An improved micropropagation protocol for stone pine (Pinus pinea L.)

A protocol for micropropagation from isolated cotyledons from Pinus pinea L. has been developed. Major improvements are the increase in rooting rates and the description for the first time of a successful continuous multiplication procedure. For bud induction, isolated cotyledons were cultured on 1/2 LP with 44.4 mu M BA during 4 days. Shoot development was obtained by transfer to 1/2 LP hormone-free medium with activated charcoal. Since an additional 20 weeks were required for shoot elongation and rooting, at least 70 plantlets can be expected per seed after 29 weeks based on a 70% success rate. Besides, shoots can be micropropagated successively by subculturing on 1/2 LPC.