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Screening of valid reference genes for real-time RT-PCR data normalization in Hevea brasiliensis and expression validation of a sucrose transporter gene HbSUT3

文献类型： 外文期刊

作者： Li, Heping 1 ; Qin, Yunxia 1 ; Xiao, Xiaohu 1 ; Tang, Chaorong 1 ;

作者机构： 1.Chinese Acad Trop Agr Sci, Key Lab Rubber Biol, Minist Agr, Danzhou 571737, Hainan, Peoples R China

2.Chinese Acad Trop Agr Sci, Rubber Res Inst, Danzhou 571737, Hainan, Peoples R China

3.Hainan Univ, Coll Agr, Danzhou 571737, Hainan, Peoples R China

关键词： Real-time RT-PCR;Gene expression;Normalization;Reference gene;Sucrose transporter HbSUT3;Hevea brasiliensis

期刊名称：PLANT SCIENCE （ 影响因子：4.729； 五年影响因子：5.132 ）

ISSN： 0168-9452

年卷期： 2011 年 181 卷 2 期

页码：

收录情况： SCI

摘要： Real-time RT-PCR (RT-qPCR) is a sensitive and precise method of quantifying gene expression, however, suitable reference genes are required. Here, a systematic reference gene screening was performed by RT-qPCR on 22 candidate genes in Hevea brasiliensis. Two ubiquitin-protein ligases (UBC2a and UBC4) were the most stable when all samples were analyzed together. A mitosis protein (YLS8)and a eukaryotic translation initiation factor (eIF1Aa)were the most stable in response to tapping. UBC2b and UBC1 were the most stable among different genotypes. UBC2b and a DEAD box RNA helicase (RH2b) were the most stable across individual trees. YLS8 and RH8 were most stably expressed in hormone-treated samples. Expression of the candidate reference genes varied significantly across different tissues, and at least four genes (RH2b, RH8, UBC2a and elF2) were needed for expression normalization. In addition, examination of relative expression of a sucrose transporter HbSUT3 in different RNA samples demonstrated the importance of additional reference genes to ensure accurate quantitative expression analysis. Overall, our work serves as a guide for selection of reference genes in RT-qPCR gene expression studies in H. brasiliensis. Published by Elsevier Ireland Ltd.

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