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目的:黑木耳具有丰富营养价值和重要的药用价值,分析和了解黑木耳遗传结构对其种质资源保护和遗传性状改良具有重要的意义。方法:对黑木耳7个菌株rDNA ITS区序列(含5.8S rDNA)进行PCR扩增并测序,对测序结果进行生物信息学分析及RNA二级预测和分析。结果:ITS区序列长度约为650bp,去除两端测序质量不好的区域后长度为571bp,GC含量为49.46~50.70%,整个ITS区共有5个变异位点,其中有3个简约信息位点。N-J系统进化树中8个黑木耳聚为一支,木耳属另外4个种聚在另一支,ITS序列分支与传统的分类结果基本一致。不同种间5.8S rDNA区RNA二级结构基本一致,表现出属的特异性,ITS1、ITS 2区结构表现种间差异。结论:rDNA ITS区序列一、二级结构相结合可为黑木耳分类鉴定提供有用的分子结构信息。
Objective: Black fungus has rich nutritional value and important medicinal value. It is of great significance to analyze and understand the genetic structure of black fungus for the conservation of germplasm resources and the improvement of genetic traits. Methods: The sequences of rDNA ITS region (including 5.8S rDNA) of 7 strains of black fungus were amplified by PCR and sequenced. Bioinformatic analysis and RNA secondary prediction and analysis of sequencing results were performed. Results: The length of ITS region was about 650bp. The length of ITS region was 571bp, the GC content was 49.46% ~ 50.70%. There were 5 variation sites in ITS region, of which 3 parsimony informative sites . In the N-J phylogenetic tree, eight black fungi were clustered together, while the other four species belonging to genus Agaric were clustered in the other one. ITS branching was basically the same as the traditional classification. The RNA secondary structure of 5.8S rDNA regions of different species was basically the same, showing the genus specificity, ITS1, ITS2 structure showed interspecific differences. Conclusion: The combination of primary and secondary structure of rDNA ITS sequence can provide useful molecular structure information for the classification of black fungus.