The tetrazolium dyes MTS and XTT provide new quantitative assays for superoxide and superoxide dismutase

Article

Sutherland, Mark W. and Learmonth, Barbara A.. 1997. "The tetrazolium dyes MTS and XTT provide new quantitative assays for superoxide and superoxide dismutase." Free Radical Research. 27 (3), pp. 283-289. https://doi.org/10.3109/10715769709065766
Article Title

The tetrazolium dyes MTS and XTT provide new quantitative assays for superoxide and superoxide dismutase

ERA Journal ID2219
Article CategoryArticle
AuthorsSutherland, Mark W. (Author) and Learmonth, Barbara A. (Author)
Journal TitleFree Radical Research
Journal Citation27 (3), pp. 283-289
Number of Pages7
Year1997
PublisherTaylor & Francis
Place of PublicationNetherlands
ISSN1029-2470
1071-5762
Digital Object Identifier (DOI)https://doi.org/10.3109/10715769709065766
Web Address (URL)https://www.tandfonline.com/doi/abs/10.3109/10715769709065766
Abstract

The tetrazolium dyes MTS and XTT were reduced to their soluble formazans by superoxide radical anions (O2_) produced by the oxidation of xanthine by xanthine oxidase under standard conditions. These reactions were compared to the well-known reductions of NBT and cytochrome c by the xanthine/xanthine oxidase system. Reduction of the dyes was completely inhibited by superoxide dismutase (SOD). Rate constants for the reaction of MTS and XTT with O2_: were estimated at 1.3 times .1 times 105 M-1s-1 and 8.6 times .8 times 104 M-1s-1 respectively. The stable MTS and XTT formazans have high extinction coefficients in the visible range which enable sensitive detection and quantification of superoxide radicals, avoiding some of the problems inherent in assays based on production of the insoluble NBT formazan. MTS and XTT have considerable potential both for the quantitative assay of radical production in living tissues and for the assay of superoxide dismutase activity in tissue extracts. Implications for the interpretation of cell culture growth assays which employ these dyes are discussed.

KeywordsMTS; NBT; XlT; cytochrome c; superoxide assay; tetrazolium dyes
ANZSRC Field of Research 2020319999. Other biological sciences not elsewhere classified
310105. Cellular interactions (incl. adhesion, matrix, cell wall)
340102. Bioassays
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Knight, N. L., Platz, G. J., Lehmensiek, A. and Sutherland, M. W.. 2010. "An investigation of genetic variation among Australian isolates of Bipolaris sorokiniana from different cereal tissues and comparison of their abilities to cause spot blotch on barley." Australasian Plant Pathology. 39 (3), pp. 207-216. https://doi.org/10.1071/AP09082
Genomic regions associated with common root rot resistance in the barley variety Delta
Lehmensiek, A., Bovill, J., Sutherland, M. W. and McNamara, R. B.. 2010. "Genomic regions associated with common root rot resistance in the barley variety Delta." Australasian Plant Pathology. 39 (3), pp. 241-246. https://doi.org/10.1071/AP09089
Population structure of South African and Australian Pyrenophora teres isolates
Lehmensiek, A., Bester-van der Merwe, A. E., Sutherland, M. W., Platz, G., Kriel, W. M., Potgieter, G. F. and Prins, R.. 2010. "Population structure of South African and Australian Pyrenophora teres isolates." Plant Pathology. 59 (3), pp. 504-515. https://doi.org/10.1111/j.1365-3059.2009.02231.x
Pyramiding QTL increases seedling resistance to crown rot (Fusarium pseudograminearum) of wheat (Triticum aestivum)
Bovill, W. D., Horne, M., Herde, D., Davis, M., Wildermuth, G. B. and Sutherland, M. W.. 2010. "Pyramiding QTL increases seedling resistance to crown rot (Fusarium pseudograminearum) of wheat (Triticum aestivum)." Theoretical and Applied Genetics: international journal of plant breeding research. 121 (1), pp. 127-136. https://doi.org/10.1007/s00122-010-1296-7
Phosphate transporter expression in the soybean AM symbiosis
Dearnaley, John, Learmonth, Barbara A. and Bougoure, Damian S.. 2002. "Phosphate transporter expression in the soybean AM symbiosis." 7th International Mycological Congress (IMC7). Oslo, Norway 11 - 17 Aug 2002 Australia.
The generation of oxygen radicals during host plant responses to infection
Sutherland, Mark W.. 1991. "The generation of oxygen radicals during host plant responses to infection." Physiological and Molecular Plant Pathology. 39 (1), pp. 71-93.
Defence gene expression in single mycorrhizal roots of soybean
Dearnaley, John and Learmonth, Barbara A.. 2003. "Defence gene expression in single mycorrhizal roots of soybean." 4th International Conference on Mycorrhizae (ICOM4) . Montreal, Canada 10 - 15 Aug 2003 Québec, Canada.
Genes involved in hypersensitive cell death responses during fusarium crown rot infection in wheat
Petrisko, Jill E., Sutherland, Mark W. and Windes, Juliet M.. 2009. "Genes involved in hypersensitive cell death responses during fusarium crown rot infection in wheat." Guest, David (ed.) 17th Biennial Australasian Plant Pathology Society Conference (APPS 2009): Plant Health Management: An Integrated Approach. Newcastle, Australia 29 Sep - 01 Oct 2009 Toowoomba, Australia.
Bipolaris sorokiniana resistance loci identified on chromosomes 3HS and 7HS in three related barley
Sutherland, M. W., Lehmensiek, A., Leist, C., Platz, G. and Usher, T. R.. 2009. "Bipolaris sorokiniana resistance loci identified on chromosomes 3HS and 7HS in three related barley." Poulsen, David (ed.) ABTS 2009: Barley - The Grain for Gain. Sunshine Coast, Australia 13 - 16 Sep 2009 Indooroopilly, Australia.
Chromosome composition in an F2 hexaploid x durum cross analysed by DArT markers and MCFISH
Eberhard, F. S., Zhang, P., Lehmensiek, A., Sutherland, M. W., Hare, R. and Simpfendorfer, S.. 2008. "Chromosome composition in an F2 hexaploid x durum cross analysed by DArT markers and MCFISH." Appels, Rudi, Eastwood, Russell, Lagudah, Evans, Mackay, Michael, McIntyre, Lynne, Sharp, Peter and Langridge, Peter (ed.) 11th International Wheat Genetics Sympoium (IWGS 2008). Brisbane, Australia 24 - 29 Aug 2008 Sydney, Australia.
The application of molecular markers for partial resistance against fusarium crown rot in hexaploid and tetraploid wheats
Sutherland, Mark, Bovill, W. D., Horne, M., Lehmensiek, A., Eberhard, F., Percy, C., Wildermuth, G. B., Simpfendorfer, S. and Hare, R.. 2008. "The application of molecular markers for partial resistance against fusarium crown rot in hexaploid and tetraploid wheats." 10th International Fusarium and Fusarium Genomics Workshop 2008. Sardinia, Italy 30 Aug - 02 Sep 2008 Sardinia,Italy.
Segregation of glutenins in wheatxmaize-derived doubled haploid wheat populations
Kammholz, Stephen J., Grams, Raechelle A., Banks, Phillip and Sutherland, Mark W.. 1998. "Segregation of glutenins in wheatxmaize-derived doubled haploid wheat populations." Australian Journal of Agricultural Research. 49 (8), pp. 1253-1259. https://doi.org/10.1071/A98032
Use of a new tetrazolium-based assay to study the production of superoxide radicals by tobacco cell cultures challenged with avirulent zoospores of Phytopthora parasitica var nicotianae
Able, Amanda J., Guest, David I. and Sutherland, Mark 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 Phytopthora parasitica var nicotianae." Plant Physiology. 117 (2), pp. 491-499.
Improving the efficiency of haploid wheat production mediated by wide crossing
Kammholz, Stephen J., Sutherland, Mark W. and Banks, Phillip. 1996. "Improving the efficiency of haploid wheat production mediated by wide crossing." Sabrao Journal. 28, pp. 37-46.
Hydrogen peroxide yields during the incompatible interaction of tobacco suspension cells inoculated with Phytophthora nicotiannae
Able, Amanda J., Guest, David I. and Sutherland, Mark W.. 2000. "Hydrogen peroxide yields during the incompatible interaction of tobacco suspension cells inoculated with Phytophthora nicotiannae." Plant Physiology. 124 (2), pp. 899-910.
Relationship between transembrane ion movements, production of reactive oxygen species and the hypersensitive response during the challenge of tobacco suspension cells by zoospores of Phytophthora nicotianae
Able, Amanda J., Guest, David I. and Sutherland, Mark W.. 2001. "Relationship between transembrane ion movements, production of reactive oxygen species and the hypersensitive response during the challenge of tobacco suspension cells by zoospores of Phytophthora nicotianae." Physiological and Molecular Plant Pathology. 58 (5), pp. 189-198. https://doi.org/10.1006/pmpp.2001.0328
High and low pre-inoculation temperatures decrease the effectiveness of the Lr20 and Sr15 rust resistance genes in wheat
Ramage, R. A and Sutherland, Mark W.. 1995. "High and low pre-inoculation temperatures decrease the effectiveness of the Lr20 and Sr15 rust resistance genes in wheat." Plant Pathology. 44 (5), pp. 772-778. https://doi.org/10.1111/j.1365-3059.1995.tb02734.x
Mapping QTLs for polyphenol oxidase activity in a DH population from common wheat
Zhang, Li-Ping, Ge, Xiu-Xiu, He, Zhong-Hu, Wang, De-Sen, Yan, Jun, Xia, Xian-Chun and Sutherland, Mark W.. 2005. "Mapping QTLs for polyphenol oxidase activity in a DH population from common wheat." Zuowu Xuebao. 31 (1), pp. 7-10.
Biological methods: a novel course in undergraduate biology
Kotiw, Michael, Learmonth, Robert P. and Sutherland, Mark W.. 1999. "Biological methods: a novel course in undergraduate biology." Biochemical Education. 27 (3), pp. 131-134. https://doi.org/10.1016/S0307-4412(99)00002-3
Mapping markers for priority traits in the northern grains region of Australia
Mace, E., Sutherland, Mark, Lehmensiek, Anke, Bovill, Bill, Bovill, Jessica, Collard, Bert, Daggard, Grant and Christopher, M. J.. 2006. "Mapping markers for priority traits in the northern grains region of Australia." ITMI Workshop and ACPFG Genomics Symposium (2006). Victor Harbour, Australia 27 - 31 Aug 2006
Genetic control of dormancy in white-grained wheats of diverse origin
Mares, Daryl, Mrva, Kolumbina, Cheong, Judy, Williams, Kevin, Cavallaro, Benedette, Storlie, Eric, Sutherland, Mark and Zou, Y.. 2004. "Genetic control of dormancy in white-grained wheats of diverse origin." Black, C. K., Panozzo, J. F. and Rebetzke, G. J. (ed.) 54th Australian Cereal Chemistry Conference (CEREALS 2004). Canberra, Australia 21 - 24 Sep 2004 Melbourne, Australia.
Identification of novel QTL for resistance to crown rot in the doubled haploid wheat population 'W21MMT70' x 'Mendos'
Bovill, W. D., Ma, W., Ritter, K., Collard, B. C. Y., Davis, M., Wildermuth, G. B. and Sutherland, M. W.. 2006. "Identification of novel QTL for resistance to crown rot in the doubled haploid wheat population 'W21MMT70' x 'Mendos' ." Plant Breeding. 125 (6), pp. 538-543. https://doi.org/10.1111/j.1439-0523.2006.01251.x
Effectiveness of partial resistance sources in hexaploid wheats against fusarium pseudograminearum isolates
Percy, C. D., Wildermuth, G. B. and Sutherland, Mark W.. 2005. "Effectiveness of partial resistance sources in hexaploid wheats against fusarium pseudograminearum isolates." Cahill, David (ed.) 15th Biennial Australasian Plant Pathology Society Conference. Geelong, Australia 26 - 29 Sep 2005 Geelong, Australia.
Strategies for fine mapping QTLs for crown rot resistance in wheats
Bovill, W. D., Collard, B. C. Y., Lehmensiek, Anke, Wildermuth, G. B. and Sutherland, Mark W.. 2005. "Strategies for fine mapping QTLs for crown rot resistance in wheats." Cahill, David (ed.) 15th Biennial Australasian Plant Pathology Society Conference. Geelong, Australia 26 - 29 Sep 2005 Geelong.
Excel table of genotyping data of the markers on the A and B genome chromosomes of eight hexaploid/tetraploid populations
Martin, Anke, Sutherland, Mark W. and Eberhard, Susette. 2007. Excel table of genotyping data of the markers on the A and B genome chromosomes of eight hexaploid/tetraploid populations. Toowoomba. University of Southern Queensland. https://doi.org/10.26192/q37y8
Targetting AFLP-DNA markers to specific traits and chromosome regions
Campbell, A. W., Daggard, G., Békés, F., Pedler, A., Sutherland, M. W and Appels, R.. 2001. "Targetting AFLP-DNA markers to specific traits and chromosome regions ." Australian Journal of Agricultural Research. 52 (11-12), pp. 1153-1160. https://doi.org/10.1071/ar01028