菌根真菌——赭丝膜伞原生质体分离与再生研究

振荡培养48h的赭丝膜伞（Cortinariusrussus）菌丝体经0．5％流基乙醇预处理后，离心法收集，以06mol／LMgSO_4作高渗稳定剂，PH5．8，采用纤维素酶、离析酶和浸解酶联合处理，在三个温度（24℃、31℃、35℃）下经11～16h反应均可获得较高的原生质体产量，最高产量达8×10~7个／g鲜重菌丝。分离得到的原生质体表现了快速的再生能力，在MMNC固体培养基上可达到20％以上的再生频率。文章较详细地研究了酶系组成、酶解温度和时间、菌丝菌龄等多个因素对C。russus原生质体分离及再生的影响。

ISOLATION AND REGENERATION OF PROTOPLAST FROM ECOTOMYCORRHIZAL FUNGUS CORTINARIUS RUSSUS

A method is described for isolation of protoplasts from Cortinariusrussus, a species of ecotomycorrhizal fungus. The 48hrs-cultured mycelials werepretreated with 0.5% mercaptoethanol, gathered with cenhfugal method, then treatedwith a combinahon of cellulase Onozuka R-10, lysing enzyme and Macerozyme in PH5.8 citrate buffer, and 0.6mmol / L MgSO4 as the osmotic stabilizer. High protoplastyields(5- 8 ×107 / g fwt mycelials) were obtained within 11-16hrs of incubation at35t, 31℃ or 24℃. The isolated protoplasts possessed a rapid regeneration ability,and the regeneration frequency reached at least 20% on a solid MMNC medium. Theeffects of several factors on isolation and generation of protoplasts are discussed.