Open Access
Use of recombinant iron-superoxide dismutase as a marker of nitrative stress
(Elservier, 2008-04-20) Larrainzar Rodríguez, Estíbaliz; Urarte Rodríguez, Estíbaliz; Auzmendi, Iñigo; Ariz Arnedo, Idoia; Arrese-Igor Sánchez, César; González García, Esther; Morán Juez, José Fernando; Ciencias del Medio Natural; Natura Ingurunearen Zientziak; IdAB. Instituto de Agrobiotecnología / Agrobioteknologiako Institutua; Gobierno de Navarra / Nafarroako Gobernua, 57/2007
Superoxide dismutases (SODs; EC 1.15.1.1) are a group of metalloenzymes which are essential to protect cells under aerobic conditions. In biological systems, it has been reported that SODs and other proteins are susceptible to be attacked by peroxynitrite (ONOO-) which can be originated from the reaction of nitric oxide with superoxide radical. ONOO- is a strong oxidant molecule capable of nitrating peptides and proteins at the phenyl side chain of the tyrosine residues. In the present work, bovine serum albumin (BSA) and recombinant iron¿superoxide dismutase from the plant cowpea (Vu_FeSOD) are used as target molecules to estimate ONOO- production. The method employs the compound SIN-1, which simultaneously generates -NO and O2- in aerobic aqueous solutions. First, assay conditions were optimized incubating BSA with different concentrations of SIN-1, and at a later stage, the effect on the tyrosine nitration and catalytic activity of Vu_FeSOD was examined by in-gel activity and spectrophotometric assays. Both BSA and Vu_FeSOD are nitrated in a dose-dependent manner, and, at least in BSA nitration, the reaction seems to be metal catalyzed.
Open Access
Learning plant biodiversity in nature: the use of the citizen–science platform iNaturalist as a collaborative tool in secondary education
(MDPI, 2021) Echeverría Obanos, Andrés; Ariz Arnedo, Idoia; Moreno Echeverría, Judit; Peralta de Andrés, Francisco Javier; González García, Esther; Zientziak; Institute for Multidisciplinary Research in Applied Biology - IMAB; Ciencias; Gobierno de Navarra / Nafarroako Gobernua, CENEDUCA3/2019
Biodiversity is a concept of great scientific interest and social value studied in different subjects of the secondary education curriculum. Citizen–science programs may contribute to increasing the engagement of students when studying biodiversity. This work aimed to explore the use of the citizen–science platform iNaturalist as a complement of the elaboration of herbaria in an outdoor activity for 4th course 16-year-old students in the Basaula Reserve. The platform iNaturalist was chosen for its suitability to develop collaborative projects in an educational context. The Basaula project was created and 122 students were trained to record plant species in an outdoor activity. A total of 32 species were recorded, among them the most abundant were beech (Fagus sylvatica) and holm oak (Quercus ilex). The students positively evaluated their experience, highlighting its adequacy to record biodiversity data and make a virtual herbarium. Students valued the innovative character of iNaturalist and its usefulness for research but also the opportunity to integrate mobile devices in school education. We concluded that iNaturalist is a valuable tool to carry out collaborative projects dealing with biodiversity in secondary education.
Open Access
Drought stress causes a reduction in the biosynthesis of ascorbic acid in soybean plants
(Frontiers Media, 2017) Seminario Huárriz, Amaia; Song, Li; Zulet González, Amaia; Nguyen, Henry T.; González García, Esther; Larrainzar Rodríguez, Estíbaliz; Ciencias del Medio Natural; Natura Ingurunearen Zientziak; Gobierno de Navarra / Nafarroako Gobernua, 2016/PI013; Universidad Pública de Navarra / Nafarroako Unibertsitate Publikoa, 1287/2011
Drought provokes a number of physiological changes in plants including oxidative damage. Ascorbic acid (AsA), also known as vitamin C, is one of the most abundant water-soluble antioxidant compound present in plant tissues. However, little is known on the regulation of AsA biosynthesis under drought stress conditions. In the current work we analyze the effects of water deficit on the biosynthesis of AsA by measuring its content, in vivo biosynthesis and the expression level of genes in the Smirnoff-Wheeler pathway in one of the major legume crop, soybean (Glycine max L. Merr). Since the pathway has not been described in legumes, we first searched for the putative orthologous genes in the soybean genome. We observed a significant genetic redundancy, with multiple genes encoding each step in the pathway. Based on RNA-seq analysis, expression of the complete pathway was detected not only in leaves but also in root tissue. Putative paralogous genes presented differential expression patterns in response to drought, suggesting the existence of functional specialization mechanisms. We found a correlation between the levels of AsA and GalLDH biosynthetic rates in leaves of drought-stressed soybean plants. However, the levels of GalLDH transcripts did not show significant differences under water deficit conditions. Among the other known regulators of the pathway, only the expression of VTC1 genes correlated with the observed decline in AsA in leaves.
Open Access
Drought stress provokes the down-regulation of methionine and ethylene biosynthesis pathways in Medicago truncatula roots and nodules
(Wiley, 2014) Larrainzar Rodríguez, Estíbaliz; Molenaar, Johanna A.; Wienkoop, Stefanie; Gil Quintana, Erena; Alibert, Bénédicte; Limami, Anis M.; Arrese-Igor Sánchez, César; González García, Esther; Ciencias del Medio Natural; Natura Ingurunearen Zientziak; Universidad Pública de Navarra / Nafarroako Unibertsitate Publikoa, 735/2008
Symbiotic nitrogen fixation is one of the first physiological processes inhibited in legume plants under water-deficit conditions. Despite the progress made in the last decades, the molecular mechanisms behind this regulation are not fully understood yet. Recent proteomic work carried out in the model legume Medicago truncatula provided the first indications of a possible involvement of nodule methionine (Met) biosynthesis and related pathways in response to waterdeficit conditions. To better understand this involvement, the drought-induced changes in expression and content of enzymes involved in the biosynthesis of Met, S-adenosyl-Lmethionine (SAM) and ethylene in M. truncatula root and nodules were analyzed using targeted approaches. Nitrogenfixing plants were subjected to a progressive water deficit and a subsequent recovery period. Besides the physiological characterization of the plants,the content of total sulphur,sulphate and main S-containing metabolites was measured. Results presented here show that S availability is not a limiting factor in the drought-induced decline of nitrogen fixation rates in M. truncatula plants and provide evidences for a downregulation of the Met and ethylene biosynthesis pathways in roots and nodules in response to water-deficit conditions.
Open Access
Split‐root systems applied to the study of the legume‐rhizobial symbiosis: what have we learned?
(Wiley, 2014) Larrainzar Rodríguez, Estíbaliz; Gil Quintana, Erena; Arrese-Igor Sánchez, César; González García, Esther; Marino Bilbao, Daniel; Ciencias del Medio Natural; Natura Ingurunearen Zientziak
Split-root system (SRS) approaches allow the differential treatment of separate and independent root systems, while sharing a common aerial part. As such, SRS is a useful tool for the discrimination of systemic (shoot origin) versus local (root/nodule origin) regulation mechanisms. This type of approach is particularly useful when studying the complex regulatory mechanisms governing the symbiosis established between legumes and Rhizobium bacteria. The current work provides an overview of the main insights gained from the application of SRS approaches to understand how nodule number (nodulation autoregulation) and nitrogen fixation are controlled both under non-stressful conditions and in response to a variety of stresses. Nodule number appears to be mainly controlled at the systemic level through a signal which is produced by nodule/root tissue, translocated to the shoot, and transmitted back to the root system, involving shoot Leu-rich repeat receptor-like kinases. In contrast, both local and systemic mechanisms have been shown to operate for the regulation of nitrogenase activity in nodules. Under drought and heavy metal stress, the regulation is mostly local, whereas the application of exogenous nitrogen seems to exert a regulation of nitrogen fixation both at the local and systemic levels.
Open Access
Editorial: Drought stress in legumes
(Frontiers Media, 2022) Furlan, Ana Laura; González García, Esther; Roy Choudhury, Swarup; Signorelli, Santiago; Institute for Multidisciplinary Research in Applied Biology - IMAB; Gobierno de Navarra / Nafarroako Gobernua; Universidad Pública de Navarra / Nafarroako Unibertsitate Publikoa
Plants are challenged by diverse environmental constraints, among which drought stress is increasingly important. Meteorological models predict an increase in the areas prone to drought in the future. Legumes are important sources of fiber, oils, and protein, constituting an essential amenity in the global economy. Additionally, legumes contribute to nitrogen input in the biosphere due to their ability to establish symbiotic interactions with diazotrophs, collectively named rhizobia. Therefore, efforts to decipher the molecular, metabolic, physiological, and agronomic responses are crucial contributing novel strategies to aid drought tolerance in legumes. This Research Topic contains articles either providing new findings or discussing the latest research concerning drought research in legumes, including one mini-review on soybean tolerance to drought (Arya et al.) and seven original research papers dealing with strategies to confer drought tolerance such as priming (Zhou et al.); studies on intraspecific variation in traits associated with drought tolerance (Prince et al.); the analysis of water use efficiency under terminal drought (Polania et al.); and the contribution of a legume dehydrin to drought tolerance (Sun et al.); the functional characterization of a LOX gene family (Mou et al.); the functional analysis of a soybean APETALA2/ETHYLENE RESPONSIVE FACTOR (AP2/ERF) (Wang et al.); and a study on AP2/ERF gene family in a tolerant desert legume (Zhao et al.).
Open Access
Strategies to aply water-deficit stress: similarities and disparities at the whole plant metabolism level in medicago truncatula
(MDPI, 2021) Castañeda Presa, Verónica; González García, Esther; Ciencias; Zientziak; Universidad Pública de Navarra / Nafarroako Unibertsitate Publikoa
Water-deficit stresses such as drought and salinity are the most important factors limiting crop productivity. Hence, understanding the plant responses to these stresses is key for the improvement of their tolerance and yield. In this study M. truncatula plants were subjected to 250 mM NaCl as well as reduced irrigation (No-W) and 250 g/L polyethylene glycol (PEG)-6000 to induce salinity and drought stress, respectively, provoking a drop to −1.7 MPa in leaf water potential. The whole plant physiology and metabolism was explored by characterizing the stress responses at root, phloem sap and leaf organ level. PEG treatment led to some typical responses of plants to drought stress, but in addition to PEG uptake, an important impairment of nutrient uptake and a different regulation of carbon metabolism could be observed compared to No-W plants. No-W plants showed an important redistribution of antioxidants and assimilates to the root tissue, with a distinctive increase in root proline degradation and alkaline invertase activity. On the contrary, salinity provoked an increase in leaf starch and isocitrate dehydrogenase activity, suggesting key roles in the plant response to this stress. Overall, results suggest higher protection of salt-stressed shoots and non-irrigated roots through different mechanisms, including the regulation of proline and carbon metabolism, while discarding PEG as safe mimicker of drought. This raises the need to understand the effect at the whole plant level of the different strategies employed to apply water-deficit stress.
Open Access
Functional analysis of the taproot and fibrous roots of Medicago truncatula: sucrose and proline catabolism primary response to water deficit
(Elsevier, 2019) Castañeda Presa, Verónica; Peña, Marlon de la; Azcárate Górriz, Lidia; Aranjuelo Michelena, Iker; González García, Esther; Ciencias; Zientziak; Universidad Pública de Navarra / Nafarroako Unibertsitate Publikoa
Root performance represents a target factor conditioning plant development under drought conditions. Moreover, recent root phenotyping studies remark relevant differences on functionality of the different root types. However, despite its relevance, the performance of different types of roots such as primary/taproot (tapR) and lateral/fibrous roots (fibR) under water stress conditions is largely unknown. In the current study, the impact of water stress on target C and N metabolism (namely sucrose and proline) processes were characterized in tapR and fibR of Medicago truncatula plants exposed to different water stress severity regimes (moderate versus severe). While both root types exhibit some common responses to face water stress, the study highlighted important physiological and metabolic differences between them. The tapR proved to have an essential role on carbon and nitrogen partitioning rather than just on storage. Moreover, this root type showed a higher resilience towards water deficit stress. Sucrose metabolization at sucrose synthase level was early blocked in this tissue together with a selective accumulation of some amino acids such as proline and branched chain amino adds, which may act as alternative carbon sources under water deficit stress conditions. The decline in respiration, despite the over-accumulation of carbon compounds, suggests a modulation at sucrose cleavage level by sucrose synthase and invertase. These data not only provide new information on the carbon and nitrogen metabolism modulation upon water deficit stress but also on the different role, physiology, and metabolism of the taproot and fibrous roots. In addition, obtained results highlight the fact that both root types show distinct performance under water deficit stress; this factor can be of great relevance to improve breeding programs for increasing root efficiency under adverse conditions.
Open Access
Local inhibition of nitrogen fixation and nodule metabolism in drought-stressed soybean
(Oxford University Press, 2013) Gil Quintana, Erena; Larrainzar Rodríguez, Estíbaliz; Seminario Huárriz, Amaia; Díaz Leal, Juan Luis; Alamillo, Josefa M.; Pineda, Manuel; Arrese-Igor Sánchez, César; Wienkoop, Stefanie; González García, Esther; Ciencias del Medio Natural; Natura Ingurunearen Zientziak; Universidad Pública de Navarra / Nafarroako Unibertsitate Publikoa: 735/2008; Universidad Pública de Navarra / Nafarroako Unibertsitate Publikoa: 134/2012
Drought stress is a major factor limiting symbiotic nitrogen fixation (NF) in soybean crop production. However, the regulatory mechanisms involved in this inhibition are still controversial. Soybean plants were symbiotically grown in a split-root system (SRS), which allowed for half of the root system to be irrigated at field capacity while the other half remained water deprived. NF declined in the water-deprived root system while nitrogenase activity was maintained at control values in the well-watered half. Concomitantly, amino acids and ureides accumulated in the water-deprived belowground organs regardless of transpiration rates. Ureide accumulation was found to be related to the decline in their degradation activities rather than increased biosynthesis. Finally, proteomic analysis suggests that plant carbon metabolism, protein synthesis, amino acid metabolism, and cell growth are among the processes most altered in soybean nodules under drought stress. Results presented here support the hypothesis of a local regulation of NF taking place in soybean and downplay the role of ureides in the inhibition of NF
Open Access
Proyecto río Arga: ciencia ciudadana, biodiversidad & educación
(2022) González García, Esther; Peralta de Andrés, Francisco Javier; Ciencias; Zientziak; Gobierno de Navarra / Nafarroako Gobernua, CENEDUCA3-2019
El objetivo del proyecto ARGA, desarrollado en la plataforma iNaturalist (https://www.inaturalist.org/projects/arga), es reunir observaciones de las especies presentes en el río Arga a su paso por la Cuenca de Pamplona. Cualquier persona puede participar, aunque en su creación se ha pensado en su utilización principalmente por estudiantes de Enseñanza Secundaria. Este proyecto se lleva a cabo en colaboración con la Mancomunidad de la Comarca de Pamplona a través del programa Mancoeduca, que ofrece actividades de educación ambiental dirigidas a centros de educación secundaria en el entorno del río Arga.