A Scale-up System for Lowbush Blueberry Micropropagation Using a Bioreactor

Abstract

In an attempt to improve the micropropagation protocol for lowbush blueberry (Vaccinium angustifolium Ait.), a protocol using a bioreactor system combined with a semisolid gelled medium has been developed. Cultures of cultivar Fundy and two wild clones (‘NB1’ and ‘QB1’) were established in vitro on a gelled modified cranberry basal medium (BM) containing 5 μM zeatin or 10 μM N6-[2-isopentenyl]adenine. Multiple shoots were obtained within 8 weeks by transferring zeatin-induced shoots from the gelled BM to a bioreactor containing liquid BM with 1 to 4 μM zeatin. Genotypes differed significantly with respect to multiplication rate in liquid and gelled BM containing 1 μM zeatin with ‘NB1’ producing 8.5 ± 1.1 and 2.9 ± 0.3 shoots per explant in liquid and gelled media, respectively, after one subculture followed by ‘QB1’ (7.1 ± 0.6 and 2.6 ± 0.4 shoots per explant, respectively) and ‘Fundy’ (5.8 ± 0.4 and 2.0 ± 0.2 shoots per explant, respectively). With subculture, there was an increase of shoot multiplication rate for all genotypes. Bioreactor- and gelled medium-proliferated shoots were treated with 39.4 mm indole-3-butyric acid powder, rooted in a 2 peat:1 perlite (v/v) medium, plantlets acclimatized, and eventually established in the greenhouse with 64% to 74% rooting of microshoots and 90% to 99% survival of rooted shoots. Results obtained suggested the possibility of large-scale multiplication of lowbush blueberry shoots in bioreactors.