蛇足石杉1株产生物碱内生真菌的分离鉴定
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摘要
【目的】从蛇足石杉[Huperzia serrata(Thunb.)Trev.]植株中分离内生真菌以及筛选能产生物碱的菌株,并对产生物碱菌株进行鉴定。【方法】采用内生真菌常用的方法从蛇足石杉健康植株中分离获得内生真菌,采用生物碱沉淀法和薄层色谱法(TLC)对分离得到的内生真菌的代谢产物进行检测,分析是否产生物碱,并应用形态学、分子生物学和生物信息学等方法对其进行鉴定和系统发育分析。【结果】从蛇足石杉茎中分离的内生真菌,筛选出1株产生物碱菌株LYS081,初步鉴定为木霉属(Trichoderma)真菌。【结论】蛇足石杉内生真菌LYS081能产生生物碱,在天然产物活性研究和资源开发以及内生真菌的研究中具有重要的意义。
【Objective】The endophytic fungi and the strains that could produce alkaloids were isolated and identified from Huperzia serrata. 【Method】The endophytic fungi were isolated from Huperzia serrata by means of alkaloid precipitation and TLC, the metabolites of the isolated endophytic fungi were detected, the alkaloid production was analyzed, and morphological, molecular biology and bioinformatics methods were used to identify and systematize them. 【Result】A strain of alkaloid-producing strain LYS081 was isolated from the stem of Huperzia serrata and identified as Trichoderma fungus. 【Conclusion】The endophytic fungus LYS081 isolated from Huperzia serrata can produce alkaloids, which is of great significance in the research of natural product activity, resource development and endophytic fungi.
【Objective】The endophytic fungi and the strains that could produce alkaloids were isolated and identified from Huperzia serrata. 【Method】The endophytic fungi were isolated from Huperzia serrata by means of alkaloid precipitation and TLC, the metabolites of the isolated endophytic fungi were detected, the alkaloid production was analyzed, and morphological, molecular biology and bioinformatics methods were used to identify and systematize them. 【Result】A strain of alkaloid-producing strain LYS081 was isolated from the stem of Huperzia serrata and identified as Trichoderma fungus. 【Conclusion】The endophytic fungus LYS081 isolated from Huperzia serrata can produce alkaloids, which is of great significance in the research of natural product activity, resource development and endophytic fungi.
引文
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[14]韩文霞,宋涛,李伟泽,等. 产石杉碱甲新资源研究方法探索[J]. 海南医学,2015,26(15):2275.
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[16]闵长莉,汪学军. 蛇足石杉产石杉碱甲内生真菌的分离鉴定[J]. 天然产物研究与开发,2013,25(5):590-593.
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[18]邵力平.真菌分类学[M]. 北京:中国林业出版社,1984.
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[21]Weindling R. Trichoderma lignorum as a parasite of other soil fungi[J]. Phytopathology, 1932, 22: 837-845.
[22]Gajera H, Domadiya R, Patel S, et al. Molecular mechanism of Trichoderma as bio-control agents against phytopathogen system a review[J]. Curr Res Microbiol Biotechnol, 2013, 1(4): 133-142.
[23]李向东,王云强,王卉,等. 铁皮石斛软腐病的生物防治[J]. 中国药学杂志,2013,48(19):1669-1673.
[24]魏洪璇,陈向东,汪辉,等. 哈茨木霉的分离纯化、鉴定及发酵液活性的初步研究[J].药物生物技术,2017,24(3):206-210.
[25]CHENG L, ZHOUX J. Review on anticancermechanism ofsome plant alkaloids[J]. Chin TraditHerb Drugs(中草药), 2004,35(2): 216-221.
[26]张弘弛,刘瑞,周凤,等. 无花果内生真菌 FL10 中吲哚二酮哌嗪类生物碱的研究[J]. 中国实验方剂学杂志,2013,19(18):95-98.
[27]Ahimsa-Müller M A, Markert A, Hellwig S, et al. Clavicipitaceous fungi associated with ergoline alkaloid containing convolvulaceae[J]. J Nat Prod, 2007, 70(12): 1955-1960.
[28]Zhu D,Wang J,Zeng Q,et al. A novel endophytic huperzine A producing fungus,Shiraia sp. Slf14,isolated from Huperzia serrata[J]. J Appl Microbiol,2010,109: 1469.
[2]Strobel G, Stierle A, Stierle D. Taxol and taxane production by Taxomycetes andreanae,an endophytic fungus of Pacific Yew[J]. Science, 1993, 260(5105): 214-216.
[3]李建秀,周凤琴,张照荣,等. 山东药用植物志[M]. 西安:西安交通大学出版社,2013.
[4]王月娥,岳冬贤,唐希灿. 石杉碱甲的抗胆碱酯酶作用[J].中国药理学报,1986,7(2):110-113.
[5]张磊,万谦宏,高文远. 石杉碱甲的研究进展[J]. 中草药,2005,36(9):1422-1425.
[6]Yu H Y,Sun Y M,Yang Y J. Advances in studies on Huperzia serrata[J]. Chinese Traditional Herbal Drugs, 2001, 32(3): 279-281.
[7]王瑞萍,赵政凯,胡黎立. 石杉碱甲对血管性痴呆患者认知功能及生活能力的影响:36例6个月随访[J]. 中国临床康复,2004,8(19):3892.
[8]陈婕,朱文炳,汪昕,等. 石杉碱甲片剂(双益平)治疗记忆障碍的临床研究[J]. 中国临床医学,2000,7(1):63.
[9]廖建湘,陈黎,黄铁栓,等. 石杉碱甲治疗小儿语言发育迟缓的初步观察[J]. 儿科药学杂志,2002,8(1):26.
[10]林立,陈榕,陈学水,等. 精神发育迟滞的综合治疗(附20例石杉碱甲片治疗观察)[J]. 海峡药学,1999,11(增刊2):31.
[11]夏强,刘才群. 石杉碱甲治疗重症肌无力临床观察[J]. 实用医学杂志,2002,19(1): 31.
[12]杨晓阳,张宏颖. 石杉碱甲治疗脑卒中后尿失禁的疗效观察[J]. 实用神经疾病杂志,2004,7(5):77.
[13]王子文,张航向,任平. 哈伯因治疗阿尔茨海默病的临床对比研究[J]. 医学信息, 2000,13(8):462.
[14]韩文霞,宋涛,李伟泽,等. 产石杉碱甲新资源研究方法探索[J]. 海南医学,2015,26(15):2275.
[15]Ward J,Caprio V. A radical mediated approach to the core structure of huperzine A[J]. Tetrahedron Lett,2006,47: 553.
[16]闵长莉,汪学军. 蛇足石杉产石杉碱甲内生真菌的分离鉴定[J]. 天然产物研究与开发,2013,25(5):590-593.
[17]Zhang Z B,Zeng Q G,Yan R M,et al. Endophytic fungus Cladosporium cladosporioides LF70 from Huperzia serrata produces huperzine A[J]. World J Microbiol Biotechnol,2010,doi:10.1007/s11274-010-0476-6.
[18]邵力平.真菌分类学[M]. 北京:中国林业出版社,1984.
[19]魏景超. 真菌鉴定手册[M]. 上海:上海科学技术出版社,1979.
[20]H. L. Barnett,Barry B. Hunter. Illustrated Genera of Imperfect Fungi[M]. APS Press,Fourth Edition,1999.
[21]Weindling R. Trichoderma lignorum as a parasite of other soil fungi[J]. Phytopathology, 1932, 22: 837-845.
[22]Gajera H, Domadiya R, Patel S, et al. Molecular mechanism of Trichoderma as bio-control agents against phytopathogen system a review[J]. Curr Res Microbiol Biotechnol, 2013, 1(4): 133-142.
[23]李向东,王云强,王卉,等. 铁皮石斛软腐病的生物防治[J]. 中国药学杂志,2013,48(19):1669-1673.
[24]魏洪璇,陈向东,汪辉,等. 哈茨木霉的分离纯化、鉴定及发酵液活性的初步研究[J].药物生物技术,2017,24(3):206-210.
[25]CHENG L, ZHOUX J. Review on anticancermechanism ofsome plant alkaloids[J]. Chin TraditHerb Drugs(中草药), 2004,35(2): 216-221.
[26]张弘弛,刘瑞,周凤,等. 无花果内生真菌 FL10 中吲哚二酮哌嗪类生物碱的研究[J]. 中国实验方剂学杂志,2013,19(18):95-98.
[27]Ahimsa-Müller M A, Markert A, Hellwig S, et al. Clavicipitaceous fungi associated with ergoline alkaloid containing convolvulaceae[J]. J Nat Prod, 2007, 70(12): 1955-1960.
[28]Zhu D,Wang J,Zeng Q,et al. A novel endophytic huperzine A producing fungus,Shiraia sp. Slf14,isolated from Huperzia serrata[J]. J Appl Microbiol,2010,109: 1469.