75 años como referente de la investigación agraria y medioambiental española en condiciones de clima mediterráneo [Sitio Web]

1 .pdf con imagen de acceso al “website”, su url y los créditos relacionados con su creación y diseño.-- Créditos: Organización, Estación Experimental de Aula Dei (EEAD-CSIC); Dirección, Jesús Val Falcón; Coordinación, Ana Álvarez-Fernandez, Jorge Álvaro-Fuentes, Ernesto Igartua; Contenido, Anunciación Abadía, Javier Abadía, Carlos Albiñana, Miguel Alfonso, Arancha Arbeloa, Raúl Arbués, Isabel Armillas, Manuel Becana, Santiago Beguería, Carmen Castañeda, Ana Castillo, José Cavero, Bruno Contreras, Azahara Díaz, Edgar García, Elena García, Juan Manuel Gascuñana, Leticia Gaspar, Yolanda Gogorcena, Juan Herrero, Victoria Lafuente, María Victoria López, Juan Antonio Marín, José Martínez, José Carlos Martínez-Giménez, Ana Pilar Mata, Manuel Matamoros, Pierre Mignard, María Ángeles Moreno, Paula Murillo, Ana Navas, Antonio Pérez, Rafael Picorel, María Pilar Vallés, Irene Villar, Inmaculada Yruela, Nery Zapata, Isabel Zarazaga; Diseño y programación: DigitalWorks (Juanjo Ascaso y Asun Dieste); Vídeo, Delegación del CSIC en Aragón (Sara Gutiérrez y Yolanda Hernáiz); Fotografía, Archivo EEAD-CSIC, Anunciación Abadía, Jorge Álvaro-Fuentes, Arancha Arbeloa, Juanjo Ascaso, Santiago Beguería, Elena García, Ernesto Igartua, Ignasi Iglesias, José Manuel Lasa, José Carlos Martínez-Giménez, Pierre Mignard, María Ángeles Moreno, Rubén Sancho, Kosana Suvocarev, María Pilar Vallés, Nery Zapata."Sitio web" de nueva creación y conmemorativo del 75 Aniversario de la EEAD-CSIC que contiene: 1) Foto esférica de su personal en activo; 2) Recopilación de sus hitos históricos más destacados, en orden cronológico; 3) Un vídeo con participación de su personal y muestra de algunas de sus instalaciones; 4) Un mapa con la distribución geográfica de los egresado del Instituto; 5) Algunas fotos, destacando las tomadas a su personal en las celebraciones del 25 y 50 Aniversarios de la EEAD-CSIC.Presentado durante la "Jornada. 75 Aniversario EEAD-CSIC (Zaragoza, Patio de la Infanta. 30 octubre 2019)".Financiación: CSIC, Vicepresidencia Adjunta de Organización y Cultura Científica.N

Estructura y función de hemoglobinas atípicas de leguminosas modelo

187 p.- Figs. © The author. Under Creative Commons License Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0).El objetivo general de la tesis doctoral ha consistido en la caracterización bioquímica y funcional de hemoglobinas de las leguminosas modelo Medicago truncatula y Lotus japonicus. La tesis se ha centrado, por una parte, en el estudio de dos hemoglobinas de M. truncatula (MtGlb1-2 y MtLb3) y una hemoglobina de L. japonicus (LjGlb2-1). Los resultados obtenidos muestran que MtGlb1-2 tiene unas características inusuales a las descritas hasta el momento, presentando una reactividad muy elevada a diferentes ligandos de relevancia biológica. Estas características inusuales nos llevan a proponer que MtGlb1-2 puede actuar secuestrando o produciendo NO, dependiendo de la disponibilidad de oxígeno en el tejido. Además, hemos estudiado y comparado, tanto a nivel bioquímico como a nivel funcional, MtLb3 y LjGlb2-1, que presentan unas características intermedias entre las leghemoglobinas y las hemoglobinas de clase 2. El estudio y compresión de las leguminosas en simbiosis con bacterias del suelo presenta numerosas ventajas agrícolas ya que son capaces de fijar el nitrógeno atmosférico en una forma asimilable por las plantas (biofertilización).En primer lugar, me gustaría agradecer al Ministerio de Ciencia e Innovación, por financiar estos cuatro años con un contrato FPI (BES-2015-073059), asociado al proyecto AGL2014-53717-R, así como al proyecto AGL2017-85775-R por el contrato que disfruto en la actualidad. Ambos proyectos han sido cofinanciados por el Fondo Europeo de Desarrollo Regional (FEDER) de la Unión Europea.Peer reviewe

In vitro regeneration of two Populus hybrid clones. The role of pectin domains in cell processes underlying shoot organogenesis induction

26 Pags.- 6 Tabls.- 7 Figs. The definitive version is available at: https://link.springer.com/journal/10535An efficient plant regeneration protocol has been established for two commercial Populus hybrid clones, MC (Populus × euramericana) and UNAL (Populus × interamericana). The culture of internode segments on Murashige and Skoog (MS) medium with 0.5 μM α-naphthalene acetic acid (NAA) and 4 μM N6-benzyladenine for 7 weeks (2 weeks in absence of activated charcoal and 5 weeks in its presence) resulted in the highest frequency of shoot regeneration (100 % for MC and 82 % for UNAL). All regenerated shoots longer than 2 cm rooted on half-strength MS medium, independent of the addition of 0.1 μM NAA. Nevertheless, shoots developed better-formed roots in NAA-free medium, which had a positive effect on the acclimatization of plants. In order to know the cellular processes underlying in vitro shoot organogenesis, a histological study was made in UNAL internode-explants. Results revealed that in vitro culture caused swelling around the cut-off zones in all explants, but only those undergoing organogenesis formed proliferation centers under subepidermal cells, which led to formation of bud primordia. Moreover, in vivo tissues and explants with different in vitro response showed different immunolabelling patterns when they were treated with fluorescentmonoclonal antibodies directed to several pectin-polysaccharides of the cell wall. Results allow us to assign a predominant role of homogalacturonan with a low degree of methyl-esterification in the initiation of bud primordia, a role of β-1,4-D-galactan side chains of rhamnogalacturonan-I in the cellular differentiation, ra ole of α-1,5-L-arabinan side chains of rhamnogalacturonan-I and of homogalacturonan with a high degree of methyl-esterification in cell division and growth.Peer reviewe

Characterization of Lotus japonicus mutant plants deficient in phytoglobins

1 Pag.In plants, symbiotic hemoglobins occur only in the nodules of legumes (leghemoglobins; Lbs), actinorhizal plants, and Parasponia. These hemoglobins transport a low, steady concentration of O2 to the N2-fixing bacteroids. In contrast, nonsymbiotic hemoglobins or phytoglobins (Glbs) are expressed in most plant organs, from seeds to flowers and fruits (Bustos-Sanmamed et al., 2011). Based on phylogenetic analyses and biochemical properties, Glbs can be grouped in three classes. The genome of Lotus japonicus encodes five Glbs: two class 1 (Glb1-1 and Glb1-2), one class 2 (Glb2), and two class 3 (Glb3-1 and Glb3-2). The functions of Glbs are largely unknown. To gain information, we phenotyped glb knockout mutants from the LORE1 collection (Malolepszy et al., 2016). Phenotypes were examined in nodulated plants grown in plates for 4-5 weeks on Jensen medium. The plants were then transferred to pots and the phenotypes were analyzed again at 8 weeks and at the flowering and fruiting stages. Nodulated plants of the glb1-1 mutant were smaller and had fewer nodules than the WT. The glb1-2 plants showed no growth alterations but delayed flowering, although they had many flowers that did not produce pods. The glb2 mutant showed a reduction in size and nodule number, as well as a delay in flowering. Finally, the glb3-2 mutants showed reduced shoot and root weights, delayed flowering, and enhanced production of flowers and pods.I.V. is the recipient of a contract (BES2015-073059) from the Ministry of Economy and Competitiveness (MINECO). This work was funded by grant AGL2017-85775-R to M.B. and M.C.R. from MINECO - European Regional Development Fund.Peer reviewe

Unusually Fast bis-Histidyl Coordination in a Plant Hemoglobin

13 Pags.- 4 Figs.- 2 Tabls- 1 Suppl File. © 2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license.The recently identified nonsymbiotic hemoglobin gene MtGlb1-2 of the legume Medicago truncatula possesses unique properties as it generates four alternative splice forms encoding proteins with one or two heme domains. Here we investigate the ligand binding kinetics of MtGlb1-2.1 and MtGlb1-2.4, bearing two hemes and one heme, respectively. Unexpectedly, the overall time-course of ligand rebinding was unusually fast. Thus, we complemented nanosecond laser flash photolysis kinetics with data collected with a hybrid femtosecond–nanosecond pump–probe setup. Most photodissociated ligands are rebound geminately within a few nanoseconds, which leads to rates of the bimolecular rebinding to pentacoordinate species in the 108 M−1s−1 range. Binding of the distal histidine to the heme competes with CO rebinding with extremely high rates (kh ~ 105 s−1). Histidine dissociation from the heme occurs with comparable rates, thus resulting in moderate equilibrium binding constants (KH ~ 1). The rate constants for ligation and deligation of distal histidine to the heme are the highest reported for any plant or vertebrate globin. The combination of microscopic rates results in unusually high overall ligand binding rate constants, a fact that contributes to explaining at the mechanistic level the extremely high reactivity of these proteins toward the physiological ligands oxygen, nitric oxide and nitrite.This research was funded by the Spanish Agencia Estatal de Investigación (AEI) grant AGL2017-85775-R, co-funded by the European Regional Development Fund, and by Government of Aragón (group A09_17R). The APC was funded by AEI and, in part, by the CSIC Open Access Publication Support Initiative through its Unit of Information Resources for Research (URICI-CSIC). G.C. acknowledges support from the PRIN 2017 Project 201795SBA3—HARVESTPeer reviewe

Hemoglobins in the legume–Rhizobium symbiosis

13 Pags.- 7 Figs. ©2020 The Authors. Under LicenceCC BY-NC-NDLegume nodules have two types of hemoglobins: symbiotic or leghemoglobins (Lbs) and nonsymbiotic or phytoglobins (Glbs). The latter are categorized into three phylogenetic classes differing in heme coordination and O2 affinity. This review is focused on the roles of Lbs and Glbs in the symbiosis of rhizobia with crop legumes and the model legumes for indeterminate (Medicago truncatula) and determinate (Lotus japonicus) nodulation. Only two hemoglobin functions are well established in nodules: Lbs deliver O2 to the bacteroids and act as O2 buffers, preventing nitrogenase inactivation; and Glb1‐1 modulates nitric oxide concentration during symbiosis, from the early stage, avoiding the plant’s defense response, to nodule senescence. Here, we critically examine early and recent results, update and correct the information on Lbs and Glbs with the latest genome versions, provide novel expression data and identify targets for future research. Crucial unresolved questions include the expression of multiple Lbs in nodules, their presence in the nuclei and in uninfected nodule cells, and, intriguingly, their expression in nonsymbiotic tissues. RNA‐sequencing data analysis shows that Lbs are expressed as early as a few hours after inoculation and that their mRNAs are also detectable in roots and pods, which clearly suggests that these heme proteins play additional roles unrelated to nitrogen fixation. Likewise, issues awaiting investigation are the functions of other Glbs in nodules, the spatiotemporal expression profiles of Lbs and Glbs at the mRNA and protein levels, and the molecular mechanisms underlying their regulation during nodule development and in response to stress and hormones.This work has been funded by the Spanish Ministry of Science and Innovation-European Regional Development Fund (grants AGL2017-85775-R and RTI2018-094623-B-C22) and by Government of Aragon, Spain (groupA09_17R).EL is a Ramon y Cajal fellow (RYC2018-023867-I) and IV is a Formacionde Personal Investigador fellow (BES-2015-073059).Peer reviewe

Subcellular localization and nitric oxide-scavenging activity of plant hemoglobins

1 Pag.Symbioc hemoglobins provide O2 to N2-fixing bacteria within legume nodules. However, the funcons of nonsymbioc hemoglobins or phytoglobins (Glbs) are less defined. Three Glb classes can be disnguished based on phylogenec and biochemical analyses and may coexist in plant ssues: class 1 Glbs have extreme O2 affinity and are induced by hypoxia; class 2 Glbs have moderate O2 affinity and are precursors of leghemoglobins; and class 3 have low O2 affinity and high sequence homology with bacterial truncated hemoglobins. Immunogold labeling combined with confocal microscopy of Glbs tagged with GFP at the C-terminus was used to determine the subcellular localizaons of Glbs in the model plants Arabidopsis and Lotus japonicus. To this end, we used overexpressing and knockout or silenced lines of Arabidopsis, performed quantave immunolabeling, and monitored the GFP-tagged proteins in leaf cells and protoplasts.This work was funded by grant AGL2017-85775-R from the Spanish Ministry of Economy, Industry and Competitiveness/European Regional Development Fund.Peer reviewe

Biodegradación de la lignocelulosa: aspectos microbiológicos, químicos y enzimáticos del ataque fúngico a la lignina

Wood is the main renewable material on Earth and is largely used as building material and in paper-pulp manufacturing. This review describes the composition of lignocellulosic materials, the different processes by which fungi are able to alter wood, including decay patterns caused by white, brown, and soft-rot fungi, and fungal staining of wood. The chemical, enzymatic, and molecular aspects of the fungal attack of lignin, which represents the key step in wood decay, are also discussed. Modern analytical techniques to investigate fungal degradation and modification of the lignin polymer are reviewed, as are the different oxidative enzymes (oxidoreductases) involved in lignin degradation. These include laccases, high redox potential ligninolytic peroxidases (lignin peroxidase, manganese peroxidase, and versatile peroxidase), and oxidases. Special emphasis is given to the reactions catalyzed, their synergistic action on lignin, and the structural bases for their unique catalytic properties. Broadening our knowledge of lignocellulose biodegradation processes should contribute to better control of wood-decaying fungi, as well as to the development of new biocatalysts of industrial interest based on these organisms and their enzymes.These studies have been partially supported by ENCE (Spain), by Spanish projects AGL2002-393 and BIO2002-1166, by EU projects QLK5-99-1357 and QLK3-99-590, and by an EUFORES(ENCE)-PDT/MEC(Uruguay) grant. Carmen Ascaso (CCMA, CSIC, Madrid) is acknowledged for low-temperature scanning-electron microscopy facilities. Klaus Piontek (ETH, Zurich) is acknowledged for solving the VP crystal structure. Lina Bettucci (Universidad de la República, Montevideo) is acknowledged for an Inocutis jamaicensis strain. M.S acknowledges MEC for a Postdoctoral Fellowship. F.J.R.-D. thanks CSIC for an I3P contract. A.G. and S.C. thank MEC for their “Ramón y Cajal” contracts.Peer reviewe