皂质芦荟细胞悬浮培养初探

Preliminary Studies on the Cell Suspension Culture of the Aloe saponaria Haw

试验初步研究了皂质芦荟细胞悬浮培养过程。比较了不同基本培养基、接种量对悬浮培养过程中细胞生长量的影响。采用正交设计方法,对影响皂质芦荟悬浮细胞生长的因素进行了优化研究。初步探讨了皂质芦荟在细胞生长周期里生理生化特性。结果表明:B5培养基有利于细胞的增殖;每50 mL培养液中加入2 g愈伤组织的接种量为宜;筛选出最适激素组合为2,4-D 3.0 mg/L+KT 0.5 mg/L,葡萄糖25 g/L pH值5.8;皂质芦荟悬浮培养的生长周期为25 d,细胞增值率曲线呈S型;在悬浮细胞生长前期,pH值明显下降,在细胞对数生长期,pH值略有升高,并趋于平缓;总酚含量、POD、SOD均出现2个峰值,但出现时间不同,其变化趋势与细胞生长趋势呈正相关,PPO活性与细胞生长趋势呈负相关。

The study focused on the suspension culture of the Aloe saponaria Haw. Compared the influence of different basic medium and inoculums amount on cell growth increment during the process of suspension culture, Optimized the effecting faclors of the Aloe saponaria Haw cell in suspension culture by orthogonal design. Discussed the characters of physiological and biochemical indices in the cell growth cycle. The results indicated, B5 medium was in favor of the cell muhiplication; lhe most suitable inoculums amount was 2 g callus per 50 mL liquid medium ; the most suitable hormone combinations was 2,4-D 3.0 mg/L＋KT 0. 5 mg/L, glucose 25 g/L pH 5. 8;the suspension culture growth cycle of the Aloe saponaria Haw was 25 days, the.cell multiplication graph was in shape of ＇S＇; in the previous of suspension cuhure cell, the pH decreased obviously, in the logarithmic growth phase of cell, the pH slightly increased and became stable; total phenolic content ,POD,SOD showed two peaks respectively , but the peaks appeared at different time , and the change trend was in direct proportion to the cell growth trend, PPO activity was in converse proportion to the cell growth trend.