Evaluation of reference genes for normalizing RT-qPCR in leaves and suspension cells of Cephalotaxus hainanensis under various stimuli
文献类型： 外文期刊
作者： Sun, Huapeng 1 ; Jiang, Xuefei 2 ; Sun, Mengli 2 ; Cong, Hanging 1 ; Qiao, Fei 1 ;
作者机构： 1.Chinese Acad Trop Agr Sci, Key Lab Crop Gene Resources & Germplasm Enhanceme, Minist Agr, Trop Crops Genet Resources Inst, Danzhou 571737, Hainan, Peoples R China
2.Hainan Univ, Inst Trop Agr & Forestry, Hainan Key Lab Sustainable Utilizat Trop Bioresou, Haikou 570228, Hainan, Peoples R China
关键词： RT-qPCR; Reference gene; C; hainanensis; Expression stability; Normalization
期刊名称：PLANT METHODS （ 影响因子：4.993； 五年影响因子：5.312 ）
ISSN： 1746-4811
年卷期： 2019 年 15 卷
页码：
收录情况： SCI
摘要： BackgroundReverse transcription quantitative real-time PCR (RT-qPCR) is a widely used approach for investigating gene expression levels in plants because of its high reproducibility, sensitivity, accuracy and rapidness. Evaluation of reference genes for normalizing RT-qPCR data is a necessary step, especially in new plant varieties. Cephalotaxus hainanensis is a precious medicinal plant belonging to the family of Cephalotaxaceae and no RT-qPCR studies have been reported on it.ResultsIn this study, 9 candidate reference genes were selected from the transcriptome data of C. hainanensis; 3 statistical algorithms (geNorm, NormFinder, BestKeeper) were applied to evaluate their expression stabilities through 180 samples under 6 stimuli treatments in leaves and leaf-derived suspension cultured cells; a comprehensive stabilities ranking was also performed by RefFinder. The results showed that suitable reference genes in C. hainanensis should be selected for normalization relative to different experimental sets. 18S showed a higher stability than other candidate reference genes which ranked at the top two suitable genes under all experimental setups in this study.ConclusionThis study is the first to evaluate the stability of reference genes in C. hainanensis and supply an important foundation to use the RT-qPCR for an accurate and far-reaching gene expression analysis in C. hainanensis.
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