Selection of appropriate reference genes in different tissues of Vaccinium dunalianum Wight by quantitative real-time PCR for gene expression studies

Abstract

Quantitative real-time polymerase chain reaction (PCR) is a powerful tool for investigating relevant changes in gene expression. At present, there are no reports on reference genes for Vaccinium dunalianum Wight (V. dunalianum Wight). Here, the expression profiles of 15 frequently used reference genes were used to screen for appropriate reference genes in different tissues of V. dunalianum Wight collected in Yunnan and Wuding, China. The cycle threshold (Ct) values of genes in different samples were compared and analyzed. Furthermore, BestKeeper, NormFinder, and geNorm software were used to evaluate the stability of each candidate reference gene. Analysis using geNorm showed that the expression levels of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and protein phosphatase 2A-2 (PP2A-2) were the most stable. In turn, the expression level of eukaryotic initiation factor-2 (EIF-4A-2) was the most stable as per analysis by NormFinder, while Bestkeeper revealed that actin-related protein-1 (ARP-1) was the most stably expressed. Comprehensive analysis and validation experiments showed that EIF-4A-2 and PP2A-2 were the most suitable reference genes for V. dunalianum Wight.