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Plant Physiol, November 2001, Vol. 127, pp. 817-831

Antioxidant Systems and O2.minus /H2O2 Production in the Apoplast of Pea Leaves. Its Relation with Salt-Induced Necrotic Lesions in Minor Veins1

José A. Hernández, Maria Angeles Ferrer, Ana Jiménez, Alfonso Ros Barceló, and Francisca Sevilla*

Department of Nutrition and Plant Physiology, Centro de Edafología y Biología Aplicada del Segura-Consejo Superior de Investigaciones Científicas, Campus de Espinardo, E-30100 Murcia, Spain (J.A.H., A.J., F.S.); and Department of Plant Biology (Plant Physiology), University of Murcia, E-30100 Murcia, Spain (M.A.F., A.R.B.)

The present work describes, for the first time, the changes that take place in the leaf apoplastic antioxidant defenses in response to NaCl stress in two pea (Pisum sativum) cultivars (cv Lincoln and cv Puget) showing different degrees of sensitivity to high NaCl concentrations. The results showed that only superoxide dismutase, and probably dehydroascorbate reductase (DHAR), were present in the leaf apoplastic space, whereas ascorbate (ASC) peroxidase, monodehydroascorbate reductase (MDHAR), and glutathione (GSH) reductase (GR) seemed to be absent. Both ASC and GSH were detected in the leaf apoplastic space and although their absolute levels did not change in response to salt stress, the ASC/dehydroascorbate and GSH to GSH oxidized form ratios decreased progressively with the severity of the stress. Apoplastic superoxide dismutase activity was induced in NaCl-treated pea cv Puget but decreased in NaCl-treated pea cv Lincoln. An increase in DHAR and GR and a decrease in ASC peroxidase, MDHAR, ASC, and GSH levels was observed in the symplast from NaCl-treated pea cv Lincoln, whereas in pea cv Puget an increase in DHAR, GR, and MDHAR occurred. The results suggest a strong interaction between both cell compartments in the control of the apoplastic ASC content in pea leaves. However, this anti-oxidative response does not seem to be sufficient to remove the harmful effects of high salinity. This finding is more evident in pea cv Lincoln, which is characterized by a greater inhibition of the growth response and by a higher rise in the apoplastic hydrogen peroxide content, O2.- production and thiobarbituric acid-reactive substances, and CO protein levels. This NaCl-induced oxidative stress in the apoplasts might be related to the appearance of highly localized O2.-/H2O2-induced necrotic lesions in the minor veins in NaCl-treated pea plants. It is possible that both the different anti-oxidative capacity and the NaCl-induced response in the apoplast and in the symplast from pea cv Puget in comparison with pea cv Lincoln contributes to a better protection of pea cv Puget against salt stress.

1 This work was supported by the Dirección General de Ensenñanza Superior e Investigación Científica, Spain (project no. PB95-004-02), by the Ministerio de Educación y Cultura, Spain (project no. PB-97-1042), by the European Union (project no. FAIR-CT-98-5020), and by the University of Murcia (postdoctoral fellowship to M.A.F.).

* Corresponding author; e-mail fsevilla{at}natura.cebas.csic.es; fax 34-968-396213.

© 2001 American Society of Plant Physiologists


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