东北红豆杉内生真菌Fusarium tricinctum J8的鉴定及抗菌活性研究  

Identification and Antimicrobial Activity of Endophytic Fungus Fusarium tricinctum J8 from Taxus Cuspidate

作　　者：傅岳峰 徐铖 魏巍[1] 崔海超 宋金凤[2] 顾成波[1,2] 袁肖寒 Fu Yuefeng(Key Laboratory of Forest Plant Ecology of Ministry of Education,Heilongjiang Provincial Key Laboratory of Ecological Utilization of Forestry-Based Active Substances,Engineering Research Center of Forest Bio-Preparation of Ministry of Education,College of Chemistry,Chemical Engineering and Resource Utilization,Northeast Forestry University,Harbin Heilongjiang 150040)

机构地区：[1]东北林业大学,化学化工与资源利用学院,森林植物生态学教育部重点实验室,林业生物制剂教育部工程研究中心,黑龙江省林源活性物质生态利用重点实验室,哈尔滨150040 [2]东北林业大学林学院,森林生态系统可持续经营教育部重点实验室,哈尔滨150040 [3]东北农业大学生命科学与生物技术研究中心,哈尔滨150030

出　　处：《黑龙江医药》2023年第3期497-501,共5页Heilongjiang Medicine journal

基　　金：黑龙江省自然科学基金联合引导项目(编号:LH2021C015);黑龙江头雁创新团队计划(林木遗传育种创新研究团队);森林生态系统可持续经营教育部重点实验室基金(编号:KFJJ2021YB01)。

摘　　要：目的:从东北红豆杉内生真菌中筛选出一株对常见致病细菌具有广谱拮抗活性的菌株。方法:以大肠杆菌、白色念珠菌、金黄色葡萄球菌、蜡样芽孢杆菌4种常见致病细菌为指示菌,采用滤纸片扩散法筛选具有广谱抗菌活性的内生真菌,基于形态观察与系统发育分析对该菌株进行种属鉴定,利用高效液相色谱(HPLC)分析该菌株不同发酵时间次级代谢产物的代谢情况,通过超高效液相色谱-四极杆-静电场轨道阱高分辨质谱(UPLC-Q-Orbitrap-MS/MS)分析该菌株可能产生的抗菌物质。结果:通过筛选获得一株广谱拮抗菌株J8,鉴定为三线镰刀菌(Fusarium tricinctum),根据HPLC色谱图对比,确定最佳培养时间为8天,通过mzCloud数据库比对分析其质谱数据,发现该菌株产生环(脯氨酸-亮氨酸)二肽、环(脯氨酸-苏氨酸)二肽、恩镰孢菌素B1、曲酸4种匹配度大于90%的抗菌物质。结论:从东北红豆杉中获得了一株对4种常见致病细菌均具有抑菌活性的菌株Fusarium tricinctum J8,为开发新型抗生素候选药物提供了菌种资源。Objective:To screen a strain with broad spectrum antagonistic activity against common pathogenic bacteria from the endophytic fungi of Taxus cuspidate.Methods:Four common pathogenic bacteria were used as indicators to screen endophytic fungus with broad-spectrum antibacterial activity by filter paper diffusion method.The specie of the strain was identified based on morphological observation and phylogenetic analysis.HPLC was used to analyze the abundance of fungal secondary metabolites in crude extracts on different days to determine the optimal fermentation days.UPLC-Q-Orbitrap-MS/MS was used to detect the possible antibacterial substances produced by the strain.Results:Endophytic fungus J8 was screened by antibacterial test and identified as Fusarium tricinctum.According to the comparison of HPLC chromatograph,the optimal culture time was determined to be 8 days.The mass spectrum data were compared and analyzed by mzCloud database,and 4 antibacterial active substances with matching degree greater than 90%were found.Conclusion:Fusarium tricinctum J8,a strain with with broad-spectrum antibacterial activity,was selected from endophytic fungi of Taxus cuspidate,which provides strain resource for the development of new antibiotic drugs.

关 键 词：东北红豆杉 内生真菌 三线镰刀菌 抗菌活性 

分 类 号：R284.1[医药卫生—中药学]