Evaluation of candidate reference genes for real time quantitative PCR normalization in pear fruit

Abstract

In this study, eight candidate reference genes including EF1α, TUB-b2, GAPDH, 18S rRNA, RP-L27P, UBI,ACT and ACT2 were selected to evaluate expression stability in pear fruit of different genotypes, tissues, developmental stages and phytohormone-treatments, using three software approaches: geNorm, NormFinder and BestKeeper. The results showed that EF1α, TUB-b2 and GAPDH had the most stable expression across all experimental conditions tested. EF1α and TUB-b2 were the most stable reference genes across all genotypes. GAPDH and EF1α were the most suitable reference genes across different tissues. For developmental stage and phytohormone-treated samples, TUB-b2 and GAPDH were the most stable reference genes.   Key words: qPCR, reference gene, pear.