为适应双孢蘑菇工业化生产需要,以控制菌丝球直径和密度、并提高菌丝体生物量为目的,实验以双孢蘑菇As2796为出发菌株,采用单因素、正交设计方法,优化其液体菌种繁育条件。结果表明,筛选的双孢蘑菇一级液体种子的优化培养基为:马铃薯80 g、红糖12 g、葡萄糖9.0 g、蛋白胨2 g、酵母膏1 g、KH2PO42g、MgSO4.7H2O 1g、VB10.01 g、水1 000 mL;优化的二级种子液体繁育条件为:转速150 r/min、培养温度25℃、初始pH6.0、接种量12%、装液量100 mL。在此条件下菌丝球直径可有效控制在1.5～2.0 mm之间,菌丝体生物量干重可达25.0 mg/mL。

In order to obtain the proper spawn fitting for industrial cultivation of Agaricus bisporus,the study was carried out to increase the mycelium biomass and control the hyphal ball diameter and density in submerged culture process.Using Agaricus bisporus As2796 as the strain,the single factor tests and orthogonal design method were employed to optimize medium composition and its liquid culture condition in the trial.The results showed that the optimum medium composition for culturing Agaricus bisporus primary spawn in liquid was as follows: potato 80 g,brown sugar 12 g,glucose 9.0 g,peptone 2 g,yeast extract 1 g,KH2PO4 2 g,MgSO4·7H2O 1 g,and VB1 0.01 g in 1 000 mL water.The best liquid culture conditions for secondary spawn were as follows: shaking speed of 150 r/min,culture temperature 25℃,initial pH 6.0,inoculation quantity 12%,and filling with 100 mL fluid in 250mL flask.Under the conditions,the hypha ball diameter could be controlled effectively in the range of 1.5 mm to 2.0 mm,and mycelium biomass yield reached 25.0 mg/mL(on a dry-weight basis).