Micropropagation of mature Calabrian pine (Pinus brutia Ten.) from fascicular buds

Dormant meristems of fascicles explanted from 10-year-old, field-grown trees of Pinus brutia Ten. were cultured in vitro. Browning of cultured fascicles was reduced by including 150 mg l(-1) sodium diethyldithiocarbamate (SDD) in a cytokinin-containing medium. The stage of development of fascicles when placed in culture affected both shoot-bud production and the degree of browning. Only fascicles at an advanced stage of development had a high rate of shoot-bud production. Fascicles cultured for 6 weeks in initiation medium containing 150 mg l(-1) SDD and then for 4 weeks in initiation medium containing 1 mg l(-1) insoluble polyvinylpolypyrrolidone showed the highest rate (23%) of shoot-bud development and the lowest rate (15%) of browning. Elongation of activated shoot buds was considerably enhanced by reducing the concentration of cytokinin in the culture medium. Only shoots >/= 20 mm in height were capable of producing further crops of buds and shoots. When treated with a combination of auxin and cytokinin, only 16% of the elongated shoots produced roots.