Activated oxygen and antioxidant defences in iron-deficient pea plants

Iron (Fe) deficiency in pea leaves caused a large decrease (44–62&) in chlorophyll a, chlorophyll b and carotenoids, and smaller decreases in soluble protein (18&) and net photosynthesis (28&). Catalase, non-specific peroxidase and ascorbate peroxidase activities declined by 51& in young Fe-deficient leaves, whereas monodehydroascorbate reductase, dehydroascorbate reductase and glutathione reductase activities remained unaffected. Ascorbate peroxidase activity was highly correlated (r²= 0. 99, P < 0. 001) with the Fe content of leaves, which allows its use as an indicator of the Fe nutritional status of the plant. Fe deficiency resulted in an increase of CuZn-superoxide dismutase but not of Mn-superoxide dismutase. The content of ascorbate decreased by only 24& and those of reduced and oxidized glutathione and vitamin E did not vary. The low-molecular-mass fraction of Fe-sufficient leaves contained 30–65 μg (g dry weight)^?1 Mn. This concentration was 15–60 times greater than that of Fe and Cu in the same fraction, and was further enhanced (1. 5- to 2. 5-fold) by Fe deficiency without causing Mn toxicity. The concentration of catalytic Fe, that is, of Fe active for free radical generation, was virtually zero and that of catalytic Cu did not change with severe Fe deficiency. Because catalytic metals mediate lipid and protein oxidation in vivo, the above findings would explain why oxidatively damaged lipids and proteins do not accumulate in Fe-deficient leaves.     

N2 Fixation (C2H2-Reducing Activity) and Leghaemoglobin Content during Nitrate- and Water-Stress-Induced Senescence of Medicago sativa Root Nodules

Nitrate and water stress were used to induce senescence in rootnodules of alfalfa (Medicago sativa L. cv. Aragon). Nodule senescencewas assessed by determinations of the nitrogenase (C₂H₂-reducing)activity, and the leghaemoglobin (LHb) and total soluble proteincontents of the nodules. Nodules responded similarly to and water stress in many respects, but there was a significant difference.All parameters of nodule activity, expressed on the basis ofnodule dry weight (DW), consistently decreased following treatmentwith or during drought; there was a significant interaction (synergism) between the inhibitory effects of and water stress on nitrogenase activity, but sucheffects were merely additive in the case of LHb content or LHb/solubleprotein ratio. However, caused the selective decay of LHb with respect to other nodular soluble proteins,whereas the decrease of LHb during water stress was due to ageneral inhibition of protein synthesis and to an increasedproteolytic activity in the nodule cytosol rather than to aspecific proteolysis of LHb. Key words: Leghaemoglobin, Medicago saliva, nitrogen fixation, root nodule senescence, water stress     

Nitrate Metabolism in Alfalfa Root Nodules under Water Stress

Three-month-old plants (vegetative stage) of alfalfa (Medicagosaliva L cv. Aragon) were supplied for one week with 1.0dm³(uniformly distributed) nutrient solutions containing 0 or 20mol m^–3 . One week after initiation of treatment, the plants were subjected to drought by withholding water. Bacteroidsand cytosol of nodules were obtained at different stages ofstress, and used for enzyme assays and for determination of, and . Proteins of bacteroids were more stable than cytosolic proteinswith respect to the detrimental effects of water stress and. Protein contents of bacteroids and cytosol were inversely related to proteolytic activitiesagainst azocasein in both nodule fractions. Specific nitrate reductase activity (NRA) and nitrite reductaseactivity (NiRA) of bacteroids from -treated plants were inhibited by c. 70% and 45%, respectively, as leafwater potential (_w) declined from –0.5 MPa (control) to–1.8 MPa. At still lower _w both activities began to increase:NRA was doubled, whereas NiRA only returned to its control level.Cytosolic NRA was strongly inhibited by drought, but the correspondingNiRA remained constant. Ammonia concentration in bacteroids and nodule cytosol keptbasically constant, whereas accumulated in the cytosol at severe stress, due to the activationof bacteroid nitrate reductase. Results indicate that nitrate and nitrite reductases of thebacteroids and the nodule cytosol act in different form: assimilatory,the cytosolic enzymes; and dissimilatory, the enzymes of bacteroidsat low _w The possibility that assimilation of also occurs in bacteroids at control or mild waterstress conditions is suggested. Key words: Assimilatory and dissimilatory reduction, bacteroids, Medicago saliva, nodule cytosol, water stress     

Effect of Nitrate on Components of Nodule Leghaemoglobins

Multiple components of leghaemoglobins (Lbs) from several legumespecies have been separated by polyacrylamide gel electrophoresis(PAGE), and identified by densitometry at 405 nm, extractionfrom gels and formation of their respective CO-ferrous complexes.Components were numbered consecutively starting from the anodeside, and their relative concentrations were calculated by densitometryin control and -treated nodules. The number of Lb components was five in Pisum sativum, four in Glycinemax, at least two in Lupinus luteus and Trifolium repens, andtwo in Vigna radiata and V. unguiculata. Only one componentwas detected in fresh nodules of Phaseolus vulgaris. Nitratesignificantly reduced the total content of Lb in all legumes,except Lupinus, the extent of reduction varying with the species.It also had a major effect on the proportions of components,causing an inversion in the relative abundance of Lb I and LbII of V. radiata: the concentration of Lb I increased from 8%to 81% with application, while total Lb only decreased by 13%. Drastic changes in proportions of Lbcomponents were not artifactual and cannot be attributed exclusivelyto different turnover rates. They should correspond to variationsin biosynthesis of the various components, probably originatingat the genetic level. Key words: Leghaemoglobin components, nitrate, nodule senescence