Abstract
Mitochondria are vital cytoplasmic organelle of eukaryotic cells responsible for oxidative energy metabolism and the synthesis of intermediates utilized in various other metabolic pathways. The functions of mitochondrion are the oxidation of organic acids by the tricarboxylic acid (TCA) cycle and the synthesis of ATP by the oxidative phosphorylation in the mitochondrial electron transport chain. The TCA cycle is composed by a set of enzymes that are essential for optimal functioning of the primary carbon metabolism in plants. The activity of each TCA cycle enzyme in plants may vary according to cell type, plant tissue, stage of plant development, and the environment. Here, we describe current methods used for the determination of the TCA cycle enzyme activities in different plant tissues.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Fernie AR, Carrari F, Sweetlove LJ (2004) Respiratory metabolism: glycolysis, the TCA cycle and mitochondrial electron transport. Curr Opin Plant Biol 7:254–261. doi:10.1016/j.pbi.2004.03.007
Araújo WL, Nunes-Nesi A, Nikoloski Z et al (2012) Metabolic control and regulation of the tricarboxylic acid cycle in photosynthetic and heterotrophic plant tissues. Plant Cell Environ 35:1–21. doi:10.1111/j.1365-3040.2011.02332.x
Millar AH, Whelan J, Soole KL et al (2011) Organization and regulation of mitochondrial respiration in plants. Annu Rev Plant Biol 62:79–104. doi:10.1146/annurev-arplant-042110-103857
Sweetlove LJ, Beard KFM, Nunes-Nesi A et al (2010) Not just a circle: flux modes in the plant TCA cycle. Trends Plant Sci 15:462–470. doi:10.1016/j.tplants.2010.05.006
Schmidtmann E, König A-C, Orwat A et al (2014) Redox regulation of Arabidopsis mitochondrial citrate synthase. Mol Plant 7:156–169. doi:10.1093/mp/sst144
Daloso DM, Müller K, Obata T et al (2015) Thioredoxin, a master regulator of the tricarboxylic acid cycle in plant mitochondria. Proc Natl Acad Sci 112. doi:10.1073/pnas.1424840112
Bocobza SE, Malitsky S, Araújo WL et al (2013) Orchestration of thiamin biosynthesis and central metabolism by combined action of the thiamin pyrophosphate riboswitch and the circadian clock in Arabidopsis. Plant Cell 25:288–307. doi:10.1105/tpc.112.106385
Araújo WL, Nunes-Nesi A, Trenkamp S et al (2008) Inhibition of 2-oxoglutarate dehydrogenase in potato tuber suggests the enzyme is limiting for respiration and confirms its importance in nitrogen assimilation. Plant Physiol 148:1782–1796. doi:10.1104/pp.108.126219
Henkes S, Sonnewald U, Badur R et al (2001) A small decrease of plastid transketolase activity in antisense tobacco transformants has dramatic effects on photosynthesis and phenylpropanoid metabolism. Plant Cell 13(3):535–552
Bunik VI, Fernie AR (2009) Metabolic control exerted by the 2-oxoglutarate dehydrogenase reaction: a cross-kingdom comparison of the crossroad between energy production and nitrogen assimilation. Biochem J 422(3):405–421
Tovar-Méndez A, Miernyk JA, Randall DD (2003) Regulation of pyruvate dehydrogenase complex activity in plant cells. Eur J Biochem 270:1043–1049. doi:10.1046/j.1432-1033.2003.03469.x
Randall DD, Rubin PM, Fenko M (1977) Plant pyruvate dehydrogenase complex purification, characterization and regulation by metabolites and phosphorylation. Biochim Biophys Acta 485:336–349. doi:10.1016/0005-2744(77)90169-3
Randall DD, Miernyk J (2012) 10 the mitochondrial pyruvate dehydrogenase complex. Enzymes of primary. Metabolism 3:175
Gibon Y, Blaesing OE, Hannemann J et al (2004) A robot-based platform to measure multiple enzyme activities in arabidopsis using a set of cycling assays: comparison of changes of enzyme activities and transcript levels during diurnal cycles and in prolonged darkness. Plant Cell 16:3304–3325. doi:10.1105/tpc.104.025973
Cox GF, Davies DD (1967) Nicotinamide–adenine dinucleotide-specific isocitrate dehydrogenase from pea mitochondria: purification and properties. Biochem J 105(2):729–734
Araújo WL, Nunes-Nesi A, Osorio S et al (2011) Antisense inhibition of the iron-sulphur subunit of succinate dehydrogenase enhances photosynthesis and growth in tomato via an organic acid–mediated effect on stomatal aperture. Plant Cell 23:600–627. doi:10.1105/tpc.110.081224
Jenner HL, Winning BM, Millar AH et al (2001) NAD malic enzyme and the control of carbohydrate metabolism in potato tubers. Plant Physiol 126(3):1139–1149
Studart-Guimarães C, Gibon Y, Frankel N et al (2005) Identification and characterisation of the α and β subunits of succinyl coa ligase of tomato. Plant Mol Biol 59:781–791. doi:10.1007/s11103-005-1004-1
Voll LM, Zell MB, Engelsdorf T et al (2012) Loss of cytosolic NADP-malic enzyme 2 in Arabidopsis thaliana is associated with enhanced susceptibility to Colletotrichum higginsianum. New Phytol 195:189–202. doi:10.1111/j.1469-8137.2012.04129.x
Murcha MW, Whelan J (2015) Isolation of intact mitochondria from the model plant species Arabidopsis thaliana and Oryza sativa. In: Whelan J, Murcha WM (eds) Plant mitochondria: methods and protocols. Springer New York, New York, NY, pp 1–12. doi:10.1007/978-1-4939-2639-8_1
Sweetlove LJ, Taylor NL, Leaver CJ (2007) Isolation of intact, functional mitochondria from the model plant Arabidopsis thaliana. In: Leister D, Herrmann JM (eds) Mitochondria: practical protocols. Humana Press, Totowa, NJ, pp 125–136. doi:10.1007/978-1-59745-365-3_9
Tronconi MA, Fahnenstich H, Gerrard Weehler MC et al (2008) Arabidopsis NAD-malic enzyme functions as a homodimer and heterodimer and has a major impact on nocturnal metabolism. Plant Physiol 146:1540–1552. doi:10.1104/pp.107.114975
Acknowledgements
Financial support was provided by Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq- to W.L.A.), Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG), and Max Planck Society to A.N.N. and W.L.A. Research fellowships granted by CNPq to A.N.N. and W.L.A. as well as scholarship granted by the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) to R.P.O.G. are also gratefully acknowledged.
Conflict of interest : The authors declare that they have no conflict of interest.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2017 Springer Science+Business Media LLC
About this protocol
Cite this protocol
Omena-Garcia, R.P., Araújo, W.L., Gibon, Y., Fernie, A.R., Nunes-Nesi, A. (2017). Measurement of Tricarboxylic Acid Cycle Enzyme Activities in Plants. In: Jagadis Gupta, K. (eds) Plant Respiration and Internal Oxygen. Methods in Molecular Biology, vol 1670. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7292-0_14
Download citation
DOI: https://doi.org/10.1007/978-1-4939-7292-0_14
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-7291-3
Online ISBN: 978-1-4939-7292-0
eBook Packages: Springer Protocols