期刊论文详细信息
BMC Complementary and Alternative Medicine
Antimicrobial activities of endophytic fungi obtained from the arid zone invasive plant Opuntia dillenii and the isolation of equisetin, from endophytic Fusarium sp.
Raymond J. Andersen2  David E. Williams2  E. Dilip de Silva3  Pamoda B. Ratnaweera1 
[1] Department of Science and Technology, Uva Wellassa University, Badulla, Sri Lanka;Department of Chemistry and Earth, Ocean and Atmospheric Science, University of British Columbia, Vancouver, Canada;Department of Chemistry, University of Colombo, Colombo 03, Sri Lanka
关键词: Fusarium sp.;    Arid ecosystem;    Equisetin;    Opuntia dillenii;    Invasive plants;    Endophytic fungi;   
Others  :  1220067
DOI  :  10.1186/s12906-015-0722-4
 received in 2014-12-22, accepted in 2015-06-12,  发布年份 2015
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【 摘 要 】

Background

Opuntia dillenii is an invasive plant well established in the harsh South-Eastern arid zone of Sri Lanka. Evidence suggests it is likely that the endophytic fungal populations of O. dillenii assist the host in overcoming biotic and abiotic stress by producing biologically active metabolites. With this in mind there is potential to discover novel natural products with useful biological activities from this hitherto poorly investigated source. Consequently, an investigation of the antimicrobial activities of the endophytes of O. dillenii, that occupies a unique ecological niche, may well provide useful leads in the discovery of new pharmaceuticals.

Methods

Endophytic fungi were isolated from the surface sterilized cladodes and flowers of O. dillenii using several nutrient media and the antimicrobial activities were evaluated against three Gram-positive and two Gram-negative bacteria and Candida albicans. The two most bioactive fungi were identified by colony morphology and DNA sequencing. The secondary metabolite of the endophyte Fusarium sp. exhibiting the best activity was isolated via bioassay guided chromatography. The chemical structure was elucidated from the ESIMS and NMR spectroscopic data obtained for the active metabolite. The minimum inhibitory concentrations (MICs) of the active compound were determined.

Results

Eight endophytic fungi were isolated from O. dillenii and all except one showed antibacterial activities against at least one of the test bacteria. All extracts were inactive against C. albicans. The most bioactive fungus was identified as Fusarium sp. and the second most active as Aspergillus niger. The structure of the major antibacterial compound of the Fusarium sp. was shown to be the tetramic acid derivative, equisetin. The MIC’s for equisetin were 8 μg mL −1against Bacillus subtilis, 16 μg mL −1against Staphylococcus aureus and Methicillin Resistant Staphylococcus aureus (MRSA).

Conclusions

O. dillenii, harbors several endophytic fungi capable of producing antimicrobial substances with selective antibacterial properties. By producing biologically active secondary metabolites, such as equisetin isolated from the endophytic Fusarium sp., the endophytic fungal population may be assisting the host to successfully withstand stressful environmental conditions. Further investigations on the secondary metabolites produced by these endophytes may provide additional drug leads.

【 授权许可】

   
2015 Ratanweera et al.

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【 参考文献 】
  • [1]Rodriguez RJ, White Jr JF, Arnold AE, Redman RS. Fungal endophytes:diversity and functional roles. New Phytol. 2009;182:314–330.
  • [2]Zhang HW, Song YC, Tan RX. Biology and chemistry of endophytes. Nat Prod Rep. 2006; 23:753-71.
  • [3]Yadav M, Yadav A, Kumar S, Sharma D, Yadav JP. Evaluation of in vitro antimicrobial potential of endophytic fungi isolated from Eugenia Jambolana Lam. Int J Pharm Pharm Sci. 2014; 6:208-11.
  • [4]Tan RX, Zou WX. Endophytes: a rich source of functional metabolites. Nat Prod Rep. 2001; 18:448-59.
  • [5]Yu H, Zhang L, Li L, Zheng C, Guo L, Li W, Sun P, Qin L. Recent developments and future prospects of antimicrobial metabolites produced by endophytes. Microbiol Res. 2010; 165:437-49.
  • [6]Arid zone programme, A note on the climate and vegetation of the arid zone of Ceylon. United Nations Educational Scientific and Cultural Organization, 1951, UNESCO/NS/AZ/56, Paris; 1951.
  • [7]Ranwala S. Flora of Sri Lanka. Training course for national tourist guides, conducted by the Tourist Board, Sri Lanka, Department of Plant Science, University of Colombo; 2012.
  • [8]Invasive Species Advisory Committee (ISAC): Invasive species definition clarification and guidance white paper. The National Invasive Species Council (NISC), 2006. http://www.invasivespeciesinfo.gov/docs/council/isacdef.pdf, accessed date: 15 June 2015.
  • [9]The 2007 Red List of Threatened Fauna and Flora of Sri Lanka. The World Conservation Union (IUCN) and Ministry of Environment and Natural Resources, Colombo, Sri Lanka; 2007.
  • [10]Alwis SMDAU, Dayawansa PN, How R, Padmalal UKGK, Singhakumara BMP, Weerakoon D, Wijesinghe MR, editors. Biodiversity Baseline Survey. In: Bundala National Park Consultancy Services Report prepared by Green MJB. Sri Lanka Protected Areas Management and Wildlife Conservation Project (PAM&WCP/CONSULT/02/BDBS), Department of Wildlife Conservation, Ministry of Environment and Natural Resources, Colombo; 2008: p.46.
  • [11]Beeson CFC. Prickly pear and cochineal insects. Indian Forester. 1934; 60:203-5.
  • [12]Gunatilaka AAL. Natural products from plant-associated microorganisms: Distribution, Structural Diversity, bioactivity and implication of their occurrence. J Nat Prod. 2006; 69(3):509-26.
  • [13]Newcombe G, Shipunov A, Eigenbrode SD, Raghavendra AKH, Ding H, Anderson AL, Menjivar R, Craeford M, Schwarzlander M. Endophytes influence protection and growth of an invasive plant. Commun Integr Biol. 2009; 2(1):29-31.
  • [14]Weerasooriya A. Cactaceae. In: A Revised Handbook to the Flora of Ceylon, Volume XIII. Dassanayake MD, Clayton WD, editors. Oxford & IBH Publishing CO. PVT. LTD, New Delhi, Calcutta; 1999: p.41-4.
  • [15]Ratnaweera PB, Williams DE, de Silva ED, Wijesundera RLC, Dalisay DS, Andersen RJ. Helvolic acid, an antibacterial nortriterpenoid from a fungal endophyte, Xylaria sp. of orchid Anoectochilus setaceus endemic to Sri Lanka. Mycology. 2014; 5(1):23-8.
  • [16]National Committee for Clinical Laboratory Standards. Performance standards for antimicrobial susceptibility tests, approved standard, document M2-A8: NCCLS, 8th ed. Wayne (PA); 2003.
  • [17]Kariyawasam GK, Mithrasena YJPK, Fernando THPS, Wijesundera RLC, Wijesundera WSS. A new cost effective method for extracting genomic DNA from fungi. In: Abstracts of papers, 5th Annual sessions of Institute of Biochemistry, Molecular Biology and Biotechnology Colombo. Institute of Biochemistry, Molecular Biology and Biotechnology, Colombo; 2012: p.49.
  • [18]NCBI GenBank. [http://. http://blast. ncbi.nlm.nih.gov/Blast.cgi webcite
  • [19]National Committee for Clinical Laboratory Standards. Performance standards for antimicrobial susceptibility testing, twelfth information supplement, M100-S12: NCCLS. Wayne (PA); 2002.
  • [20]de Freitas SE, Marcon J, Luvizotto DM, Quecine MC, Tsui S, Pereira JO, Pizzirani-Kleiner AA, Azevedo JL. Endophytic fungi from the Amazonian plant Paullinia cupanaand from Olea europaea isolated using cassava as an alternative starch media source. Springerplus. 2013; 2:579. BioMed Central Full Text
  • [21]O’Donnell K, Sutton DA, Rinaldi MG, Gueidan C, Crous PW, Geiser DM. Novel multilocus sequence typing scheme reveals high genetic diversity of human pathogenic members of the Fusarium incarnatum-F. equiseti and F. chlamydosporium species complexes within the United States. J Clin Microbiol. 2009; 47(12):3851-61.
  • [22]Lv XC, Huang ZQ, Zhang W, Rao PF, Ni L. Identification and characterization of filamentous fungi isolated from fermentation starters for Hong Qu glutinous rice wine brewing. J Gen Appl Microbiol. 2012; 58(1):33-42.
  • [23]Ferreira IRCG, Wolff T, Scheinvar LA, Castro FAV, Pereira MD, Almeida RV, Valente LMM. Investigation of new and/or bioactive metabolites from endophytic fungi associated with Opuntia moncantha (Cactacease). In Proceedings of 22nd Symposium of Medicinal Plants of Brazil. 2012;18–21. http://www.ufrgs.br/spmb2012/Trabalhos/3430_1337084576_Resumo.SPMB.2012.fungos_endof%C3%ADticos.pdf Accessed date: 15 June 2015.
  • [24]Fisher PJ, Sutton BC, Petrini LE, Petrini O. Fungal endophytes from Opuntia stricta: a first report. Nova Hedwig. 1994; 59:195-200.
  • [25]Philips NJ, Goodwin JT, Fraiman A, Cole RJ, Lynn DG. Characterization of the Fusarium Toxin Equisetin: The use of Phenylboronates in structure assignment. J Am Chem Soc. 1989; 3(21):8223-31.
  • [26]Burmeister HR, Bennett GA, Vesonder RF, Hesseltine CW. Antibiotic produced by Fusarium equiseti NRRL 5537. Antimicrob Agents Chemother. 1974; 5(6):634-9.
  • [27]Singh SB, Zink DL, Goetz MA, Dombrowski AW, Polishhook JD, Hazuda DJ. Equisetin and a novel opposite stereochemical homolog Phomasetin, two fungal metabolites as inhibitors of HIV-1 Integrase. Tetrahedron Lett. 1998; 39:2243-6.
  • [28]Yuan Y, Tang J, Leng D, Hu S, Yong JWH, Chen X. Invasive plant promotes its arbuscular Mycorrhizal symbioses and competitiveness through its secondary metabolite: indirect evidence from activated carbon. PLoS One. 2014; 9(5):1-9.
  • [29]Yang Q, Ye W, Liao F, Yin X. Effects of allelochemical on seed germination. Chin J Ecol. 2005; 24:1459-65.
  • [30]Alford ER, Perry LG, Qin B, Vivanco JM, Paschke MW. A putative allelopathic agents of Russian knapweed occurs in invaded soils. Soil Biol Biochem. 2007; 39:1812-5.
  • [31]Shipunov A, Newcombe G, Raghavendra AKH, Andersen CL. Hidden diversity of endophytic fungi in an invasive plant. Am J Bot. 2008; 95(9):1096-108.
  • [32]Aschehoug ET, Metlen KL, Callaway RM, Newcombe G. Fungal endophytes directly increase the competitive effects of an invasive forb. Ecology. 2012; 93(1):3-8.
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