Micropropagation and in vitro studies of Pinus patula Scheide et Deppe.
For the South African forestry industry, the patula pine (Pinus patula) is the most commercially important softwood species. A pine clonal programme has yet to be fully implemented in this country and at present much effort is being made to establish clonal plantings of selected trees. In order to accomplish this, it is essential that satisfactory commercially viable propagation technologies be developed for this species. This study examined the possibilities and constraints of three different in vitro systems for mass propagation of rare and important P. patula material. Seed germination and sterilisation techniques were developed for adventitious bud and somatic embryogenesis experimentation. Adventitious buds were initiated from excised 'mature P. patula embryos cultured on LM medium containing 5 mg 1-1 BA. Although, between 50 and 60% of the embryo explants produced adventitious buds, only 3-5 buds per explant actually developed further to form distinct shoots. The adventitious shoots elongated slowly (±8 mm in 2 months) on LM medium, containing 10 g 1-1 activated charcoal. Axillary buds were induced on 10 week-old juvenile shoots, after the development of an effective surface sterilisation procedure, using 0.02% HgCL2. The effect of removing the apex and trimming the needles on bud induction was significant. Dwarf shoots elongated at a rate of 25 mm in 5 weeks. Rooting studies conducted on juvenile P. patula shoots indicated that the most effective treatment was wounding the shoot base and placing the shoot in composted bark growing medium, under a greenhouse mist regime. Rooting percentages were low (50%). Included in this study is the first successful production of somatic pro-embryos from mature Pinus patula embryos. Calli were produced on LM induction medium containing 2 mg 1-1 2,4-D. Cultures were first placed in the dark for 4 weeks and then transferred to a 16 h photoperiod for a further 2 weeks, after which Stage 1 embryogenic cells were observed. When calli were placed on LM maturation medium, containing 12 mg 1-1 ABA, for a further 6-8 weeks, pro-embryo structures (maximum of 7 pro-embryos per callus) were detected embedded In the callus mass. Hence, investigations into the development of protocols for the micropropagation of Pinus patula, were undertaken. Two major constraints for applying in vitro techniques to the commercial production of pine were identified: the poor yield of shoots and pro-embryos and the length of time taken for plantlets to be produced. This study, however, provides some fundamental knowledge and background work required by tree breeders who wish to implement biotechnological techniques in the selection and improvement of P. patula genotypes.