药用植物半夏细胞悬浮培养和植株再生体系建立研究

System of Calli Cell Suspension Cultural and Its Plant Regenerstion and Its Influencing Factors from Medicinal Pinellia ternata Briet.

为了建立甘肃产药用植物半夏的细胞悬浮培养和植株再生体系,通过对其不同外质体筛选、愈伤组织诱导和悬浮培养的愈伤组织类型、激素水平、起始密度、震荡速率等影响因素以及细胞生长状态、生长曲线和pH值变化等指标的研究。结果表明:以叶柄为外质体,选择"松散型"愈伤组织和2×105个/mL的起始培养密度,在附加2,4-D 2.0 mg/L+BA 0.5 mg/L+NAA0.5 mg/L的1/2 MS培养基上培养,温度(23±2)℃,转速90 rpm,pH值6.0,黑暗培养为最适悬浮培养体系;经检测细胞生长曲线呈"S"型,培养液pH随着细胞的生长先上升后下降,18 d时细胞基数达到最大值;悬浮继代培养后,通过降低或停止震荡转速,在添加BA 0.5 mg/L+NAA0.5 mg/L的MS液体和固体培养基上可诱导分化出芽和根,进一步形成完整的植株。

The systems of cell suspension cultures and efficient plant regeneration of Medicinal Pinellia ternata Briet. were studied as the objective in this paper. Some effecting factors and index were studied, such as choosing of explants, inducing of calli, type of call i, primary cultural density, shake rate and cell character, growth status, growth curve, pH value et al. The results showed that stem of leaf as explants and ＂loose＂calli were choosed in the experiment. The most suitable condition of cell suspension cultures was found to be 1/2 MS medium supplemented with 2,4-D 2. 0 mg/L ＋BA 0.5mg/L＋NAA 0. 5mg/L, primary cultural density： 2× 10^5个/ml, temperature： 23±2℃, shake rate： 90rpm, pH value：6. 0 and dark cultures. Suspension cell growth curves were like ＂S＂. The pH value cultured mediums were increased at first, and then reduced gradually. Cell number approached its maximum value in 18d. After subculture, calli was induced to regenerated shoots and roots by reducing and stopping shake rate on medium MS supplemented with BA 0. 5mg/L and NAA 0. 5mg/L. Further, Complete plants were obtained gradually.