【摘要】 研究桦褐孔菌原生质体制备与再生条件。采用正交试验法确定复合酶比例,用单因素试验测定不同等渗液、酶解时间、桦褐孔菌菌龄、酶解温度、再生培养基对桦褐孔菌原生质体制备与再生的影响。结果表明:混合酶(纤维素酶5mg/mL、蜗牛酶15mg/mL、溶壁酶15mg/mL、崩溃酶15mg/mL)、菌龄为7d的菌丝,采用0.6mol/LMgSO4为渗透压稳定液,酶解温度35℃,酶解时间5.5h,有利于原生质体的制备。就原生质体再生而言,酶解时间为3.5h制得的原生质体,以MgSO4作为渗透压稳定液,以完全再生培养基(CYM)和麦芽酵母葡萄糖再生培养基(GYM)有利于原生质体再生,再生率最高为0.13%。更多还原

【Abstract】 The conditions for protoplast preparation and regeneration from the mycelia of Inonotus obliquus were investigated.The optimal amounts of four enzymes were explored by orthogonal array design and other conditions were studied by single factor method.The results showed that the mycelium harvesting after 7-day culture,adding 5 mg/mL cellulase,15 mg/mL snailase,15 mg/mL lywallzyme and 15 mg/mL drislease,using 0.6 mol/L MgSO4 as the osmotic stabilizer,digestion at 35 ℃ for 5.5 h were found to be best for the preparation of protoplasts.The optimal conditions for protoplast regeneration of Inonotus obliquus were CYM or GYM as the medium,0.6 mol/L mannitol as the osmotic stabilizer using and digestion time of 3.5 h.No significant difference in protoplast regeneration rate was observed between CYM and GYM media.The highest protoplast regeneration rate was 0.13%.更多还原