Plant Cell Reports

, Volume 29, Issue 2, pp 153–161 | Cite as

Mycotoxin Ochratoxin A-induced cell death and changes in oxidative metabolism of Arabidopsis thaliana

Original Paper


We evaluated the phytotoxicity of mycotoxin ochratoxin A (OTA) from Aspergillus and Penicillium strains on Arabidopsis thaliana. The results demonstrate that the growth of Arabidopsis thaliana on media containing OTA was inhibited significantly. Moreover, OTA induced necrotic lesions in detached leaves, which are reminiscent of hypersensitive response lesions that are activated during plant–pathogen interactions and other abiotic stress factors. From our study, we can see that OTA exposure stimulated a biphasic oxidative burst in the leaves, resulting in the generation of hydrogen peroxide (H2O2) and superoxide anion radicals (O2·−) and in the concomitant down-regulation of antioxidant enzyme defense responses and up-regulation of lipid peroxidation. These results suggested that OTA damage might result from reactive oxygen species pathways. Our experiments provide a useful model plant system for research on OTA-induced plant cell death.


Arabidopsis thaliana Ochratoxin A Cell death Reactive oxygen species 



Ascorbate peroxidases








Fumonisin B1


2′,7′-Dichlorodihydrofluorescein diacetate


Hydrogen peroxide


Hypersensitive response




2-(N-morpholino) ethanesulphonic acid


Murashige and Skoog


Nitroblue tetrazolium


Superoxide anion radicals


Ochratoxin A


Programmed cell death




Pathogenesis related


Reactive oxygen species


Superoxide dismutase

Supplementary material

299_2009_808_MOESM1_ESM.doc (150 kb)
Supplementary material 1 (DOC 149 kb)


  1. Abarca ML, Bragulat MR, Sastella G, Cabanes FJ (1994) Ochratoxin A production by strains of Aspergillus niger var niger. Appl Environ Microbiol 60:2650–2652PubMedGoogle Scholar
  2. Asai T, Stone JM, Heard JE, Kovtun Y, Yorgey P, Sheen J, Ausubel FM (2000) Fumonisin B1-induced cell death in Arabidopsis protoplasts requires jasmonate-, ethylene-, and salicylate-dependent signaling pathways. Plant Cell 12:1823–1835CrossRefPubMedGoogle Scholar
  3. Babior BM, Benna JE, Chanock SJ, Smith RM (1997) The NADPH oxidase of leukocytes: the respiratory burst oxidase. In: Scandalios JG (ed) Oxidative stress and the molecular biology of antioxidant defenses. Cold Spring Harbour Press, Plainview, pp 737–783Google Scholar
  4. Boesch-Saadatmandi C, Loboda A, Jozkowicz A, Huebbe P, Blank R, Wolffram S, Dulak J, Rimbach G (2008) Effect of ochratoxin A on redox-regulated transcription factors, antioxidant enzymes and glutathione-S-transferase in cultured kidney tubulus cells. Food Chem Toxicol 46:2665–2671CrossRefPubMedGoogle Scholar
  5. Bowling SA, Clarke JD, Liu YD, Klessig DF, Dong XN (1997) The cpr5 mutant of Arabidopsis expresses both NPR1-dependent and NPR1-independent resistance. Plant Cell 9:1573–1584CrossRefPubMedGoogle Scholar
  6. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein–dye binding. Anal Biochem 72:248–254CrossRefPubMedGoogle Scholar
  7. Breisegem FV, Dat JF (2006) Reactive oxygen species in plant cell death. Plant Physiol 141:384–390CrossRefGoogle Scholar
  8. Ciegler A, Fennell DJ, Mintzlaff HJ, Leistner L (1972) Ochratoxin synthesis by Penicillium species. Naturwissenschaften 59:365–366CrossRefPubMedGoogle Scholar
  9. Clark HA, Snedeker SM (2006) Ochratoxin A: its cancer risk and potential for exposure. J Toxicol Environ Health B Crit Rev 9:265–296CrossRefPubMedGoogle Scholar
  10. Costa S, Utan A, Cervellati R, Speroni E, Guerra MC (2007) Catechins: natural free-radical scavengers against ochratoxin A-induced cell damage in a pig kidney cell line (LLC-PK1). Food Chem Toxicol 45:1910–1917CrossRefPubMedGoogle Scholar
  11. Dietrich RA, Richberg MH, Schmidt R, Dean C, Dangl JL (1997) A novel zinc finger protein is encoded by the Arabidopsis LSD1 gene and functions as a negative regulator of plant cell death. Cell 88:685–694CrossRefPubMedGoogle Scholar
  12. Gechev TS, Gadjev IZ, Hille J (2004) An extensive microarray analysis of AAL-toxin-induced cell death in Arabidopsis thaliana brings new insights into the complexity of programmed cell death in plants. Cell Mol Life Sci 61:1185–1197CrossRefPubMedGoogle Scholar
  13. Gechev TS, Breusegem FV, Stone JM, Denev I, Laloi C (2006) Reactive oxygen species as signals that modulate plant stress responses and programmed cell death. BioEssays 28:1091–1101CrossRefPubMedGoogle Scholar
  14. Greenberg JT, Guo A, Klessig DF, Ausubel FM (1994) Programmed cell death in plants: a pathogen-triggered response activated coordinately with multiple defense functions. Cell 77:551–563CrossRefPubMedGoogle Scholar
  15. Hoeberichts FA, Woltering EJ (2002) Multiple mediators of plant programmed cell death: interplay of conserved cell death mechanisms and plant-specific regulators. BioEssays 25:47–57CrossRefGoogle Scholar
  16. Horie Y (1995) Productivity of ochratoxin A of Aspergillus carbonarius in Aspergillus section Nigri. Nippon King-akukai Kaiho 36:73–76Google Scholar
  17. Jabs T, Dietrich RA, Dang JL (1996) Initiation of runaway cell death in an Arabidopsis mutant by extracellular superoxide. Science 273:1853–1856CrossRefPubMedGoogle Scholar
  18. Kamp HG, Eisenbrand G, Schlatter J, Würth K, Janzowski C (2005) Ochratoxin A: induction of (oxidative) DNA damage, cytotoxicity and apoptosis in mammalian cell lines and primary cells. Toxicology 206:413–425CrossRefPubMedGoogle Scholar
  19. Liang ZH (2008) Detection of ochratoxin A and analysis of ochratoxingenic-fungi in foodstuff. Dissertation, China Agriculture UniversityGoogle Scholar
  20. Mahalingam R, Jambunathan N, Gunjan SK, Faustin E, Weng H, Ayoubi P (2006) Analysis of oxidative signaling induced by ozone in Arabidopsis thaliana. Plant Cell Environ 29:1357–1371CrossRefPubMedGoogle Scholar
  21. Maksymiec W, Krupa Z (2006) The effects of short-term exposition to Cd, excess Cu ions and jasmonate on oxidative stress appearing in Arabidopsis thaliana. Environ Exp Bot 57:187–194CrossRefGoogle Scholar
  22. Masuda D, Ishida M, Yamaguchi K, Yamaguchi I, Kimura M, Nishiuchi T (2007) Phytotoxic effects of trichothecenes on the growth and morphology of Arabidopsis thaliana. J Exp Bot 58:1617–1626CrossRefPubMedGoogle Scholar
  23. Mitterbauer R, Adam G (2002) Saccharomyces cerevisae and Arabidopsis thaliana: useful model systems for the identification of molecular mechanisms involved in resistance of plants to toxins. Eur J Plant Pathol 108:699–703CrossRefGoogle Scholar
  24. Nagy NE, Dalen LS, Jones DL, Swensen B, Fossdal CG, Eldhuset TD (2004) Cytological and enzymatic responses to aluminium stress in root tips of Norway spruce seedlings. New Phytol 163:595–607CrossRefGoogle Scholar
  25. Nakano Y, Asada K (1981) Hydrogen peroxide is scavenged by ascorbate-specific peroxidase in spinach chloroplasts. Plant Cell Physiol 22:867–880Google Scholar
  26. Overmyer K, Brosché M, Pellinen R, Kuittinen T, Tuominen H, Ahlfors R, Keinänen M, Saarma M, Scheel D, Kangasjärvi J (2005) Ozone-induced programmed cell death in the Arabidopsis radical-induced cell death1 mutant. Plant Physiol 137:1092–1104CrossRefPubMedGoogle Scholar
  27. Paciolla C, Dipierro N, Mulè G, Logrieco A, Dipierro S (2004) The mycotoxins beauvericin and T-2 induce cell death and alteration to the ascorbate metabolism in tomato protoplasts. Physiol Mol Plant Pathol 65:49–56CrossRefGoogle Scholar
  28. Petrik J, Žanić-Grubišić T, Barišić K, Pepeljnjak S, Radić B, Ferenčić Ž, Čepelak I (2003) Apoptosis and oxidatice stress induced by ochratoxin A in rat kidney. Arch Toxicol 77:585–693CrossRefGoogle Scholar
  29. Romero-Puertas MC, Rodríguez-serrano M, Corpas FJ, Gómez M, Del Río LA, Sandalio LM (2004) Cadmium-induced subcellular accumulation of O2·− and H2O2 in pea leaves. Plant Cell Environ 27:1122–1134CrossRefGoogle Scholar
  30. Schraudner M, Moeder W, Wiese C, Camp WV, Inzé D, Langebartels C, Sandermann H Jr (1998) Ozone-induced oxidative burst in the ozone biomonitor plant, tobacco Bel W3. Plant J 16:235–245CrossRefGoogle Scholar
  31. Schwerdt G, Freudinger R, Mildenberger S, Silbernagl S, Gekle M (1999) The nephrotoxin ochratoxin A induces apoptosis in cultured human proximal tubule cells. Cell Biol Toxicol 15:405–415CrossRefPubMedGoogle Scholar
  32. Schwerdt G, Holzinger H, Sauvant C, Königs M, Humpf HU, Gekle M (2007) Long-term effects of ochratoxin A on fibrosis and cell death in human proximal tubule or fibroblast cells in primary culture. Toxicology 232:57–67CrossRefPubMedGoogle Scholar
  33. Schwerdt G, Holzinger H, Königs M, Humpf HU, Gekle M (2009) Effect of ochratoxin A on cell survival and collagen homeostasis in human mesangial cells in primary culture. Food Chem Toxicol 27:209–213Google Scholar
  34. Stone JM, Heard JE, Asai T, Ausubel FM (2000) Simulation of fungal-mediated cell death by fumonisin B1 and selection of fumonisin B1-resistant (fbr) Arabidopsis mutants. Plant Cell 12:1811–1822CrossRefPubMedGoogle Scholar
  35. Van Der Merwe KJ, Steyne PS, Fourie L, Scott DB, Theron JJ (1965) Ochratoxin A, a toxic metabolite produced by Aspergillus ochraceus. Nature 205:1112–1113CrossRefPubMedGoogle Scholar
  36. WHO (2001) Safety evaluation of certain mycotoxins in food, fifty-sixth meeting of the joint FAO/WHO expert committee on food additives. WHO Food Addit Ser 47:706Google Scholar
  37. Wohlgemuth H, Mittelstrass K, Kschieschan S, Bender J, Weigel HJ, Overmyer K, Kangasjärvi J, Sandermann H, Langebartels C (2002) Activation of an oxidative burst is a general feature of sensitive plants exposed to the air pollutant ozone. Plant Cell Environ 25:717–726CrossRefGoogle Scholar
  38. Xu WT, Peng XL, Luo YB, Wang JA, Guo X, Huang KL (2009) Physiological and biochemical responses of gragefruit seed extract dip on ‘Redglobe’ grape. LWT-Food Sci Technol 42:471–476CrossRefGoogle Scholar
  39. Zhang X, Boesch-Saadatmandi C, Lou Y, Wolffram S, Huebbe P, Rimbach G (2009) Ochratoxin A induces apoptosis in neuronal cells. Genes Nutr 4:41–48CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  1. 1.Laboratory of Fruit Physiology and Molecular Biology, College of Food Science and Nutritional EngineeringChina Agricultural UniversityBeijingChina
  2. 2.Supervision and Testing Center of Agricultural Products QualityMinistry of AgricultureBeijingChina

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