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Role of light and CO2 fixation in the control of nitrate-reductase activity in barley leaves

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Abstract

Nitrate reductase (NR, NADH:nitrate oxidoreductase, EC 1.6.6.1) from barley (Hordeum vulgare L. cv. Hassan) leaves was inactivated during a light-dark transition, losing approx. 50% of activity after 30 min of darkness. The dark inactivation was reversed by illumination of the seedlings, the kinetics of reactivation being similar to those of inactivation. High extractable NR activity and significant differences between illuminated and darkened leaves were observed in media containing EDTA and inorganic phosphate (Pi). Addition of Ca2+ ions during extraction and assay decreased NR activity from illuminated and darkened leaves, enhancing the light-dark difference. While no clear correlation could be found between irradiance and NR activity, a hyperbolic correlation appeared between extractable NR activity and in-vivo rates of CO2 fixation, indicating that NR activation follows saturation kinetics with respect to CO2 fixation. Furthermore, hexoses and hexose-phosphates fed to the leaves via the transpiration stream protected against the dark-inactivation of NR. The results indicate that carbon-assimilation products are regulatory factors of NR activity in barley leaves, mediating both the light-dark modulation of NR and its dependence upon CO2 fixation.

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Abbreviations

NR:

NADH-nitrate reductase

Pi :

inorganic phosphate

PPFD:

photosynthetic photon flux density

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Authors and Affiliations

  1. Departamento de Biología Vegetal y Ecología, Universidad de Sevilla, Apdo. 1095, E-41080, Sevilla, Spain

    Alfonso de Cires & Angel de la Torre

  2. Institute de Bioquímica Vegetal y Fotosíntesis, Universidad de Sevilla-CSIC, Apdo. 1095, E-41080, Sevilla, Spain

    Begoña Delgado & Catalina Lara

Authors
  1. Alfonso de Cires
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  2. Angel de la Torre
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  3. Begoña Delgado
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  4. Catalina Lara
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Additional information

This work was supported by grants from Dirección General de Investigación Científica y Técnica (PB88-0019 and PB91-0611) and Plan Andaluz de Investigación (gr. 4038 and 3101), Spain. We are grateful to Professor M.G. Guerrero (Sevilla, Spain) for fruitful discussions and a critical reading of the manuscript, and to Professors H.W. Heldt (Göttingen, FRG) and W.M. Kaiser (Würzburg, FRG) for discussing and making available prepublished data. Thanks are also due to P. Pérez de León for skillful secretarial assistance. A.C. is a recipient of a fellowship from Fundación Cámara.

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de Cires, A., de la Torre, A., Delgado, B. et al. Role of light and CO2 fixation in the control of nitrate-reductase activity in barley leaves. Planta 190, 277–283 (1993). https://doi.org/10.1007/BF00196622

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  • DOI: https://doi.org/10.1007/BF00196622

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