Abstract
Reactive oxygen species (ROS) are involved in many signalling pathways and numerous stress responses in plants. Consequently, it is important to be able to identify and localize ROS in vivo to evaluate their roles in signalling. A number of probes that have a high affinity for specific ROS and that are effectively taken up by cells and tissues are commercially available. Applications to intact leaves of singlet oxygen sensor green (SOSG), nitroblue tetrazolium (NBT), di-amino benzidine (DAB) and Amplex Red to detect singlet oxygen, superoxide and hydrogen peroxide are described. Imaging of the probes in the cells and tissues of leaves allows sites of ROS production to be identified.
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References
Foyer, C.H. and Noctor, G. (2003) Redox sensing and signalling associated with reactive oxygen species in chloroplasts, peroxisomes and mitochondria. Physiol. Plant. 119,355–364.
Apel, Kand . Hirt, H. (2004) Reactive oxygen species: Metabolism, oxidative stress, and signal transduction. Ann. Rev. Plant Biol. 55, 373–399.
Molecular Probes (2004) Product Information. www.probes.invitrogen.com/media/pis/mp36002.pdf?id=mp36002
Flohe, L and . Otting, F. (1984) Superoxide dismutase assays. Methods Enzymol. 105, 93–104.
Bielski, B.H.J., Shiue, G.G and ., Bajuk, S. (1980) Reduction of nitro blue tétrazolium by C02- and 02- radicals. J. Phys. Chem. 84, 830–833.
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.
Fryer, M.J., Oxborough, K., Mullineaux, P.M. and , Baker, N.R (2002) Imaging of photo-oxidative stress responses in leaves. J. Exp. Bot. 53, 1249–1254.
Thordal-Christensen, H., Zhang, Z., Wei, Y. and , Collinge, D.B. (1997) Subcellular localisation of H202 in plants. H202 accumulation in papillae and hypersensitive response during the barley-powdery mildew interaction. Plant J. 11,1187–1194.
Orozoco-Cardenas, M. and Ryan, C.A. (1999) Hydrogen peroxide is generated systemically in plant leaves by wounding and systemin via the octadecanoid pathway. Proc. Natl. Acad. Sci. USA96, 6553–6557.
Mohanty, J.G., Jaffe, J.S., Schulman, E.S., and Raible, D.G. (1997) A highly sensitive fluorescent micro-assay of H202 release from activated human leukocytes using a dihydroxyphenoxazine derivative./. Immunol. Methods 202,133–141.
Zhou, M., Diwu, Z., Panchuk-Voloshina, N., and Haugland, RP. (1997) A stable nonfluorescent derivative of resorufin for the fluorometric determination of trace hydrogen peroxide: applications in detecting the activity of phagocyte NADPH oxidase and other oxidases. Anal. Biochem. 253,162–168.
Abramoff, M.D., Magalhaes, P.J., and Ram, S.J. (2004) Image processing with Image]. Biophotonics Int. July, 36-42.
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Driever, S., Fryer, M., Mullineaux, P., Baker, N. (2009). Imaging of Reactive Oxygen Species In vivo. In: Pfannschmidt, T. (eds) Plant Signal Transduction. Methods in Molecular Biology, vol 479. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-59745-289-2_7
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DOI: https://doi.org/10.1007/978-1-59745-289-2_7
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-58829-943-7
Online ISBN: 978-1-59745-289-2
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