苎麻原生质体培养及植株再生

PROTOPLAST CULTURE AND PLANT REGENERATION OF RAMIE

用苎麻（Ｂｏｅｈｍ ｅｒｉａ ｎｉｖｅａ）子叶诱导愈伤组织并建立悬浮细胞系． 用４．５％ 纤维素酶、０．８％ 离析酶、０．８％ 半纤维素酶的混合酶液分离悬浮培养细胞，可得到２×１０６ 个／ｇ ｆｒ．ｗｔ的原生质体．这些原生质体以海藻酸钠包埋方式培养在附加２，４－Ｄ ０．５ ｍ ｇ／Ｌ、ＫＴ ０．５ ｍ ｇ／Ｌ 的ＫＭ８ｐ 培养基中，５０ ｄ 左右可形成肉眼可见的小愈伤组织．愈伤组织经过扩增，在不同的分化培养基上可诱导芽、根的形成，再生出完整植株．

Calli were induced and suspension cell lines were established from cotyledones of ramie (Boehmeria nivea). Protoplasts (2×10 6/g fr.wt) were isolated from suspension cell cultures in enzyme mixture solution containing 4.5% cellulase Onozuka R 10 and 0.8% Macerozyme R 10 , 0.8% hemicellulase. When cultivated on KM8p medium containing 2,4 D 0.5 mg/L , KT 0.5 mg/L with alginate embedding method, they grew vigorously and produced microcalli within fifty days. After subcultured, the protoplast derived calli produced shoots and roots on different differentiation media, then complete plants were formed. Protoplasts from cotyledones divided only several times.