Luo J P, Jia J F, Gu Y H
School of Life Sciences, University of Science and Technology of China, Hefei 230026.
Shi Yan Sheng Wu Xue Bao. 1999 Dec;32(4):401-8.
Protoplasts from 4-day-old embryogenic cell suspension cultures of Astragalus adsurgens, when cultured in KM8P medium which ammonium concentration was reduced to 2.5 mmol/L and supplemented with 0.5 mg/L NAA, 1.0 mg/L 2, 4-D, 0.7 mg/L BA and 0.4 mol/L glucose, underwent cell sustained divisions and formed cell colonies at a frequency of 16%-20%. Preplasmolysis or low temperature treatment of suspension cells prior to enzyme incubation enhanced colony formation. Following proliferation on MS medium containing 1.0 mg/L 2, 4-D and 0.5 mg/L BA, cell colonies were cultured on MS medium containing 0.1 mg/L NAA and 1.0 mg/L BA, where approximately 40% of colonies produced somatic embryos ranging in number from 20 to 40 per colony. No significant decrease was found in the potential of somatic embryogenesis when protoplast colonies were obtained from long-term cell suspensions. On hormone-free 1/2 MS medium, somatic embryos developed into intact plants, which showed normal morphology and stable chromosome number.
斜茎黄芪4日龄胚性细胞悬浮培养物的原生质体，在铵浓度降至2.5 mmol/L并添加0.5 mg/L萘乙酸、1.0 mg/L 2,4-二氯苯氧乙酸、0.7 mg/L苄氨基腺嘌呤和0.4 mol/L葡萄糖的KM8P培养基中培养时，进行细胞持续分裂并以16%-20%的频率形成细胞集落。在酶解前对悬浮细胞进行质壁分离前处理或低温处理可提高集落形成率。在含有1.0 mg/L 2,4-二氯苯氧乙酸和0.5 mg/L苄氨基腺嘌呤的MS培养基上增殖后，将细胞集落在含有0.1 mg/L萘乙酸和1.0 mg/L苄氨基腺嘌呤的MS培养基上培养，约40%的集落产生体细胞胚，每个集落的体细胞胚数量为20至40个。当从长期细胞悬浮液中获得原生质体集落时，体细胞胚胎发生潜力没有显著下降。在无激素的1/2 MS培养基上，体细胞胚发育成完整植株，这些植株形态正常，染色体数目稳定。
