Controlling food-contaminating fungi by targeting their antioxidative stress-response system with natural phenolic compounds

  • Jong H. Kim
  • Noreen Mahoney
  • Kathleen L. Chan
  • Russell J. Molyneux
  • Bruce C. Campbell
Applied Microbial and Cell Physiology


The antioxidative stress-response system is essential to fungi for tolerating exposure to phenolic compounds. We show how this system can be targeted to improve fungal control by using compounds that inhibit the fungal mitochondrial respiratory chain. Targeting mitochondrial superoxide dismutase with selected phenolic acid derivatives (e.g., vanillyl acetone) resulted in a 100- to 1,000-fold greater sensitivity to strobilurin or carboxin fungicides. This synergism is significantly greater with strobilurin than with carboxin, suggesting that complex III of the mitochondrial respiratory chain is a better target than complex II for fungal control, using phenolics. These results show certain natural compounds are effective synergists to commercial fungicides and can be used for improving control of food-contaminating pathogens. These results suggest that the use of such compounds for fungal control can reduce environmental and health risks associated with commercial fungicides, lower cost for control, and the probability for development of resistance.



This research was conducted under USDA-ARS project no. 5325-42000-032-00D.


  1. Ammar H, Michaelis G, Lisowsky T (2000) A screen of yeast respiratory mutants for sensitivity against the mycotoxin citrinin identifies the vacuolar ATPase as an essential factor for the toxicity mechanism. Curr Genet 37:277–284CrossRefGoogle Scholar
  2. Beekrum S, Govinden R, Padayachee T, Odhav B (2003) Naturally occurring phenols: a detoxification strategy for fumonisin B1. Food Addit Contam 20:490–493CrossRefGoogle Scholar
  3. Clemons KV, Miller TK, Selitrennikoff CP, Stevens DA (2002) fos-1, a putative histidine kinase as a virulence factor for systemic aspergillosis. Med Mycol 40:259–262CrossRefGoogle Scholar
  4. Demasi APD, Pereira GAG, Netto LES (2001) Cytosolic thioredoxin peroxidase I is essential for the antioxidant defense of yeast with dysfunctional mitochondria. FEBS Lett 509:430–434CrossRefGoogle Scholar
  5. Dietz KJ, Tavakoli N, Kluge C, Mimura T, Sharma SS, Harris GC, Chardonnens AN, Golldack D (2001) Significance of the V-type ATPase for the adaptation to stressful growth conditions and its regulation on the molecular and biochemical level. J Exp Bot 52:1969–1980CrossRefGoogle Scholar
  6. Florianowicz T (1998) Penicillium expansum growth and production of patulin in the presence of benzoic acid and its derivatives. Acta Microbiol Pol 47:45–53Google Scholar
  7. Garrido E, Voss U, Muller P, Castillo-Lluva S, Kahmann R, Perez-Martin J (2004) The induction of sexual development and virulence in the smut fungus Ustilago maydis depends on Crk1, a novel MAPK protein. Genes Dev 18:3117–3130CrossRefGoogle Scholar
  8. Hamilton AJ, Holdom MD (1999) Antioxidant systems in the pathogenic fungi of man and their role in virulence. Med Mycol 37:375–389CrossRefGoogle Scholar
  9. Hamilton CA, Taylor GJ, Good AG (2002) Vacuolar H(+)-ATPase, but not mitochondrial F(1)F(0)-ATPase, is required for NaCl tolerance in Saccharomyces cerevisiae. FEMS Microbiol Lett 208:227–232CrossRefGoogle Scholar
  10. Hnatova M, Gbelska Y, Obernauerova M, Subikova V, Subik J (2003) Cross-resistance to strobilurin fungicides in mitochondrial and nuclear mutants of Saccharomyces cerevisiae. Folia Microbiol (Praha) 48:496–500Google Scholar
  11. Kim JH, Mahoney N, Chan K, Molyneux RJ, Campbell BC (2004) Identification of phenolics for control of Aspergillus flavus using Saccharomyces cerevisiae in a model target-gene bioassay. J Agric Food Chem 52:7814–7821CrossRefGoogle Scholar
  12. Kim JH, Campbell BC, Yu J, Mahoney N, Chan K, Molyneux RJ, Bhatnagar D, Cleveland TE (2005) Examination of fungal stress response genes using Saccharomyces cervisiae as a model system: targeting genes affecting aflatoxin biosynthesis by Aspergillus flavus Link. Appl Microbiol Biotechnol 67:807–815CrossRefGoogle Scholar
  13. Kupferwasser LI, Yeaman MR, Nast CC, Kupferwasser D, Xiong YQ, Palma M, Cheung AL, Bayer AS (2003) Salicylic acid attenuates virulence in endovascular infections by targeting global regulatory pathways in Staphylococcus aureus. J Clin Invest 112:222–233Google Scholar
  14. Lopez-Malo A, Alzamora SM, Palou E (2002) Aspergillus flavus dose–response curves to selected natural and synthetic antimicrobials. Int J Food Microbiol 73:213–218CrossRefGoogle Scholar
  15. Mahoney N, Molyneux R (2004) Phytochemical inhibition of aflatoxigenicity in Aspergillus flavus by constituents of walnut (Juglans regia). J Agric Food Chem 52:1882–1889CrossRefGoogle Scholar
  16. Nelson N (1992) The vacuolar H(+)-ATPase—one of the most fundamental ion pumps in nature. J Exp Biol 172:19–27Google Scholar
  17. Parsons AB, Brost RL, Ding H, Li Z, Zhang C, Sheikh B, Brown GW, Kane PM, Hughes TR, Boone C (2004) Integration of chemical-genetic and genetic interaction data links bioactive compounds to cellular target pathways. Nat Biotechnol 22:62–69CrossRefGoogle Scholar
  18. Tawata S, Taira S, Kobamoto N, Zhu J, Ishihara M, Toyama S (1996) Synthesis and antifungal activity of cinnamic acid esters. Biosci Biotechnol Biochem 60:909–910CrossRefGoogle Scholar
  19. Tucker CL, Fields S (2004) Quantitative genome-wide analysis of yeast deletion strain sensitivities to oxidative and chemical stress. Compar Funct Genom 5:216–224CrossRefGoogle Scholar
  20. Winzeler EA, Shoemaker DD, Astromoff A et al (1999) Functional characterization of the S. cerevisiae genome by gene deletion and parallel analysis. Science 285:901–906CrossRefGoogle Scholar
  21. Yamauchi J, Takayanagi N, Komeda K, Takano Y, Okuno T (2004) cAMP-pKA signaling regulates multiple steps of fungal infection cooperatively with Cmk1 MAP kinase in Colletotrichum lagenarium. Mol Plant Microbe Interact 17:1355–1365CrossRefGoogle Scholar
  22. Zelko IN, Mariani TJ, Folz RJ (2002) Superoxide dismutase multigene family: a comparison of the CuZn-SOD (SOD1), Mn-SOD (SOD2), and EC-SOD (SOD3) gene structures, evolution, and expression. Free Radic Biol Med 33:337–349CrossRefGoogle Scholar
  23. Zheng D, Olaya G, Koller W (2000) Characterization of laboratory mutants of Venturia inaequalis resistant to the strobilurin-related fungicide kresoxim-methyl. Curr Genet 38:148–155CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2006

Authors and Affiliations

  • Jong H. Kim
    • 1
  • Noreen Mahoney
    • 1
  • Kathleen L. Chan
    • 1
  • Russell J. Molyneux
    • 1
  • Bruce C. Campbell
    • 1
  1. 1.Plant Mycotoxin Research UnitWestern Regional Research Center, USDA-ARSAlbanyUSA

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