攀枝花块菌菌丝体的分离及种属鉴定

Mycelium Isolation and Species Identification of Panzhihua Truffle

目的:分离攀西地区野生块菌组织,获得块菌的纯培养菌丝体,并用分子生物学方法对萌发菌丝体进行种属鉴定。方法:用收集自攀枝花地区的野生块菌进行组织培养,对萌发的菌丝体进行ITS序列测定并构建系统发育树,以鉴定其种属分类。结果:无菌块菌组织在PDA培养基中培养1周后开始萌发白色菌丝,经组织化学切片染色观察发现萌发的菌丝源自组织块的体细胞而非子囊孢子。萌发菌丝的r DNA-ITS序列分别与印度块菌(Tuber indicum,JQ638991.1)、会东块菌(Tuber umbilicatum,FJ797880.1)的参考序列高度同源,可确认组织培养成功获得块菌的菌丝体。结论:通过组织分离培养获得了攀枝花地区块菌的纯培养菌丝体。

To obtain pure mycelia from cultured truffle tissues, and to identify the species of germinated mycelia based on their rDNA -ITS sequences. [ Methods ] The sterile tissues were separated from the wild truffles collected from Panzhihua region, and cultured on PDA medium. The species of germinated mycelia were identified using their rDNA-ITS sequences. [ Results ] White germinated mycelia were obtained after one week＇s culture on PDA medium. HE staining found that the germinated mycelia were derived from somatic tissue rather than ascospores. The rDNA - ITS sequences of germinated mycelia were highly homologous with the reference sequences of Tuber indicum （JQ638991.1） and Tuber umbilicatum （FJ797880. 1）,which indicated that the truffle mycelia were successfully obtained. [ Conclusion ] The pure truffle mycelia were obtained from the separated sterile truffle tissue.