麦角菌和雀稗麦角菌原生质体的形成与再生

本文报道了快速制备麦角菌(Claviceps purpurea)和雀稗麦角菌(Claviceps paspali)原生质体的方法及影响原生质体形成的若干因素。用适当方法培养菌丝体,利用商品溶壁酶制剂(10mg/ml),以0.7mol/l KCl为原生质体高渗液,28℃为酶解温度,处理时间仅需1—2h,菌丝体细胞壁即被彻底消化而释放出大量原生质体。在适当的固体再生培养基上,两种麦角菌原生质体的再生频率分别为7.7%和13.2%左右。培养基中添加25mg/l脱氧胆酸钠有利于形成易于计数和分离的小菌落。本文还就CaCl_2,MgCl_2及低温保藏(3—4℃,30天)对于原生质体稳定性与再生的影响作了初步探讨,并证实溶壁酶对原生质体有明显的破坏作用。

PROTOPLAST FORMATION AND REGENERATION OF CLAVICEPS PURPUREA AND CLAVICEPS PASPALI

This report describes a fast and reliable method of obtaining proto- plasts from Claviceps purpurea and Claviceps paspali.Factors affecting the release of the protoplasts were investigated.Using commercial enzyme preparation Lywallzyme (10 mg/ml) in the presence of 0.7 mol/l KCl at 28℃ for 1—2 hours,all the hyphae,which had been cul- tured by a proper means,were easily transformed into protoplasts.The protoplast regenera- tion frequencies of C.purpurea and C.paspali were about 7.7% and 13.2%,respectively.Ad- dition of sodium deoxycholate (25 mg/l) to the agar reversion medium induced the formation of small colonies which were more easily counted and isolated.The effects of CaCl_2,MgCl_2 and low temperature storage (3—4℃ for 30 days) on the stability and regeneration of the protoplasts were also discussed,and the protoplast damage caused by Lywallzyme was detected.