Slash pine (Pinus elliottii Engelm.) somatic embryogenesis II. Maturation of somatic embryos and plant regeneration

Maturation of slash pine (Pinus elliottii Engelm.) somatic embryos was achieved using two protocols, each starting with a different agar incubation step to deplete plant growth regulators (PGRs) used in previous cultural steps. Strength of maturation medium (single vs. double) was found important in the first protocol to develop normal, mature embryos. In the second protocol, abscisic acid (ABA) concentrations (0, 15 and 30 M) and carbohydrate sources were tested for embryo maturation. Thirty M ABA and 6% maltose were deemed the best combination. Embryo germination was accomplished in a continuously lighted environment and embryos receiving a cold pretreatment (4 °C in darkness for 16 days) germinated better than embryos which did not receive cold pretreatment. With a survival rate of 33% after acclimation in a mist system, more than 25 plants from somatic embryos have been established in a greenhouse. Incompletely germinated embryos (lacking roots) were rooted via adventitious rooting techniques and subsequently established in the greenhouse. All established plants obtained from somatic embryogenesis appear normal in morphology.