Abstract
Pinus patula Schiede et Deppe is one of the most important softwood species for pulp production by the South African forestry industry. This study was aimed at developing a protocol for the genetic transformation of Pinus patula embryogenic tissue. This was achieved via the introduction of a bar-GUS cassette under the control of the ubiquitin promoter, through biolistic transfer. A stepwise selection was obtained using MSG3 maintenance medium supplemented with BASTA® herbicide at 1 mg l−1 followed by 3 mg l−1 active ingredient. Transgene delivery was more efficient when the medium was supplemented with 0.25 M sorbitol. Expression of positive histochemical GUS activity in embryonal heads was observed. Positive PCR analysis of both GUS and bar transgenes (40% and 47% transformation efficiency, respectively) indicated successful genetic modification of P. patula embryonal suspensor masses by the pAHC25 plasmid. This indicates that P. patula is amenable to gene transfer.
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