马蹄金悬浮细胞系的建立与优化

Establishment and Optimization of Suspension Cell System in Creeping Dichondra (Dichondra repens Forst.)

以马蹄金下胚轴愈伤组织为外植体,采用均匀设计探讨了不同生长调节剂和培养基对细胞悬浮培养的影响,建立了马蹄金悬浮细胞系.结果表明,不同生长调节剂组合的培养基对马蹄金悬浮细胞的干重和生长速度有显著影响,以B5培养基作悬浮培养的基本培养基效果较好,用B5+0.5 mg/L 2,4-D+0.1 mg/L6-BA+0.5 mg/L KT+2.0 mg/L α-NAA作为悬浮培养基,对悬浮细胞的干重增长和生长速度效果较好,经固化培养后,愈伤组织生长良好.悬浮细胞生长的时间进程是,起始培养的第6天前,细胞增殖十分缓慢;第6～12天期间生长迅速;第14天后基本停止生长.培养基的pH值迅速下降时是细胞增殖缓慢的时期;当培养基的pH值开始升高时,细胞增殖也显著加快.

The effects of different growth regulators and media on the establishment of cell suspension culture system were studied with the uniform design and the callus induction from hypocotyls of creeping dichondra （Dichondra repens Forst. ） were used as explants. The results indicated that the kinds of growth regulator combinations had significant effect on weight and growth speed of the suspension cell of creeping dichondra. It showed that the optimal medium which could induce suspension cell culture was B5 medium ahd good growth of suspension cell culture were acquired on B5 medium contained 0. 5 mg/L 2,4-D, 0. 1 mg/L 6-BA, 0. 5 mg/L KT and 2.0 mg/L α-NAA. The callus could growth good after transferring solid medium. The schedule of suspension cell growth showed that cell grow lagged in 0-6 days and enhanced greatly in 6-12 days and almost ceased after 14 days of cultivation. The pH value was changing during the culture period. As the suspension cell grew slowly, the pH value went down, and vice versus.