Shoots were regenerated from in vitro leaf tissues of two genotypes of Viburnum dentatum, a popular shrub species for landscape use. Adventitious shoots were induced when leaf tissues were cultured on woody plant medium (WPM) supplemented with either benzyladenine (BA) or thidiazuron (TDZ). Effects of cytokinin concentration, indole-3-butyric acid (IBA), and dark treatment on shoot regeneration were investigated. Dark treatment for the first 4 weeks of leaf explants cultured in the regeneration medium significantly increased the frequency of regeneration. The highest frequency of shoot regeneration (70%) for 'Synnesvedt' was obtained when leaf tissues were cultured in the medium with 40 lM BA or 8 lM TDZ with 4 weeks dark treatment. The highest frequency of shoot regeneration (90%) for 'MN34' was found in the 4 lM TDZ medium with 4 weeks dark treatment. Addition of IBA significantly enhanced shoot regeneration. Ethyl methanesulfonate (EMS) treatment inhibited callus proliferation, particularly in the early stage of callus recovery; however, no significant difference in shoot regeneration among different treatments was observed, indicating that the inhibitory effect of EMS was minimal after calluses re-acquired their capacity to grow and regenerate in the regular medium. Regenerated shoots ([1.5 cm) were rooted in the half-strength MS medium containing 5-10 lM IBA or naphthalene acetic acid (NAA). Rooted plants were transferred to the potting medium and grown in the greenhouse.

Two Buddleia cultivars, B. davidii ‘Potters Purple’ and Buddleia ‘Lochinch’, were transformed using Agrobacterium tumefaciens strain EHA105 harboring the binary vector pBI121 carrying the neomycin phosphotransferase gene and β-glucuronidase gene (uidA). Transgenic plants were recovered from the Agrobacterium-infected leaf tissues through organogenesis in the selection medium (woody plant medium containing 250 mg·L−1 cefotaxime plus 500 mg·L−1 carbenicillin plus 40 mg·L−1 kanamycin). The rate of shoot regeneration from transformed leaf tissues increased from 5.7% to 32% through extending cocultivation time from 3 to 9 days. Integration of marker genes was verified with polymerase chain reaction (PCR) and Southern blot analysis. Southern blot analysis confirmed that one to three copies of transgenes were integrated into the buddleia genome. This transformation system could be used for improvement of buddleia or other related species. Chemical names used: 6-benzyladenine (BA), naphthalene acetic acid (NAA), acetosyringone (AS), 5-Bromo-4-chloro-3-indoxyl-beta-D-glucuronide cyclohexylammonium (X-Glu), cefotaxime (Cef), carbenicillin (Carb), kanamycin (Km).