曼地亚红豆杉愈伤组织诱导和继代培养研究

以曼地亚红豆杉(Taxusmedia var.hickss)嫩茎和嫩叶为外植体,进行愈伤组织诱导和继代培养研究。愈伤组织诱导以MS为基本培养基,继代培养以MS和B5为基本培养基,分别添加不同浓度和组合的细胞分裂素(6-BA、TDZ、KT、2-ip)、生长素类激素(NAA、2,4-D),和水解乳蛋白、活性碳等抗褐化试剂,考察其对愈伤组织诱导和继代培养的影响。结果表明,曼地亚红豆杉嫩茎有很强的分生能力,比嫩叶更易于诱导愈伤;在培养基MS TDZ0.002mg/L NAA1.0mg/L、MS TDZ0.002mg/L 2,4-D1.0mg/L、MS TDZ0.002mg/L NAA1.0mg/L 2,4-D1.0mg/L和MS KT1.0mg/L 2,4-D1.0mg/L NAA1.0mg/L上,嫩茎愈伤的诱导率达100%,生长迅速,品质良好,利于继代培养;培养基B5 KT1.0mg/L 2,4-D2.0mg/L NAA1.0mg/L适用于愈伤继代,每30d增殖倍数5.2倍;培养基中添加活性碳500mg/L、维生素C 1000mg/L或水解乳蛋白1000mg/L对抑制材料褐化有一定的效果。

Callus Induction and Subculture of Taxus media var.hickss

The tender stems and leaves of Taxus media var.hickss were used as explants.The basic medium MS was used for callus induction,MS and B5 for callus subculture respectively.Different concentrations and combinations of cytokinin(6-BA,TDZ,KT,2-ip),auxin(NAA and 2,4-D) and darkening inhibitors(activated charcoal,vitamin C and lactalbumin hydrolysate) were added to the basic media.The results indicated that the tender stems had strong merisis ability,and more effective than leaf in inducing callus.The rate of callus induction was 100% when the tender stems were cultured in MS TDZ0.002 mg/L NAA1.0 mg/L,MS TDZ0.002 mg/L 2,4-D1.0 mg/L,MS TDZ0.002 mg/L NAA1.0 mg/L 2,4D1.0 mg/L and MS KT1.0 mg/L 2,4-D1.0 mg/L NAA1.0 mg/L.These callus grew rapidly and had a fine quality which was good for next subculture.The medium B5 KT1.0 mg/L 2,4-D2.0 mg/L NAA1.0 mg/L was optimum for callus subculture,and the proliferation coefficient was 5.2 every 30d.Subculture media with activated charcoal 500 mg/L,vitamin C 1000 mg/L or lactalbumin hydrolysate 1000 mg/L could inhibit the darkening of callus to some extent.