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大兴安岭北方森林凋落物真菌及其抗菌化合物
Litter Fungi in the Boreal Forests and Their Antibacterial Compounds in the Greater Khingan Mountains
为了研究大兴安岭森林凋落物真菌的抗菌活性,首先利用颗粒涂布平板法分离真菌,通过扩增与比对内部转录间隔区序列初步鉴定分离到的真菌。采用双平板打孔药剂扩散法检测抗菌活性,使用层析法分离目标菌株单体化合物。通过波谱学分析确定其化学结构,使用96孔板液体稀释法检测单体化合物的最低抑菌浓度。共分离得到88株真菌,其隶属于57个属、74个分类单元,其中18株真菌内部转录间隔区序列相似性≤97%。其中6株真菌的乙酸乙酯提取物具有抗菌活性,2株真菌KNFL008和KNFL040抗菌活性较强。经活性追踪分离,共得到4个单体化合物,分别为化合物A（1,3-2-羟基-5-十五烷基苯）、化合物B（麦角固醇）、化合物C（2-氨基-3H-苯氧嗪-3-酮）和化合物D（2-乙酰氨基-3H-苯氧嗪-3-酮）。对化合物A、B、C进行了最低抑菌浓度的测定,为3.13~200 μg/mL。化合物A和D为首次在真菌提取物中分离。本研究为进一步开发利用大兴安岭凋落物真菌资源打下了基础。
To study the antimicrobial activity of forest litter fungi in the Greater Khingan Mountains, firstly, fungi were isolated by a method of spreading particles on plates, and the isolated fungi were preliminarily identified by the amplification and alignment of internal transcribed spacer sequences. The antibacterial activities were detected by a two-layer-plate diffusion method and monomeric compounds of targeted strains were separated by chromatography. Spectroscopic analyses were used to determine the chemical structures of the monomeric compounds. A 96-well plate liquid dilution method was used to detect their minimum inhibitory concentration. A total of 88 fungi were isolated, belonging to 57 genera and 74 taxa. Of them, the internal transcribed spacer sequence similarities of 18 fungi were not higher than 97%. Among them, the ethyl acetate extracts of 6 fungi had antibacterial activities, and the two fungal strains KNFL008 and KNFL040 had stronger antibacterial activities than the others. By a bioassay-guided separation, four monomeric compounds, A (1,3-2-hydroxy-5-pentaalkylbenzene), B (ergosterol), C (2-amino-3h-phenoxyzin-3-ketone) and D (2-acetylamino-3h-phenoxyzin-3-ketone), were separated. The minimum inhibitory concentration of compound A, B and C was tested as 3.13-200 μg/mL. Compound A and D were firstly isolated from fungal extracts. This study lays a foundation for the further resource development and utilization of litter fungi in the Greater Khingan Mountains.
大兴安岭 / 凋落物 / 真菌 / 抗菌活性 / 次级代谢产物 / 化合物分离 {{custom_keyword}} /
the Greater Khingan Mountains / litter / fungus / antibacterial activity / secondary metabolites / compound separation {{custom_keyword}} /
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