title: Evaluation of reference genes for gene expression studies in radish (Raphanus sativus L.) using quantitative real-time PCR.
abstract: Real-time quantitative reverse transcription PCR (RT-qPCR) is a rapid and reliable method for gene expression studies. Normalization based on reference genes can increase the reliability of this technique; however, recent studies have shown that almost no single reference gene is universal for all possible experimental conditions. In this study, eight frequently used reference genes were investigated, including Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), Actin2/7 (ACT), Tubulin alpha-5 (TUA), Tubulin beta-1 (TUB), 18S ribosomal RNA (18SrRNA), RNA polymerase-II transcription factor (RPII), Elongation factor 1-b (EF-1b) and Translation elongation factor 2 (TEF2). Expression stability of candidate reference genes was examined across 27 radish samples, representing a range of tissue types, cultivars, photoperiodic and vernalization treatments, and developmental stages. The eight genes in these sample pools displayed a wide range of Ct values and were variably expressed. Two statistical software packages, geNorm and NormFinder showed that TEF2, RPII and ACT appeared to be relatively stable and therefore the most suitable for use as reference genes. These results facilitate selection of desirable reference genes for accurate gene expression studies in radish.
authors: National Key Laboratory of Crop Genetics and Germplasm Enhancement, Key Laboratory of Biology and Genetic Improvement of Horticultural Crops (East China), Ministry of Agriculture of PR China, College of Horticulture, Nanjing Agricultural University, Nanjing 210095, PR China.
authors: Yuanyuan
authors: Y
authors: Xu
authors: Xianwen
authors: X
authors: Zhu
authors: Yiqin
authors: Y
authors: Gong
authors: Liang
authors: L
authors: Xu
authors: Yan
authors: Y
authors: Wang
authors: Liwang
authors: L
authors: Liu
mesh_terms: D017931:DNA Primers / Q000737:chemistry / Q000235:genetics; D015870:Gene Expression; D018506:Gene Expression Regulation, Plant; D017343:Genes, Plant; D031224:Raphanus / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction / Q000592:standards*; D012015:Reference Standards
chemical_list: D017931:DNA Primers