千层塔内生真菌的分离与鉴定

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、尖孢镰刀茵和盾壳霉。

Isolation and Identification of Endophytic Fungi of Huperzia Serrata

Objective： The research aimed to provided the foundation for seeking the endophytic fungi of the medicinal value in Hu.perzia serrata. Methods： A stem segment from Huperzia serrata was surface sterilized by successive submersion in 70% ethanol, 10% mercuric chloride, and then was placed on a potato dextrose agar （PDA） plate to isolate endophytic fungi. The outgrowing fungi strains were successive subcultured to obtain pure cultures. The purified strains were identified by morphology together with similarity of internal transcribed spacer （ITS） sequence. Results： A total of 4 endophytic fungi strains were isolated from Huperzia serrata. Strain I was compared with Cladosporium cladosporioides （Fres.） de Vries with the similar morphological character and ITS region. The results indicated that this endophyte was identified as Cladosporium cladosporioides（Fres. ） de Vries. Strain II was identified as Penicillium chrysogenum Thorn based on the analyses of morphological character and ITS region. Strain III was identified as Fusarium oxysporum Schlechte. emend. Snyd.＆ Hans based on the analyses of morphological character and ITS region. StrainlV was identified as Coniothyrium sporulosum （W. Gams ＆ Domsch） van der Aa based on the analyses of morphological character and ITS region. Conclusion： A total of 4 endophytic fungi were isolated from Huperzia serrata. The further molecular and morphological identification indicated that these fungal isolates belonged to Cladosporium cladosporioides （Fres.） de Vries,Penicillium chrysogenum Thom,Fusarium oxysporum Schlechte. emend. Snyd.＆ Hans and Coniothyrium sporulosum （W. Gams ＆ Domsch） van der Aa respectively.