姬松茸原生质体再生条件的优化

Optimization of regeneration condition of protoplast of Agaricus blazei Murill

为提高姬松茸原生质体的再生率,对姬松茸原生质体再生的条件进行了优化.结果表明,将培养5 d的姬松茸种子液点种在覆以玻璃纸的菌丝生长培养基上,26℃培养5 d,用2.0%的裂解酶以pH6.0,0.8 mol/L NaCl为稳定液,在30℃下酶解6 h,可制备得到菌落数为1.8×107 cfu/mL的原生质体液.将原生质体悬液经G2过滤器过滤除菌后在GM上再生,原生质体再生率可达8.53%,残片率2.43%.

The best regeneration condition of protoplast of Agaricus blazei Murill is： The seeds of Agaricus blazei Murill which had been cultured for 5 days were inoculated on the hypha growtheal substrate by dot method which was covered with cellophane. The yield of protoplast was 1.8×10^7 cfu/mL after being cultured 5 days at 26 ℃ and zymolysis involved for 6 hours at 30 ℃ with 2.0% of lyase plus 0.8 mol/L of NaC1 （pH6.0） as stabilizing buffer. The suspending liquid of protoplast was regenerated on GM substrate which was purified through G2 filter. The rate of regeneration was 8.53% and the rate of residual was 2.43%.