培养基成分对怀牛膝愈伤组织诱导及牛膝多糖含量的影响

目的:研究培养基成分对怀牛膝愈伤组织诱导形成及牛膝多糖含量的影响。方法:以怀牛膝无菌苗的叶片、茎段为材料,采用正交设计研究基本培养基,碳源,2,4-二氯苯氧乙酸(2,4-D)、6-苄基腺嘌呤(6-BA)、N-苯基-N′-噻二唑-5-脲(TDZ)、水解酪蛋白(CH)对愈伤组织诱导和多糖含量的影响。结果:诱导叶片愈伤组织形成的适宜培养基为B₅+2mg·L^-12,4-D+0.5mg·L^-16-BA+30g·L^-1葡萄糖+1g·L^-1CH,诱导茎段愈伤组织形成的适宜培养基为B₅+2mg·L^-12,4-D+1mg·L^-16-BA+30g·L^-1葡萄糖+0.5g·L^-1CH;有利于叶片愈伤组织中多糖含量提高的培养基是LS+1mg·L^-16-BA+1mg·L^-1TDZ+30g·L^-1蔗糖,有利于茎段愈伤组织中多糖含量提高的培养基是LS+1mg.L^-12,4-D+0.5mg·L^-16-BA+1mg·L^-1TDZ+30g·L^-1葡萄糖。结论:初步建立了怀牛膝叶片、茎段诱导愈伤组织的适宜培养条件,为通过植物组织细胞培养途径生产牛膝多糖提供了依据。

Effect of medium components on callus induction and contents of polysaccharides in calli from Achyranthes bidentata

Objective:To study the effect of medium components on the callus induction and the contents of polysaccharides in calli from Achyranthes bidentata.Method:Leaves and stems were selected as explants.The effects of six kinds of factors including basal culture medium,carbon source,2,4-D,6-BA,TDZ,CH on the callus induction and the contents of ABPS in calli on the high growth point were studied by orthogonal design method.The data were analyzed with range analysis and variance analysis.Result:To leaf,the optimal medium of callus induction was B5 with 2 mg·L^-1 2,4-D,0.5 mg·L^-1 6-BA,30 g·L^-1 glucose and 1 g·L^-1 CH;to stem,the optimal one was B5 with 2 mg·L^-12,4-D,1 mg·L^-1 6-BA,30 g·L^-1 glucose and 0.5 g·L^-1 CH.In order to obtain higher contents ABPS,to leaf calli,the optimal medium was LS with 1 mg·L^-1 6-BA,1 mg·L^-1 TDZ and 30 g·L^-1 sucrose;to stem calli,the optimal one was LS with 1 mg·L^-1 2,4-D,0.5 mg·L^-1 6-BA,1 mg·L^-1 TDZ and 30 g·L^-1 glucose.Conclusion:The optimal media of callus induction were established with stems and leaves of A.bidentata as explants and with a view to an industrial production of polysaccharides by tissue and cell culture.