Validation of reference genes for real-time quantitative PCR normalization in soybean developmental and germinating seeds

文献类型： 外文期刊

第一作者： Li, Qing

作者： Li, Qing;Fan, Cheng-Ming;Zhang, Xiao-Mei;Fu, Yong-Fu

作者机构：

关键词： Soybean;Seed development;Seed germination;RT-qPCR;Normalization;Reference gene

期刊名称：PLANT CELL REPORTS （ 影响因子：4.57； 五年影响因子：4.463 ）

ISSN： 0721-7714

年卷期： 2012 年 31 卷 10 期

页码：

收录情况： SCI

摘要： Most of traditional reference genes chosen for real-time quantitative PCR normalization were assumed to be ubiquitously and constitutively expressed in vegetative tissues. However, seeds show distinct transcriptomes compared with the vegetative tissues. Therefore, there is a need for re-validation of reference genes in samples of seed development and germination, especially for soybean seeds. In this study, we aimed at identifying reference genes suitable for the quantification of gene expression level in soybean seeds. In order to identify the best reference genes for soybean seeds, 18 putative reference genes were tested with various methods in different seed samples. We combined the outputs of both geNorm and NormFinder to assess the expression stability of these genes. The reference genes identified as optimums for seed development were TUA5 and UKN2, whereas for seed germination they were novel reference genes Glyma05g37470 and Glyma08g28550. Furthermore, for total seed samples it was necessary to combine four genes of Glyma05g37470, Glyma08g28550, Glyma18g04130 and CYP for normalization. Key message We identified several reference genes that stably expressed in soybean seed developmental and germinating processes.

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