BUSCADOR RECOLECTA

Ammonium is an important nitrogen (N) source for living organisms, a key metabolite for pH control, and a potent cytotoxic compound. Ammonium is transported by the widespread AMT-Mep-Rh membrane proteins, and despite their significance in physiological processes, the nature of substrate translocation (NH3/NH4+) by the distinct members of this family is still a matter of controversy. Using Saccharomyces cerevisiae cells expressing representative AMT-Mep-Rh ammonium carriers and taking advantage of the natural chemical-physical property of the N isotopic signature linked to NH4+/NH3 conversion, this study shows that only cells expressing AMT-Mep-Rh proteins were depleted in 15N relative to 14N when compared to the external ammonium source. We observed 15N depletion over a wide range of external pH, indicating its independence of NH3 formation in solution. On the basis of inhibitor studies, ammonium transport by nonspecific cation channels did not show isotope fractionation but competition with K+. We propose that kinetic N isotope fractionation is a common feature of AMT-Mep-Rh–type proteins, which favor 14N over 15N, owing to the dissociation of NH4+ into NH3 + H+ in the protein, leading to 15N depletion in the cell and allowing NH3 passage or NH3/H+ cotransport. This deprotonation mechanism explains these proteins’ essential functions in environments under a low NH4+/K+ ratio, allowing organisms to specifically scavenge NH4+. We show that 15N isotope fractionation may be used in vivo not only to determine the molecular species being transported by ammonium transport proteins, but also to track ammonium toxicity and associated amino acids excretion.

Accessing different nitrogen (N) sources involves a profound adaptation of plant metabolism. In this study, a quantitative proteomic approach was used to further understand how the model plant Arabidopsis thaliana adjusts to different N sources when grown exclusively under nitrate or ammonium nutrition. Proteome data evidenced that glucosinolate metabolism was differentially regulated by the N source and that both TGG1 and TGG2 myrosinases were more abundant under ammonium nutrition, which is generally considered to be a stressful situation. Moreover, Arabidopsis plants displayed glucosinolate accumulation and induced myrosinase activity under ammonium nutrition. Interestingly, these results were also confirmed in the economically important crop broccoli (Brassica oleracea var. italica). Moreover, these metabolic changes were correlated in Arabidopsis with the differential expression of genes from the aliphatic glucosinolate metabolic pathway. This study underlines the importance of nitrogen nutrition and the potential of using ammonium as the N source in order to stimulate glucosinolate metabolism, which may have important applications not only in terms of reducing pesticide use, but also for increasing plants’ nutritional value., This research was financially supported by the Basque Government (IT932-16), the Spanish Ministry of Economy and Competitiveness (AGL2015-64582-C3-1-R and BIO2014-56271-R co-funded by FEDER), and the
People Programme (Marie Curie Actions) of the European Union’s Seventh Framework Programme (FP7/2007–2013) under REA grant agreement number 334019. Navarrabiomed Proteomics Unit is a member of ProteoRed, PRB2-ISCIII, and is supported by grant PT13/0001, of the PE I+D+I 2013-2016 funded by ISCIII and FEDER.

Los inhibidores de la nitrificación (IN) pueden ser bien productos biológicos producidos por las plantas o de síntesis química, estos últimos son añadidos a los fertilizantes granulados, solubles o líquidos y del mismo modo que los biológicos, son capaces de inhibir el crecimiento de los microrganismos nitrificantes. Los inhibidores procedentes de la síntesis química se han venido utilizando desde hace décadas en la agricultura y han sido desarrollados para tratar de disminuir las pérdidas de N de los agrosistemas (lixiviado de nitrato y emisiones de óxidos de nitrógeno), aumentando la eficiencia del uso del N de los fertilizantes aplicados a los cultivos. Actualmente la diciandiamida (DCD) y el 3,4-dimetilpirazol fosfato (DMPP) son los IN comercializados más utilizados y estudiados. Un nuevo producto ha sido desarrollado recientemente, el ácido 2- (N-3,4-dimetil-1H-pirazol-1-il) succínico, y que pertenece al mismo grupo de los dimetilpirazoles. Aunque su eficiencia es similar al DMPP, hasta la fecha hay pocos estudios sobre esta nueva molécula y su modo de acción no está del todo resuelto. En este trabajo se ha estudiado el efecto de los IN derivados de dimetilpirazol, así como la nutrición amoniacal sobre la fisiología de las plantas, dado que el empleo de IN modifica la presencia de las formas de nitrógeno disponibles en el suelo a favor de una mayor presencia de amonio. En nuestro estudio hemos observado que ambos IN derivados de dimetilpirazol, DMPP y DMPSA pueden ser absorbidos y translocados de la raíz hacia la parte aérea, aunque únicamente se ha observado afectación de las plantas con el DMPP a dosis muy elevadas. Para intentar dilucidar el posible modo de acción del DMPP sobre los procesos fisiológicos y metabólicos de la planta, se realizó un estudio con las herramientas moleculares (transcriptómica y proteómica), apoyados con estudios fisiológicos para así tener un mejor conocimiento de los mecanismos y rutas fisiológicas de las plantas que pudieran estar afectadas por el inhibidor.
Dado que los IN favorecen la presencia de amonio en el suelo durante mayor tiempo tras la aplicación de fertilizantes de base amoniacal hemos realizado un estudio del efecto de diferentes ratios de NO3- y NH4+ en plantas de guisante y espinaca representativas de especies tolerantes y sensibles a la nutrición amoniacal. Hemos constatado que el mantenimiento del nivel de amonio en los tejidos es un factor importante en la respuesta a su tolerancia a esta fuente nitrogenada en las plantas., Este trabajo ha sido financiado por el proyecto AGL2015-64582-C3-1-R del Ministerio Economía y Competitividad., Programa Oficial de Doctorado en Agrobiología Ambiental (RD 1393/2007), Ingurumen Agrobiologiako Doktoretza Programa Ofiziala (ED 1393/2007)

This paper presents a three-year pilot experience of a new municipal waste management system developed in Navarre, Spain that integrates composting and hens. The aim of this new system is to motivate the general public to participate more in waste prevention programs. The Composter-Henhouse (CH) is a compact facility comprised of a henhouse and three composters. This is shared by 30 families who provide the organic part of their kitchen waste to feed the hens. Hens help speed up the composting process by depositing their droppings and turning the organic residue into compost. This study assesses the CH in terms of treatment capacity, the technical adequacy of the composting process, the quality and safety of the compost obtained and some social aspects. Over three years, the CH has managed nearly 16.5 tons of organic waste and produced approximately 5600 kg of compost and more than 6000 high-quality fresh eggs. No problems or nuisances have been reported and the level of animal welfare has been very high. The follow up of the composting process (temperature, volume reduction and compost maturity) and a physicochemical and microbiological analysis of the compost have ensured the proper management of the process. The level of involvement and user satisfaction has been outstanding and the project has presented clear social benefits., The study was financially supported by the Spanish Ministry of Science and Technology (projects AGL2012-37815-C05-05 and AGL2015-64582-C3-1-R).

Ammonium is an important nitrogen (N) source for living organisms, a key metabolite for pH control, and a potent cytotoxic compound. Ammonium is transported by the widespread AMT-Mep-Rh membrane proteins, and despite their significance in physiological processes, the nature of substrate translocation (NH3/NH4+) by the distinct members of this family is still a matter of controversy. Using Saccharomyces cerevisiae cells expressing representative AMT-Mep-Rh ammonium carriers and taking advantage of the natural chemical-physical property of the N isotopic signature linked to NH4+/NH3 conversion, this study shows that only cells expressing AMT-Mep-Rh proteins were depleted in N-15 relative to N-14 when compared to the external ammonium source. We observed N-15 depletion over a wide range of external pH, indicating its independence of NH3 formation in solution. On the basis of inhibitor studies, ammonium transport by nonspecific cation channels did not show isotope fractionation but competition with K+. We propose that kinetic N isotope fractionation is a common feature of AMT-Mep-Rh-type proteins, which favor N-14 over N-15, owing to the dissociation of NH4+ into NH3+ H+ in the protein, leading to N-15 depletion in the cell and allowing NH3 passage or NH3/H+ cotransport. This deprotonation mechanism explains these proteins' essential functions in environments under a low NH4+/K+ ratio, allowing organisms to specifically scavenge NH4+. We show that N-15 isotope fractionation may be used in vivo not only to determine the molecular species being transported by ammonium transport proteins, but also to track ammonium toxicity and associated amino acids excretion., I. A. was supported by a postdoctoral fellowship from the Government of Navarra, Spain (Anabasid outgoing Programme, 2011) and by a postdoctoral fellowship from the Portuguese Fundaçao para a Ciencia e a Tecnologia (SFRH/BPD/90436/2012). A.M.M. is a senior research associate of the Belgian Fonds de la Recherche Scientifique Fonds de la Recherche Scientifique-FNRS (grants CDR J017617F, PDR T011515F, and ARC) and a WELBIO investigator, and M.B. is a scientific research worker supported by WELBIO. This work was also developed in the context of the following projects: PTDC/BIA-BEC/099323/2008 and PTDC/AGR-PRO/115888/2009 to cE3c and FCUL, UID/DTP/04138/2013 to iMed. ULisboa, and AGL2015-64582-C3-1-R and AGL2012-37815-C05-05 to UPNa.