Abstract
Evidence is emerging that reactive oxygen species (ROS) and antioxidants, together with plant hormones and other reactive species, such as reactive nitrogen species, are part of signalling networks pertinent to plant stress responses, cell division, and cell death. Consequently, they play pivotal roles in the regulation of seed development and maturation, germination and dormancy, seedling establishment, and seed ageing. Importantly, ROS, although essentially required at low concentrations, must be kept under stringent control by antioxidants. If the balance between pro- and antioxidative processes is lost and ROS production prevails, oxidative stress is the result, which can induce cell death and ultimately seed death. This chapter offers a variety of protocols for the determination of ROS, antioxidants, and stress markers aimed at enabling the reader to quantify these compounds. Protocols are also described to visualize ROS and localize the sites of ROS production, hoping to stimulate more research into ROS signalling and antioxidant control in key physiological and biochemical processes in seeds.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Bailly, C., El-Maarouf-Bouteau, H., and Corbineau, F. (2008) From intracellular signalling networks to cell death: the dual role of reactive oxygen species in seed physiology. C. R. Biol. 331, 806–14.
Bailly, C. (2004) Active oxygen species and antioxidants in seed biology. Seed Sci. Res. 14, 93–107.
Desikan, R., A-H-Mackerness, S., Hancock, J.T., and Neill, S.J. (2001) Regulation of the Arabidopsis transcriptome by oxidative stress. Plant Physiol. 127, 159–72.
Queval, G., Hager, J., Gakière, B., and Noctor, G. (2008) Why are literature data for H2O2 contents so variable? A discussion of potential difficulties in the quantitative assay of leaf extracts. J Exp Bot. 59, 135–46.
O’Kane, D., Gill, V., Boyd, P., and Burdon, R. (1996) Chilling, oxidative stress and antioxidant responses in Arabidopsis thaliana callus. Planta 198, 371–7.
Elstner, E.F. and Heupel, A. (1976) Inhibition of nitrite formation from hydroxylammoniumchloride: a simple assay for superoxide dismutase. Anal. Biochem. 70, 616–20.
Warm, E. and Laties, G.G. (1982) Quantification of hydrogen peroxide in plant extracts by the chemiluminescence reaction with luminol. Phytochem. 4, 827–31.
Gay, C. and Gebicki, J. M. (2000) A critical evaluation of the effect of sorbitol on the ferric-xylenol orange hydroperoxide assay. Anal. Biochem. 284, 217–220.
Misra, H. R. and Fridovich, I. (1972) The univalent reduction of oxygen by reduced flavins and quinones. J. Biol. Chem. 247, 188–92.
Able, A. J., Guest, D. I., and Sutherland, M. W. (1998) Use of a new tetrazolium-based assay to study the production of superoxide radicals by tobacco cell cultures challenged with avirulent zoospores of Phytophthora parasitica var nicotianae. Plant Physiol. 117, 491–9.
Sutherland, M. W., and Learmonth, B. A. (1997) The tetrazolium dyes MTS and XTT provide new quantitative assays for superoxide and superoxide dismutase. Free Radical Res. 27, 283–9.
Bestwick, C.S., Brown, I.R., Benneth, M.H.R., and Mansfield, J.W. (1997) Localization of hydrogen peroxide accumulation during the hypersensitive reaction of lettuce cells to Pseudomonas syringae pv. phaseolicola. Plant Cell 9, 209–21.
Barcelo, A.R. (1998) Hydrogen peroxide production is a general property of the lignifying xylem from vascular plants. Ann. Bot. 82, 97–103.
Maffei, M.E., Mithöfer, A., Arimura, G., Uchtenhagen, H., Bossi, S., Bertea, C.M., Cucuzza, L.S., Novero, M., Volpe, V., Quadro, S., and Boland, W. (2006) Effects of feeding Spodoptera littoralis on lima bean leaves. III. Membrane depolarization and involvement of hydrogen peroxide. Plant Physiol. 140, 1022–35.
Beyer, W.F., and Fridovich, I. (1987) Assaying for superoxide dismutase activity: some large consequences of minor changes in conditions. Anal. Biochem. 161, 559–66.
Schöpfer, P., Plachy, C., and Frahry, G. (2001) Release of reactive oxygen intermediates (superoxide radicals, hydrogen peroxide, and hydroxyl radicals) and peroxidase in germinating radish seeds controlled by light, gibberellin, and abscisic acid. Plant Physiol. 125, 1591–1602.
Tarpey, M.M., Wink, D.A., and Grisham, M.B. (2004). Methods for detection of reactive metabolites of oxygen and nitrogen: In vitro and in vivo considerations. Am. J. Physiol. Regul. Integr. Comp. Physiol. 286, R431-44.
Giannopolitis, C.N., and Ries, S.K. (1977) Superoxide dismutases I. Occurrence in higher plants. Plant Physiol. 59, 309–14.
Clairbone, A. (1985) Catalase activity. In: Greenwald R.A. (ed.), Handbook of Methods for Oxygen Radical Research. CRC Press, Boca Raton, FL., pp. 283–4.
Esterbauer, H., and Grill, D. (1978) Seasonal variation of glutathione and glutathione reductase in needles of Picea abies. Plant Physiol. 61, 119–21.
Nakano, Y., and Asada, K. (1981) Hydrogen peroxide is scavenged by ascorbate-specific peroxidase in spinach chloroplasts. Plant Cell Physiol. 22, 867–80.
Bailly, C., Leymarie, J., Lehner, A., Rousseau, S., Côme, D., and Corbineau F. (2004) Catalase activity and expression in developing sunflower seeds as related to drying. J Exp. Bot. 55, 475–83.
Kranner, I., and Grill, D. (1996) Determination of glutathione and glutathione disulfide in lichens: a comparison of frequently used methods. Phytochem. Anal. 7, 24–8.
Kranner, I., Birtić, S., Anderson, K.M., and Pritchard, H.W. (2006) Glutathione half-cell reduction potential: a universal stress marker and modulator of programmed cell death? Free Radic. Biol. Med. 40, 2155–65.
Tausz, M., Kranner, I., and Grill, D. (1996) Simultaneous determination of ascorbic-acid and dehydroascorbic-acid in plant materials by high performance liquid chromatography. Phytochem. Anal. 7, 69–72.
Pfeifhofer, H.W., Willfurth, R., Zorn, M., and Kranner, I. (2002) Analysis of chlorophylls, carotenoids, and tocopherols in lichens. In: Kranner I., Beckett R. und Varma A. (eds) Protocols in Lichenology. Culturing, Biochemistry, Ecophysiology and Use in Biomonitoring, Springer Verlag, Berlin, pp. 363–78.
Heath, R.L., and Parker L. (1968) Photoperoxidation in isolated chloroplasts. I. Kinetics and stoichiometry of fatty acid peroxidation. Arch. Bioch. Bioph. 125, 189–98.
Gidrol, X., Serghini, H., Noubhani, A., Mocquot, B., and Mazliak, P. (1989) Biochemical-changes induced by accelerated aging in sunflower seeds.1. Lipid-peroxidation and membrane damage. Physiol Plant. 76, 591–7.
Schafer, F.Q., and Buettner, G. R. (2001) Redox environment of the cell as viewed through the redox state of the glutathione disulfide/glutathione couple. Free Radic. Biol. Med. 30, 1191–1212.
Acknowledgements
We thank Dr. Thomas Roach and Dr. Farida Minibayeva for useful discussions on the epinephrine, XTT, and xylenol orange assays. The Royal Botanic Gardens, Kew, grant-in-aid from Defra, and the Millennium Seed Bank Project supported by the Millennium Commission, The Wellcome Trust, Orange Plc., and Defra.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2011 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Bailly, C., Kranner, I. (2011). Analyses of Reactive Oxygen Species and Antioxidants in Relation to Seed Longevity and Germination. In: Kermode, A. (eds) Seed Dormancy. Methods in Molecular Biology, vol 773. Humana Press. https://doi.org/10.1007/978-1-61779-231-1_20
Download citation
DOI: https://doi.org/10.1007/978-1-61779-231-1_20
Published:
Publisher Name: Humana Press
Print ISBN: 978-1-61779-230-4
Online ISBN: 978-1-61779-231-1
eBook Packages: Springer Protocols