松茸菌丝体的纯培养及其鉴定

The Pure Cultivation of Tricholoma matsutake and It’s ITS Identification

采用组织分离法,对采自云南中甸、丽江、武定、巍山4个地方的松茸进行了分离培养,并利用感官及ITS测序对所培养的菌丝进行了鉴定。结果表明,利用斜面培养基很好地诱导出了菌丝体母种,诱导率达75%;而将分离培养得到的菌丝体进行液体培养,菌丝生长良好,且能进行转扩和较大规模培养;液体培养的菌丝体仍具有松茸的特殊气味,证明可能为松茸的纯培养物;经ITS测序比较,采自中甸、丽江、武定、巍山的松茸子实体和其液体培养的菌丝体,其遗传相似度分别为0.9970、1、0.9980、0.9980,表明取自不同地点的子实体与其相应的培养物在种质上是一致的。

In this study,the mycelia of Tricholoma matsutake from Zhongdian,Lijiang,Wuding and Weishan were cultivated by the method of tissue separation and identified by the method of sense organ distinguishing and ITS sequencing.The results showed that the mycelia of maternal strain were well induced and the rate of induction is 75% on the slant medium of PDA＋ yeast extract paste（0.5%）.The mycelia gained above could grow very well and be cultivated in larger scale on liquid culture medium.The mycelia of liquid culture had special smell as which Tricholoma matsutake had,and this showed it were likely to be pure culture from Tricholoma matsutake.By ITS sequencing,the genetic identities,between mycelia of liquid culture and fruiting body from Zhongdian,Lijiang,Wuding and Weishan,were 0.997,1.00,0.998 and 0.998 respectively,that implied the mycelia were indeed from their corresponding fruiting body.