胡萝卜悬浮细胞原生质体的培养研究

STUDY ON THE CULTURE OF PROTOPLASTS FROM CARROT SUSPENSION CELL

继代培养 4～ 5d后的胡萝卜悬浮细胞 ,经酶解获得大量原生质体 ,并在不同的培养基上培养 ,探讨了影响原生质体培养的多种因素。实验表明 :①原生质体的培养密度在 5× 1 0 4 mL～ 5× 1 0 5 mL范围内均可 ,以 2 5× 1 0 5 mL为佳 ;②对于细胞的分裂、细胞团的生长 ,以改良的B5培养最好 ,N6培养其次 ,MS较差 ;③适当降低原生质体培养基中的甘露醇浓度 ,有利于细胞的分裂和生长 ;④在本实验所涉及的培养方法之中 ,以液体浅层培养为优。

Large number of protoplasts were isolated enzymatically from 4 to 5 days old carrot suspension cells and cultured in different media.Cultured protoplasts could divide.Various factors affecting the protoplast culture were disocussed.The results showed:①culture densities ranging from 5×10 4/mL to 5×10 5/mL were suitable for carrot protoplasts,with 2 5×10 5mL being the best;②for carrot protoplast division and cell cluster growth,modified B 5 medium was the best,followed by N 6 medium,MS medium was not suitable;③properly lowering the concentration of mannitol in the protoplast culture medium was beneficial to the division and growth of cells;④among the culture methods involved in the experiment,thin layer liquid culture was the most suitable one.