Validation of Reference Genes in a Population of Blueberry Plants Regenerated in Colchicine.

For the first time we report the validation of reference genes in plants from a population of blueberry () clones cultured in vitro on a colchicine-supplemented medium. Nodal segment explants of the cultivar Duke were regenerated by organogenesis under different periods of colchicine 1 mg/L exposure (1, 2, 3, 5, 30 days). The clones selected for the study showed variability for phenotypic traits after 2 years of adaptation to field conditions, compared to plants of the donor genotype that were regenerated on a medium without colchicine. () and ( and ) were used as reference genomes for primer design. The results show that colchicine treatments can cause genomic changes in blueberry plants. At the molecular level, exposure of plants to colchicine in early periods could promote an increase in gene expression of specific genes such as and RPOB. However, prolonged exposure (30 days) could decrease gene expression of the genes studied. For qPCR assays, the primers designed for and genes showed high efficiency. In addition, the , , and genes showed high stability and could be recommended as potential reference genes for gene expression assays in Vaccinium.