Open Access
¿Están los bosques mixtos pirenaicos de pino silvestre y haya en el camino hacia la saturación por nitrógeno?
(Asociación Española de Ecología Terrestre, 2017) Blanco Vaca, Juan Antonio; San Emeterio Garciandía, Leticia; González de Andrés, Ester; Imbert Rodríguez, Bosco; Larrainzar Rodríguez, Estíbaliz; Peralta de Andrés, Francisco Javier; Lo, Yueh-Hsin; Castillo Martínez, Federico; Ciencias del Medio Natural; Natura Ingurunearen Zientziak
Las actividades humanas causan altos niveles de deposición atmosférica crónica de N que pueden estar trastornando el ciclo del N en los bosques de los Pirineos occidentales. Para probar esta hipótesis, se han investigado los efectos de la deposición de N atmosférico en el ciclo de N en dos bosques mixtos de pino silvestre y haya en Navarra. Un bosque está situado a 1350 m de altitud y tiene un clima continental, mientras que el otro está situado a 650 m y tiene un clima mediterráneo húmedo. Pruebas preliminares indicaron una fijación biológica de N 2 atmosférico indetectable, así como la casi nula presencia de plantas con simbiontes fijadores como en la actividad de fijadores libres. Por lo tanto se asumió que la principal entrada de N en estos bosques es la deposición atmosférica. Se estimó la dependencia de la productividad de estos ecosistemas de la deposición por medio del modelo ecológico FORECAST, calibrado para estos sitios. Se simularon seis escenarios con tasas de deposición en un rango de 5 a 30 kg ha -1 año -1 . Los resultados indicaron que la productividad de estos bosques es dependiente de la deposición de N, pero indicios de saturación por N (aumento de lixiviación y carencia de aumento de productividad) indican que pueden saturarse a partir de 20-25 kg N ha -1 año -1 , unos 5-10 kg N ha -1 año -1 por encima de los niveles observados actualmente.
Open Access
Is N-feedback involved in the inhibition of nitrogen fixation in drought-stressed Medicago truncatula?
(Oxford University Press, 2013) Gil Quintana, Erena; Larrainzar Rodríguez, Estíbaliz; Arrese-Igor Sánchez, César; González García, Esther; Ciencias del Medio Natural; Natura Ingurunearen Zientziak; Universidad Pública de Navarra / Nafarroako Unibertsitate Publikoa: 735/2008
Drought stress is a major factor limiting nitrogen fixation (NF) in crop production. However, the regulatory mechanism involved and the origin of the inhibition, whether local or systemic, is still controversial and so far scarcely studied in temperate forage legumes. Medicago truncatula plants were symbiotically grown with a split-root system and exposed to gradual water deprivation. Physiological parameters, NF activity, and amino acid content were measured. The partial drought treatment inhibited NF in the nodules directly exposed to drought stress. Concomitantly, in the droughted below-ground organs, amino acids accumulated prior to any drop in evapotranspiration (ET). It is concluded that drought exerts a local inhibition of NF and drives an overall accumulation of amino acids in diverse plant organs which is independent of the decrease in ET. The general increase in the majority of single amino acids in the whole plant questions the commonly accepted concept of a single amino acid acting as an N-feedback signal.
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
Physiological responses of legume nodules to drought
(Global Science Books, 2011) Arrese-Igor Sánchez, César; González García, Esther; Marino Bilbao, Daniel; Ladrera Fernández, Rubén; Larrainzar Rodríguez, Estíbaliz; Gil Quintana, Erena; Ciencias del Medio Natural; Natura Ingurunearen Zientziak
Legumes include important agricultural crops, as their high protein content is of primary importance for human food and animal feed. In addition, the ability of most of them to establish symbiotic relationships with soil bacteria allows them to obtain their N requirements from nitrogen fixation in nodules and, therefore, avoids the use of nitrogen fertilizers. Thus, legumes are also essential to improve the soil fertility and quality of agricultural lands and to reclaim eroded or barren areas, making them crucial for agricultural and environmental sustainability. However, legume nitrogen fixation in crop species is very sensitive to environmental constraints and drought, in particular. The present contribution reviews our current knowledge on the processes involved in this inhibition, with particular emphasis on oxygen, nitrogen and carbon physiology. Emerging aspects such as oxidative damage, C/N interactions and sulphur metabolism together with future prospects are also discussed.
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
A novel Enterococcus-based nanofertilizer promotes seedling growth and vigor in wheat (Triticum aestivum L.)
(MDPI, 2024-10-14) Batool, Salma; Safdar, Maryam; Naseem, Saira; Sami, Abdul; Saleem, Rahman Shah Zaib; Larrainzar Rodríguez, Estíbaliz; Shahid, Izzah; Ciencias; Zientziak; Institute for Multidisciplinary Research in Applied Biology - IMAB
Excessive use of chemical fertilizers poses significant environmental and health concerns. Microbial-based biofertilizers are increasingly being promoted as safe alternatives. However, they have limitations such as gaining farmers¿ trust, the need for technical expertise, and the variable performance of microbes in the field. The development of nanobiofertilizers as agro-stimulants and agro-protective agents for climate-smart and sustainable agriculture could overcome these limitations. In the present study, auxin-producing Enterococcus sp. SR9, based on its plant growth-promoting traits, was selected for the microbe-assisted synthesis of silver nanoparticles (AgNPs). These microbial-nanoparticles SR9AgNPs were characterized using UV/Vis spectrophotometry, scanning electron microscopy, and a size analyzer. To test the efficacy of SR9AgNPs compared to treatment with the SR9 isolate alone, the germination rates of cucumber (Cucumis sativus), tomato (Solanum lycopersicum), and wheat (Triticum aestivum L.) seeds were analyzed. The data revealed that seeds simultaneously treated with SR9AgNPs and SR9 showed better germination rates than untreated control plants. In the case of vigor, wheat showed the most positive response to the nanoparticle treatment, with a higher vigor index than the other crops analyzed. The toxicity assessment of SR9AgNPs demonstrated no apparent toxicity at a concentration of 100 ppm, resulting in the highest germination and biomass gain in wheat seedlings. This work represents the first step in the characterization of microbial-assisted SR9AgNPs and encourages future studies to extend these conclusions to other relevant crops under field conditions.
Open Access
A proteomic view on the role of legume symbiotic interactions
(Frontiers Media, 2017) Larrainzar Rodríguez, Estíbaliz; Wienkoop, Stefanie; Ciencias del Medio Natural; Natura Ingurunearen Zientziak
Legume plants are key elements in sustainable agriculture and represent a significant source of plant-based protein for humans and animal feed worldwide. One specific feature of the family is the ability to establish nitrogen-fixing symbiosis with Rhizobium bacteria. Additionally, like most vascular flowering plants, legumes are able to form a mutualistic endosymbiosis with arbuscular mycorrhizal (AM) fungi. These beneficial associations can enhance the plant resistance to biotic and abiotic stresses. Understanding how symbiotic interactions influence and increase plant stress tolerance are relevant questions toward maintaining crop yield and food safety in the scope of climate change. Proteomics offers numerous tools for the identification of proteins involved in such responses, allowing the study of sub-cellular localization and turnover regulation, as well as the discovery of post-translational modifications (PTMs). The current work reviews the progress made during the last decades in the field of proteomics applied to the study of the legume-Rhizobium and -AM symbioses, and highlights their influence on the plant responses to pathogens and abiotic stresses. We further discuss future perspectives and new experimental approaches that are likely to have a significant impact on the field including peptidomics, mass spectrometric imaging, and quantitative proteomics.
Open Access
The CCAAT box-binding transcription factor NF-YA1 controls rhizobial infection
(Oxford University Press, 2014) Laporte, Philippe; Lepage, Agnes; Fournier, Joëlle; Catrice, Olivier; Larrainzar Rodríguez, Estíbaliz; Ciencias del Medio Natural; Natura Ingurunearen Zientziak
Symbiosis between legume plants and soil rhizobia culminates in the formation of a novel root organ, the 'nodule', containing bacteria differentiated as facultative nitrogen-fixing organelles. MtNF-YA1 is a Medicago truncatula CCAAT box-binding transcription factor (TF), formerly called HAP2-1, highly expressed in mature nodules and required for nodule meristem function and persistence. Here a role for MtNF-YA1 during early nodule development is demonstrated. Detailed expression analysis based on RNA sequencing, quantitiative real-time PCR (qRT-PCR), as well as promoter–β-glucuronidase (GUS) fusions reveal that MtNF-YA1 is first induced at the onset of symbiotic development during preparation for, and initiation and progression of, symbiotic infection. Moreover, using a new knock-out mutant, Mtnf-ya1-1, it is shown that MtNF-YA1 controls infection thread (IT) progression from initial root infection through colonization of nodule tissues. Extensive confocal and electronic microscopic observations suggest that the bulbous and erratic IT growth phenotypes observed in Mtnf-ya1-1 could be a consequence of the fact that walls of ITs in this mutant are thinner and less coherent than in the wild type. It is proposed that MtNF-YA1 controls rhizobial infection progression by regulating the formation and the wall of ITs.
Open Access
Three classes of hemoglobins are required for optimal vegetative and reproductive growth of Lotus japonicus: genetic and biochemical characterization of LjGlb2-1
(Oxford University Press, 2021) Villar, Irene; Rubio, María Carmen; Calvo Begueria, Laura; Pérez Rontomé, Carmen; Larrainzar Rodríguez, Estíbaliz; Wilson, Michael T.; Sandal, Niels; Mur, Luis A.; Wang, Longlong; Reeder, Brandon; Duanmu, Deqiang; Uchiumi, Toshiki; Stougaard, Jens; Becana, Manuel; Zientziak; Institute for Multidisciplinary Research in Applied Biology - IMAB; Ciencias
Legumes express two major types of hemoglobins, namely symbiotic (leghemoglobins) and non-symbiotic (phytoglobins), with the latter being categorized into three classes according to phylogeny and biochemistry. Using knockout mutants, we show that all three phytoglobin classes are required for optimal vegetative and reproductive development of Lotus japonicus. The mutants of two class 1 phytoglobins showed different phenotypes: Ljglb1-1 plants were smaller and had relatively more pods, whereas Ljglb1-2 plants had no distinctive vegetative phenotype and produced relatively fewer pods. Non-nodulated plants lacking LjGlb2-1 showed delayed growth and alterations in the leaf metabolome linked to amino acid processing, fermentative and respiratory pathways, and hormonal balance. The leaves of mutant plants accumulated salicylic acid and contained relatively less methyl jasmonic acid, suggesting crosstalk between LjGlb2-1 and the signaling pathways of both hormones. Based on the expression of LjGlb2-1 in leaves, the alterations of flowering and fruiting of nodulated Ljglb2-1 plants, the developmental and biochemical phenotypes of the mutant fed on ammonium nitrate, and the heme coordination and reactivity of the protein toward nitric oxide, we conclude that LjGlb2-1 is not a leghemoglobin but an unusual class 2 phytoglobin. For comparison, we have also characterized a close relative of LjGlb2-1 in Medicago truncatula, MtLb3, and conclude that this is an atypical leghemoglobin.
Open Access
Nodule carbohydrate catabolism is enhanced in the Medicago truncatula A17-Sinorhizobium medicae WSM419 symbiosis
(Frontiers Media, 2014) Larrainzar Rodríguez, Estíbaliz; Gil Quintana, Erena; Seminario Huárriz, Amaia; Arrese-Igor Sánchez, César; González García, Esther; Ciencias del Medio Natural; Natura Ingurunearen Zientziak; Universidad Pública de Navarra / Nafarroako Unibertsitate Publikoa
The symbiotic association between Medicago truncatula and Sinorhizobium meliloti is a well-established model system in the legume–Rhizobium community. Despite its wide use, the symbiotic efficiency of this model has been recently questioned and an alternative microsymbiont, S. medicae, has been proposed. However, little is known about the physiological mechanisms behind the higher symbiotic efficiency of S. medicae WSM419. In the present study, we inoculated M. truncatula Jemalong A17 with either S. medicae WSM419 or S. meliloti 2011 and compared plant growth, photosynthesis, N2-fixation rates, and plant nodule carbon and nitrogen metabolic activities in the two systems. M. truncatula plants in symbiosis with S. medicae showed increased biomass and photosynthesis rates per plant. Plants grown in symbiosis with S. medicae WSM419 also showed higher N2-fixation rates, which were correlated with a larger nodule biomass, while nodule number was similar in both systems. In terms of plant nodule metabolism, M. truncatula–S. medicae WSM419 nodules showed increased sucrose-catabolic activity, mostly associated with sucrose synthase, accompanied by a reduced starch content, whereas nitrogen-assimilation activities were comparable to those measured in nodules infected with S. meliloti 2011. Taken together, these results suggest that S. medicae WSM419 is able to enhance plant carbon catabolism in M. truncatula nodules, which allows for the maintaining of high symbiotic N2-fixation rates, better growth and improved general plant performance.