铁皮石斛原球茎与活体真菌液体悬浮共培养研究

目的:建立铁皮石斛原球茎与活体真菌MF24液体悬浮共培养体系,为原球茎产业化生产提供科学依据。方法:研究原球茎的培养体系是否适用于MF24真菌的生长,对光照和黑暗条件下单独培养的原球茎及与真菌共培养的原球茎的生长状况进行研究,测定生物量及增殖倍数,用HPLC分析比较原球茎中11个特征峰面积的变化。结果:MF24真菌在培养原球茎的6,7-V液体培养基中能正常生长,以1 500 lx光照8～10 h.d-1会延缓真菌的生长速度。原球茎在光照或暗培养条件下都能正常生长,在原球茎培养初期加入MF24真菌,两者相互抑制生长。在原球茎的生长稳定期接入5块直径9 mm真菌菌块,对原球茎的生长和化学成分的种类无显著影响,但光照条件下共培养5 d的原球茎中有10个化合物含量比对照降低13.64%～138.47%,暗培养条件下共培养5 d的原球茎中有7个化合物含量比对照提高0.71%～12.82%,4个化合物含量比对照小幅降低3.03%～14.14%。结论:在适宜的条件下,活体真菌MF24可与铁皮石斛原球茎共培养,并影响其次生代谢水平。

Fluid suspension co-culture of Dendrobium officinale protocorm and living fungus

Objective: To set up a system for fluid suspension co-culture of Dendrobium officinale protocorm and living fungus MF24,so as to provide certain scientific evidence for industrial production of protocorm. Method: Whether the protocorm culture system was suitable for the normal growth of MF24 fungus were studied,the growth of protocorm cultured alone and co-cultured with the fungus were researched under light and dark culture conditions,the biomass and proliferation times were determined,and HPLC method was used to analyze and compare the changes of 11 characteristic peak areas in D. officinale protocorm. Result: The MF24 fungus could grow normally in the 6,7-V liquid medium used to culture the protocorm,and when it was cultured by 8-10 hours per day under 1500 lx ,the growth rate of the fungus was slowed. Protocorm could grow normally in light and dark culture conditions,and add the MF24 fungus in the early cultivation stages of protocorm,both inhibit the growth of each other. In the protocorm for the growth stability to add 5 diameter 9 mm fungi block,the protocorm growth and chemical composition type had no significant effect. However,under illumination,co-cultured for 5 days protocorm of which 10 compounds content decreased 13.64% to 138.47%,in dark conditions,co-cultured for 5 days protocorm of which 7 compounds increased by 0.71% to 12.82%,and 4 compounds slightly reduced by 3.03% to 14.14% compared with the control. Conclusion: Under the appropriate condition,living fungus MF24 could co-culture with the D. officinale protocorm,and affected the latter's secondary metabolite levels.