抗癌植物曼地亚红豆杉离体诱导培养及褐化抑制

Callus induction, subculture, and browning inhibition of the anticancer plant Taxus media

摘要:

为了获得抗癌植物红豆杉更为有效的细胞培养参数，本研究以曼地亚红豆杉带芽茎段为外植体，利用植物组织培养技术和正交试验对其愈伤组织诱导和继代培养条件进行优化，并对培养过程中出现的褐化问题进行探讨。结果表明在黑暗条件下，B₅+0.25 mg/L 2,4-二氯苯氧乙酸(2,4-dichlorophenoxyacetic acid,2,4-D)+1.5 mg/L萘乙酸(naphthaleneacetic acid,NAA)+0.5 mg/L激动素(kinetin,KT)是愈伤组织诱导最适培养基，表现为诱导时间短、诱导率高(86.7%)；最适继代培养基为B₅+0.5 mg/L 2,4-D+2.0 mg/L NAA+1.5 mg/L KT；在愈伤组织生长第10天进行继代培养的愈伤组织增殖倍数最高；活性炭能够最大限度减轻褐变的发生，其最佳浓度为0.8 g/L。本研究将为曼地亚红豆杉细胞悬浮培养及通过细胞培养生产紫杉醇奠定基础。

Abstract:

In order to obtain more effective cell culture parameters of Taxus anticancer plants, we optimized the callus induction and subculture conditions of the explants (stem segments with buds) of the anticancer medicinal plant Taxus media by using the plant tissue culture technology and orthogonal test. Furthermore, we studied the method to inhibit browning in the culture. The results indicated that the optimal conditions for inducing callus was culture in the medium composed of B₅+0.25 mg/L 2,4-D+1.5 mg/L NAA+0.5 mg/L KT in the dark, which showed short induction time and a high induction rate (86.7%). The formula of the optimal medium for subculture was B₅+0.5 mg/L 2,4-D+2.0 mg/L NAA+1.5 mg/L KT.The proliferation multiple of callus cultured by subculture on the 10th day of callus growth was the highest. Activated carbon inhibited the browning in callus subculture, with the optimal inhibitory concentration of 0.8 g/L. The results of this study lay a foundation for the production of taxol by suspension culture of T. media cells.