糙皮侧耳原生质体制备与再生条件

Isolation and Regeneration of Protoplasts from Edible Fungus Pleurotus ostreatus

原生质体制备是通过体杂交或基因转导进行种质改良的基础。探讨了糙皮侧耳原生质体制备及再生条件。用ＭＹＧ培养菌丝，以甘露醇为稳渗剂，用１．５％溶壁酶Ｌｙｗａｌｌｚｙｍｅ进行酶解脱壁，在菌龄７天，酶液ｐＨＷＦＨＧ４．５，温度２８℃，酶解４ｈｒ的条件下原生质体产量最高（２０×１０＾６／ｍｌ，０．１ｇ湿菌丝），其再生率大于８％，菌丝培养前对接种物进行予切碎处理可进一步使产量提高约２倍，而再生率不并显著下降。

Preparation and culture of protoplasts are basic techniques for genetic improvement of plants and fungi through somatic cell fusion or gene transfer.Conditions for preparing the protoplasts from edible fungus Pleurotus ostreahs were examined.The highest production of protoplasts with plating efficiency of more than 8% was abtained with the enzyme mixture of pH 4.5 containing 1.5% Lywallzyme,0.6mol/L mannitol as osmotic stabilizer, and incubated at 28℃,from 7-day-age mycelia cultured in MYG medium.Shredding the inocula before mycelium incubation increased by 2 times the production of protoplasts. with plating efficiency not markedly affected.The protoplast production was lower at pH5.5 or pH6.5 than at pH4.5,but plating efficiency was increased at the former two pH value.