Factors Affecting Plant Regeneration from Leaf Tissues of Buddleia Species

Abstract

The effects of genotype, basal medium, plant growth regulator (PGR), dark treatment, and antibiotics on shoot regeneration of two Buddleia cultivars, B. davidii ‘Potters Purple’ and Buddleia ‘Lochinch’, were investigated. In vitro shoots were regenerated from leaf tissues in either Murashige and Skoog (MS) or woody plant medium (WPM) media supplemented with benzyladenine (BA). In general, more shoots were regenerated in WPM medium than in MS medium. Dark treatment for 3 to 5 weeks dramatically increased shoot regeneration. Addition of indole-3-butyric acid (IBA) or naphthalene acetic acid (NAA) significantly enhanced the regeneration rate and shoots of each explant. The maximum regeneration rate (100%) of B. davidii ‘Potters Purple’ was achieved when cultured in WPM containing 5 μm BA plus 5 μm IBA. The maximum regeneration rate (98.4%) of Buddleia ‘Lochinch’ was found in WPM supplemented with 20 μm BA plus 4 μm IBA. Carbenicillin at 250 to 500 mg·L−1 and cefotaxime at 125 to 250 mg·L−1, individually or combined, promoted shoot regeneration. Interactions between genotype and medium or PGRs were found. In vitro shoots were easily rooted in half-strength MS medium with or without NAA. Rooted plants were transferred to potting mix and grown in the greenhouse. This research will facilitate genetic improvement and fast propagation of Buddleia species using biotechnology.