Identification and Testing of Reference Genes for qRT-PCR Analysis During Pear Fruit Development

Abstract

Background: Quantitative real-time PCR (qRT-PCR) is currently one of the most reliable and improved tools for analyzing gene expression. Various studies have shown that housekeeping genes was varied with cultivars, tissues and treatment. The reliable and stable reference genes were necessarily identified and evaluated according to different experimental requirements. Result: In this study, 10 candidate reference genes were initially screened based on the transcriptome sequencing data of four pear fruit development stages of three different pear cultivars, including a candidate housekeeping gene PbrTUB. Furthermore, we ranked the expression stability of 10 candidate reference genes using algorithms GeNorm, NormFinder, BestKeeper and ReFinder. Finally, the result showed that Pbr028511, Pbr038418 and Pbr041114 were the most stable reference gene in Cuiguan, Housui and Xueqing fruit, respectively. Concludion: Thee results provide a valuable resource that serve as significant reference for gene function explorations and molecular mechanism studies in fruit development and ripening of different pear cultivars.