Antioxidant responses of peanut roots exposed to realistic groundwater doses of arsenate: Identification of glutathione S-transferase as a suitable biomarker for metalloid toxicity.

Arsenic (As)-polluted groundwater constitutes a serious problem for peanut plants, as roots can accumulate the metalloid in their edible parts. Characterization of stress responses to As may help to detect potential risks and identify mechanisms of tolerance, being the induction of oxidative stress a key feature. Fifteen-day old peanut plants were treated with arsenate in order to characterize the oxidative stress indexes and antioxidant response of the legume under realistic groundwater doses of the metalloid. Superoxide anion (O2-) and hydrogen peroxide (H2O2) histochemical staining along with the activities of NADPH oxidase, superoxide dismutase (SOD), catalase (CAT) and thiol (glutathione and thioredoxins) metabolism were determined in roots. Results showed that at 20 µM H2AsO4-, peanut growth was reduced and the root architecture was altered. O2- and H2O2 accumulated at the root epidermis, while lipid peroxidation, NADPH oxidase, SOD, CAT and glutathione S-transferase (GST) activities augmented. These variables increased with increasing As concentration (100 µM) while glutathione reductase (GR) and glutathione peroxidase/peroxiredoxin (GPX/PRX) were significantly decreased. These findings demonstrated that the metalloid induced physiological and biochemical alterations, being the NADPH oxidase enzyme implicated in the oxidative burst. Additionally, the strong induction of GST activity, even at the lowest H2AsO4- doses studied, can be exploited as suitable biomarker of As toxicity in peanut plants, which may help to detect risks of As accumulation and select tolerant cultivars.

Influence of cadmium on the symbiotic interaction established between peanut (Arachis hypogaea L.) and sensitive or tolerant bradyrhizobial strains.

Heavy metals in soil are known to affect rhizobia-legume interaction reducing not only rhizobia viability, but also nitrogen fixation. In this work, we have compared the response of the symbiotic interaction established between the peanut (Arachis hypogaea L.) and a sensitive (Bradyrhizobium sp. SEMIA6144) or a tolerant (Bradyrhizobium sp. NLH25) strain to Cd under exposure to this metal. The addition of 10 µM Cd reduced nodulation and nitrogen content in both symbiotic associations, being the interaction established with the sensitive strain more affected than that with the tolerant one. Plants inoculated with the sensitive strain accumulated more Cd than those inoculated with the tolerant strain. Nodules showed an increase in reactive oxygen species (ROS) production when exposed to Cd. The histological structure of the nodules exposed to Cd revealed a deposit of unknown material on the cortex and a significant reduction in the infection zone diameter in both strains, and a greater number of uninfected cells in those nodules occupied by the sensitive strain. In conclusion, Cd negatively impacts on peanut-bradyrhizobia interaction, irrespective of the tolerance of the strains to this metal. However, the inoculation of peanut with Bradyrhizobium sp. NLH25 results in a better symbiotic interaction suggesting that the tolerance observed in this strain could limit Cd accumulation by the plant.

Importance of glutathione in the nodulation process of peanut (Arachis hypogaea).

GSH appears to be essential for proper development of the root nodules during the symbiotic association of legume-rhizobia in which the entry of rhizobia involves the formation of infection threads. In the particular case of peanut-rhizobia symbiosis, the entry of rhizobia occurs by the mechanism of infection called 'crack entry', i.e. entry at the point of emergence of lateral roots. We have previously shown the role of GSH content of Bradyrhizobium sp. SEMIA 6144 during the symbiotic association with peanut using a GSH-deficient mutant obtained by disruption of the gshA gene, encoding gamma-glutamylcysteine synthetase (gamma-GCS), which was able to induce nodules in peanut roots without alterations in the symbiotic phenotype. To investigate the role of the peanut GSH content in the symbiosis, the compound L-buthionine-sulfoximine (BSO), a specific inhibitor of gamma-GCS in plants, was used. There were no differences in the plant growth and the typical anatomic structure of the peanut roots when the plants grew in the Fahraeus medium either in presence or in absence of 0.1 mM BSO. However, the GSH content was reduced by 51% after treatment with BSO. The BSO-treated plants inoculated with wild-type or mutant strains of Bradyrhizobium sp. showed a significant reduction in the number and dry weight of nodules, suggesting that GSH content could play an important role in the nodulation process of root peanut with Bradyrhizobium sp.

Changes in ligno-suberization of cell walls of tomato hairy roots produced by salt treatment: the relationship with the release of a basic peroxidase.

A highly basic peroxidase isoenzyme was shown to be released to the culture medium of tomato (Lycopersicon esculentum) hairy roots grown in Murashige-Skoog (MS) liquid medium when it was supplemented with 100 mM NaCl. In this paper we demonstrate that this enzyme is ionically bound to cell walls and that the release was a consequence of the continuous agitation of the tissue in a high ionic strength medium with salt addition. In order to establish the physiological role of this isoenzyme we partially purified it, and we analysed its kinetic properties as coniferyl alcohol peroxidase. The peroxidase isoenzyme showed a high catalytic efficiency for this substrate, which suggests that it would be associated with the ligno-suberization process. To confirm the involvement of this isoenzyme in that process, we studied the pattern of ligno-suberization of the tissue under different conditions of growth. Our results suggest that this basic peroxidase would be indeed involved in ligno-suberization since its leakage from cell walls, induced by 100 mM NaCl in liquid MS, caused less ligno-suberization of exo and endodermis. On the contrary, more ligno-suberization was seen in cell walls when the hairy roots were grown in a salt-supplemented MS solid medium without contact with it, a condition in which the release of the isoenzyme would be avoided. Thus, through the changes produced by the release of the enzyme from its site of action, we could demonstrate the physiological role of this peroxidase in the processing of root cell walls, being part of control mechanisms of ion and water fluxes through the root.