烟管菌T1原生质体制备及菌丝再生条件优化

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/m L的溶壁酶于30℃下酶解3 h,获得原产量最高,达1.18×10~6个/m L;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。

Protoplast Preparation and Hyphae Regeneration Condition Optimization of Bjerkandera adusta T1

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.