海岛棉子叶原生质体游离和再生愈伤的获得

Isolation and Calli Regeneration in Cotyledon Protoplasts of Cotton(Gossypium barbadense L.)

对１０个海岛棉品种的子叶进行了原生质体的游离和培养。培养３～５天的无菌苗子叶经１２～１６小时的酶解，获得了一定数量的原生质体。不同的酶液浓度组成对原生质体的产量有较大的影响。纯化的原生质体在修改的Ｋ３培养基上进行培养，３～５天后再生细胞开始分裂，２个星期后形成小细胞团，ｌ个月后获得了小愈伤组织。

Protoplasts were isolated from cotyledons of 3 to 5 days’old sterile seedings ofGossy pium barbadense. By Enzyming with in 12 to 16 hours,separated and purificated,theprotoplasts were then cultured in modified K3 medium which supplemented with 0.1～0.5mg/L 2,4一D and 0.5～1.0mg/L 2iP, The mixed enzyme solution was composed with l%～2%cellulase(R一10),0.5%～1.O% hemicellulase (Sigina)and 0.5%～1.0%malerozyme(R一10), pH5.8. Protoplasts regencrated cell walls in 2～3days in culture.The first cell divi-sion was apparent after 3～5days and the second cell division was apparent after 5～8days。Cell clusters of l0～100 cells could be seen in two weeks.Microcalli were then obtained afterarnonth.Most microcalli browned and died in late culture.Protoplast yield and viability,celldivision and colonv,Callus formation and propagation were influenced by genotype,age,growth condition of the sterile seeding,enzyme mixture and concentration, and culture medi-um.