玉蕈原生质体制备条件研究

Study on the Preparation Condition for Protoplast of Hypsizygus marmoreus

[目的]探索了玉蕈原生质体的制备与再生的适宜条件。[方法]采用斜面、平板培养基和液体培养基,研究了玉蕈菌丝体酶解时间（1-5 h）、酶解温度（25℃）和菌龄（4-7 d）以及渗稳剂种类（甘露醇、蔗糖、KCl、MgSO4）对玉蕈原生质体制备与再生的影响,优化了玉蕈原生质体制备与再生的条件。[结果]玉蕈原生质体制备与再生的适宜条件是：渗稳剂为0.6 mol/L MgSO4.7H2O,玉蕈菌丝体酶解时间2h,酶解温度25℃,菌龄5 d。在此条件下,玉蕈原生质体的产量最高,达16.7×10^6个/ml。[结论]该研究为利用原生质体技术进行玉蕈育种奠定了基础。

[Objective]The study aimed to explore the suitable condition for the preparation and regeneration of Hypsizygus marmoreus protoplast.[Method]The slant,plate and liquid media was used to study the effect of enzymolysis time（1～5 h）,enzymolysis temp.（25 ℃） and mushroom age（4～7 d） and kinds of osmotic stabilizer（mannitol,cane sugar,KCl,MgSO4） on the preparation and regeneration of H.marmoreus protoplast.The condition for the preparation and regeneration of H.marmoreus protoplast was optimized.[Result]The suitable condition for the preparation and regeneration of H.marmoreus protoplast was that the osmotic stabilizer was 0.6 mol/L MgSO4·7H2O,the enzymolysis time of mycelium was 2 h,the enzymolysis temp.was 25 ℃ and the mushroom age was 5 d,under which,the yield of H.marmoreus protoplast was highest,reaching 16.7×10^6/ml.[Conclusion]The study lays foundation for the breeding of H.marmoreus by using protoplast technology.