Abstract
Reactive oxygen species (ROS) are formed in biological systems as part of normal metabolism. Adverse environmental factors like drought stress result in increased levels of ROS that are detrimental to the plant (1, 2). To avoid damage caused by these excess ROS, plants have developed elaborate mechanisms to manage them at sustainable levels. Enzymes play an important role in lowering the ROS levels and helping avoid oxidative stress. Superoxide dismutase, catalase, ascorbate peroxidase, and glutathione reductase play a vital role in combating oxidative stress. Measuring these enzyme activities spectrophotometrically provides researchers an easy and precise way to study and understand an important part of the defense against oxidative stress. In this chapter we provide details of the assays we used to determine the enzyme activities spectrophotometrically. Antioxidant enzyme responses to moderate water-deficit stress were studied. All enzyme assays were conducted using wheat leaf tissue.
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References
Asada, K. (1999) The water–water cycle in chloroplasts: scavenging of active oxygens and dissipation of excess photons. Annu Rev Plant Physiol Plant Mol Biol 50, 601–639.
Borsani, O., DÃaz, P., Agius, M.F., Valpuesta, V., and Monza, J. (2001) Water stress generates an oxidative stress through the induction of a specific Cu/Zn superoxide dismutase in Lotus corniculatus leaves. Plant Sci 161, 757–763.
Allen, R.D. (1995) Dissection of oxidative stress tolerance using transgenic plants. Plant Physiol 107, 1049–1054.
Khanna-Chopra, R. and Selote, D.S. (2007) Acclimation to drought stress generates oxidative stress tolerance in drought-resistant than susceptible wheat cultivar under field conditions. Environ Exp Bot 60, 276–283.
Loggini, B., Scartazza, A., Brugnoli, E., and Navari-Izzo, F. (1999) Antioxidative defense system, pigment composition, and photosynthetic efficiency in two wheat cultivars subjected to drought. Plant Physiol 119, 1091–1100.
Abebe, T., Guenzi, A.C., Martin, B., and Cushman, J.C. (2003) Tolerance of mannitol-accumulating transgenic wheat to water stress and salinity. Plant Physiol 131, 1748–1755.
Shen, B., Jensen, R.G., and Bohnert, H.J. (1997) Mannitol protects against oxidation by hydroxyl radicals. Plant Physiol 115, 527–532.
Wang, F.-Z., Wang, Q.-B., Kwon, S.-Y., Kwak, S.-S., and Su, W.-A. (2005) Enhanced drought tolerance of transgenic rice plants expressing a pea manganese superoxide dismutase. J Plant Physiol 62, 465.
Yoshimura, K., Yabuta, Y., Ishikawa, T., and Shigeoka, S. (2000) Expression of spinach ascorbate peroxidase isoenzymes in response to oxidative stresses. Plant Physiol 123, 223–234.
Schaedle, M. and Bassham, J.A. (1977) Chloroplast glutathione reductase. Plant Physiol 59, 1011–1012.
Beyer, W.F. and Fridovich, I. (1987) Assaying for superoxide dismutase activity: some large consequences of minor changes in conditions. Anal Biochem 161, 559–566.
Giannopolitis, C.N. and Ries, S.K. (1977) Superoxide dismutases: I. Occurrence in higher plants. Plant Physiol 59, 309–314.
Aebi, H. and Lester, P. (1984) Catalase in vitro. Meth Enzymol,121–126.
Nakano, Y. and Asada, K. (1981) Hydrogen peroxide is scavenged by ascorbate-specific peroxidase in spinach chloroplasts. Plant Cell Physiol 22, 867–280.
Smith, I.K., Vierheller, T.L., and Thorne, C.A. (1988) Assay of glutathione reductase in crude tissue homogenates using 5,5ʹ-dithiobis(2-nitrobenzoic acid). Anal Biochem 175, 408–413.
Acknowledgments
The authors would like to thank Dr. Kalpalatha Melmaiee and Shraddha Vadvalkar for valuable suggestions and help in the critical review of the chapter.
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Elavarthi, S., Martin, B. (2010). Spectrophotometric Assays for Antioxidant Enzymes in Plants. In: Sunkar, R. (eds) Plant Stress Tolerance. Methods in Molecular Biology, vol 639. Humana Press. https://doi.org/10.1007/978-1-60761-702-0_16
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DOI: https://doi.org/10.1007/978-1-60761-702-0_16
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