Evaluation of reference genes for transcript normalization in fruit and vegetative tissues.

Unlabelled: Quantitative real-time PCR (RT-qPCR) is used extensively in gene expression studies. For adequate comparisons, the identification and use of reliable reference genes are crucial. Nevertheless, the availability of such genes in strawberry species is limited and has yet to be described for the Chilean strawberry, . In this study, the expression dynamics of a set of 10 candidate reference genes were analyzed in various vegetative tissues (root, runners, stem, leaf, and flower), and fruits at different ripening stages or subjected to different hormonal treatments (ABA, auxin). The expression stability of candidate genes was examined by a series of algorithms, such as geNorm, NormFinder, BestKeeper, and ΔCt, for comparisons and rankings. Finally, by using RefFinder, a comprehensive and comparative ranking of the four methods was achieved. The results highlight that the expression stability of candidate reference genes fluctuates depending on tissue type, fruit stage, and hormonal treatment. As reference genes, the use of and is recommended for vegetative tissues; and for fruit ripening stages; and for fruit subjected to ABA and NDGA treatments; and for fruit under AUXIN and TIBA treatments; and and when all fruit stages and hormonal treatments are compared. If just one reference gene is employed as a normalizer, should be chosen as it is the most stable internal control in most conditions. Therefore, the present study delivers a set of reliable reference genes for RT-qPCR expression analysis in tissues and fruits subjected to several hormonal treatments.

Supplementary Information: The online version contains supplementary material available at 10.1007/s12298-022-01227-y.