An improved micropropagation protocol for maritime pine (Pinus pinaster Ait.) isolated cotyledons

In this work an improved micropropagation protocol for Pinus pinaster Ait. using isolated cotyledons is presented. The variables studied during bud induction were embryo preculture, type and concentration of cytokinin and influence of illumination culture conditions. Shoot development and proliferation was obtained by transfer to hormone-free medium with activated charcoal. This protocol also describes a procedure for successful continuous multiplication using axillary shoots and brachyblasts formed during shoot development. Based on 86 per cent rooting success rate, at least 70 plantlets can be expected per seed after 16 weeks.