Somatic embryogenesis and plantlet regeneration from cultured shoots and cotyledons of seedlings from stored seeds of black and white sprees (<i>Picea Mariana</i> and <i>Picea glauca</i>)

Abstract

Embryogenic callus was induced from cultured explants of 12- to 30-day seedlings germinated from 10- and 4-year stored seeds of black and white spruce, respectively. Plantlets were regenerated from a 12-day black spruce seedling derived embryogenic callus. This is the oldest conifer seedling material shown to give rise to embryogenic callus, and the first demonstration in spruce that seedling-derived embryogenic calli are competent of plantlet regeneration. The effect of a cytokinin pretreatment on embryogenic callus induction from 12-day black spruce seedling shoot explants was investigated. Although an induction frequency of 18% occurred after 2 weeks preculture of shoot explants on medium containing 4.5 μM benzyladenine, the cytokinin pretreatment was not significantly superior to controls given no pretreatment (12%). Age of explant influenced embryogenic induction frequencies from black and white spruce. Explants from 21-day black spruce seedlings produced embryogenic calli at a frequency of 5%. In comparison, shoot explants from 12- and 30-day white spruce seedlings given no benzyladenine pretreatment displayed induction frequencies of 38 and 18%, respectively.