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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。
OBJECTIVE: To establish a liquid suspension co-culture system of protocorm-like Dendrobium officinale with live fungi MF24 to provide a scientific basis for the industrialization of protocorm-like bodies. Methods: Whether the culture system of protocorm-like bodies was suitable for the growth of MF24 fungi was studied. The growth of protocorm-like bodies and protocorms co-cultured with fungi under light and dark conditions were studied. The biomass and multiplication rate were determined. Analysis of prothallome in the 11 characteristic peak area changes. Results: The MF24 fungi grew normally in the 6,7-V liquid medium of protocorm. The growth of fungi was delayed by irradiation at 1 500 lx for 8-10 h. Protocorms can grow normally under light or dark culture conditions, adding MF24 fungi in the early stage of protocorm growth, the two inhibit each other growth. There were no significant effects on the growth and the chemical composition of the protocorms when 5 bacterial fungi with a diameter of 9 mm were introduced into the stable growth phase of the protocorm. However, the contents of 10 compounds in the protocorms co-cultured for 5 days under light were higher than those of the control Decreased by 13.64% -138.47%. Compared with the control, the content of 7 compounds in protocorm increased by 0.71% -12.82% and the content of 4 compounds decreased by 3.03% -14.14% compared with the control. CONCLUSION: Under appropriate conditions, MF24 of live fungi can co-culture with protocorm-like bodies of Dendrobium candidum and affect their secondary metabolites.