selection of reliable reference genes for gene expression studies using real-time pcr in tung tree during seed development选择可靠的参考基因的基因表达研究使用实时pcr在东树种子发展.pdfVIP

Selection of Reliable Reference Genes for Gene Expression Studies Using Real-Time PCR in Tung Tree during Seed Development 1,2 1,3 1,2 2 2 1,2 Xiaojiao Han , Mengzhu Lu , Yicun Chen , Zhiyong Zhan , Qinqin Cui , Yangdong Wang * 1 State Key Laboratory of Tree Genetics and Breeding, Chinese Academy of Forestry, Beijing, People’s Republic of China, 2 Research Institute of Subtropical Forestry, Chinese Academy of Forestry, Fuyang, People’s Republic of China, 3 Research Institute of Forestry, Chinese Academy of Forestry, Beijing, People’s Republic of China Abstract Quantitative real-time PCR (RT-qPCR) has become an accurate and widely used technique to analyze expression levels of selected genes. It is very necessary to select appropriate reference genes for gene expression normalization. In the present study, we assessed the expression stability of 11 reference genes including eight traditional housekeeping genes and three novel genes in different tissues/organs and developing seeds from four cultivars of tung tree. All 11 reference genes showed a wide range of Ct values in all samples, indicating that they differently expressed. Three softwares – geNorm, NormFinder and BestKeeper – were used to determine the stability of these references except for ALB (2S albumin), which presented a little divergence. The results from the three softwares showed that ACT7 (Actin7a), UBQ (Ubiquitin), GAPDH (glyceraldehyde- 3-phosphate dehydrogenase) and EF1a (elongation factor 1-a) were the most stable reference genes across all of the tested tung samples and tung developing seeds, while ALB (2S albumin) was unsuitable as internal controls. ACT7, EF1b (elongation factor1