Diwan Renuka, Malpathak Nutan
Department of Botany, University of Pune, Pune, Maharashtra 411007, India.
N Biotechnol. 2008 Jun;25(1):85-91. doi: 10.1016/j.nbt.2008.02.002. Epub 2008 Apr 23.
Wide applications of Ruta graveolens L. in pharmaceutical industry has led to increased interest in large-scale plant production, with emphasis on use of in vitro cultures. Earlier reports describe use of in vitro germinated seedlings for raising shoot cultures and not regeneration. There is only a single regeneration protocol of R. graveolens; however, it employs conventional labour intensive techniques deterring automation. The aim of present investigation was to establish a cost effective protocol for large-scale plant production. We report for the first time a one-step protocol with improved regeneration efficiency for multiple shoots induction employing liquid culture systems. Effect of polyamines (putrescine and spermine) on growth and furanocoumarin was studied. Addition of spermine enhanced the number of multiple shoots formed (2.5-fold) and reduced the time taken by half. Spermine addition resulted in 1.47-fold in furanocoumarin production. The selected shoot line, RS2 was successfully scaled up to 5L in culture vessels, with 1.53-fold increase in biomass without affecting the productivity of these cultures. This proves to be a commercially feasible alternative to bioreactors for large-scale biomass and furanocoumarin production.
白藓在制药行业的广泛应用引发了人们对大规模植物生产的兴趣，尤其是对体外培养的关注。早期报道描述了使用体外萌发的幼苗来培养芽苗，但并非用于再生。白藓只有一种再生方案；然而，它采用的是传统的劳动密集型技术，不利于自动化操作。本研究的目的是建立一种具有成本效益的大规模植物生产方案。我们首次报道了一种采用液体培养系统提高多芽诱导再生效率的一步法方案。研究了多胺（腐胺和精胺）对生长和呋喃香豆素的影响。添加精胺可使形成的多芽数量增加（2.5倍），并将所需时间减半。添加精胺使呋喃香豆素产量提高了1.47倍。所选的芽苗系RS2在培养容器中成功扩大到5升，生物量增加了1.53倍，且不影响这些培养物的生产力。这被证明是一种在商业上可行的替代生物反应器的方法，可用于大规模生产生物量和呋喃香豆素。
