文献类型：期刊文献

英文题名：Selection of reference genes for quantitative real-time PCR in six oil-tea camellia based on RNA-seq

作者：Zhou, C. F.[1] Lin, P.[1] Yao, X. H.[1] Wang, K. L.[1] Chang, J.[1] Han, X. J.[1]

第一作者：Zhou, C. F.

通信作者：Zhou, CF[1]

机构：[1]Chinese Acad Forestry, Res Inst Subtrop Forestry, Hangzhou 311400, Zhejiang, Peoples R China

年份：2013

卷号：47

期号：6

起止页码：836-851

外文期刊名：MOLECULAR BIOLOGY

收录：;Scopus(收录号：2-s2.0-84890088232);WOS:【SCI-EXPANDED(收录号:WOS:000328217400010)】；

基金：This work was supported by the Creation of High Yield and Quality New Oil-tea camellia Germplasm ( no. 2009BADB1B01) and the Research and Development of Crucial Technology for Oil-tea camellia Industrial Upgrading ( no. 2009BADB1B00).

语种：英文

外文关键词：reference genes; real-time PCR; oil-tea camellia

摘要：qRT-PCR is becoming a routine tool in molecular biology to study gene expression. It is necessary to find stable reference genes when performing qRT-PCR. The expression of genes cloned in oil-tea camellia currently cannot be accurately analyzed due to a lack of suitable reference genes. We collected different tissues (including roots, stems, leaves, flowers and seeds) from six oil-tea camellia species to determine stable reference genes. Five novel and ten traditional reference gene sequences were selected from the RNA-seq database of Camellia oleifera Abel seeds and specific PCR Primers were designed for each. Cycle threshold (C (t)) data were obtained from each reaction for all samples. Three different software tools, geNorm, NormFinder and Best-Keeper were applied to calculate the expression stability of the candidate reference genes according to the C (t) values. The results were similar between the three software packages, and indicated that the traditional genes TUB alpha-3, ACT7 alpha and the novel gene CESA were relatively stable in all species and tissues. However, no genes were sufficiently stable across all species and tissues, thus the optimal number of reference genes required for accurate normalization varied from 2 to 6. Finally, the relative expression of squalene synthase (SQS) and squalene epoxidase (SQE) genes related to important ingredients squalene and tea saponin in oil-tea camellia seeds were compared by using stable to less stable reference genes. The comparison results validated the selection of reference genes in the current study. In summary, for the different tissues of six oil-tea camellia species different optimal numbers of suitable reference genes were found.