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目的:为从千层塔中分离具有药用价值的内生真菌奠定基础。方法:新鲜千层塔茎段,经酒精和升汞消毒后,接种于PDA平板培养基上进行内生真菌的分离、纯化;根据菌落形态和孢子等形态特征,结合核糖体基因居间序列(ITS序列)进行菌株鉴定。结果:从千层塔的茎中分离出4株内生真菌。内生真菌Ⅰ菌落形态和孢子特征与枝状枝孢霉属的特征相符合,ITS序列与GenBank中多条属于枝状枝孢霉的ITS序列相似,鉴定该菌株属于枝状枝孢霉;内生真菌Ⅱ菌落形态和孢子特征与黄青霉的特征相符合,鉴定该菌株属于黄青霉;内生真菌Ⅲ菌落形态和孢子特征与尖孢镰刀菌的特征相符合,ITS序列与GenBank中多条属于尖孢镰刀菌的ITS序列相似程度高,鉴定该菌株属于尖孢镰刀菌;内生真菌Ⅳ菌落形态与盾壳霉相似,ITS序列与GenBank中6条属于盾壳霉的ITS序列具有较高的相似性,鉴定该菌株属于盾壳霉。结论:从千层塔中分离和鉴定出4株内生真菌,分别属于枝状枝孢霉、黄青霉、尖孢镰刀菌和盾壳霉。
Objective: To lay the foundation for separation of endophytic fungi with medicinal value from melaleuca. Methods: The stems of fresh Melaleuca tower were sterilized with alcohol and mercuric chloride and then inoculated on PDA plate medium for the isolation and purification of endophytic fungi. According to the morphological characteristics of colonies and spores, Sequence) for strain identification. Results: Four strains of endophytic fungi were isolated from the stems of Melaleuca. Endophytic fungi Ⅰ colony morphology and spore characteristics consistent with the characteristics of Mycobacterium Cladosporium, ITS sequences and GenBank belonging to several Cladosporium Cladosporium ITS sequence similar to the strain identified as Mycobacterium Cladosporium; within The morphological and spore characteristics of virulent strains Ⅱ were consistent with those of P. chrysogenum. The strain was identified as belonging to the genus Penicillium chrysogenum. The morphology and spore characteristics of the endophytic fungi Ⅲ were consistent with the characteristics of Fusarium oxysporum. ITS sequences were similar to those of GenBank The ITS sequence similarity of Fusarium oxysporum was high, and the strain was identified as Fusarium oxysporum. The colony morphology of endophytic Fungus Ⅳ was similar to that of C. conidia, and ITS sequences were similar to 6 ITS sequences of C. jecorvae in GenBank High similarity, the strain identified as belonging to the mold cell mold. Conclusion: Four strains of endophytic fungi were isolated and identified from Melaleuca, which belong to Cladosporium fulvum, Penicillium chrysogenum, Fusarium oxysporum and Pyrenia spp. Respectively.