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  • 卢护木,詹振宇,李蜜,罗双宇,高程海,罗小卫,刘永宏.涠洲岛珊瑚礁海洋真菌的分离鉴定及其抗MRSA活性筛选[J].广西科学,2020,27(5):520-525.    [点击复制]
  • LU Humu,ZHAN Zhenyu,LI Mi,LUO Shuangyu,GAO Chenghai,LUO Xiaowei,LIU Yonghong.Isolation,Identification,and Anti-MRSA Activity Screening of Marine Symbiotic Fungi Derived from the Weizhou Island Coral Reef[J].Guangxi Sciences,2020,27(5):520-525.   [点击复制]
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涠洲岛珊瑚礁海洋真菌的分离鉴定及其抗MRSA活性筛选
卢护木, 詹振宇, 李蜜, 罗双宇, 高程海, 罗小卫, 刘永宏
0
(广西中医药大学海洋药物研究院, 广西南宁 530200)
摘要:
本研究从广西涠洲岛礁栖珊瑚、海绵和海藻等样品中挖掘海洋真菌资源,旨在筛选发现潜在的具有抗耐甲氧西林金黄色葡萄球菌(MRSA)活性的菌株。研究首先采用稀释分离法从采集的珊瑚、海绵和海藻等样品中分离纯化出真菌,基于内转录间隔区(ITS)序列和系统发育树分析对菌种进行鉴定;然后采用大米培养基对分离得到的单一菌株进行小量发酵,利用乙酸乙酯提取浓缩获得提取物;最后采用薄层色谱(TLC)、高效液相色谱(HPLC-DAD)等方法分析菌株提取物中代谢产物的丰富度,并采用滤纸片琼脂扩散法对菌株粗提物进行抗MRSA活性初步筛选。结果从样品中分离获得40株海洋真菌,以曲霉属(Aspergillus)和木霉属(Trichoderma)为主,其中A.terreus MF01、A.flavus MF07、T.asperellum HP01、Fusarium equiseti HP08和A.unguis HP17等菌株具有丰富的代谢产物及抗MRSA活性。本研究发现的菌株MF01、MF07、HP01、HP08和HP17具有潜在的抗MRSA活性,为后续开展活性代谢产物研究提供基础资料。
关键词:  涠洲岛  珊瑚礁  海洋真菌  分离鉴定  抗MRSA活性
DOI:10.13656/j.cnki.gxkx.20201214.011
基金项目:广西高校中青年教师科研基础能力提升项目(2020KY07032),广西自然科学基金项目(2020GXNSFBA159001),广西壮族自治区大学生创新创业训练计划项目(202010600093),广西中医药大学2019年博士科研启动基金项目(2019BS021),广西中医药大学海洋药物研究院团队科研专项(2018ZD005),广西中医药大学岐黄工程高层次人才团队培育项目(2018006)和广西八桂学者专项(05019055)资助。
Isolation,Identification,and Anti-MRSA Activity Screening of Marine Symbiotic Fungi Derived from the Weizhou Island Coral Reef
LU Humu, ZHAN Zhenyu, LI Mi, LUO Shuangyu, GAO Chenghai, LUO Xiaowei, LIU Yonghong
(Institute of Marine Drugs, Guangxi University of Chinese Medicine, Nanning, Guangxi, 530200, China)
Abstract:
The objective of this study was to isolate fungal strains from the samples of reef-dwelling coral,sponge,and algae collected from the Weizhou Island Coral Reef and further to discover the potential bioactive strains by anti-methicillin resistant Staphylococcus aureus(MRSA) activity screening.Firstly,the fungi were isolated and purified from these coral,sponge,and algae by dilution separation method,and were preliminarily identified by analysis of the internal transcriptional spacer (ITS) sequence and phylogenetic tree.Then the isolated strains were subjected to the small-scale fermentations based on solid rice culture medium,and the extract was obtained by extraction and concentration with ethyl acetate.Finally,Thin Layer Chromatography (TLC) and High-Performance Liquid Chromatography (HPLC-DAD) and other methods were used to analyze the abundance of metabolites in the strain extract.And the anti-MRSA activity of the crude extracts of the strains was preliminarily screened by filter paper agar diffusion method.Forty fungal strains were isolated and purified from the above samples,while Aspergillus and Trichoderma were found to be the predominant genera.The strains A.terreus MF01,A.flavus MF07,T.asperellum HP01,Fusarium equiseti HP08 and A.unguis HP17 were found with abundant metabolites and anti-MRSA activities.The strains of MF01,MF07,HP01,HP08 and HP17 have potential anti-MRSA activities,which provide basic information for subsequent studies on active metabolites.
Key words:  Weizhou Island  coral reef  marine fungi  isolation and identification  anti-MRSA activity

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