烟管菌T1原生质体制备及菌丝再生条件优化

摘要

[目的]研究烟管菌T1(Bjerkandera adusta)原生质体制备及菌丝再生的最佳条件.[方法]研究菌龄、渗透压稳定剂种类和浓度、pH、酶浓度及酶解时间对原生质体得率的影响,并研究再生培养基种类对菌丝再生的影响.[结果]在 PDA 液体培养基中,以28 ℃摇床培养3 d 的菌丝最适于该菌原生质体制备;用0.5 mol/L KCl配制的含有15 mg/mL的溶壁酶于30 ℃下酶解3 h,获得原产量最高,达1.18×106个/Ml;RM1培养基再生率最高,达1.22%.[结论]该研究优化了原生质体制备及菌丝再生条件,为进一步研究烟管菌噬藻分子机制提供理论依据.%[Objective]To study protoplasts preparation and hyphae regeneration conditions of Bjerkandera adusta.[Method] The effect of different culture conditions (hyphae age,stabilizer and stabilizer concentration,pH,enzymatic concenfration,enzymatic hydrolysis time) on protoplasts preparation and hyphae regeneration conditions of Bjerkandera adusta was studied.[Result] The fungal mycelia cultured in the liquid PDA medium at 28 ℃ for 3 days was the most suitable for protoplast preparation.15 mg/mL soluble wall enzyme in 0.5 mol /L KCl hydrolyzing for 4 hours could generate as many as 1.18×106 protoplasts/mL.The RM1 medium was the preferred regeneration medium on which the regeneration rate was as high as 1.22%.[Conclusion] The study optimize the conditions of protoplasts preparation and regeneration,and provide theory basis for further study of molecular mechanism of Bjerkandera adusta.