采用固定化培养法模拟东北红豆杉(Taxus cuspidate)和美丽镰刀菌(Fusarium mairei K178)的共生环境,对其相互作用进行初步研究以期提高各自紫杉醇产量。共培养条件为:海藻酸钠浓度2%(M/V),Zcuspidate细胞包埋量0.15g/ml,CaCl2浓度4%(M/V),凝胶直径为3~3.5mm,钙化0.5h,以40ml接种量接入装液量为70ml/250ml摇瓶的改良MS培养基,25℃、160r/min振荡黑暗培养,第9d接入10%(V/V)对数期的F.mairei,10d后收获。结果表明,F.mairei对rcuspidate细胞的生长有强烈抑制作用,能诱导紫杉醇等次生代谢物向胞外分泌;Fmairei与rcuspidate的相互作用对共生培养物的紫杉醇产量也有显著影响。
The interaction mechanism between Taxus cuspidate and the endophytic fungi Fusarium mairei K178 was studied preliminarily for their respective paclitaxel productivity improvement by immobilized culturing,which imitated their symbiotic environment under the condition of maintaining their biological activity. The co-cultivation condition established were : the concentration of sodium alginate solution was 2% ( M/V ), the entrapped suspension plant cells density was 0. 15g/ml, the concentration of calcium chloride solution was 4% (M/V) , the diameter of the beads was ranged from 3 to 3.5 mm, 0. 5h calcification curing, transferred 40ml into the 70ml amended MS medium in 250ml flask, then the immobilizing cells were placed on a rotary shaker at 25 ℃, dark and 160 r / min. The inoculation of F. mairei in log phase was 10% ( V / V) in the 9th day of cultivation, harvested after 10 days. The results showed that F. mairei can induce strong growth inhibition of T.cuspidate, can induce the extracellular secretion of paclitaxel and other secondary metabolites. The paclitaxel production of the co-cultivation had been influenced by the interaction significantly. Suggest that this model can indeed show something about the relationship between Taxus cells and their endophytic fungi which produce the same secondary metabolites.