Advertisement

Applied Microbiology and Biotechnology

, Volume 98, Issue 16, pp 7173–7183 | Cite as

Quorum quenching is an antivirulence strategy employed by endophytic bacteria

  • Parijat Kusari
  • Souvik Kusari
  • Marc Lamshöft
  • Selahaddin Sezgin
  • Michael Spiteller
  • Oliver Kayser
Applied microbial and cell physiology

Abstract

Bacteria predominantly use quorum sensing to regulate a plethora of physiological activities such as cell-cell crosstalk, mutualism, virulence, competence, biofilm formation, and antibiotic resistance. In this study, we investigated how certain potent endophytic bacteria harbored in Cannabis sativa L. plants use quorum quenching as an antivirulence strategy to disrupt the cell-to-cell quorum sensing signals in the biosensor strain, Chromobacterium violaceum. We used a combination of high-performance liquid chromatography high-resolution mass spectrometry (HPLC-ESI-HRMSn) and matrix-assisted laser desorption ionization imaging high-resolution mass spectrometry (MALDI-imaging-HRMS) to first quantify and visualize the spatial distribution of the quorum sensing molecules in the biosensor strain, C. violaceum. We then showed, both quantitatively and visually in high spatial resolution, how selected endophytic bacteria of C. sativa can selectively and differentially quench the quorum sensing molecules of C. violaceum. This study provides fundamental insights into the antivirulence strategies used by endophytes in order to survive in their ecological niches. Such defense mechanisms are evolved in order to thwart the plethora of pathogens invading associated host plants in a manner that prevents the pathogens from developing resistance against the plant/endophyte bioactive secondary metabolites. This work also provides evidence towards utilizing endophytes as tools for biological control of bacterial phytopathogens. In continuation, such insights would even afford new concepts and strategies in the future for combating drug resistant bacteria by quorum-inhibiting clinical therapies.

Keywords

Bacterial endophytes Quorum quenching N-acylated homoserine lactones Cannabis sativaHigh-resolution mass spectrometry MALDI imaging high-resolution mass spectrometry Microbe-microbe interaction Microbe-plant interaction Phytopathology 

Notes

Acknowledgments

This research was funded by the Ministry of Innovation, Science and Research of the German Federal State North Rhine-Westphalia (NRW) and TU Dortmund by a scholarship to P.K. from the CLIB-Graduate Cluster Industrial Biotechnology. We thank the German Research Foundation (DFG) for financing the MALDI imaging high-resolution mass spectrometers. We are thankful to Federal Institute for Drugs and Medical Devices (Bundesinstituts für Arzneimittel und Medizinprodukte, BfArM), Bonn, Germany for granting us the necessary permissions for working with Cannabis plants (BtM number 458 49 89). S.K. was a Visiting Researcher at the Department of Plant Sciences, University of Oxford, South Parks Road, Oxford OX1 15 3RB, UK, during part of the work and preparation of the manuscript (Apr 2013 to Mar 2014). S.K. gratefully acknowledges M.S. for approving and authorizing, Dr. Gail M. Preston for hosting, and TU Dortmund for supporting his stay at the University of Oxford.

Conflict of Interest

The authors declare no conflict of interest.

References

  1. Aly AH, Debbab A, Proksch P (2013) Fungal endophytes—secret producers of bioactive plant metabolites. Pharmazie 68:499–505PubMedGoogle Scholar
  2. Amara N, Krom BP, Kaufmann GF, Meijiler MM (2011) Macromolecular inhibition of quorum sensing: enzymes, antibodies, and beyond. Chem Rev 111:195–208PubMedCrossRefGoogle Scholar
  3. Amaral L, Martins A, Spengler G, Molnar J (2014) Efflux pumps of Gram-negative bacteria: what they do, how they do it, with what and how to deal with them. Front Pharmacol 4:168. doi: 10.3389/fphar.2013.00168 PubMedCentralPubMedCrossRefGoogle Scholar
  4. Bjarnholt N, Li B, D’Alvise J, Janfelt C (2014) Mass spectrometry imaging of plant metabolites—principles and possibilities. Nat Prod Rep. doi: 10.1039/c3np70100j PubMedGoogle Scholar
  5. Burmølle M, Thomsen TR, Fazli M, Dige I, Christensen L, Homøe P, Tvede M, Nyvad B, Tolker-Nielsen T, Givskov M, Moser C, Kirketerp-Møller K, Johansen HK, Høiby N, Jensen P, Sørensen SJ, Bjarnsholt T (2010) Biofilms in chronic infections—a matter of opportunity—monospecies biofilms in multispecies infections. FEMS Immmunol Med Microbiol 59:324–336Google Scholar
  6. Choo JH, Rukayadi Y, Hwang J-K (2006) Inhibition of bacterial quorum sensing by vanilla extract. Lett Appl Microbiol 42:637–641PubMedGoogle Scholar
  7. Christiaen SEA, Matthijs N, Zhang X-H, Nelis HJ, Bossier P, Coenye T (2014) Bacteria that inhibit quorum sensing decrease biofilm formation and virulence in Pseudomonas aeruginosa PAO1. Pathog Dis 70:271–279PubMedCrossRefGoogle Scholar
  8. Claessen D, Rozen DE, Kuipers OP, Søgaard-Andersen L, van Wezel GP (2014) Bacterial solutions to multicellularity: a tale of biofilms, filaments and fruiting bodies. Nat Rev Microbiol. doi: 10.1038/nrmicro3178 PubMedGoogle Scholar
  9. Clatworthy AE, Pierson E, Hung DT (2007) Targeting virulence: a new paradigm for antimicrobial therapy. Nat Chem Biol 3:541–548PubMedCrossRefGoogle Scholar
  10. Cornforth DM, Popat R, McNally L, Gurney J, Scott-Phillips TC, Ivens A, Diggle SP, Brown SP (2014) Combinatorial quorum sensing allows bacteria to resolve their social and physical environment. Proc Natl Acad Sci U S A 111:4280–4284PubMedCentralPubMedCrossRefGoogle Scholar
  11. Galloway WRJD, Hodgkinson JT, Bowden SD, Welch M, Spring DR (2011) Quorum sensing in Gram-negative bacteria: small-molecule modulation of AHL and AI-2 quorum sensing pathways. Chem Rev 111:28–67PubMedCrossRefGoogle Scholar
  12. Grotenhermen F, Müller-Vahl K (2012) The therapeutic potential of Cannabis and cannabinoids. Dtsch Arztebl Int 109:495–501PubMedCentralPubMedGoogle Scholar
  13. Hosni T, Moretti C, Devescovi G, Suarez-Moreno ZR, Fatmi MB, Guarnaccia C, Pongor S, Onofri A, Buonaurio R, Venturi V (2011) Sharing of quorum-sensing signals and role of interspecies communities in a bacterial plant disease. ISME J 5:1857–1870PubMedCentralPubMedCrossRefGoogle Scholar
  14. Kharwar RN, Mishra A, Gond SK, Stierle A, Stierle D (2011) Anticancer compounds derived from fungal endophytes: their importance and future challenges. Nat Prod Rep 28:1208–1228PubMedCrossRefGoogle Scholar
  15. Kim S-H, Park H-D (2013) Ginger extract inhibits biofilm formation by Pseudomonas aeruginosa PA14. PLoS ONE 8:e76106PubMedCentralPubMedCrossRefGoogle Scholar
  16. Kusari S, Spiteller M (2011) Are we ready for industrial production of bioactive plant secondary metabolites utilizing endophytes? Nat Prod Rep 28:1203–1207PubMedCrossRefGoogle Scholar
  17. Kusari S, Zühlke S, Spiteller M (2009a) An endophytic fungus from Camptotheca acuminata that produces camptothecin and analogues. J Nat Prod 72:2–7PubMedCrossRefGoogle Scholar
  18. Kusari S, Lamshöft M, Spiteller M (2009b) Aspergillus fumigatus Fresenius, an endophytic fungus from Juniperus communis L. Horstmann as a novel source of the anticancer pro-drug deoxypodophyllotoxin. J Appl Microbiol 107:1019–1030PubMedCrossRefGoogle Scholar
  19. Kusari S, Hertweck C, Spiteller M (2012) Chemical ecology of endophytic fungi: origins of secondary metabolites. Chem Biol 19:792–798PubMedCrossRefGoogle Scholar
  20. Kusari P, Kusari S, Spiteller M, Kayser O (2013) Endophytic fungi harbored in Cannabis sativa L.: diversity and potential as biocontrol agents against host plant-specific phytopathogens. Fungal Divers 60:137–151CrossRefGoogle Scholar
  21. Kusari S, Singh S, Jayabaskaran C (2014) Biotechnological potential of plant-associated endophytic fungi: hope versus hype. Trends Biotechnol. doi: 10.1016/j.tibtech.2014.03.009 Google Scholar
  22. Lane DJ (1991) 16S/23S rRNA sequencing. In: Stackebrandt E, Goodfellow M (eds) Nucleic acid techniques in bacterial systematics. Wiley, New York, pp 115–175Google Scholar
  23. LaSarre B, Federle MJ (2013) Exploiting quorum sensing to confuse bacterial pathogens. Microbiol Mol Biol Rev 77:73–111PubMedCentralPubMedCrossRefGoogle Scholar
  24. Lau YY, Sulaiman J, Chen JW, Yin W-F, Chan K-G (2013) Quorum sensing activity of Enterobacter asburiae isolated from lettuce leaves. Sensors 13:14189–14199PubMedCentralPubMedCrossRefGoogle Scholar
  25. Limsuwan S, Voravuthikunchai SP (2008) Boesenbergia pandurata (Roxb.) Schltr., Eleutherine americana Merr. and Rhodomyrtus tomentosa (Aiton) Hassk. as antibiofilm producing and antiquorum sensing in Streptococcus pyogenes. FEMS Immunol Med Microbiol 53:429–436PubMedCrossRefGoogle Scholar
  26. Márquez LM, Redman RS, Rodriguez RJ, Roossinck MJ (2007) A virus in a fungus in a plant: three‑way symbiosis required for thermal tolerance. Science 315:513–515PubMedCrossRefGoogle Scholar
  27. McClean KH, Winson MK, Fish L, Taylor A, Chhabra SR, Camara M, Daykin M, Lamb JH, Swift S, Bycroft BW, Stewart GSAB, Williams P (1997) Quorum sensing and Chromobacterium violeceum: exploitation of violacein production and inhibition for the detection of N-acyl homoserine lactones. Microbiology 143:3703–3711PubMedCrossRefGoogle Scholar
  28. Morohoshi T, Fukamachi K, Kato M, Kato N, Ikeda T (2010) Regulation of the violacein biosynthetic gene cluster by acylhomoserine lactone-mediated quorum sensing in Chromobacterium violaceum ATCC 12472. Biosci Biotechnol Biochem 74:2116–2119PubMedCrossRefGoogle Scholar
  29. Morohoshi T, Tokita K, Ito S, Saito Y, Maeda S, Kato N, Ikeda T (2013) Inhibition of quorum sensing in gram-negative bacteria by alkylamine-modified cyclodextrins. J Biosci Bioeng 116:175–179PubMedCrossRefGoogle Scholar
  30. O’Loughlin CT, Miller LC, Siryaporn A, Drescher K, Semmelhack MF, Bassler BL (2013) A quorum-sensing inhibitor blocks Pseudomonas aeruginosa virulence and biofilm formation. Proc Natl Acad Sci U S A 110:17981–17986PubMedCentralPubMedCrossRefGoogle Scholar
  31. Ou CC, Lu TM, Tsai JJ, Yen JH, Chen HW, Lin MY (2009) Antioxidative effect of lactic acid bacteria: intact cells vs. intracellular extracts. J Food Drug Anal 17:209–216Google Scholar
  32. Porras-Alfaro A, Bayman P (2011) Hidden fungi, emergent properties: endophytes and microbiomes. Annu Rev Phytopathol 49:291–315PubMedCrossRefGoogle Scholar
  33. Rasko DA, Sperandio V (2010) Anti-virulence strategies to combat bacteria-mediated disease. Nat Rev Drug Discov 9:117–128PubMedCrossRefGoogle Scholar
  34. Rishi P, Preet S, Kaur P (2011) Effect of L. plantarum cell-free extract and co-trimoxazole against Salmonella typhimurium: a possible adjunct therapy. Ann Clin Microbiol Antimicrob 10:9PubMedCentralPubMedCrossRefGoogle Scholar
  35. Rompp A, Spengler B (2013) Mass spectrometry imaging with high resolution in mass and space. Histochem Cell Biol 139:759–783PubMedCentralPubMedCrossRefGoogle Scholar
  36. Safari M, Amache R, Esmaeilishirazifard E, Keshavarz T (2014) Microbial metabolism of quorum-sensing molecules acyl-homoserine lactones, γ-heptalactone and other lactones. Appl Microbiol Biotechnol 98:3401–3412PubMedCrossRefGoogle Scholar
  37. Samoilova Z, Muzyka N, Lepekhina E, Oktyabrsky O, Smirnova G (2014) Medicinal plant extracts can variously modify biofilm formation in Escherichia coli. A Van Leeuw J Microb 105:709–722CrossRefGoogle Scholar
  38. Schafhauser J, Lepine F, McKay G, Ahlgren HG, Khakimova M, Nguyen D (2014) The stringent response modulates 4-hydroxy-2-alkylquinoline biosynthesis and quorum-sensing hierarchy in Pseudomonas aeruginosa. J Bacteriol 196:1641–1650PubMedCrossRefGoogle Scholar
  39. Schulz B, Guske S, Dammann U, Boyle C (1998) Endophyte-host interactions II. Defining symbiosis of the endophyte-host interaction. Symbiosis 25:213–227Google Scholar
  40. Shih C-J, Chen P-Y, Liaw C-C, Lai Y-M, Yang Y-L (2014) Bringing microbial interactions to light using imaging mass spectrometry. Nat Prod Rep. doi: 10.1039/c3np70091g PubMedGoogle Scholar
  41. Sifri CD (2008) Quorum sensing: bacteria talk sense. Clin Infect Dis 47:1070–1076PubMedCrossRefGoogle Scholar
  42. Taura F, Sirikantaramas S, Shoyamaa Y, Shoyamaa Y, Morimotoa S (2007) Phytocannabinoids in Cannabis sativa: recent studies on biosynthetic enzymes. Chem Biodivers 4:1649–1663PubMedCrossRefGoogle Scholar
  43. Teplitski M, Mathesius U, Rumbaugh KP (2011) Perception and degradation of N-acyl homoserine lactone quorum sensing signals by mammalian and plant cells. Chem Rev 111:100–116PubMedCrossRefGoogle Scholar
  44. Thenmozhi R, Nithyanand P, Rathna J, Pandian SK (2009) Antibiofillm activity of coral-associated bacteria against different clinical M serotypes of Streptococcus pyogenes. FEMS Immunol Med Microbiol 57:284–294PubMedCrossRefGoogle Scholar
  45. Zhao Q, Mutukumira A, Lee SJ, Maddox I, Shu Q (2011) Functional properties of free and encapsulated Lactobacillus reuteri DPC16 during and after passage through a simulated gastrointestinal tract. World J Microbiol Biotechnol 28:61–70PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2014

Authors and Affiliations

  • Parijat Kusari
    • 1
  • Souvik Kusari
    • 2
  • Marc Lamshöft
    • 2
  • Selahaddin Sezgin
    • 2
  • Michael Spiteller
    • 2
  • Oliver Kayser
    • 1
  1. 1.Department of Biochemical and Chemical EngineeringTU DortmundDortmundGermany
  2. 2.Institute of Environmental Research (INFU), Department of Chemistry and Chemical BiologyTU DortmundDortmundGermany

Personalised recommendations