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Modified alternative oxidase expression results in different reactive oxygen species contents in Arabidopsis cell culture but not in whole plants

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Biologia Plantarum

Abstract

Alternative oxidase (AOX) transfers electrons from ubiquinone to oxygen in the respiratory chain of plant mitochondria. It is widely accepted that AOX functions as a mechanism decreasing the formation of reactive oxygen species (ROS) produced during respiratory electron transport. However, there are no experimental data to provide unambiguous proof of this hypothesis. We have studied growth characteristics, ROS content, and stress sensitivity in Arabidopsis transgenic lines with reduced or increased levels of AOX. We demonstrated that AOX-deficient plants grown in soil had an extended reproductive phase. Changes in AOX activity did not affect ROS content or stress sensitivity in the whole plants. However in the suspension culture, cells overexpressing AOX had significantly lower ROS content, whereas the AOX-deficient cells had higher ROS contents compared to the wild-type (WT) cells. Prooxidant treatment led to the increase in ROS content and to the reduction of viability more in the cells overexpressing AOX than in WT and AOX-deficient cells. Thus, we demonstrated that differences in the metabolism of whole plants and cultured cells might affect AOX functioning.

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Abbreviations

AOX:

alternative oxidase

DAB:

3,3′-diaminobenzidine

DCF-DA:

2′,7′-dichlorofluorescein diacetate

ETC:

electron transport chain

NBT:

nitroblue tetrazolium

ROS:

reactive oxygen species

WT:

wild-type

References

  • Amirsadeghi, S., Robson, C.A., McDonald, A.E., Vanlerberghe, G.C.: Changes in plant mitochondrial electron transport alter cellular levels of reactive oxygen species and susceptibility to cell death signaling molecules. — Plant Cell Physiol. 47: 1509–1519, 2006.

    Article  PubMed  CAS  Google Scholar 

  • Baker, C.J., Mock, N.M.: An improved method for monitoring cell death in cell suspension and leaf disc assays using Evans blue. — Plant Cell Tissue Organ Cult. 39: 7–12, 1994.

    Article  Google Scholar 

  • Boyes, D.C., Zayed, A.M., Ascenzi, R., McCaskill, A.J., Hoffman, N.E., Davies, K.R., Gorlach, J.: Growth stagebased phenotypic analysis of Arabidopsis: a model for high throughput functional genomics in plants. — Plant Cell 13: 1499–1510, 2001.

    PubMed  CAS  Google Scholar 

  • Clifton, R., Lister, R., Parker, K.L., Sappl, P.G., Elhafez, D., Millar, A.H., Day, D.A., Whelan, J.: Stress-induced coexpression of alternative respiratory chain components in Arabidopsis thaliana. — Plant mol. Biol. 58: 193–212, 2005.

    Article  PubMed  CAS  Google Scholar 

  • Clifton, R., Millar, A.H., Whelan, J.: 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, 2006.

    Article  PubMed  CAS  Google Scholar 

  • Doyle, J.J., Doyle, J.L.: A rapid isolation program for small quantities of fresh leaf tissue. — Phytochem. Bull. 19: 11–15, 1987.

    Google Scholar 

  • Ferreira, A.L., Arrabaca, J.D., Vaz-Pinto, V., Lima-Costa, M.E.: Induction of alternative oxidase chain under salt stress conditions. — Biol. Plant. 52: 66–71, 2008.

    Article  CAS  Google Scholar 

  • Fiorani, F., Umbach, A.L., Siedow, J.N.: The alternative oxidase of plant mitochondria is involved in the acclimation of shoot growth at low temperature: a study of Arabidopsis AOX1a transgenic plants. — Plant Physiol. 139: 1795–1805, 2005.

    Article  PubMed  CAS  Google Scholar 

  • Garnik, E.Y., Tarasenko, V.I., Kobzev, V.F., Konstantinov, Y.M.: Differential expression of maize mitochondrial genes as dependent on mitochondria redox state. — Russ. J. Plant Physiol. 53: 463–468, 2006.

    Article  CAS  Google Scholar 

  • Gilliland, A., Singh, D.P., Hayward, J.M., Moore, C.A., Murphy, A.M., York, C.J., Slator, J., Carr, J.P.: Genetic modification of alternative respiration has differential effects on antimycin A-induced versus salicylic acidinduced resistance to tobacco mosaic virus. — Plant Physiol. 132: 1518–1528, 2003.

    Article  PubMed  CAS  Google Scholar 

  • Giraud, E., Ho, L.H.M., Clifton, R., Carroll, A., Estavillo, G., Tan, Y.F., Howell, K.A., Ivanova, A., Pogson, B.J., Millar, A.H., Whelan, J.: The absence of ALTERNATIVE OXIDASE1a in Arabidopsis results in acute sensitivity to combined light and drought stress. — Plant Physiol. 147: 595–610, 2008.

    Article  PubMed  CAS  Google Scholar 

  • Hu, W.H., Xiao, Y.A., Zeng, J.J., Hu, X.H.: Photosynthesis, respiration and antioxidant enzymes in pepper leaves under drought and heat stresses. — Biol. Plant. 54: 761–765, 2010.

    Article  CAS  Google Scholar 

  • Karpova, O.V., Kuzmin, E.V., Elthon, T.E., Newton, K.J.: Differential expression of alternative oxidase genes in maize mitochondrial mutants. — Plant Cell 14: 3271–3284, 2002.

    Article  PubMed  CAS  Google Scholar 

  • Lee, B.H., Lee, H., Xiong, L., Zhu, J.K.: A mitochondrial complex I defect impairs cold-regulated nuclear gene expression. — Plant Cell. 14: 1235–1251, 2002.

    Article  PubMed  CAS  Google Scholar 

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

    Article  PubMed  CAS  Google Scholar 

  • Meyer, E.H., Tomaz, T., Carroll, A.J., Estavillo, G., Delannoy, E., Tanz, S.K., Small, I.D., Pogson, B.J., Millar, A.H.: Remodeled respiration in ndufs4 with low phosphorylation efficiency suppresses Arabidopsis germination and growth and alters control of metabolism at night. — Plant Physiol. 151: 603–619, 2009.

    Article  PubMed  CAS  Google Scholar 

  • Myouga, F., Hosoda, C., Umezawa, T., Iizumi, H., Kuromori, T., Motohashi, R., Shono, Y., Nagata, N., Ikeuchi, M., Shinozaki, K.: A heterocomplex of iron superoxide dismutases defends chloroplast nucleoids against oxidative stress and is essential for chloroplast development in Arabidopsis. — Plant Cell 20: 3148–3162, 2008.

    Article  PubMed  CAS  Google Scholar 

  • Moller, I.M.: Plant mitochondria and oxidative stress: electron transport, NADPH turnover, and metabolism of reactive oxygen species. — Annu. Rev. Plant Physiol. Plant mol. Biol. 52: 561–591, 2001.

    Article  PubMed  CAS  Google Scholar 

  • Noctor, G., Dutilleul, C., De Paepe, R., Foyer, C.H.: Use of mitochondrial electron transport mutants to evaluate the effects of redox state on photosynthesis, stress tolerance and the integration of carbon/nitrogen metabolism. — J. exp. Bot. 55: 49–57, 2004.

    Article  PubMed  CAS  Google Scholar 

  • Pasqualini, S., Paolocci, F., Borgogni, A., Morettini, R., Ederli, L.: The overexpression of an alternative oxidase gene triggers ozone sensitivity in tobacco plants. — Plant Cell Environ. 30: 1545–1556, 2007.

    Article  PubMed  CAS  Google Scholar 

  • Ramel, F., Sulmon, C., Bogard, M., Coue, I., Gouesbet, G.: Differential dynamics of reactive oxygen species and antioxidative mechanisms during atrazine injury and sucrose-induced tolerance in Arabidopsis thaliana plantlets. — BMC Plant Biol. 9: 28, 2009.

    Article  PubMed  Google Scholar 

  • Robson, C.A., Vanlerberghe, G.C.: Transgenic plant cells lacking mitochondrial alternative oxidase have increased susceptibility to mitochondria-dependent and -independent pathways of programmed cell death. — Plant Physiol. 129: 1908–1920, 2002.

    Article  PubMed  CAS  Google Scholar 

  • Strodtkotter, I., Padmasree, K., Dinakar, C., Speth, B., Niazi, P.S., Wojtera, J., Voss, I., Do, P.T., Nunes-Nesi, A., Fernie, A.R., Linke, V., Raghavendra, A.S., Scheibe, R.: Induction of the AOX1D isoform of alternative oxidase in A. thaliana T-DNA insertion lines lacking isoform AOX1A is insufficient to optimize photosynthesis when treated with antimycin A. — Mol. Plants 2: 284–297, 2009.

    Article  Google Scholar 

  • Tarasenko, V.I., Garnik, E.Y., Konstantinov, Y.M.: Characterization of Arabidopsis mutant with inactivated gene coding for Fe-S subunit of mitochondrial respiratory chain complex I. — Russ. J. Plant Physiol. 57: 392–400, 2010.

    Article  CAS  Google Scholar 

  • Tarasenko, V.I., Garnik, E.Y., Shmakov, V.N., Konstantinov, Y.M.: Induction of Arabidopsis gdh2 gene expression during changes in redox state of the mitochondrial respiratory chain. — Biochemistry (Moscow) 74: 47–53, 2009.

    Article  CAS  Google Scholar 

  • Umbach, A.L., Fiorani, F., Siedow, J.N.: Characterization of transformed Arabidopsis with altered alternative oxidase levels and analysis of effects on reactive oxygen species in tissue. — Plant Physiol. 139: 1806–1820, 2005.

    Article  PubMed  CAS  Google Scholar 

  • Vanlerberghe, G.C., Cvetkovska, M., Wang, J.: Is the maintenance of homeostatic mitochondrial signaling during stress a physiological role for alternative oxidase. — Physiol. Plant. 137: 392–406, 2009.

    Article  PubMed  CAS  Google Scholar 

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

    Article  PubMed  CAS  Google Scholar 

  • Vanlerberghe, G.C., Robson, C.A., Yip, J.Y.H.: Induction of mitochondrial alternative oxidase in response to a cell signal pathway down-regulating the cytochrome pathway prevents programmed cell death. — Plant Physiol. 129: 1829–1842, 2002.

    Article  PubMed  CAS  Google Scholar 

  • Vanlerberghe, G.C., Vanlerberghe, A.E., McIntosh, L.: Molecular genetic alteration of plant respiration: silencing and overexpression of alternative oxidase in transgenic tobacco. — Plant Physiol. 106: 1503–1510, 1994.

    PubMed  CAS  Google Scholar 

  • Watanabe, C.K., Hachiya, T., Terashima, I., Noguchi, K.: The lack of alternative oxidase at low temperature leads to a disruption of the balance in carbon and nitrogen metabolism, and to an up-regulation of antioxidant defense systems in Arabidopsis thaliana leaves. — Plant Cell Environ. 31: 1190–1202, 2008.

    Article  PubMed  CAS  Google Scholar 

  • Yoshida, K., Terashima, I., Noguchi, K.: Up-regulation of mitochondrial alternative oxidase concomitant with chloroplast over-reduction by excess light. — Plant Cell Physiol. 48: 606–614, 2007.

    Article  PubMed  CAS  Google Scholar 

  • Yoshida, K., Watanabe, C., Kato, Y., Sakamoto, W., Noguchi, K.: Influence of chloroplastic photo-oxidative stress on mitochondrial alternative oxidase capacity and respiratory properties: a case study with Arabidopsis yellow variegated 2. — Plant Cell Physiol. 49: 592–603, 2008.

    Article  PubMed  CAS  Google Scholar 

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Correspondence to V. I. Tarasenko.

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Acknowledgements: The study was financially supported by the Integration project SD RAS № 98.

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Tarasenko, V.I., Garnik, E.Y., Shmakov, V.N. et al. Modified alternative oxidase expression results in different reactive oxygen species contents in Arabidopsis cell culture but not in whole plants. Biol Plant 56, 635–640 (2012). https://doi.org/10.1007/s10535-012-0115-1

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  • DOI: https://doi.org/10.1007/s10535-012-0115-1

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