De novo shoot organogenesis of Pinus eldarica Medw. in vitro. I. Reproducible regeneration from long-term callus cultures

Seedling-derived explants of the Afghan pine, Pinus eldarica, were cultured in a triplicate experiment to produce callus that was serially subcultured for up to three years. Callus was removed at various times and induced to regenerate shoots by de novo organogenesis. The shoot regeneration process involved the identification of four discrete developmental steps, each requiring a separate cultural manipulation. In one case a regenerated shoot was induced to root following an auxin pulse treatment. Induction and limited development of buds in callus derived from mature-tree explants was also achieved. This is the first reproducible system for shoot regeneration from long-term callus cultures of a conifer.