瑞香狼毒细胞悬浮培养及黄酮积累的研究  

A study on cell suspension culture and flavonoids accumulation of Stellera chamaejasme

作　　者：王文星[1] 安琪[1] 汪莹[1] 王猛[1] 曹成有[1] 

机构地区：[1]东北大学理学院生物技术研究所,辽宁沈阳110004

出　　处：《草业学报》2010年第6期132-139,共8页Acta Prataculturae Sinica

基　　金：国家"十一五"科技支撑计划项目(2006BAD26B04);国家大学生创新性实验计划项目(080105)资助

摘　　要：以瑞香狼毒顶芽、叶片和茎段为外植体,考察不同植物生长调节剂对愈伤组织诱导率及生长状态的影响。由疏松型的愈伤组织建立稳定的瑞香狼毒细胞悬浮培养体系,采用正交试验法初步研究碳源种类和植物生长调节剂对细胞生长和黄酮积累的影响。结果表明,在附加了2,4-D 1.0 mg/L+NAA 0.1 mg/L的MS培养基上诱导茎段可获得适于悬浮培养的疏松型愈伤组织,诱导率达到90%。正交试验中各因素对细胞生长和黄酮含量的影响效应均为2,4-D>碳源>NAA>6-BA;较高浓度的2,4-D、NAA和6-BA均有利于细胞生长和黄酮积累;蔗糖最有利于细胞生长和黄酮积累,其次是葡萄糖和果糖;适于细胞生长和黄酮积累的培养基为:MS+2,4-D 2.0 mg/L+NAA1.0 mg/L+6-BA 0.5 mg/L+蔗糖30 g/L。细胞生长与黄酮积累呈极显著正相关(R2=0.967),P<0.01,这属于生长偶联型的次生代谢模型。A cell suspension culture system was established using cultures of loose calli of Stellera charnaejasme to investigate the effects of different plant growth regulators on callus induction rate and growth state from explants such as buds, leaves and stem segments of S. chamaejasme, and to study the effects of carbon species and plant growth regulators on cell growth and flavonoids accumulation using orthogonal tests. The loose calli suitable for suspension culture can be induced from stem segments on MS with 2,4-D 1.0 mg/L＋NAA 0. 1 mg/L, and the induction rate can reach 90%. The effects of factors in the orthogonal test on cell growth and flavonoids content are consistent and in the order 2,4-D〉carbon source〉NAA〉6-BA. Higher concentrations of 2,4 D, NAA or 6-BA are more conducive to cell growth and flavonoids accumulation. Sucrose is most conducive to cell growth and flavonoids accumulation, followed by glucose and fructose. The optimum medium for cell growth and flavonoids accumulation is MS＋2,4-D 2.0 mg/L＋NAA 1.0 mg/L＋6-BA 0.5 mg/L＋surcrose 30 g/L. There is a significant positive correlation between cell growth and flavonoids content （R2 =0. 967, P〈0 0.01）, belonging to growth-coupling-type of the secondary metabolism model.

关 键 词：瑞香狼毒 细胞悬浮培养 黄酮 碳源 植物生长调节剂 

分 类 号：S567.239[农业科学—中草药栽培]