Slash pine (Pinus elliottii Engelm.) somatic embryogenesis I. Initiation of embryogenic cultures from immature zygotic embryos

Immature female gametophytes and enclosed zygotic embryos of slash pine (Pinus elliottii Engelm.) collected at various developmental stages were cultured either on eight initiation media supplemented with different plant growth regulator (PGR) combinations or on LPM medium solidified with two brands of agar. Embryonal-suspensor cell masses extruded from gametophyte explants infrequently produced white or translucent mucilaginous embryogenic cultures. Variation in extrusion frequency was observed across seed sources (families), explant collection dates, agar types and culture media. Increased explant maturity (as related to collection dates) reduced cell extrusion rates. Four types of extrusions were grouped according to their morphology. One type of extrusion produced the greatest number of established and long-surviving embryogenic cultures. This extrusion type (type I) was associated with an extrusion condition termed empty in which the entirety of the embryonal-suspensor mass was extruded out of the gametophyte. Three other extrusion types (types II, III and IV) showed less culture establishment and survival.