Tissue Culture of Propagation of Mature Trees of Prunus serotina Ehrh. I. Establishment, Multiplication, and Rooting in Vitro

Bud tissue explants were obtained from winter buds of grafted Prunus serotina Ehrh. trees growing in a seed orchard in Pennsylvania and from three 50-year-old P. serotina trees growing in central New York. Twigs were collected and explants prepared in late winter just as the buds were beginning to swell. Explants were established and elongated on Murashige and Skoog inorganic salt medium (MS), supplemented with 100 mg/l I-inositol, 0.4 mg/l thiamine-HCl, 1.0 mg/1 N-6-benzylaminopurine (BAP), 0.1 mg/l gibberellic acid (GA3), 0.1 mg/l indole-3-butyric acid (IBA), 2 percent sucrose, and 0.7 percent phytagar. Multiple shoots were induced on the same medium but with 0.75 mg/l BAP, 0.2 mg/l GA3, 0.01 mg/l IBA, and 3 percent sucrose. A five-fold shoot multiplication rate was obtained every 3 to 4 weeks. Root formation was induced in vitro after shoots had grown for 14 days on MS medium supplemented with 100 mg/l I-inositol, 0.4 mg/l thiamine-HCl, 1.0 mg/l IBA, 3 percent sucrose, 0.7 percent phytagar on a 16-hour photoperiod. Rooting could be enhanced by incubating cultures in continuous darkness. The addition of rutin or quercetin to the rooting medium increased rooting in the light. Plantlets were successfully transferred to soil, following establishment, multiplication, and rooting in vitro . Forest Sci. 31:201-208.