利用气升式生物反应器培养铁皮石斛原球茎  

Cultivation of protocorms of Dendrobium candidum in air-lift bioreactors

作　　者：姚睿[1] 朴炫春[1] 李铁军[1] 邵春绘[1] 廉美兰[1] 

机构地区：[1]延边大学长白山生物资源与功能分子教育部重点实验室,吉林延吉133002

出　　处：《中国中药杂志》2012年第24期3763-3767,共5页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(31160079)

摘　　要：目的:探明影响反应器内铁皮石斛原球茎生长及物质合成的因素,为铁皮石斛原材料的大量生产提供一种新方法。方法:利用3 L气升式生物反应器,以组培原球茎为材料,研究了接种量、光照强度和通气量对原球茎增殖生长和多糖及石斛碱积累的影响。结果:当接种量为10 g.L-1时,培养30 d后原球茎颜色深绿,生长健壮。多糖含量在接种量处理间无差异,但石斛碱含量有差异(接种量10 g.L-1处理最高),多糖和石斛碱生产量在10 g.L-1接种密度中最高。光照强度1 600lx条件下原球茎生长最旺盛,光照对铁皮石斛原球茎多糖的积累起到促进作用,但对石斛碱的积累则有抑制作用,多糖含量和生产量在1 600,2 400 lx光照处理下好于暗条件处理;石斛碱的含量虽然在暗处理中最高,但因原球茎生长不佳,石斛碱的生产量在1 600 lx的光照强度处理中出现最大值。通气量为0.2(空气体积/培养体积/min)时原球茎生长健壮,颜色鲜绿,生长优于0.1,0.3处理,且多糖和石斛碱含量和产量均达最大值。结论:在工作体积为2 L的气升式球型生物反应器内,接入10 g.L-1原球茎外植体,光照强度调节为1 600 lx,通气量为0.2有利于原球茎生长和多糖及石斛碱的生产,铁皮石斛原球茎生物反应器培养是大量快速生产多糖和石斛碱的有效途径。Objective： To explore the factors affecting the growth of protocorms of Dendrobium candidum and substance synthesis in a reactor, in order to provide a new method for mass production of raw materials of D. candidum. Method： Protocorms in vitro were used as experimental materials to study the effect of inoculum volume, light intensity and air volume on the growth of proto- corms of D. candidum and the accumulation of polysaccharide and dendrobine in a 3 L-air lift balloon type bioreactor. Result： After 30 days of cultivation in a bioreactor, protocorms became dark green and grew well at the inoculum volume of 10 g ·L^-1. The polysaccharide content in protocorms showed no difference at various inoculum volumes; whereas the dendrobine content showed differences（with the highest treatment at the inoculum volume of 10 g ·L^-1 ） , particularly the productions of polysaccharide and alkaloid reached the maximum at the inoculum volume of 10 g ·L^-1 The condition of 1 600 lx of light intensity was the most favorable for the growth of protocorms. Though light played a role of improving the accumulation of poiysaccharide in protocorms of D. candidum, it could inhibit the accumulation of dendrobine. Polysaccharide content and production were better under light conditions of 1 600 and 2 400 lx than dark conditions. Despite the maximum dendrobine content in dark conditions, the dendrobine production showed the maximum in the light condition of 1 600 lx due to poor growth of protocorms. Protocorms grew well and became dark green at the air volume of 0. 2 vvm （ air volume culture volume per minute）, which was better than at 0. 1 and 0. 3, with maximum polysaccharide and dendrobine contents and productions. Conclusion： In a 3 L-air lift balloon type bioreactor with a working volume of 2 L, the conditions of 10 inoculum volume, 1 600 lx light intensity and 0. 2 air volume were favorable for the growth of protocorms and the production of dendrobine. This demon- strates that the cultivation of D. candidum and subst

关 键 词：接种量 光照强度 通气量 多糖 石斛碱 

分 类 号：S567.239[农业科学—中草药栽培]