In vitro regeneration of plantlets from mature embryos of Pinus ayacahuite

• Published in: Tree physiology Vol. 17; no. 12; pp. 787 - 796
• Main Authors: Saborio, F, Dvorak, W.S, Donahue, J.K, Thorpe, T.A
• Format: Journal Article
• Language: English
• Published: Canada 01.12.1997
• Subjects: buds; culture media; dose response; embryo (plant); explants; genetic variation; genotype; growth; in vitro culture; methodology; micropropagation; organogenesis; Pinus ayacahuite; plant growth substances; provenance; regenerative ability; roots; shoots
• ISSN: 0829-318X; 1758-4469
• DOI: 10.1093/treephys/17.12.787

A plantlet regeneration protocol was developed for Pinus ayacahuite var. ayacahuite (Ehrenb.). Embryos from mature seeds from ten provenances were cultured in a 16-h photoperiod for 3 days on a medium containing 30 mM sucrose and 0.7% agar. Cotyledons from these embryos were subcultured onto MCM medium (Bornman 1983) supplemented with 50 micromolar N(6)-benzyladenine and 90 mM sucrose for 2 weeks. Bud development and shoot elongation were maximized by subculturing the explants on half strength AE medium (von Arnold and Ericksson 1981), supplemented with 60 mM sucrose and 0.05% activated charcoal every 30 days. Seed source had a significant effect on the responses of the embryos to the bud induction protocol. For the provenance with the best response to bud induction, about 79% of the cultured cotyledons formed buds, and each cotyledon formed a mean of 9.1 buds, so that about 70 shoots could be induced from each seed. The best rooting response (40% rooting) was obtained by treating the shoots for 8 h with 100 micromolar naphthalene acetic acid.