Evaluation of reference genes for normalizing RT-qPCR in leaves and suspension cells of Cephalotaxus hainanensis under various stimuli

Reverse 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...

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Bibliographic Details
Published in:Plant methods 2019-03, Vol.15 (1), p.31-31, Article 31
Main Authors: Sun, Huapeng, Jiang, Xuefei, Sun, Mengli, Cong, Hanqing, Qiao, Fei
Format: Article
Language:English
Subjects:
Algorithms
Biochemistry
Biosynthesis
C. hainanensis
Cephalotaxus
Cephalotaxus hainanensis
Conifers
Evaluation
Expression stability
Flowers & plants
Gene expression
Genes
Genetic aspects
Genetic engineering
Herbal medicine
Leaves
Leukemia
Medicinal plants
Metabolites
Normalization
Normalizing (statistics)
Physiological aspects
Raw materials
Reference gene
Reproducibility
Reverse transcription
RT-qPCR
Sensitivity analysis
Stability analysis
Statistical methods
Stimuli
Transcription (Genetics)
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Summary:Reverse 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. is a precious medicinal plant belonging to the family of Cephalotaxaceae and no RT-qPCR studies have been reported on it. In this study, 9 candidate reference genes were selected from the transcriptome data of ; 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 should be selected for normalization relative to different experimental sets. showed a higher stability than other candidate reference genes which ranked at the top two suitable genes under all experimental setups in this study. This study is the first to evaluate the stability of reference genes in and supply an important foundation to use the RT-qPCR for an accurate and far-reaching gene expression analysis in .
ISSN:1746-4811
1746-4811
DOI:10.1186/s13007-019-0415-y