Alginate Encapsulation Technique and Cryogenic Procedures for Long-Term Storage of the Tropical Forest Tree Guazuma crinita Mart. In Vitro Cultures
Long-term storage of the tropical forest tree Guazuma crinita Mart. Following alginate encapsulation technique and cryogenic procedures was attempted. Shoot tips excised from in vitro plantlets were encapsulated in calcium alginate beads and stored on different substrates at 12, 20, and 25℃. Viability was retained when encapsulated shoot tips were stored on substrate containing only water solidified with 1% (w/v) agar. Percentage of viability 12 months after storage was 90% at 25℃ and 70% at 20℃. For cryogenic experiments, shoot tip, nodal segment, root tip. And adventitious bud cluster explants were stored in liquid nitrogen following 4 cryopreservation methods: (1) simple freezing, (2) rapid freezing, (3) slow pre-freezing. and (4) encapsulation/dehydration method. High survival rates (about 80%) were achieved when the adventitious bud cluster explants were exposed to a cryoprotectant mix solution containing (w/V), 25% glycerol, 15% sucrose, 15% ethylene glycol, 13% dimethyl sulfoxide, and 2% polyethylene glycol(4,000), at 25℃ for 15-60 min prior to storage in liquid nitrogen.
|作成者||Emilio MARUYAMA Katsuaki ISHII Isao KINOSHITA|