人参胚愈伤组织诱导和悬浮培养的研究

Study on Suspension Culture and Induction of Callus from Panax ginseng Embryos

[目的]建立一种从人参胚诱导愈伤组织并进行悬浮培养的方法,考察这种悬浮细胞的生长和人参皂苷积累情况。[方法]以人参胚为外植体,在添加了2,4-D3mg/L、NAA2mg/L和6-BA0.5mg/L的MS培养基上诱导愈伤组织。选取经8次继代培养的松散的愈伤组织在含有同样激素的液体培养基中进行悬浮培养。继代悬浮培养5次后测定悬浮细胞生长曲线,用高效液相色谱(HPLC)技术分析细胞中所含的人参皂苷。[结果]有活性的人参胚愈伤组织的诱导率为100%。用这种愈伤组织建立的悬浮培养体系生物量在21d内可增加1.5倍,悬浮培养物至少含有6种人参皂苷,其中Rb1、Rg和Re的含量较高,分别为细胞干重的0.44%、0.35%和0.37%。[结论]由人参胚诱导的愈伤细胞进行悬浮培养比较容易,生长良好,含有较丰富的人参皂苷。

[Objective]The research aimed to establish a method for suspension culture and induction of callus from Panax ginseng embryos,and investigate growth and ginsenoside accumulation of the suspension culture cells. [Method]The calli were induced from P. ginseng embryos in the MS medium added with 2. 4 -D （3 mg/L）,NAA （2 mg/L） and 6-BA （0. 5 mg/L）. Suspension cultures of the calli were established by putting the loose calli subcultured 8 times into liquid media with the same plant hormone as the media used to induce calli. The growth curve was described and six ginsenisides in the cells were analyzed by HPLC after the cells were subcultured 5 times. [Result]The induction rate of calli from the active embryos was 100%. The biomass of suspension cultures of the calli increased 1. 5 -fold in 21 -day cultivation. Six ginsenoside individ-uals were detected in the suspension cell cultures. Ginsenoside Rb1,Rg and Re in the cells were higher levels and 0. 44% ,0. 35% and 0. 37% of the dry cell weight,respectively. [Conclusion]Calli induction from P. ginseng embryos and suspension culture of the calli were easy. Suspension cultures of the calli grew fast and contained many ginsenoside individuals.