Change Cluster (C3), University of Technology Sydney, 15 Broadway, Ultimo, 2007, NSW, Australia.
authors.forename: S
authors.initials: S
authors.lastname: Sinutok
authors.affiliation: TropWATER - Centre for Tropical Water and Aquatic Ecosystem Research, James Cook University, 1-88 McGregor Road, Smithfield, 4878, QLD, Australia.
authors.forename: C V
authors.initials: CV
authors.lastname: Bryant
authors.affiliation: TropWATER - Centre for Tropical Water and Aquatic Ecosystem Research, James Cook University, 1-88 McGregor Road, Smithfield, 4878, QLD, Australia.
authors.forename: P H
authors.initials: PH
authors.lastname: York
authors.affiliation: TropWATER - Centre for Tropical Water and Aquatic Ecosystem Research, James Cook University, 1-88 McGregor Road, Smithfield, 4878, QLD, Australia.
authors.forename: M A
authors.initials: MA
authors.lastname: Rasheed
authors.affiliation: Plant Functional Biology and Climate Change Cluster (C3), University of Technology Sydney, 15 Broadway, Ultimo, 2007, NSW, Australia.
authors.forename: P J
authors.initials: PJ
authors.lastname: Ralph
mesh_terms: D000222:Adaptation, Physiological / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics; D017753:Ecosystem; D018506:Gene Expression Regulation, Plant; D020043:Genes, Essential; D017343:Genes, Plant; D000076249:Heat Shock Transcription Factors; D006360:Heat-Shock Proteins / Q000235:genetics; D008027:Light; D045331:Photosystem I Protein Complex / Q000235:genetics; D010940:Plant Proteins / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction / Q000592:standards*; D012015:Reference Standards; D012984:Software; D013312:Stress, Physiological; D014157:Transcription Factors / Q000235:genetics; D035962:Zosteraceae / Q000235:genetics*
chemical_list: D004268:DNA-Binding Proteins; D000076249:Heat Shock Transcription Factors; D006360:Heat-Shock Proteins; D045331:Photosystem I Protein Complex; D010940:Plant Proteins; D014157:Transcription Factors; C079741:photosystem I, psaE subunit