In vitro regeneration of plantlets from embryonic and seedling explants of Engelmann spruce (Picea engelmannii Parry)

Abstract

A protocol has been developed for the in vitro production of plantlets of Engelmann spruce. Embryos and various parts of Engelmann spruce seedlings formed multiple shoots when cultured on defined media containing a cytokinin. The site and time of occurrence of the shoot buds, as well as their number, differed in the various explants. The frequency of shoot-forming explants was influenced by the salt formulation used, the type and concentrations of cytokinins and their mode of application. Development of buds was achieved by transferring the explants to basal medium containing no growth regulators. Elongation of shoots was stimulated by reducing the concentration of salts and sugar, addition of activated charcoal and transferral to increased photoperiod and lower temperature regimes. Maximum rooting was induced by giving a pulse of high concentration of indolebutyric acid to the shoots. The roots developed within 8-10 weeks and the regenerated plantlets were transferred to soil under non-sterile conditions.