红豆杉离体细胞四倍体的诱导

Tetraploid induction of in vitro Taxus chinensis cell

关键词:  红豆杉细胞  秋水仙素  四倍体  染色体  紫杉醇

Key words:  Taxus chinensis cell  colchicine  tetraplont  chromatosome  taxol

摘要:

细胞大规模培养被认为是目前生产紫杉醇最有希望的替代途径之一,获得更高产的细胞系,可降低细胞大规模培养生产紫杉醇的成本,加速其产业化进程。药用植物多倍体与二倍体相比具有根,茎,叶和花果的巨型性,抗逆性强,次生代谢产物含量提高等特性。本研究通过同步化培养和秋水仙素诱导处理,成功建立了红豆杉四倍体细胞系并且经过7个周期的继代证明了该细胞系的稳定性。结果显示,用浓度为750 mg/L的秋水仙素处理3 d可以得到稳定传代的四倍体细胞系,其四倍体细胞比例稳定在62.5%。进一步对其次生代谢产物紫杉醇的检测显示该四倍体细胞比对照二倍体细胞具有更高的合成紫杉醇的能力。

Abstract:

Large-scale culture of Taxus cell is considered to be one of the most promising alternative methods to produce taxol. Heavy producing cell line can cut down the cost of taxol production and promote the process of industrialization. Compared with diploid,multiploid medicinal plant has the characteristics of giantism,which express in roots,stems,leaves,flowers and fruits,strong antireversion force and high contents of secondary metabolites. This research established tetraploid T.chinensis cell line by the methods of synchronization culture and inducing treatment of colchicine. Through 7 cycles subcultures,stability was demonstrated. Results showed that treating the T.chinensis cells with 750 mg/L colchicine could get stable tetraploid cell line whose tetraploid cell rate maintained in 62.5%. Further to detect taxol,its secondary metabolite,tetraploid cell line had stronger ability to produce taxol compared with diploid cell line.