三叶青松散型和致密型愈伤组织悬浮培养及黄酮积累地比较研究.PDFVIP

中草药 Chinese Traditional and Herbal Drugs 第 43 卷 第 3 期 2012 年 3 月 • 577 • 三叶青松散型和致密型愈伤组织悬浮培养及黄酮积累的比较研究 1 2 1 1 1* 彭 昕 ，张 剑 ，何军邀 ，范三微 ，凌庆枝 1. 浙江医药高等专科学校，浙江 宁波 315100 2. 中国药科大学，江苏 南京 210009 摘 要：目的 比较三叶青松散型和致密型愈伤组织悬浮培养细胞生长及其次生代谢产物黄酮的积累情况。方法 将三叶青 叶诱导的愈伤组织在不同外源植物激素种类和配比的培养基上多次继代培养，筛选获得松散型和致密型愈伤组织，分别建立 相应悬浮细胞体系，研究了两种悬浮培养细胞生长和总黄酮积累的动态。结果 三叶青松散型愈伤组织悬浮培养的继代周期 为 8 d，增加质量倍数最高达到 3.68 倍，黄酮积累的最大值 11.7 mg/g，较最初接种时的浓度增加 34.5%，该悬浮系继代4～ 5 次后生命力衰退且褐化加重；而致密型愈伤组织悬浮培养的继代周期为 20 d ，增加质量倍数最高达到2.65 倍，黄酮积累的 最大值 40.9 mg/g ，较最初接种时的浓度增加212.2% ，该悬浮系继代多次细胞无明显变化。结论 与松散型愈伤组织悬浮培 养相比，致密型愈伤组织悬浮培养更有利于继代培养和次生代谢产物的积累。 关键词：三叶青；松散型；致密型；愈伤组织；悬浮培养；黄酮 中图分类号：R282.2 文献标志码：A 文章编号：0253 - 2670(2012)03 - 0577 - 04 Comparison on accumulation of flavonoids in loose and compact callus suspension cell culture of Tetrastigma hemsleyanum 1 2 1 1 1 PENG Xin , ZHANG Jian , HE Jun-yao , FAN San-wei , LING Qing-zhi 1. Zhejiang Pharmaceutical College, Ningbo 315100, China 2. China Pharmaceutical University, Nanjing 210009, China Abstract: Objective To analyze and compare the cell growth and accumulation of its secondary metabolites, flavonoids, in loose and compact calluses suspension cell culture of Tetrastigma hemsleyanum. Methods The calluses induced from the leaves of T. hemsleyanum were subcultured in different culture media, to screen loose and compact calluses, then established two suspension cell systems. The time courses of cell growth and yields of flavonoids were studied. Results Suitable culture period of loose callus suspension cell was 8 d, however, the suitable culture period of compact callus suspension cell was 20 d. The highest multiple proliferation was 3.68, h