Selection of reference genes for quantitative real-time PCR in Cocos nucifera during abiotic stress

Xia, Wei, Liu, Zheng, Yang, Yaodong, Xiao, Yong, Mason, Annaliese S., Zhao, Songlin and Ma, Zilong (2014) Selection of reference genes for quantitative real-time PCR in Cocos nucifera during abiotic stress. Botany, 92 3: 179-186. doi:10.1139/cjb-2013-0212


Author Xia, Wei
Liu, Zheng
Yang, Yaodong
Xiao, Yong
Mason, Annaliese S.
Zhao, Songlin
Ma, Zilong
Title Selection of reference genes for quantitative real-time PCR in Cocos nucifera during abiotic stress
Journal name Botany   Check publisher's open access policy
ISSN 1916-2790
1916-2804
Publication date 2014
Year available 2013
Sub-type Article (original research)
DOI 10.1139/cjb-2013-0212
Open Access Status
Volume 92
Issue 3
Start page 179
End page 186
Total pages 8
Place of publication Ottawa, ON, Canada
Publisher N R C Research Press
Collection year 2014
Language eng
Formatted abstract
Reverse transcription quantitative real-time polymerase chain reaction (PCR) is a widely used and reproducible
method for studying gene expression changes. However, its accuracy and reliability is highly dependent on the normalization
step. Cocos nucifera L., a perennial palm with a long productive life span, is frequently exposed to soil and atmospheric drought
and other stresses. In applying gene expression analysis to understand coconut stress responses, validation of suitable reference genes is an important first step. In this study, seven putative reference genes were identified from coconut transcriptome data. The stability of these putative reference genes was assessed in a diverse set of 18 coconut samples subject to cold, drought, and high-salinity treatment and representing different endosperm developmental stages. Using statistical algorithms geNorm, NormFinder, and BestKeeper, eEF1- and UBC10 genes were identified as stable reference genes in all stress treatments and endosperm development stages, consistent with validated reference genes in rice (Oryza sativa L.), potato (Solanum tuberosum L.), and other species. Further validation of analyzed reference genes by normalization of a C-repeat binding factor (CBF)-like gene in cold-treatment samples indicated that the less stable reference genes produced different results from the more stable reference genes, with weaker variation or higher expression levels of a target gene. Our results will be beneficial for further research on molecular mechanisms of stress resistance.
Keyword Abiotic stresses
Cocos nucifera
Normalization
Reference gene
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: School of Agriculture and Food Sciences
Official 2014 Collection
 
Versions
Version Filter Type
Citation counts: TR Web of Science Citation Count  Cited 3 times in Thomson Reuters Web of Science Article | Citations
Scopus Citation Count Cited 5 times in Scopus Article | Citations
Google Scholar Search Google Scholar
Created: Tue, 15 Apr 2014, 01:26:34 EST by System User on behalf of School of Agriculture and Food Sciences