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为了研究卷边桩菇[Paxillus involutus(Batsch)Fr.]不同分离部位、不同培养基对菌丝生长的影响,并鉴别卷边桩菇组织分离株的真伪,对采集的野生卷边桩菇子实体进行了菌种分离,并对获得的菌丝进行了rDNA ITS序列分析。结果表明:A3是卷边桩菇菌丝分离较适宜的培养基配方,菌盖与菌柄连接处为分离的最佳部位。对卷边桩菇分离株rDNA ITS片段进行PCR扩增,扩增产物长度为724bp,登录NCBI与GenBank中已知的菌种进行Blastn比对,序列相似性在99%~93%之间,下载相似性高的菌种的ITS序列,通过ClustalW进行序列比对,MEGA5.2构建系统发育树,通过树图分析,鉴定出分离的菌株为卷边桩菇纯菌种。
In order to study the effects of different culture media on the mycelial growth of Paxillus involutus (Batsch) Fr. And to identify the authenticity of the tissue culture of C. curvatus, The fruiting body was isolated from the bacteria, and the obtained mycelium was analyzed by rDNA ITS sequence. The results showed that A3 was the best medium for the separation of Agkistrodon acutus, and the best place for separation was the junction of cap and stipe. The ITS region of rDNA ITS fragment was amplified by PCR. The amplified product was 724bp in length. Blastn alignment was performed between NCBI and known strains in GenBank. The sequence similarity was between 99% and 93% The ITS sequences of the highly similar strains were aligned by ClustalW. The phylogenetic tree was constructed by MEGA5.2. Through the tree analysis, the isolated strains were identified as pure strains of Curvularia crium.