Identification and Validation of Reliable Reference Genes for Gene Expression Studies in Koelreuteria paniculata

Genes

Bin Guo¹  Kai Gao¹  Wasif Ullah Khan¹  Juan Li¹  Xiong Yang¹  Ruqian Wu¹  Xinmin An¹  Sai Huang¹  Ting Guo¹ 

[1] Beijing Advanced Innovation Center for Tree Breeding by Molecular Design, National Engineering Research Center of Tree breeding and Ecological Restoration, College of Biological Sciences and Technology, Beijing Forestry University, Beijing 100083, China;

关键词: Koelreuteria paniculata;    RT-qPCR;    reference genes;    plant tissues;    embryo developmental stages;   

DOI  :  10.3390/genes13050714

来源: DOAJ

【 摘 要 】

RT-qPCR is considered a rapid and reliable technique for analyzing gene expression. This technique is commonly used to analyze the expression of various genes at diverse transcriptional levels in different samples. However, few studies have characterized ornamental Koelreuteria species for reliable reference genes. In this study, eight reference genes were evaluated as controls in RT-qPCR with SYBR green to quantify gene expression in different Koelreuteria paniculata samples. All selected reference genes showed a broad range of C_t values in all samples, which was supportive of their variable expression. Our results showed significant variation in the stable expression of K. paniculata genes. Sample data, analyzed using geNorm, NormFinder, and BestKeeper, showed that phospholipase (PLA2) and β-actin (ACT) were the most suitable and statistically reliable reference genes, whereas ribosomal protein L13 (RPL13) and elongation factor 1-α (EF1α) were less stable and unsuitable for use as internal controls. To compare gene expression levels, two or more reference genes should be used for data normalization. Thus, the stability and expression of both PLA2 and ACT were believed to provide better normalization and quantification of the transcript levels for gene expression studies in K. paniculata.

【 授权许可】

Unknown