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Involvement of alternative oxidase in the regulation of growth, development, and resistance to oxidative stress of Sclerotinia sclerotiorum

Journal of Microbiology volume 50, pages 594–602 (2012)Cite this article

Abstract

Sclerotinia sclerotiorum is a cosmopolitan, filamentous, fungal pathogen that can cause serious disease in many kinds of crops. Alternative oxidase is the terminal oxidase of the alternative mitochondrial respiratory pathway in fungi and higher plants. We report the presence of this alternative pathway respiration and demonstrate its expression in two isolates of S. sclerotiorum under unstressed, normal culture conditions. Application of salicylhydroxamic acid, a specific inhibitor of alternative oxidase, severely inhibited the mycelial growth of S. sclerotiorum both on potato dextrose agar plates and in liquid culture media. Inhibition of alternative oxidase could influence the growth pattern of S. sclerotiorum, as salicylhydroxamic acid treatment induced obvious aerial mycelia growing on potato dextrose agar plates. Under the treatment with salicylhydroxamic acid, S. sclerotiorum formed sclerotia much more slowly than the control. Treatment with hydrogen peroxide in millimolar concentrations greatly decreased the growth rate of mycelia and delayed the formation of sclerotia in both tested S. sclerotiorum isolates. As well, this treatment obviously increased their alternative pathway respiration and the levels of both mRNA and protein of the alternative oxidase. These results indicate that alternative oxidase is involved in the regulation of growth, development, and resistance to oxidative stress of S. sclerotiorum.

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References

  • Akhter, S., McDade, H.C., Gorlach, J.M., Heinrich, G., Cox, G.M., and Perfect, J.R. 2003. Role of alternative oxidase gene in pathogenesis of Cryptococcus neoformans. Infect. Immun. 71, 5794–5802.

    PubMed  Article  CAS  Google Scholar 

  • Avila-Adame, C. and Koller, W. 2002. Disruption of the alternative oxidase gene in Magnaporthe grisea and its impact on host infection. Mol. Plant-Microbe Interact. 15, 493–500.

    PubMed  Article  CAS  Google Scholar 

  • Avila-Adame, C. and Koller, W. 2003. Impact of alternative respiration and target-site mutations on responses of germinating conidia of Magnaporthe grisea to Qo-inhibiting fungicides. Pest Manag. Sci. 59, 303–309.

    PubMed  Article  CAS  Google Scholar 

  • Bardin, S.D. and Huang, H.C. 2001. Research on biology and control of Sclerotinia diseases in Canada. Can. J. Plant Pathol. 23, 88–98.

    Article  Google Scholar 

  • Boland, G.J. and Hall, R. 1994. Index of plant hosts of Sclerotinia sclerotiorum. Can. J. Plant Pathol. 16, 93–108.

    Article  Google Scholar 

  • Bolton, M.D., Thomma, B.P.H.J., and Nelson, B.D. 2006. Sclerotinia sclerotiorum (Lib.) de Bary: biology and molecular traits of a cosmopolitan pathogen. Mol. Plant Pathol. 7, 1–16.

    PubMed  Article  CAS  Google Scholar 

  • Bradford, M.M. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72, 248–254.

    PubMed  Article  CAS  Google Scholar 

  • Chivasa, S. and Carr, J.P. 1998. Cyanide restores N gene-mediated resistance to tobacco mosaic virus in transgenic tobacco expressing salicylic acid hydroxylase. Plant Cell 10, 1489–1498.

    PubMed  CAS  Google Scholar 

  • Chivasa, S., Murphy, A.M., Naylor, M., and Carr, J.P. 1997. Salicylic acid interferes with tobacco mosaic virus replication via a novel salicylhydroxamic acid-sensitive mechanism. Plant Cell 9, 547–557.

    PubMed  CAS  Google Scholar 

  • Clifton, R., Millar, A.H., and Whelan, J. 2006. Alternative oxidases in Arabidopsis: a comparative analysis of differential expression in the gene family provides new insights into function of non-phosphorylating bypasses. Biochim. Biophys. Acta. 1757, 730–741.

    PubMed  Article  CAS  Google Scholar 

  • Day, D.A., Whelan, J., Millar, A.H., Siedow, J.N., and Wiskich, J.T. 1995. Regulation of the alternative oxidase in plants and fungi. Aust. J. Plant Physiol. 22, 494–509.

    Google Scholar 

  • Fu, L.J., Shi, K., Gu, M., Zhou, Y.H., Dong, D.K., Liang, W.S., Song, F.M., and Yu, J.Q. 2010. Systemic induction and role of mitochondrial alternative oxidase and nitric oxide in a compatible tomato-tobacco mosaic virus interaction. Mol. Plant-Microbe Interact. 23, 39–48.

    PubMed  Article  Google Scholar 

  • Ishii, H., Fountaine, J., Chung, W.H., Kansako, M., Nishimura, K., Takahashi, K., and Oshima, M. 2009. Characterisation of QoI-resistant field isolates of Botrytis cinerea from citrus and strawberry. Pest Manag. Sci. 65, 916–922.

    PubMed  Article  CAS  Google Scholar 

  • Joseph-Horne, T., Babij, J., Wood, P.M., Hollomon, D., and Sessions, R.B. 2000. New sequence data enable modelling of the fungal alternative oxidase and explain an absence of regulation by pyruvate. FEBS Lett. 481, 141–146.

    PubMed  Article  CAS  Google Scholar 

  • Joseph-Horne, T., Hollomon, D.W., and Wood, P.M. 2001. Fungal respiration: a fusion of standard and alternative components. Biochim. Biophys. Acta. 1504, 179–195.

    PubMed  Article  CAS  Google Scholar 

  • Juarez, O., Guerra, G., Velazquez, I., Flores-Herrera, O., Rivera-Perez, R.E., and Pardo, J.P. 2006. The physiologic role of alternative oxidase in Ustilago maydis. FEBS J. 273, 4603–4615.

    PubMed  Article  CAS  Google Scholar 

  • Lambowitz, A.M., Sabourin, J.R., Bertland, H., Nickels, R., and McIntosh, L. 1989. Immunological identification of the alternative oxidase of Neurospora crassa mitochondria. Mol. Cell. Biol. 9, 1362–1364.

    PubMed  CAS  Google Scholar 

  • Li, G.Q., Huang, H.C., Miao, H.J., Erickson, R.S., Jiang, D.H., and Xiao, Y.N. 2006. Biological control of sclerotinia diseases of rapeseed by aerial applications of the mycoparasite Coniothyrium minitans. Eur. J. Plant Pathol. 114, 345–355.

    Article  Google Scholar 

  • Liang, W.S. and Liang, H.G. 2002. Comparison of the effects of H2O2 and salicylic acid on alternative respiratory pathway in aged potato tuber slices. Acta Botanica Sinica 44, 287–291.

    CAS  Google Scholar 

  • Liang, W.S., Liang, H.G., and Li, D.B. 2002. Induction of cyanide-resistant respiration by H2O2 in aging potato tuber slices. J. Zhejiang Univ. (Agri. and Life Sci.) 28, 525–528.

    CAS  Google Scholar 

  • Liang, W.S., Pan, J., and Liang, H.G. 2003. Activation of cyanide-resistant respiration by pyruvate in mitochondria of aged potato tuber slices. J. Plant Physiol. Mol. Biol. 29, 317–321.

    CAS  Google Scholar 

  • Livak, K.J. and Schmittgen, T.D. 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCt method. Methods 25, 402–408.

    PubMed  Article  CAS  Google Scholar 

  • Martins, V.P., Dinamarco, T.M., Soriani, F.M., Tudella, V.G., Oliveira, S.C., Goldman, G.H., Curti, C., and Uyemura, S.A. 2011. Involvement of an alternative oxidase in oxidative stress and mycelium-to-yeast differentiation in Paracoccidioides brasiliensis. Eukaryot. Cell 10, 237–248.

    PubMed  Article  CAS  Google Scholar 

  • Maxwell, D.P., Wang, Y., and McIntosh, L. 1999. The alternative oxidase lowers mitochondrial reactive oxygen production in plant cells. Proc. Natl. Acad. Sci. USA 96, 8271–8276.

    PubMed  Article  CAS  Google Scholar 

  • McDonald, A.E. 2008. Alternative oxidase: an inter-kingdom perspective on the function and regulation of this broadly distributed ‘cyanide-resistant’ terminal oxidase. Functional Plant Biol. 35, 535–552.

    Article  CAS  Google Scholar 

  • McIntosh, L. 1994. Molecular biology of the alternative oxidase. Plant Physiol. 105, 781–786.

    PubMed  Article  CAS  Google Scholar 

  • Minagawa, N., Koga, S., Nakano, M., Sakajo, S., and Yoshimoto, A. 1992. Possible involvement of superoxide anion in the induction of cyanide-resistant respiration in Hansenula anomala. FEBS Lett. 302, 217–219.

    PubMed  Article  CAS  Google Scholar 

  • Minagawa, N. and Yoshimoto, A. 1987. The induction of cyanide-resistant respiration in Hansenula anomala. J. Biochem. 101, 1141–1146.

    PubMed  CAS  Google Scholar 

  • Papapostolou, I. and Georgiou, C.D. 2010a. Hydrogen peroxide is involved in the sclerotial differentiation of filamentous phytopathogenic fungi. J. Appl. Microbiol. 109, 1929–1936.

    PubMed  Article  CAS  Google Scholar 

  • Papapostolou, I. and Georgiou, C.D. 2010b. Superoxide radical induces sclerotial differentiation in filamentous phytopathogenic fungi: a superoxide dismutase mimetics study. Microbiology 156, 960–966.

    PubMed  Article  CAS  Google Scholar 

  • Purvis, A.C. 1997. Role of the alternative oxidase in limiting superoxide production by plant mitochondria. Physiol. Plant. 100, 165–170.

    Article  CAS  Google Scholar 

  • Rachmilevitch, S., Xu, Y., Gonzalez-Meler, M.A., Huang, B., and Lambers, H. 2007. Cytochrome and alternative pathway activity in roots of thermal and non-thermal Agrostis species in response to high soil temperature. Physiol. Plant. 129, 163–174.

    Article  CAS  Google Scholar 

  • Rhoads, D.M., Umbach, A., Sweet, C.R., Lennon, A.M., Rauch, G.S., and Siedow, J.N. 1998. Regulation of the cyanide-resistant alternative oxidase of plant mitochondria. Identification of the cysteine residue involved in α-keto acid stimulation and intersubunit disulfide bond formation. J. Biol. Chem. 273, 30750–30756.

    PubMed  Article  CAS  Google Scholar 

  • Ribas-Carbo, M., Aroca, R., Gonzalez-Meler, M.A., Irigoyen, J.J., and Sanchez-Diaz, M. 2000. The electron partitioning between the cytochrome and alternative respiratory pathways during chilling recovery in two cultivars of maize differing in chilling sensitivity. Plant Physiol. 122, 199–204.

    PubMed  Article  CAS  Google Scholar 

  • Ribas-Carbo, M., Taylor, N.L., Giles, L., Busquets, S., Finnegan, P.M., Day, D.A., Lambers, H., Medrano, H., Berry, J.A., and Flexas, J. 2005. Effects of water stress on respiration in soybean leaves. Plant Physiol. 139, 466–473.

    PubMed  Article  CAS  Google Scholar 

  • Rizhsky, L., Liang, H., and Mittler, R. 2002. The combined effect of drought stress and heat shock on gene expression in tobacco. Plant Physiol. 130, 1143–1151.

    PubMed  Article  CAS  Google Scholar 

  • Umbach, A. and Siedow, J.N. 1993. Covalent and noncovalent dimers of the cyanide-resistant alternative oxidase protein in higher plant mitochondria and their relationship to enzyme activity. Plant Physiol. 103, 845–854.

    PubMed  CAS  Google Scholar 

  • Umbach, A. and Siedow, J.N. 2000. The cyanide-resistant alternative oxidases from the fungi Pichia stipitis and Neurospora crassa are monomeric and lack regulatory feature of plant enzyme. Arch. Biochem. Biophy. 378, 234–245.

    Article  CAS  Google Scholar 

  • Vanlerberghe, G.C. 1997. Alternative oxidase: from gene to function. Annu. Rev. Plant Physiol. Plant Mol. Biol. 48, 703–734.

    PubMed  Article  CAS  Google Scholar 

  • Wagner, A.M. 1995. A role for active oxygen species as second messengers in the induction of alternative oxidase gene expression in Petunia hybrida cells. FEBS Lett. 368, 339–342.

    PubMed  Article  CAS  Google Scholar 

  • Young, C.S., Clarkson, J.P., Smith, J.A., Watling, M., Phelps, K., and Whipps, J.M. 2004. Environmental conditions influencing Sclerotinia sclerotiorum infection and disease development in lettuce. Plant Pathol. 53, 387–397.

    Article  Google Scholar 

  • Yukioka, H., Inagaki, S., Tanaka, R., Katoh, K., Miki, N., Mizutani, A., and Masuko, M. 1998. Transcriptional activation of the alternative oxidase gene of the fungus Magnaporthe grisea by a respiratory-inhibiting fungicide and hydrogen peroxide. Biochim. Biophys. Acta. 1442, 161–169.

    PubMed  Article  CAS  Google Scholar 

  • Zhang, L. 2006. Application of SPSS 13.0 in biological statistics. Xiamen University Press, Xiamen, China.

    Google Scholar 

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Affiliations

  1. Institute of Biotechnology, College of Agriculture and Biotechnology, Zijingang Campus, Zhejiang University, Hangzhou, 310058, P. R. China

    Ting Xu, Fei Yao, Wu-Sheng Liang & Zheng-Yi Wang

  2. State Key Laboratory of Rice Biology, Hangzhou, 310058, P. R. China

    Ting Xu, Fei Yao, Wu-Sheng Liang & Zheng-Yi Wang

  3. Hybrid Rapeseed Research Center of Shaanxi Province, Dali, 715105, P. R. China

    Yong-Hong Li, Dian-Rong Li & Hao Wang

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  1. Ting Xu
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  2. Fei Yao
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  3. Wu-Sheng Liang
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  5. Dian-Rong Li
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  6. Hao Wang
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  7. Zheng-Yi Wang
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Correspondence to Wu-Sheng Liang.

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Xu, T., Yao, F., Liang, WS. et al. Involvement of alternative oxidase in the regulation of growth, development, and resistance to oxidative stress of Sclerotinia sclerotiorum . J Microbiol. 50, 594–602 (2012). https://doi.org/10.1007/s12275-012-2015-7

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  • DOI: https://doi.org/10.1007/s12275-012-2015-7

Keywords

  • Sclerotinia sclerotiorum
  • alternative oxidase
  • mitochondrial respiratory chain
  • hydrogen peroxide
  • oxidative stress