[Objective] The research aimed to establish the high frequency regeneration system of Atropa belladonna L. and screen the kanamycin （Kan） resistance. [Method] The leaves and axillary buds of Atropa belladonna L. wer...[Objective] The research aimed to establish the high frequency regeneration system of Atropa belladonna L. and screen the kanamycin （Kan） resistance. [Method] The leaves and axillary buds of Atropa belladonna L. were as the explants,the influences of different ratios of 6-BA and NAA in the medium on the adventitious bud differentiation and the sensibility of leaf on Kan were studied. [Result] MS＋4.5 mg/L 6-BA＋0.2 mg/L NAA was the optimum medium of leaf adventitious bud differentiation,and the differentiation ratio of adventitious bud reached 100%. The number of adventitious bud differentiation on 1. 0 cm ×1. 0 cm leaf block averagely reached 5.85. MS＋3.0 mg/L 6-BA＋0.1 mg/L NAA was the optimum medium of axillary bud adventitious bud differentiation,and the differentiation ratio of adventitious bud reached 100%. The average number of adventitious bud differentiation in every axillary bud was during 4-8. The optimum screening concentration for genetic transformation of Atropa belladonna L. leaf was 400.0 mg/L of Kan. [Conclusion] The research laid the foundation for the rapid propagation of Atropa belladonna L. aseptic seedling and the genetic transformation based on the leaf disc cocultivation.展开更多
目的:对颠茄口服溶液原质量标准中的鉴别和含量测定方法进行改进。方法:制备HPLC特征图谱来鉴别颠茄口服溶液;用HPLC双波长法同时测定颠茄口服溶液中东莨菪内酯和硫酸天仙子胺的含量。指纹图谱采用Waters Sun Fire C_(18)(250mm×4....目的:对颠茄口服溶液原质量标准中的鉴别和含量测定方法进行改进。方法:制备HPLC特征图谱来鉴别颠茄口服溶液;用HPLC双波长法同时测定颠茄口服溶液中东莨菪内酯和硫酸天仙子胺的含量。指纹图谱采用Waters Sun Fire C_(18)(250mm×4.6 mm,5μm)色谱柱,流动相为甲醇和0.05%磷酸溶液梯度洗脱,流速为1.0 ml·min^(-1),检测波长344 nm,柱温30℃;东莨菪内酯和硫酸天仙子胺的含量测定采用相同的C_(18)色谱柱,流动相为10 mmol·L^(-1)庚烷磺酸钠溶液(用冰醋酸调pH至3.3)-无水乙醇-乙腈(68.75∶6.25∶25),流速为1.0 ml·min^(-1),检测波长为344 nm和210 nm,柱温30℃。结果:颠茄口服溶液的特征图谱和颠茄酊原料的特征图谱一致,各特征峰和参照峰(东莨菪内酯峰)的保留时间及相对峰面积符合中国药典要求。在含量测定条件下,东莨菪内酯和硫酸天仙子胺与相邻的杂质峰完全分离,两者的浓度分别在5.168~103.360μg·ml^(-1)(r=1.000 0)和50.560~758.400μg·ml^(-1)(r=0.999 9)范围内线性关系良好;平均回收率分别为101.79%(RSD=1.05%,n=6)、100.92%(RSD=0.97%,n=6)。结论:改进后的质量控制方法,鉴别试验专属性强,含量测定选择两个指标性成分,更能全面控制颠茄口服溶液的质量,测定方法操作简便,准确,可为其质量控制提供可靠的方法。展开更多
基金Supported by National High Technology Research Development Plan Item (2010AA100503)~~
文摘[Objective] The research aimed to establish the high frequency regeneration system of Atropa belladonna L. and screen the kanamycin （Kan） resistance. [Method] The leaves and axillary buds of Atropa belladonna L. were as the explants,the influences of different ratios of 6-BA and NAA in the medium on the adventitious bud differentiation and the sensibility of leaf on Kan were studied. [Result] MS＋4.5 mg/L 6-BA＋0.2 mg/L NAA was the optimum medium of leaf adventitious bud differentiation,and the differentiation ratio of adventitious bud reached 100%. The number of adventitious bud differentiation on 1. 0 cm ×1. 0 cm leaf block averagely reached 5.85. MS＋3.0 mg/L 6-BA＋0.1 mg/L NAA was the optimum medium of axillary bud adventitious bud differentiation,and the differentiation ratio of adventitious bud reached 100%. The average number of adventitious bud differentiation in every axillary bud was during 4-8. The optimum screening concentration for genetic transformation of Atropa belladonna L. leaf was 400.0 mg/L of Kan. [Conclusion] The research laid the foundation for the rapid propagation of Atropa belladonna L. aseptic seedling and the genetic transformation based on the leaf disc cocultivation.
