灰紫香蘑的分离纯化及ITS序列鉴定

[目的]对采自内蒙古根河地区的灰紫香蘑进行组织分离与鉴定.[方法]分别选取菌盖处、菌褶与菌柄交界处和菌柄处进行组织分离,将分离物进行ITS序列分析,计算遗传距离,并采用邻接法构建NJ系统发育树.[结果]菌褶与菌柄交界处为最佳分离部位,菌丝生长速度快、洁白细密、污染率低;其次,子实体自然风干3d后再进行分离可有效降低菌丝污染程度;最后分离物经ITS序列测定,系统发育分析证实其为灰紫香蘑.[结论]该方法获得了灰紫香蘑的纯培养菌株,为进一步开发和利用灰紫香蘑提供了科学依据.

Isolation and Identification of Lepista glaucocana by ITS Gene Sequence

[Objective] To isolate,cultivate and accurately identify Lepista glaucocana which was collected from Genhe district in Inner Mongolia.[Method] Different tissues from pileus,stipe,junction of pileus and stipe were isolated,then the isolate was identified by internal transcribed spacer （ITS）,the genetic distance was calculated,and a NJ systematic tree was established by the neighbor-joining method.[Result] The results showed that junction of pileus and stipe is the best isolation position,which mycelia with fast growth and low pollution; Secondly,it is lower pollution when airdry the fruiting body for 3 days; At last,the isolate was identified asL.glaucocana analyzed by phylogenetic relationship of ITS gene sequences.[Conclusion] The pure strain was obtained which can provide a scientific basis for development and use of L.glaucocana.