Institute of Genetics and Cytology, Northeast Normal University, 130024, Changchu, People's Republic of China.
Plant Cell Rep. 1996 Nov;16(1-2):92-6. doi: 10.1007/BF01275458.
A study was undertaken to develop a protoplast regeneration system for pinellia. A yield of 19 29 x 10(5) protoplasts/g F. W. could be obtained from cell suspension cultures incubated in a digestion enzyme solution with 2% cellulase Onzuka R-10, 10% pectinase (Sigma), 0.01% pectolyase Y23. K8P and modified MS media were used to culture protoplasts in: a) liquid, b) liquid-solid double layer, or c) agarose embedded protoplast culture. The former two were conducive to colony formation from protoplast-derived cells. The frequency of cell division was about 8% after 3 days in culture. Gradually adding fresh medium of lower osmotic pressure into the medium for protoplast culture favored cell division. Calli (1-2 mm in diameter) formed after 30-40 days in culture. The calli transferred onto medium supplemented with KT (0.5 mg 1(-1)) and NAA (0.2 mg 1)(-1)) could regenerate plants after 40-50 days. Of 47 plantlets transplanted into plots, 29 flowered and were fertile.
一项研究旨在为半夏开发原生质体再生系统。通过在含有 2%纤维素酶 Onzuka R-10、10%果胶酶（Sigma）、0.01%果胶酶 Y23 的消化酶溶液中孵育细胞悬浮培养物，可以获得 19 29 x 10(5)个原生质体/gFW。使用改良 MS 培养基在以下条件下培养原生质体：a）液体，b）液体-固体双层，或 c）琼脂糖包埋原生质体培养。前两种方法有利于原生质体衍生细胞的集落形成。培养 3 天后，细胞分裂频率约为 8%。逐渐向原生质体培养的培养基中添加低渗透压的新鲜培养基有利于细胞分裂。培养 30-40 天后形成 1-2 毫米直径的愈伤组织。将愈伤组织转移到补充有 KT（0.5 mg 1(-1))和 NAA（0.2 mg 1(-1))的培养基上，经过 40-50 天可以再生植物。在移植到田间的 47 株植物中，有 29 株开花并具有生育能力。
