文献类型：期刊文献

英文题名：Identification and Validation of Reliable Reference Genes for Gene Expression Studies in Koelreuteria paniculata

作者：Gao, Kai[1,2] Khan, Wasif Ullah[1] Li, Juan[1] Huang, Sai[1] Yang, Xiong[1] Guo, Ting[1] Guo, Bin[1,3] Wu, Ruqian[1] An, Xinmin[1]

第一作者：高凯;Gao, Kai

通信作者：An, XM[1]

机构：[1]Beijing Forestry Univ, Coll Biol Sci & Technol, Beijing Adv Innovat Ctr Tree Breeding Mol Design, Natl Engn Res Ctr Tree Breeding & Ecol Restorat, Beijing 100083, Peoples R China;[2]Chinese Acad Forestry, Res Inst Subtrop Forestry, Hangzhou 311400, Peoples R China;[3]Shanxi Acad Forestry & Grassland Sci, Taiyuan 030012, Peoples R China

年份：2022

卷号：13

期号：5

外文期刊名：GENES

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000804357000001)】；

基金：This work was supported by the Natural Science Foundation of China (31870652), the Opening Foundation of Key Laboratory of Urban Agriculture (North China), Ministry of Agriculture, P. R. China (kf2018012), the project from Science and Technology Development Center of State Forestry Administration (KJZXSA2018030).

语种：英文

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

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 beta-actin (ACT) were the most suitable and statistically reliable reference genes, whereas ribosomal protein L13 (RPL13) and elongation factor 1-alpha (EF1 alpha) 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.