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

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。

Comparison on accumulation of flavonoids in loose and compact callus suspension cell culture of Tetrastigma hemsleyanum

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,however,the highest multiple proliferation of compact callus suspension cell was 2.65.The highest content of flavonoids was 11.7 mg/g,increased by 34.5%,compact callus suspension cell was 40.9 mg/g,increased by 212.2%,loose callus suspension cell had a high level of browning and varietal vitality decline after 4-5 subcultures.There was no significant changes in the subculture of the multicell suspension cultures.Conclusion Comparing with loose callus,the compact callus suspension cell shows a remarkable high content of flavonoids,and is preferable for subculture.