Function of C1A barley peptidases and their inhibitors (cystatins) in barley seed germination : Funcion de las peptidasas C1A de cebada y sus inhibidores (cistatinas) en la germinacion de la semilla

Plant proteolysis is a metabolic process where specific enzymes called peptidases degrade proteins. In plants, this complex process involves broad metabolic networks and different sub-cellular compartments. Several types of peptidases take part in the proteolytic process, mainly cysteine-, serine-, aspartyl- and metallo- peptidases. Among the cysteine-peptidases, the papain-like or C1A peptidases (family C1, clan CA) are extensively present in land plants and are classified into catepsins L-, B-, H- and Flike. The catalytic mechanism of these C1A peptidases is highly conserved and involves the three amino acids Cys, His and Asn in the catalytic triad, and a Gln residue which seems essential for maintaining an active enzyme conformation. These proteins are synthesized as inactive precursors, which comprise an N-terminal signal peptide, a propeptide, and the mature protein. In barley, we have identified 33 cysteine-peptidases from the papain-like family, classifying them into 8 different groups. Five of them corresponded to cathepsins L-like (5 subgroups), 1 cathepsin B-like group, 1 cathepsin F-like group and 1 cathepsin H-like group. Besides, C1A peptidases are the specific targets of the plant proteinaceous inhibitors known as phytocystatins (PhyCys). The cystatin inhibitory mechanism is produced by a tight and reversible interaction with their target enzymes. In barley, the cystatin gene family is comprised by 13 members. In this work we have tried to elucidate the role of the C1A cysteine-peptidases and their specific inhibitors (cystatins) in the germination process of the barley grain. Therefore, we selected a representative member of each group/subgroup of C1A peptidases (1 cathepsin B-like, 1 cathepsin F-like, 1 cathepsin H-like and 5 cathepsins L-like). The molecular characterization of the cysteine-peptidases was done and the peptidase-inhibitor interaction was analyzed in vitro and in vivo. A study in the structural basis for specificity of pro-peptide/enzyme interaction in barley C1A cysteine-peptidases has been also carried out by inhibitory assays and the modeling of the three-dimensional structures. The barley grain maturation produces the accumulation of storage proteins (prolamins) in the endosperm which are mobilized during germination to supply the required nutrients until the photosynthesis is fully established. In this work, we have demonstrated the participation of the cysteine-peptidases and their inhibitors in the degradation of the different storage protein fractions (hordeins, albumins and globulins) present in the barley grain. Besides, transgenic barley plants overexpressing or silencing cysteine-peptidases or cystatins were obtained by Agrobacterium-mediated transformation of barley immature embryos to analyze their physiological function in vivo. Preliminary assays were carried out with the T1 grains of several transgenic lines. Comparing the knock-out and the overexpressing lines with the WT, alterations in the germination process were detected and were correlated with their grain hordein content. These data will be validated with the homozygous grains that are being produced through the double haploid technique by microspore culture. Resumen La proteólisis es un proceso metabólico por el cual se lleva a cabo la degradación de las proteínas de un organismo a través de enzimas específicas llamadas proteasas. En plantas, este complejo proceso comprende un entramado de rutas metabólicas que implican, además, diferentes compartimentos subcelulares. En la proteólisis participan numerosas proteasas, principalmente cisteín-, serín-, aspartil-, y metalo-proteasas. Dentro de las cisteín-proteasas, las proteasas tipo papaína o C1A (familia C1, clan CA) están extensamente representadas en plantas terrestres, y se clasifican en catepsinas tipo L, B, H y F. El mecanismo catalítico de estas proteasas está altamente conservado y la triada catalítica formada por los aminoácidos Cys, His y Asn, y a un aminoácido Gln, que parece esencial para el mantenimiento de la conformación activa de la proteína. Las proteasas C1A se sintetizan como precursores inactivos y comprenden un péptido señal en el extremo N-terminal, un pro-péptido y la proteína madura. En cebada hemos identificado 33 cisteín-proteasas de tipo papaína y las hemos clasificado filogenéticamente en 8 grupos diferentes. Cinco de ellos pertenecen a las catepsinas tipo L (5 subgrupos), un grupo a las catepsinas tipo-B, otro a las catepsinas tipo-F y un último a las catepsinas tipo-H. Las proteasas C1A son además las dianas específicas de los inhibidores protéicos de plantas denominados fitocistatinas. El mecanismo de inhibición de las cistatinas está basado en una fuerte interacción reversible. En cebada, se conoce la familia génica completa de las cistatinas, que está formada por 13 miembros. En el presente trabajo se ha investigado el papel de las cisteín-proteasas de cebada y sus inhibidores específicos en el proceso de la germinación de la semilla. Para ello, se seleccionó una proteasa representante de cada grupo/subgrupo (1 catepsina tipo- B, 1 tipo-F, 1 tipo-H, y 5 tipo-L, una por cada subgrupo). Se ha llevado a cabo su caracterización molecular y se ha analizado la interacción enzima-inhibidor tanto in vivo como in vitro. También se han realizado estudios sobre las bases estructurales que demuestran la especificidad en la interacción enzima/propéptido en las proteasas C1A de cebada, mediante ensayos de inhibición y la predicción de modelos estructurales de la interacción. Finalmente, y dado que durante la maduración de la semilla se almacenan proteínas de reserva (prolaminas) en el endospermo que son movilizadas durante la germinación para suministrar los nutrientes necesarios hasta que la nueva planta pueda realizar la fotosíntesis, en este trabajo se ha demostrado la participación de las cisteínproteasas y sus inhibidores en la degradación de las diferentes tipos de proteínas de reserva (hordeinas, albúmins y globulinas) presentes en el grano de cebada. Además, se han obtenido plantas transgénicas de cebada que sobre-expresan o silencian cistatinas y cisteín-proteasas con el fin de analizar la función fisiológica in vivo. Se han realizado análisis preliminares en las semillas T1 de varias líneas tránsgenicas de cebada y al comparar las líneas knock-out y las líneas de sobre-expresión con las silvestres, se han detectado alteraciones en la germinación que están además correlacionadas con el contenido de hordeinas de las semillas. Estos datos serán validados en las semillas homocigotas que se están generando mediante la técnica de dobles haploides a partir del cultivo de microesporas.

Differential in vitro and in vivo effect of barley cysteine and serine protease inhibitors on phytopathogenic microorganisms

Protease inhibitors from plants have been involved in defence mechanisms against pests and pathogens. Phytocystatins and trypsin/α-amylase inhibitors are two of the best characterized protease inhibitor families in plants. In barley, thirteen cystatins (HvCPI-1 to 13) and the BTI-CMe trypsin inhibitor have been previously studied. Their capacity to inhibit pest digestive proteases, and the negative in vivo effect caused by plants expressing these inhibitors on pests support the defence function of these proteins. Barley cystatins are also able to inhibit in vitro fungal growth. However, the antifungal effect of these inhibitors in vivo had not been previously tested. Moreover, their in vitro and in vivo effect on plant pathogenous bacteria is still unknown. In order to obtain new insights on this feature, in vitro assays were made against different bacterial and fungal pathogens of plants using the trypsin inhibitor BTI-CMe and the thirteen barley cystatins. Most barley cystatins and the BTI-CMe inhibitor were able to inhibit mycelial growth but no bacterial growth. Transgenic Arabidopsis plants independently expressing the BTI-CMe inhibitor and the cystatin HvCPI-6 were tested against the same bacterial and fungal pathogens. Neither the HvCPI-6 expressing transgenic plants nor the BTI-CMe ones were more resistant to plant pathogen fungi and bacteria than control Arabidopsis plants. The differences observed between the in vitro and in planta assays against phytopathogenic fungi are discussed

El Hayedo de Montejo, mucho más que hayas

El Hayedo de Montejo, uno de los bosques más emblemáticos de la Comunidad de Madrid, es también uno de los mejor conocidos y más estudiados gracias a un convenio de colaboración entre la Comunidad de Madrid y la Escuela de Montes de la Universidad Politécnica de Madrid (UPM). Seguimiento del clima, inventarios forestales, estudios genéticos o ecofisiológicos y procesos de dispersión son algunos de los trabajos que vienen desarrollándose desde hace más de 17 años. Gracias a ellos descubrimos que El Hayedo de Montejo es mucho más que un bosque de hayas.

Año Internacional de los Bosques: Tópicos y realidades

Declarado “Año internacional de los bosques”, 2011 tenía como objetivo incrementar los esfuerzos para conservar las masas arbóreas del planeta y recordar los beneficios que nos proporcionan. Es el momento de despejar algunas cuestiones: ¿A qué llamamos bosques? ¿Qué evolución han tenido en nuestro país? ¿Cuál es la realidad sin tópicos ni eufemismos?

Impacts of projected maximum temperature extremes for C21 by an ensemble of regional climate models on cereal cropping systems in the Iberian Peninsula

Crops growing in the Iberian Peninsula may be subjected to damagingly high temperatures during the sensitive development periods of flowering and grain filling. Such episodes are considered important hazards and farmers may take insurance to offset their impact. Increases in value and frequency of maximum temperature have been observed in the Iberian Peninsula during the 20th century, and studies on climate change indicate the possibility of further increase by the end of the 21st century. Here, impacts of current and future high temperatures on cereal cropping systems of the Iberian Peninsula are evaluated, focusing on vulnerable development periods of winter and summer crops. Climate change scenarios obtained from an ensemble of ten Regional Climate Models (multimodel ensemble) combined with crop simulation models were used for this purpose and related uncertainty was estimated. Results reveal that higher extremes of maximum temperature represent a threat to summer-grown but not to winter-grown crops in the Iberian Peninsula. The study highlights the different vulnerability of crops in the two growing seasons and the need to account for changes in extreme temperatures in developing adaptations in cereal cropping systems. Finally, this work contributes to clarifying the causes of high-uncertainty impact projections from previous studies.

Cysteine-proteases and cystatins from barley: molecular and functional characterization in housekeeping and defense processes

Plant cysteine-proteases (CysProt) represent a well-characterized type of proteolytic enzymes that fulfill tightly regulated physiological functions (senescence and seed germination among others) and defense roles. This article is focused on the group of papain-proteases C1A (family C1, clan CA) and their inhibitors, phytocystatins (PhyCys). In particular, the protease–inhibitor interaction and their mutual participation in specific pathways throughout the plant's life are reviewed. C1A CysProt and PhyCys have been molecularly characterized, and comparative sequence analyses have identified consensus functional motifs. A correlation can be established between the number of identified CysProt and PhyCys in angiosperms. Thus, evolutionary forces may have determined a control role of cystatins on both endogenous and pest-exogenous proteases in these species. Tagging the proteases and inhibitors with fluorescence proteins revealed common patterns of subcellular localization in the endoplasmic reticulum–Golgi network in transiently transformed onion epidermal cells. Further in vivo interactions were demonstrated by bimolecular fluorescent complementation, suggesting their participation in the same physiological processes.

Lempel-Ziv complexity in schizophrenia: A MEG study

Objective The neurodevelopmental–neurodegenerative debate is a basic issue in the field of the neuropathological basis of schizophrenia (SCH). Neurophysiological techniques have been scarcely involved in such debate, but nonlinear analysis methods may contribute to it. Methods Fifteen patients (age range 23–42 years) matching DSM IV-TR criteria for SCH, and 15 sex- and age-matched control subjects (age range 23–42 years) underwent a resting-state magnetoencephalographic evaluation and Lempel–Ziv complexity (LZC) scores were calculated. Results Regression analyses indicated that LZC values were strongly dependent on age. Complexity scores increased as a function of age in controls, while SCH patients exhibited a progressive reduction of LZC values. A logistic model including LZC scores, age and the interaction of both variables allowed the classification of patients and controls with high sensitivity and specificity. Conclusions Results demonstrated that SCH patients failed to follow the “normal” process of complexity increase as a function of age. In addition, SCH patients exhibited a significant reduction of complexity scores as a function of age, thus paralleling the pattern observed in neurodegenerative diseases. Significance Our results support the notion of a progressive defect in SCH, which does not contradict the existence of a basic neurodevelopmental alteration. Highlights ► Schizophrenic patients show higher complexity values as compared to controls. ► Schizophrenic patients showed a tendency to reduced complexity values as a function of age while controls showed the opposite tendency. ► The tendency observed in schizophrenic patients parallels the tendency observed in Alzheimer disease patients.

Evolution not revolution of farming systemswill best feed and green the world

The challenge to properly feed a world population of 9.2 billion by 2050, that must be achieved on essentially currently cropped area, requires that food production be increased by 70%. This large increase can only be achieved by combinations of greater crop yields and more intensive cropping adapted to local conditions and availability of inputs. Farming systems are dynamic and continuously adapt to changing ecological, environmental and social conditions, while achieving greater production and resource-use efficiency by application of science and technology. This article argues that the solution to feed and green the world in 2050 is to support this evolution more strongly by providing farmers with necessary information, inputs, and recognition. There is no revolutionary alternative. Proposals to transform agriculture to low-input and organic systems would, because of low productiv- ity, exacerbate the challenge if applied in small part, and ensure failure if applied more widely. The challenge is, however, great. Irrigation, necessary to increase cropping intensity in many areas cannot be extended much more widely than at present, and it is uncertain if the current rate of crop yield increase can be maintained. Society needs greater recognition of the food-supply problem and must increase funding and support for agricultural research while it attends to issues of food waste and over consumption that can make valuable reductions to food demand from agriculture

Systematics of Senecio sect. Crociseris (Compositae, Senecioneae)

Se revisa taxonómicamente Senecio sect. Crociseris (Compositae, Senecioneae),una de las cuatro principales secciones del género en el ámbito de Eurasia y el norte de África. Este grupo comprende hierbas rizomatosas, distribuidas principalmente del oeste de Europa y noroeste de África al oeste asiático, y presenta la mayor diversidad en el Mediterráneo septentrional. El nuevo tratamiento taxonómico que se propone reconoce 28 especies y 8 subespecies. Se ha hecho un especial esfuerzo en el análisis de la variabilidad del gr. S. doronicum, cuya sistemática ha sido objeto de controversia durante mucho tiempo. En este grupo se aceptan tres especies: S. doronicum, S.lagascanus, y S. provincialis, la primera con tres subespecies: subsp. doronicum, subsp.longifolius, y subsp. orientalis. Con el objetivo de indagar en las relaciones de las especies dentro de la sección y de ésta con otros grupos de Senecio, se han llevado a cabo análisis filogenéticos basados en secuencias ITS y de cloroplasto. Los resultados sugieren que la sect. Crociseris es un grupo parafilético, ya que incluye las especies secuenciadas de la sect. Doria. Además, complejos patrones de algunas secuencias ITS sugieren eventos de hibridación. Como resultado del estudio taxonómico se proponen las siguientes combinaciones: Senecio doronicum subsp. longifolius(Willk.) J. Calvo, Senecio doronicum subsp. orientalis (Ten.) J. Calvo, Senecio macedonicus subsp. barckhausiaefolius (Boiss. & Heldr.) J. Calvo, y Senecio racemosus subsp. kirghisicus(DC.) J. Calvo. Por otro lado, S. ruthenensis de Francia y S. lusitanicus de Portugal han sido sinonimizados a S. lagascanus, así como S. ovatifolius, S. pisidicus y S. tmoleus de Anatolia a S. kolenatianus, S. olympicus, y S. castagneanus respectivamente, S.bertramii de Líbano a S. cilicius, y S. delbesianus de Siria a S. racemosus subsp. racemosus. Se han lectotipificado setenta y cinco nombres, y neotipificado S. pyrenaicus y S. scopolii. Senecio gerardi Godr., S. doronicum (L.) L. y S. paucifolius S.G. Gmel. se propusieron a conservar. Se incluyen claves de identificación, descripciones detalladas, mapas de distribución e ilustraciones. Nueve táxones han sido ilustrados por primera vez.

Connecting red cells in a bichromatic Voronoi diagram

Let S be a set of n + m sites, of which n are red and have weight wR, and m are blue and weigh wB. The objective of this paper is to calculate the minimum value of wR such that the union of the red Voronoi cells in the weighted Voronoi diagram of S is a connected set. The problem is solved for the multiplicatively-weighted Voronoi diagram in O((n+m)^2 log(nm)) time and for the additively-weighted Voronoi diagram in O(nmlog(nm)) time.

TIMPANO: Technology for complex Human-Machine conversational interaction with dynamic learning

El proyecto TIMPANO tiene por objetivo profundizar en el desarrollo de sistemas de comunicación oral hombre-máquina atendiendo principalmente a la capacidad de dar respuesta a múltiples requerimientos de los usuarios, como pueden ser el acceso a información, la extracción de información, o el análisis de grandes repositorios de información en audio. En el proyecto se hace especial énfasis en la adaptación dinámica de los modelos a diversos contextos, tanto de tipo acústico, como semántico o de idioma.

Outline of Europe-Japan collaborative research on concentrator photovoltaics

The Europe-Japan Collaborative Research Project on Concentrator Photovoltaics (CPV) has been initiated under support by the EC (European Commission) and NEDO (New Energy and Industrial Technology Development Organization) since June 2011. This is project (NGCPV Project; a New Generation of Concentrator PhotoVoltaic cells, modules and systems) is aiming to accelerate the move to very high efficiency and lower cost CPV technologies and to enhance widespread deployment of CPV systems. 7 organizations such as UPM, FhG-ISE Imperial College, BSQ, CEA-INES, ENEA, and PSE in Europe and 9 organizations such as TTI, Univ. Tokyo, AIST, Sharp Co. Daido Steel Co., Kobe Univ., Miyazaki Univ., Asahi Kasei Co., and Takano Co. participate in this project. The targets of this project are 1) to develop world-record efficiency CPV cells of more than 45%, 2) to develop world-record efficiency CPV modules of 35%, 3) to establish standard measurements of CPV cells and modules, 4) to install 50kW CPV system in Spain, to carry out field test of CPV system and to manage power generation of CPV systems, and 5) to develop high-efficiency and low-cost new materials and structure cells such as III-V-N, III-V-on-Si tandem, quantum dots and wells. This paper presents outline of this project and most recent results such as world record efficiency (37.9% under 1-sun) cell and high-efficiency (43.5% under 240-306 suns) concentrator cell with inverted epitaxial grown InGaP/GaAs/InGaAs 3-junction solar cells.

Assessment of insurance coverage and claims in rainfall related risks in processing tomato in Western Spain

An extension of guarantees related to rainfall-related risks in the insurance of processing tomato crops has been accompanied with a large increase in claims in Western Spain, suggesting that damages may have been underestimated in previous years. A database was built by linking agricultural insurance records, meteorological data from local weather stations, and topographic data. The risk of rainfall-related damages in processing tomato in the Extremenian Guadiana river basin (W Spain) was studied using a logistic model. Risks during the growth of the crop and at harvesting were modelled separately. First, the risk related to rainfall was modelled as a function of meteorological, terrain and management variables. The resulting models were used to identify the variables responsible for rainfall-related damages, with a view to assess the potential impact of extending insurance coverage, and to develop an index to express the suitability of the cropping system for insurance. The analyses reveal that damages at different stages of crop development correspond to different hazards. The geographic dependence of the risk influences the scale at which the model might have validity, which together with the year dependency, the possibility of implementing index based insurances is questioned.

Comportamiento mecánico de mezclas de escoria vítrea de horno alto y metacaolín activadas alcalinamente: estudio estadístico = Mechanical behaviour of alkali-activated metakaolin blended pastes: statistical study

El estudio y desarrollo de cementos alternativos y más eco-eficientes que el cemento Portland es un tema de gran impacto a nivel científico y tecnológico. Entre esos posibles cementos se encuentran los cementos alcalinos que son materiales conglomerantes obtenidos por la interacción química de materiales silico-aluminosos cálci- cos y disoluciones fuertemente alcalinas. En el presente trabajo se estudia el comportamiento mecánico y la com- posición mineralógica de mezclas de escoria vítrea de horno alto y metacaolín activadas alcalinamente con disoluciones de NaOH. El objetivo de este estudio es conocer cómo afectan parámetros tales como la relación escoria/metacaolín, la concentración de la disolución activadora y la temperatura de curado, al desarrollo re- sistente de las mezclas. A través del estudio estadístico realizado se ha podido establecer la influencia de cada variable y modelizar el comportamiento resistente de estos cementos alcalinos. Se concluye que la concentra- ción del activador y la relación escoria/metacaolín son los parámetros más relevantes.

Agroclima: Escenarios climáticos, impactos, vulnerabilidades y sistemas agrarios. Seminario sobre adaptación al cambio climático en el sector agrario. Oficina Española de Cambio climático. Secretaria General de Agricultura, Alimentación y Medio Ambiente

Modelos Globales de Clima. Modelos Regionales de Clima.