白英毛状根的培养与薯蓣皂苷元的测定.PDF

中草药 Chinese Traditional and Herbal Drugs 第 47 卷 第 7 期 2016 年 4 月 • 1199 • 白英毛状根的培养与薯蓣皂苷元的测定 1 1 1 1 2 2 1* 陈 宇 ，董 瑜 ，张楷燕 ，孙一铭 ，程孟琪 ，张 来 ，孙 敏 1. 西南大学生命科学学院 三峡库区生态环境教育部重点实验室，重庆 400715 2. 安顺学院 贵州省教育厅功能材料与资源化学特色重点实验室，贵州 安顺 561000 摘 要：目的 建立白英 Solanum lyratum 毛状根诱导与培养体系，筛选出薯蓣皂苷元高产的毛状根无性系。方法 利用发 根农杆菌 C58C1 感染白英外植体获得毛状根，建立白英毛状根遗传转化体系。采用 HPLC 法检测薯蓣皂苷元量。结果 菌 液浸染时间为 10 min、共培养时间为4 d 可获得最佳转化效果，毛状根诱导率为 83.33%。HPLC 检测结果表明，野生型白英 植株中叶片的薯蓣皂苷元量最高，为 1.742 mg/g。毛状根中薯蓣皂苷元的平均质量分数为4.620 mg/g ，是白英叶片的2.652 倍。结论 通过白英毛状根离体培养，可以高效率地获得薯蓣皂苷元。 关键词：白英；毛状根；薯蓣皂苷元；HPLC ；再生植株 中图分类号：R282.21 文献标志码：A 文章编号：0253 - 2670(2016)07 - 1199 - 05 DOI: 10.7501/j.issn.0253-2670.2016.07.023 Culture of hairy roots in Solanum lyratum and determination of diosgenin 1 1 1 1 2 2 1 CHEN Yu , DONG Yu , ZHANG Kai-yan , SUN Yi-ming , CHENG Meng-qi , ZHANG Lai , SUN Min 1. Key Laboratory of Eco-environments in Three Gorges Reservoir Region (MOE), SchooL of Life Sciense, Southwest University, Chongqing 400715, China 2. Special and Key Laboratory of Functional Materials and Resource Chemistry of Guizhou Provincial Education Department, Anshun University, Anshun 561000, China Abstract: Objective To establish the introduction and culture system of the hairy roots in Solanum lyratum and to screen the clone of hairy roots with more diosgenin. Methods The explants of S. lyratum were infected by Agrobacterium tumefaciens strain C58C1, to obtain the hairy roots and construct the genetic transformation system of the hairy roots in S. lyratum. HPLC was used to determine the diosgenin in the hairy roots. Results The optimum transformation results were obtained with the max inductivity of hairy roots of 83.33% during infecting time for 10 min by C58C1 and c