Benzyl adenine restores anthocyanin pigmentation in suspension cultures of wild Vaccinium pahalae

Abstract

The overriding influence of cytokinin source on flavonoid production in vitro was explored using a suspension culture system for Vaccinium pahalae. The substitution of kinetin by 20 μM benzyl adenine (BA) in the suspension culture media resulted in a three-fold increase in total anthocyanin yield, and a more rapid production during the cell culture cycle. Anthocyanin production reached a maximum after a 16–20 day interval in cultures containing an optimal kinetin concentration, but pigment accumulation peaked at only 12–16 days when BA was used as the sole cytokinin source. Unlike some other production systems which increase secondary metabolite production at the expense of cell growth, BA-supplementation promoted both increased growth and increased anthocyanin productivity. In BA-supplemented medium, cultures were not susceptible to typical osmotically-induced cell growth suppression. When, after multiple subcultures in kinetin-containing media, anthocyanin production capability was lost or diminished, productivity could be restored within 3 days after transfer of cells to a BA-supplemented medium.