Plant Physiol. Journal of Pharmacology and Experimental Therapeutics
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Stress-Induced Legume Root Nodule Senescence. Physiological, Biochemical, and Structural Alterations1

Manuel A. Matamoros, Lisa M. Baird, Pedro R. Escuredo, David A. Dalton, Frank R. Minchin, Iñaki Iturbe-Ormaetxe, Maria C. Rubio, Jose F. Moran, Anthony J. Gordon, and Manuel Becana*

Departamento de Nutrición Vegetal, Estación Experimental de Aula Dei, Consejo Superior de Investigaciones Científicas, Apdo 202, 50080 Zaragoza, Spain (M.A.M., P.R.E., I.I.-O., M.C.R., J.F.M., M.B.); Biology Department, University of San Diego, San Diego, California 92110 (L.M.B.); Biology Department, Reed College, Portland, Oregon 97202 (D.A.D.); and Institute of Grassland and Environmental Research, Plas Gogerddan, Aberystwyth SY23 3EB, United Kingdom (F.R.M., A.J.G.)

Nitrate-fed and dark-stressed bean (Phaseolus vulgaris) and pea (Pisum sativum) plants were used to study nodule senescence. In bean, 1 d of nitrate treatment caused a partially reversible decline in nitrogenase activity and an increase in O2 diffusion resistance, but minimal changes in carbon metabolites, antioxidants, and other biochemical parameters, indicating that the initial decrease in nitrogenase activity was due to O2 limitation. In pea, 1 d of dark treatment led to a 96% decline in nitrogenase activity and sucrose, indicating sugar deprivation as the primary cause of activity loss. In later stages of senescence (4 d of nitrate or 2-4 d of dark treatment), nodules showed accumulation of oxidized proteins and general ultrastructural deterioration. The major thiol tripeptides of untreated nodules were homoglutathione (72%) in bean and glutathione (89%) in pea. These predominant thiols declined by approximately 93% after 4 d of nitrate or dark treatment, but the loss of thiol content can be only ascribed in part to limited synthesis by gamma -glutamylcysteinyl, homoglutathione, and glutathione synthetases. Ascorbate peroxidase was immunolocalized primarily in the infected and parenchyma (inner cortex) nodule cells, with large decreases in senescent tissue. Ferritin was almost undetectable in untreated bean nodules, but accumulated in the plastids and amyloplasts of uninfected interstitial and parenchyma cells following 2 or 4 d of nitrate treatment, probably as a response to oxidative stress.

1   This work was supported by grant nos. PB95-0091 and PB98-0522 from the Dirección General de Enseñanza Superior e Investigación Científica (Ministry of Education and Culture, Spain) to M.B., and by fellowships from the Gobierno Vasco (M.A.M.), the European Union (I.I.-O.), and the Ministry of Education and Culture (P.R.E., M.C.R., J.F.M.).
*   Corresponding author; e-mail becana{at}eead.csic.es; fax 34-976-575620.

Plant Physiol. (1999) 121: 97-112
Copyright Clearance Center:   0032-0889/99/121//16
© 1999 American Society of Plant Physiologists




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