Selection of reference genes for RT-qPCR normalization in blueberry (Vaccinium corymbosum x angustifolium) under various abiotic stresses

As a small fruit rich in anthocyanins, blueberry (Vaccinium corymbosum x angustifolium) has become a focus of research in recent years for identifying genes related to anthocyanin transport and stress resistance mechanisms based on transcriptome sequencing. However, the lack of validated, stably expressed reference genes greatly limits the functional study of blueberry genes. Therefore, in this study, we selected 14 candidate reference genes from a blueberry transcriptome database and used three algorithms (geNorm, NormFinder and BestKeeper) to evaluate the expression stability of these genes in various organs at different fruit developmental stages under five abiotic stress conditions.EF1 alpha,EIFandTBPwere observed to be the most stable and were thus chosen as reference genes for quantitative real-time PCR. Measurement of the relative expression ofVcMATE1(European Nucleotide Archive accession number KF875433) in blueberry further verified the reliability of these reference genes, which may have great utility for determining the accuracy of gene expression analyses in future research on blueberry.