利用优化的甲基纤维素培养基与传统的平板涂布、显微操作、有限稀释和琼脂糖培养基方法,对蛹虫草菌孢子悬液(理论活孢子数量为150个)进行单孢分离,比较可分离单菌落数量、操作简易度及分离效率等参数,分析甲基纤维素半固体培养基分离真菌单孢的可行性。结果表明,甲基纤维素的优化质量浓度为23 g/L。甲基纤维素培养基、平板涂布、显微操作、有限稀释和琼脂糖培养基方法依次获得可分离单菌落96、70、37、35和26个,甲基纤维素培养基方法的操作最简单、便捷和高效。优化的甲基纤维素半固体培养基可有效分离真菌单孢,简便易行,应用潜力优于传统方法。
Taking Cordyceps militaris as an illustrative example, a novel method that opened up an optional way for the isolation of fungal single-spores based on semisolid methylcellulose medium was developed. Optimized methylcellulose medium and traditional methods (spread plate, micromanipulation, limited dilution, and agarose medium) were used to isolate single-spores of Cordyceps militaris spore suspension that theoretically contained 150 living spores. Comparing the number of partible single colonies, operation simplicity and the efficiency of single-spore isolation, the feasibility of this novel method to isolate fungal single-spores was investigated. The optimized concentration of methylcellulose in semisolid medium was 23 g/L. The number of partible single colonies obtained using the optimized methylcellulose medium, spread plate, micromanipulation, limited dilution and optimized agarose medium reached 96、70、37、35 and 26, respectively. This novel method was proved to be the simplest, most convenient and efficient among these methods. Fungal single-spores could be simply, efficiently and conveniently isolated using optimized methylcellulose medium. The applied potential of the novel method is superior to that of traditional methods.
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