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

文献类型： 外文期刊

作者： Xia, Wei 1 ; Liu, Zheng 1 ; Yang, Yaodong 1 ; Xiao, Yong 1 ; Mason, Annaliese S. 2 ; Zhao, Songlin 1 ; Ma, Zilong 4 ;

作者机构： 1.Chinese Acad Trop Agr Sci, Hainan Key Lab Trop Oil Crops Biol, Coconuts Res Inst, Wenchang 571339, Hainan, Peoples R China

2.Univ Queensland, Sch Agr & Food Sci, Brisbane, Qld 4072, Australia

3.Univ Queensland, Ctr Integrat Legume Res, Brisbane, Qld 4072, Australia

4.Chinese Acad Trop Agr Sci, Key Lab Trop Crop Biol & Genet Resources Utilizat, Minist Agr, Inst Trop Biosci & Biotechnol, Haikou 571101, Hainan, Peoples R China

关键词： Cocos nucifera;reference gene;abiotic stresses;normalization

期刊名称：BOTANY （ 影响因子：1.323； 五年影响因子：1.568 ）

ISSN： 1916-2790

年卷期： 2014 年 92 卷 3 期

页码：

收录情况： SCI

摘要： 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-alpha 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.

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