羊肚菌原生质体制备和再生

Preparation and regeneration of protoplasts from Morchella sp.

本文从菌丝培养、菌丝酶解、原生质体再生等方面研究了尖顶羊肚菌（M orchella conica）A401和粗腿羊肚菌（M.crassieps）一个菌株E903的原生质体制备与再生条件,结果表明：两供试菌种在20℃下于营养丰富的现配液体培养基③中匀速振荡培养2~4 d可得菌龄一致、活力强、量大的菌丝体,收集菌丝体洗涤吸干水分后在现配酶液①中于30℃恒温下振荡酶解3.5 h原生质体产量最高（A401为3.8×106个/100 mg.mL,E903为3.6×10^6个/100 mg.mL）,再将分离纯化的原生质体悬液单层涂布到新鲜的③号再生平板上,20℃恒温培养3~5 d后可得大量再生菌落（最高再生率A401为0.78%,E903为0.8%）。

For the Morchella sp.,preparation and regeneration conditions of protoplast were reported in this paper.The results showed that the optimal liquid medium and time for the mycelial growth of two strains were No.③ liquid medium and 2-4 days,respectively,the appropriate liquid enzymes,enzymolysis time and temperature for the mycelial digesting were No.① liquid enzymes,3.5 h and 30 ℃.On the other hand,there were a lot of regeneration colony in No.③ regeneration medium after 3-5 days.