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Selection of Reliable Reference Genes for Gene Expression Studies on Rhododendron molle G. Don

文献类型： 外文期刊

作者： Xiao, Zheng 1 ; Sun, Xiaobo 1 ; Liu, Xiaoqing 1 ; Li, Chang 1 ; He, Lisi 1 ; Chen, Shangping 1 ; Su, Jiale 1 ;

作者机构： 1.Jiangsu Acad Agr Sci, Inst Hort, Jiangsu Key Lab Hort Crop Genet Improvement, Nanjing, Jiangsu, Peoples R China

关键词： Rhododendron;qRT-PCR;flower;reference gene;normalization

期刊名称：FRONTIERS IN PLANT SCIENCE （ 影响因子：5.753； 五年影响因子：6.612 ）

ISSN： 1664-462X

年卷期： 2016 年 7 卷

页码：

收录情况： SCI

摘要： The quantitative real-time polymerase chain reaction (qRT-PCR) approach has become a widely used method to analyze expression patterns of target genes. The selection of an optimal reference gene is a prerequisite for the accurate normalization of gene expression in qRT-PCR. The present study constitutes the first systematic evaluation of potential reference genes in Rhododendron motto G. Don. Eleven candidate reference genes in different tissues and flowers at different developmental stages of R. molle were assessed using the following three software packages: GeNorm, NormFinder, and BestKeeper. The results showed that EF1-alpha (elongation factor 1-alpha), 18S (18s ribosomal RNA), and RPL3 (ribosomal protein L3) were the most stable reference genes in developing rhododendron flowers and, thus, in all of the tested samples, while tublin (TUB) was the least stable. ACT5 (actin), RPL3, 18S, and EF1-alpha were found to be the top four choices for different tissues, whereas TUB was not found to favor qRT-PCR normalization in these tissues. Three stable reference genes are recommended for the normalization of qRT-PCR data in R. molle. Furthermore, the expression profiles of RmPSY (phytoene synthase) and RmPDS (phytoene dehydrogenase) were assessed using EF1-alpha, 18S, ACT5, RPL3, and their combination as internals. Similar trends were found, but these trends varied when the least stable reference gene TUB was used. The results further prove that it is necessary to validate the stability of reference genes prior to their use for normalization under different experimental conditions. This study provides useful information for reliable qRT-PCR data normalization in gene studies of F. molle.

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