【摘要】 从继代后3～5天的欧当归幼叶胚性细胞悬浮系游离的原生质体,分别以KM8P、MS、改良MS、NT、D2、DPD及B5为基本培养基进行浅层液体培养,以改良MS进行平板培养和固液双层培养。在激素组合和浓度(2.4-D0.2mg/l,KT0.5mg/l,NAA1.0mg/l)及渗透稳定剂(0.5M甘露醇)相同的条件下,在KM8P中的植板率(40％)显著高于其它培养基中的植板率(0—15％)。在KM8P中(0.35M葡萄糖作渗透稳定剂),不同激素组合及浓度植板率不相同,上述激素组合中最高(50％)。葡萄糖作渗透稳定剂时0.3M优于0.4M,同时优于相同浓度的甘露醇和蔗糖。改良MS浅层培养和双层培养植板率几乎相同(15％),而平板培养银低(1％)。在KM8P中(激素同上,0.35M葡萄糖,CH250mg/l,CM 10ml/l),25天后原生质体形成肉眼可见的小愈伤组织。及时转移到固体愈伤组织增殖培养基上,诱导出胚性和非胚性两种愈伤组织,分别转移到分化培养基上后,两者分別通过体细胞胚胎发生和器官发生途径获得了完整再生植株。更多还原

【Abstract】 Protoplasts were prepared from embryogenic suspension cultures of Levisticum officiale Koch, after subculture of 3-5 days. The protoplasts were cultured in KM8P、modified MS、NT、D2、DPD and B5 liquid media, and also cultured in modified MS solid and liquid-solid double layer media. On condition that the composition and concentrations of phytohormones（CCP） and osmoticums were same(2, 4-D 0.2mgL^-1, KT0.5mgL^-1、 NAA 1.0mgL^-1, 0.5M mannitol), the plating efficiency（PE）in KM8P was much higher （40%） than those （0—15%） in other media; in KM8P liquid media（0.35M glucose）, PEs weren’t equal for different CCps, and the highest PE was achieved for the CCP as describled above（50%）. 0.35M glucose was, superior to 0.4M as osmoticum and it was superior to mannitol and sucrose with same concentrations. PEs were almost same in modified MS liquid and double layer cultures, but it was very low in plate culture. In KM8P (CCP as describled above, 0.35M glucose, CH 250mgL^-1, CM 10mlL^-1, pH5.8), minicalli form protoplasts were seen with naked eyes in 25 days. After being transferred to semisolid media timely. the minicalli produced embryogenic and nonembryogenic calli. After being transfered to differentiating media, they regenerated into intact plantlets by embryogenic and organogenic ways respectively, by the ways of somatic embryogenesis and organogenesis.更多还原