Abstract
The activity of NADP+-specific isocitrate dehydrogenase (NADP+-IDH, EC 1.1.1.42) was investigated during the ripening of tomato (Lycopersicon esculentum Mill.) fruit. In the breaker stage, NADP+-IDH activity declined but a substantial recovery was observed in the late ripening stages when most lycopene synthesis occurs. These changes resulted in higher NADP+-IDH activity and specific polypeptide abundance in ripe than in green fruit pericarp. Most of the enzyme corresponded to the predominant cytosolic isoform which was purified from both green and ripe fruits. Fruit NADP+-IDH seems to be a dimeric enzyme having a subunit size of 48 kDa. The K m values of the enzymes from green and ripe pericarp for NADP+, isocitrate and Mg2+ were not significantly different. The similar molecular and kinetic properties and chromatographic behaviour of the enzymes from the two kinds of tissue strongly suggest that the ripening process is not accompanied by a change in isoenzyme complement. The increase in NADP+-IDH in the late stage of ripening also suggests that this enzyme is involved in the metabolism of C6 organic acids and in glutamate accumulation in ripe tissues.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- DEAE:
-
diethylaminoethyl
- GADPH:
-
NADP+-dependent glyceraldehyde 3-phosphate dehydrogenase
- GOGAT:
-
glutamate synthase
- GS:
-
glutamine synthetase
- Mr :
-
relative molecular mass
- NAD+-IDH:
-
NAD+-specific isocitrate dehydrogenase
- NADP+ -IDH:
-
NADP+-specific isocitrate dehydrogenase
- PEPCase:
-
phosphoenolpyruvate carboxylase
References
Arnozis, P.A., Nelemans, J.A., Findenegg, G.R. (1988) Phophoenolpyruvate carboxylase activity in plants grown with either NO su−3 or NH +4 as inorganic nitrogen source. J. Plant Physiol. 132, 23–27
Bradford, M.M. (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilising the principle of protein-dye binding. Anal. Biochem. 72, 248–254
Brady, C.J. (1987) Fruit ripening. Annu. Rev. Plant. Physiol. 38, 155–178
Brouquisse, R., Nishimura, M., Gaillard, J., Douce, A. (1987) Characterization of a cytosolic aconitase in higher plant cells. Plant Physiol. 84, 1402–1430
Chen, R.D., Gadal, P. (1990a) Do the mitochondria provide the 2-oxoglutarate needed for glutamate synthesis in higher plant chloroplasts? Plant Physiol. Biochem. 28, 141–145
Chen, R.D., Gadal, P. (1990b) Structure, functions and regulation of NAD+ and NADP+ dependent isocitrate dehydrogenases in higher plants and in other organisms. Plant Physiol. Biochem 28, 411–427
Chen, R.D., Bismuth, E., Champigny, M.L., Gadal, P. (1989 a) Chromatographic and immunological evidence that chloroplastic and cytosolic pea (Pisum sativum L.) NADP-isocitrate dehydrogenase are distinct isoenzymes. Planta 178, 157–163
Chen, R.D., Le Marechal, P., Vidal, J., Jacquot, J.P., Gadal, P. (1988) Purification and comparative properties of the cytosolic isocitrate dehydrogenases (NADP) from pea (Pisum sativum) roots and green leaves. Eur. J. Biochem. 175, 565–572
Gallardo, F. (1992) Evolución de las enzimas de la asimilación de amonio durante la diferenciación de plastidios de Lycopersicon esculentum L. Ph. D. thesis, Universidad de Málaga
Gallardo, F., Cánovas, F.M. (1992) A macromolecular inhibitor of glutamine synthetase activity in tomato root extracts. Phytochemistry 31, 2267–2271
Gallardo, F., Gálvez, S., Quesada, M.A., Cánovas, F.M., Nuñez de Castro, I. (1988) Glutamine synthetase activity during the ripening of tomato. Plant Physiol. Biochem. 26, 747–752
Gallardo, F., Cantón, F.R., Garcia-Gutiérrez, A., Cánovas, F.M. (1993) Changes in photorespiratory enzymes and glutamate synthases in ripening tomatoes. Plant Physiol. Biochem. 31, 189–196
Gálvez, S., Bismuth, E., Sarda, C., Gadal, P. (1994) Purification and characterization of chloroplastic NADP-isocitrate dehydrogenase from mixotrophic tobacco cells. Comparison with the cytosolic isoenzyme. Plant Physiol. 105, 593–600
Goodenough, P.W., Prosser, I.M., Young, K. (1985) NADP-linked malic enzyme and malate metabolism in ageing tomato fruit. Phytochem. 24, 1157–1162
Grierson, D., Maunders, M.J., Slater, A., Ray, J., Bird, C.R., Schuch, W., Holdsworth, M.J., Tucker, G.A., Knapp, J.E. (1986) Gene expression during tomato ripening. Phil. Trans. R. Soc. Lond. B. 314, 399–410
Grierson, D., Slater, A., Maunders, M., Crookes, P., Tucker, G.A., Schuch, W., Edwards, K. (1985) In: Ethylene and plant development, pp. 147-161, Roberts, J.A., Tucker, G.A., eds. Butterworth, London
Gupta, V.K., Singh, R. (1988) Partial purification and characterization of NADP+-isocitrate dehydrogenase from immature pod walls of chickpea (Cicer arietinum L.). Plant Physiol. 87, 741–744
Hanning, I., Heldt, H.W. (1993) On the function of mitochondrial metabolism during photosynthesis in spinach (Spinacia oleracea L.) leaves. Plant Physiol. 103, 1147–1154
Jeffery, D., Goodenough, P.W., Weitzman, P.D.J. (1986) Enzyme activities in mitochondria isolated from ripening tomato fruit. Plant 168, 390–394
Laemmli, U.K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680–685
Laval-Martin, D., Farineau, J., Diamond, J. (1977) Light versus dark carbon metabolism in cherry tomato fruits. Plant Physiol. 60, 872–876
Leshem, Y.Y., Havely, A.H., Frenkel, C., eds. (1986) Processes and control of plant senescence. Elsevier Publishers B.V., Amsterdam
Miflin, B.J., Wallsgrove, R.M., Lea, P.J. (1981) Glutamine metabolism in higher plants. Curr. Top. Cell. Regul. 20, 1–43
Morrisey, J.H. (1981) Silver stain for proteins in polyacrylamide gels, A modified procedure with enhanced uniform sensitivity. Anal. Biochem. 117, 307–310
Ni, W., Robertson, E.F., Reeves, H.C. (1987) Purification and characterization of cytosolic NADP specific isocitrate dehydrogenase from Pisum sativum. Plant Physiol. 83, 785–788
Piechulla, B., Glick, R.E., Bahl, H., Melis, A., Gruissem, W. (1987) Changes in photosynthetic capacity and photosynthetic protein pattern during tomato fruit ripening. Plant Physiol. 84, 911–917
Randall, D.D., Givan, C.V. (1981) Subcellular location of NADP-isocitrate dehydrogenase in Pisum sativum leaves. Plant Physiol. 68, 70–73
Rasmusson, A.G., Möller, I.M. (1990) NADP-Utilizing enzymes in the matrix of plant mitochondria. Plant Physiol. 94, 1012–1018
Stevens, M.A. (1972) Citrate and malate concentrations in tomato fruits: genetic control and maturational effects. J. Am. Soc. Hort. Sci. 97, 655–685
Towbin, H., Staehelin, T., Gordon, J. (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc. Natl. Acad. Sci. USA. 76, 4350–4354
Willmer, C.M., Johnston, W.R. (1976) Carbon dioxide assimilation in some aerial plant organs and tissues. Planta 130, 33–37
Author information
Authors and Affiliations
Additional information
We thank to M. Hodges (Université Paris Sud) for the revision and criticism of the manuscript. This work was partially supported by a Grant from Direction General de Investigatión Científica y Técnica (PB 92-0423) to F.M.C. F.G. was supported by a fellowship from the Programa General de Formatión de Personal Investigador, Junta de Andalucia, and by a short-term fellowship from Ministerio de Educacion y Ciencia. S.G. is recipient of a fellowship from the Programa de Formatión de Personal Investigador en el Extranjero; Ministerio de Educación y Ciencia, Spain.
Rights and permissions
About this article
Cite this article
Gallardo, F., Gálvez, S., Gadal, P. et al. Changes in NADP+-linked isocitrate dehydrogenase during tomato fruit ripening. Planta 196, 148–154 (1995). https://doi.org/10.1007/BF00193228
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00193228