Open Access
Expression and localization of a Rhizobium-derived cambialistic superoxide dismutase in pea (Pisum sativum) nodules subjected to oxidative stress
(The American Phytopathological Society, 2011-09-07) Asensio, Aarón C.; Marino Bilbao, Daniel; James, Euan K.; Ariz Arnedo, Idoia; Arrese-Igor Sánchez, César; Aparicio Tejo, Pedro María; Arredondo-Peter, Raúl; Morán Juez, José Fernando; IdAB. Instituto de Agrobiotecnología / Agrobioteknologiako Institutua; Ciencias del Medio Natural; Natura Ingurunearen Zientziak
Two phylogenetically unrelated superoxide dismutase (SOD) families, i.e., CuZnSOD (copper and zinc SOD) and FeMn-CamSOD (iron, manganese, or cambialistic SOD), eliminate superoxide radicals in different locations within the plant cell. CuZnSOD are located within the cytosol and plastids, while the second family of SOD, which are considered to be of bacterial origin, are usually located within organelles, such as mitochondria. We have used the reactive oxygen species¿producer methylviologen (MV) to study SOD isozymes in the indeterminate nodules on pea (Pisum sativum). MV caused severe effects on nodule physiology and structure and also resulted in an increase in SOD activity. Purification and N-terminal analysis identified CamSOD from the Rhizobium leguminosarum endosymbiont as one of the most active SOD in response to the oxidative stress. Fractionation of cell extracts and immunogold labeling confirmed that the CamSOD was present in both the bacteroids and the cytosol (including the nuclei, plastids, and mitochondria) of the N-fixing cells, and also within the uninfected cortical and interstitial cells. These findings, together with previous reports of the occurrence of FeSOD in determinate nodules, indicate that FeMnCamSOD have specific functions in legumes, some of which may be related to signaling between plant and bacterial symbionts, but the occurrence of one or more particular isozymes depends upon the nodule type.
Open Access
Insights into the regulation of nitrogen fixation in pea nodules: lessons from drought, abscisic acid and increased photoassimilate availability
(EDP Sciences, 2001) González García, Esther; Gálvez, Loli; Royuela Hernando, Mercedes; Aparicio Tejo, Pedro María; Arrese-Igor Sánchez, César; Ciencias del Medio Natural; Natura Ingurunearen Zientziak
Nitrogen fixation in legume nodules has been shown to be very sensitive to drought and other environmental constraints. It has been widely assumed that this decline in nitrogen fixation was a consequence of an increase in the so-called oxygen diffusion barrier and a subsequent impairment to bacteroid respiration. However, it has been recently shown that nitrogen fixation is highly correlated with nodule sucrose synthase (SS) activity under drought and other environmental stresses. Whether this correlation reflects a causative relationship or not has not been proven yet. The evidence presented here suggests that SS controls nitrogen fixation under mild drought conditions. However, nitrogen fixation cannot be enhanced only by increasing glycolytic flux, as under these conditions nodules become oxygen limited. Abscisic acid also induces a decline in nitrogen fixation that is independent of SS. The overall results suggest the occurrence of a complex regulation of nodule nitrogen fixation involving, at least, both carbohydrate and oxygen fluxes within the nodule.
Open Access
Unravelling the mechanisms that improve photosynthetic performance of N₂-fixing pea plants exposed to elevated [CO₂]
(Elsevier, 2014) Aranjuelo Michelena, Iker; Cabrerizo Geijo, Pablo María; Aparicio Tejo, Pedro María; Arrese-Igor Sánchez, César; Natura Ingurunearen Zientziak; Ciencias del Medio Natural; IdAB. Instituto de Agrobiotecnología / Agrobioteknologiako Institutua
Although the predicted enhanced photosynthetic rates of plants exposed to elevated [CO₂] are expected to increase carbohydrate and plant growth, recent findings have shown a complex regulation of these processes. The aim of this study was to determine the effect of elevated [CO₂] on pathways leading to the main forms of leaf C storage (starch) and export (sucrose) and the implications of this increased [CO₂] on photosynthetic performance of exclusively N2 fixing plants. For this purpose, exclusively N2-fixing pea plants were exposed to elevated [CO₂] (1000 mol mol−1 versus 360 mol mol−1 CO₂). The data obtained highlighted that plants exposed to elevated [CO₂] were capable of maintaining hexose levels (involved in Rubisco down regulation) at control levels with the consequent avoidance of photosynthetic acclimation. More specifically, in plants exposed to elevated [CO₂] there was an increase in the activity of pathways involved in the main forms of leaf C storage (starch) and export (sucrose). Furthermore, the study highlighted that although starch content increased by up to 40% under elevated [CO₂], there was also an increase in the proteins and compounds involved in starch degradation. Such a finding, together with an increase in the activity of proteins involved in sucrose synthesis revealed that these plants up-regulated the sucrose synthesis pathway in order to meet the large nodule photoassimilate requirements. As a consequence, the study highlighted the relevance of controlling the activity of pathways that determine leaf cellular carbohydrate availability and how this is linked with C-demanding organs such as nodules.
Open Access
Pea plant responsiveness under elevated [CO2] is conditioned by the N source (N2 fixation versus NO3 fertilization)
(Elsevier, 2013) Aranjuelo Michelena, Iker; Cabrerizo Geijo, Pablo María; Arrese-Igor Sánchez, César; Aparicio Tejo, Pedro María; IdAB. Instituto de Agrobiotecnología / Agrobioteknologiako Institutua; Ciencias del Medio Natural; Natura Ingurunearen Zientziak
The main goal of this study was to test the effect of [CO2] on C and N management in 2different plant organs (shoots, roots and nodules) and its implication in the 3responsiveness of exclusively N2-fixing and NO3--fed plants. For this purpose, 4exclusively N2-fixingand NO3--fed (10 mM) pea (Pisum sativumL.) plants were 5exposed to elevated [CO2] (1000 mol mol-1versus360 mol mol-1CO2). Gas 6exchange analyses, together with carbohydrate, nitrogen, total soluble proteins and 7amino acids were determined in leaves, roots and nodules. The data obtained revealed 8that although exposure to elevated [CO2] increased total dry mass (DM)in both N 9treatments, photosynthetic activity was down-regulated in NO3--fed plants, whereas N2-10fixing plants were capable of maintaining enhanced photosynthetic rates under elevated 11[CO2]. In the case of N2-fixing plants, the enhanced C sink strength of nodules enabled 12the avoidance of harmful leaf carbohydrate build up. On the other hand, in NO3--fed 13plants, elevated [CO2] caused a large increase in sucrose and starch. The increase in root 14DM did not contribute to stimulation ofC sinks in these plants. Although N2fixation 15matched plant N requirementswith the consequent increase in photosynthetic rates, in 16NO3--fed plants, exposure to elevated [CO2] negatively affected N assimilationwith the 17consequent photosynthetic down-regulation.

Aparicio Tejo, Pedro María

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