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作 者 ：孔繁翔

期 刊 ：林业科学 1996年 32卷 4期 页码：342-347

关键词：黑核菌；鹅膏菌；菌根真菌；液泡分离；

Keywords:Amanita muscaria, Cenococcum geophilum, Mycorrhizal fungi, Vacuolar isolation,

摘 要 ：从菌根真菌黑核菌(Cenococcum geophilum Fr.)和鹅膏菌[Amanita muscaria (L.exFr.)Pers.exHook.]菌丝中,采用多元碱化合物诱导原生质体裂解方法,分离出细胞器──液泡。实验结果表明,在加磷的MMNC培养基中培养4—7天的菌丝经水解酶作用所释放出的原生质体具有较高的液泡化程度;液泡产率约为原生质体的5-10%;细胞质特征酶葡萄糖-6-磷酸-脱氢酶活性测定结果证明了液泡的纯度;而等量原生质体和液泡中酸性磷酸酶及α-甘露糖苷酶活性的比较表明这两种酶主要存在于液泡中,因此,它们可以作为菌根真菌黑核菌和鹅膏菌液泡的特征酶。

Abstract:The vacuoles were for the first time isolated from the mycorthizal fungi (Amanita muscaria and Cenococcum geophilum) by the polybase, DEAED-dextron, induced lysis of protoplasts. The protoplasts isolated from mycelium cultured in MMNC+P medium for 4-7 days showed considerable vacuolation.The yield of vacuole was in the range of 5-10% of protoplasts. The purity of vacuolar preparation was judged by measuring and comparing soluble cytoplasmic enzyme G6PDH, which indicated that it was not contaminated by the cytoplasm and other cell organelles. The measurement of activities of acid phosphatase and α-mannosidase in equal number of vacuoles and protoplasts revealed that almost all this two enzymes were located in the vacuole, so they might be established as the vacuolar marker enzymes for A. muscaria and C.geophilum.

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