The AtCathB3 gene, encoding a cathepsin B-like protease, is expressed during germination of Arabidopsis thaliana and transcriptionally repressed by the basic leucine zipper protein GBF1.

Protein hydrolysis plays an important role during seed germination and post-germination seedling establishment. In Arabidopsis thaliana, cathepsin B-like proteases are encoded by a gene family of three members, but only the AtCathB3 gene is highly induced upon seed germination and at the early post-germination stage. Seeds of a homozygous T-DNA insertion mutant in the AtCathB3 gene have, besides a reduced cathepsin B activity, a slower germination than the wild type. To explore the transcriptional regulation of this gene, we used a combined phylogenetic shadowing approach together with a yeast one-hybrid screening of an arrayed library of approximately 1200 transcription factor open reading frames from Arabidopsis thaliana. We identified a conserved CathB3-element in the promoters of orthologous CathB3 genes within the Brassicaceae species analysed, and, as its DNA-interacting protein, the G-Box Binding Factor1 (GBF1). Transient overexpression of GBF1 together with a PAtCathB3::uidA (β-glucuronidase) construct in tobacco plants revealed a negative effect of GBF1 on expression driven by the AtCathB3 promoter. In stable P35S::GBF1 lines, not only was the expression of the AtCathB3 gene drastically reduced, but a significant slower germination was also observed. In the homozygous knockout mutant for the GBF1 gene, the opposite effect was found. These data indicate that GBF1 is a transcriptional repressor of the AtCathB3 gene and affects the germination kinetics of Arabidopsis thaliana seeds. As AtCathB3 is also expressed during post-germination in the cotyledons, a role for the AtCathB3-like protease in reserve mobilization is also inferred.

Factores transcripcionales de la clase DOF en Brachypodium distachyon: caracterización molecular de BdDOF24 durante la germinación de las semillas

La semilla es el órgano que garantiza la propagación y continuidad evolutiva de las plantas espermatofitas y constituye un elemento indispensable en la alimentación humana y animal. La semilla de cereales acumula en el endospermo durante la maduración, mayoritariamente, almidón y proteínas de reserva. Estas reservas son hidrolizadas en la germinación por hidrolasas sintetizadas en la aleurona en respuesta a giberelinas (GA), siendo la principal fuente de energía hasta que la plántula emergente es fotosintéticamente activa. Ambas fases del desarrollo de la semilla, están reguladas por una red de factores de transcripción (TF) que unen motivos conservados en cis- en los promotores de sus genes diana. Los TFs son proteínas que han desempeñado un papel central en la evolución y en el proceso de domesticación, siendo uno de los principales mecanismos de regulación génica; en torno al 7% de los genes de plantas codifican TFs. Atendiendo al motivo de unión a DNA, éstos, se han clasificado en familias. La familia DOF (DNA binding with One Finger) participa en procesos vitales exclusivos de plantas superiores y sus ancestros cercanos (algas, musgos y helechos). En las semillas de las Triticeae (subfamilia Pooideae), se han identificado varias proteínas DOF que desempeñan un papel fundamental en la regulación de la expresión génica. Brachypodium distachyon es la primera especie de la subfamilia Pooideae cuyo genoma (272 Mbp) ha sido secuenciado. Su pequeño tamaño, ciclo de vida corto, y la posibilidad de ser transformado por Agrobacterium tumefaciens (plásmido Ti), hacen que sea el sistema modelo para el estudio de cereales de la tribu Triticeae con gran importancia agronómica mundial, como son el trigo y la cebada. En este trabajo, se han identificado 27 genes Dof en el genoma de B. distachyon y se han establecido las relaciones evolutivas entre estos genes Dof y los de cebada (subfamilia Pooideae) y de arroz (subfamilia Oryzoideae), construyendo un árbol filogenético en base al alineamiento múltiple del dominio DOF. La cebada contiene 26 genes Dof y en arroz se han anotado 30. El análisis filogenético establece cuatro grupos de genes ortólogos (MCOGs: Major Clusters of Orthologous Genes), que están validados por motivos conservados adicionales, además del dominio DOF, entre las secuencias de las proteínas de un mismo MCOG. El estudio global de expresión en diferentes órganos establece un grupo de nueve genes BdDof expresados abundantemente y/o preferencialmente en semillas. El estudio detallado de expresión de estos genes durante la maduración y germinación muestra que BdDof24, ortólogo putativo a BPBF-HvDOF24 de cebada, es el gen más abundante en las semillas en germinación de B. distachyon. La regulación transcripcional de los genes que codifican hidrolasas en la aleurona de las semillas de cereales durante la post‐germinación ha puesto de manifiesto la existencia en sus promotores de un motivo tripartito en cis- conservado GARC (GA-Responsive Complex), que unen TFs de la clase MYB-R2R3, DOF y MYBR1-SHAQKYF. En esta tesis, se ha caracterizado el gen BdCathB de Brachypodium que codifica una proteasa tipo catepsina B y es ortólogo a los genes Al21 de trigo y HvCathB de cebada, así como los TFs responsables de su regulación transcripcional BdDOF24 y BdGAMYB (ortólogo a HvGAMYB). El análisis in silico del promotor BdCathB ha identificado un motivo GARC conservado, en posición y secuencia, con sus ortólogos en trigo y cebada. La expresión de BdCathB se induce durante la germinación, así como la de los genes BdDof24 y BdGamyb. Además, los TFs BdDOF24 y BdGAMYB interaccionan en el sistema de dos híbridos de levadura e in planta en experimentos de complementación bimolecular fluorescente. En capas de aleurona de cebada, BdGAMYB activa el promotor BdCathB, mientras que BdDOF24 lo reprime; este resultado es similar al obtenido con los TFs ortólogos de cebada BPBF-HvDOF24 y HvGAMYB. Sin embargo, cuando las células de aleurona se transforman simultáneamente con los dos TFs, BdDOF24 tiene un efecto aditivo sobre la trans-activación mediada por BdGAMYB, mientras que su ortólogo BPBF-HvDOF24 produce el efecto contrario, revirtiendo el efecto de HvGAMYB sobre el promotor BdCathB. Las diferencias entre las secuencias deducidas de las proteínas BdDOF24 y BPBF-HvDOF24 podrían explicar las funciones opuestas que desempeñan en su interacción con GAMYB. Resultados preliminares con líneas de inserción de T-DNA y de sobre-expresión estable de BdGamyb, apoyan los resultados obtenidos en expresión transitoria. Además las líneas homocigotas knock-out para el gen BdGamyb presentan alteraciones en anteras y polen y no producen semillas viables. ABSTRACT The seed is the plant organ of the spermatophytes responsible for the dispersion and survival in the course of evolution. In addition, it constitutes one of the most importan elements of human food and animal feed. The main reserves accumulated in the endosperm of cereal seeds through the maturation phase of development are starch and proteins. Its degradation by hydrolases synthetized in aleurone cells in response to GA upon germination provides energy, carbon and nitrogen to the emerging seedling before it acquires complete photosynthetic capacity. Both phases of seed development are controlled by a network of transcription factors (TFs) that interact with specific cis- elements in the promoters of their target genes. TFs are proteins that have played a central role during evolution and domestication, being one of the most important regulatory mechanisms of gene expression. Around 7% of genes in plant genomes encode TFs. Based on the DNA binding motif, TFs are classified into families. The DOF (DNA binding with One Finger) family is involved in specific processes of plants and its ancestors (algae, mosses and ferns). Several DOF proteins have been described to play important roles in the regulation of genes in seeds of the Triticeae tribe (Pooideae subfamily). Brachypodium distachyon is the first member of the Pooideae subfamily to be sequenced. Its small size and compact structured genome (272 Mbp), the short life cycle, small plant size and the possibility of being transformed with Agrobacterium tumefaciens (Ti-plasmid) make Brachypodium the model system for comparative studies within cereals of the Triticeae tribe that have big economic value such as wheat and barley. In this study, 27 Dof genes have been identified in the genome of B. distachyon and the evolutionary relationships among these Dof genes and those frome barley (Pooideae subfamily) and those from rice (Oryzoideae subfamily) have been established by building a phylogenetic tree based on the multiple alignment of the DOF DNA binding domains. The barley genome (Hordeum vulgare) contains 26 Dof genes and in rice (Oryza sativa) 30 genes have been annotated. The phylogenetic analysis establishes four Major Clusters of Orthologous Genes (MCOGs) that are supported by additional conserved motives out of the DOF domain, between proteins of the same MCOG. The global expression study of BdDof genes in different organs and tissues classifies BdDof genes into two groups; nine of the 27 BdDof genes are abundantly or preferentially expressed in seeds. A more detailed expression analysis of these genes during seed maturation and germination shows that BdDof24, orholog to barley BPBF-HvDof24, is the most abundantly expressed gene in germinating seeds. Transcriptional regulation studies of genes that encode hydrolases in aleurone cells during post-germination of cereal seeds, have identified in their promoters a tripartite conserved cis- motif GARC (GA-Responsive Complex) that binds TFs of the MYB-R2R3, DOF and MYBR1-SHAQKYF families. In this thesis, the characterization of the BdCathB gene, encoding a Cathepsin B-like protease and that is ortholog to the wheat Al21 and the barley HvCathB genes, has been done and its transcriptional regulation by the TFs BdDOF24 and BdGAMYB (ortholog to HvGAMYB) studied. The in silico analysis of the BdCathB promoter sequence has identified a GARC motif. BdCathB expression is induced upon germination, as well as, those of BdDof24 and BdGamyb genes. Moreover, BdDOF24 and BdGAMYB interact in yeast (Yeast 2 Hybrid System, Y2HS) and in planta (Bimolecular Fluorecence Complementation, BiFC). In transient assays in aleurone cells, BdGAMYB activates the BdCathB promoter, whereas BdDOF24 is a transcriptional repressor, this result is similar to that obtained with the barley orthologous genes BPBF-HvDOF24 and HvGAMYB. However, when aleurone cells are simultaneously transformed with both TFs, BdDOF24 has an additive effect to the trans-activation mediated by BdGAMYB, while its ortholog BPBF-HvDOF24 produces an opposite effect by reducing the HvGAMYB activation of the BdCathB promoter. The differences among the deduced protein sequences between BdDOF24 and BPBF-HvDOF24 could explain their opposite functions in the interaction with GAMYB protein. Preliminary results of T-DNA insertion (K.O.) and stable over-expression lines of BdGamyb support the data obtained in transient expression assays. In addition, the BdGamyb homozygous T-DNA insertion (K.O.) lines have anther and pollen alterations and they do not produce viable seeds.

Arabidopsis thaliana DOF6 negatively affects germination in non-after ripened seeds and interacts with TCP14

Seed dormancy prevents seeds from germinating under environmental conditions unfavourable for plant growth and development and constitutes an evolutionary advantage. Dry storage, also known as after-ripening, gradually decreases seed dormancy by mechanisms not well understood. An Arabidopsis thaliana DOF transcription factor gene (DOF6) affecting seed germination has been characterized. The transcript levels of this gene accumulate in dry seeds and decay gradually during after-ripening and also upon seed imbibition. While constitutive over-expression of DOF6 produced aberrant growth and sterility in the plant, its over-expression induced upon seed imbibition triggered delayed germination, abscisic acid (ABA)-hypersensitive phenotypes and increased expression of the ABA biosynthetic gene ABA1 and ABA-related stress genes. Wild-type germination and gene expression were gradually restored during seed after-ripening, despite of DOF6-induced over-expression. DOF6 was found to interact in a yeast two-hybrid system andin planta with TCP14, a previously described positive regulator of seed germination. The expression of ABA1 and ABA-related stress genes was also enhanced in tcp14 knock-out mutants. Taken together, these results indicate that DOF6 negatively affects seed germination and opposes TCP14 function in the regulation of a specific set of ABA-related genes

Transcription factors of the bzip factors of the BZIP family affecting arabidopsis thaliana germination sensu stricto

During seed germination, the endosperm cell walls (CWs) suffer an important weakening process mainly driven by hydrolytic enzymes, such are endo-?- mannanases (MAN; EC. 3.2.1.78) that catalyze the cleavage of ?1?4 bonds in the mannan-polymers. In Arabidopsis thaliana seeds, endo-?-mannanase activity increases during seed imbibition, decreasing after radicle emergence1. AtMAN7 is the most highly expressed MAN gene in seeds upon germination and their transcripts are restricted to the micropylar endosperm and to the radicle tip just before radicle emergence. Mutants with a T-DNA insertion in this gene (K.O. MAN7) have a slower germination rate than the wild type (t50=34 h versus t50=25 h). To gain insight into the transcriptional regulation of the AtMAN7 gene, a bioinformatic search for conserved non-coding cis-elements (phylogenetic shadowing) within the Brassicaceae orthologous MAN7 gene promoters has been done and these conserved motives have been used as baits to look for their interacting transcription factors (TFs), using as a prey an arrayed yeast library of circa 1,200 TFs from A. thaliana. The basic leucine zipper AtbZIP44, but not its closely related ortholog AtbZIP11, has been thus identified and its regulatory function upon AtMAN7 during seed germination validated by different molecular and physiological techniques, such are RT-qPCR analyses, mRNA Fluorescence in situ Hybridization (FISH) experiments, and by the establishment of the germination kinetics of both over-expression (oex) lines and TDNA insertion mutants in AtbZIP44. The transcriptional combinatorial network through which AtbZIP44 regulates AtMAN7 gene expression during seed germination has been further explored through protein-protein interactions between AtbZIP44 and other bZIP members. In such a way, AtbZIP9 has been identified by yeast two-hybrid experiments and its physiological implication in the control of AtMAN7 expression similarly established.

Hybrid analysis of nonlinear circuits: DAE models with indices zero and one

We extend in this paper some previous results concerning the differential-algebraic index of hybrid models of electrical and electronic circuits. Specifically, we present a comprehensive index characterization which holds without passivity requirements, in contrast to previous approaches, and which applies to nonlinear circuits composed of uncoupled, one-port devices. The index conditions, which are stated in terms of the forest structure of certain digraph minors, do not depend on the specific tree chosen in the formulation of the hybrid equations. Additionally, we show how to include memristors in hybrid circuit models; in this direction, we extend the index analysis to circuits including active memristors, which have been recently used in the design of nonlinear oscillators and chaotic circuits. We also discuss the extension of these results to circuits with controlled sources, making our framework of interest in the analysis of circuits with transistors, amplifiers, and other multiterminal devices.

An Efficient Hybrid GO-PWS Algorithm to Analyze Conformal Serrated-Edge Reflectors for Millimeter-Wave Compact Range

A method to analyze parabolic reflectors with arbitrary piecewise rim is presented in this communication. This kind of reflectors, when operating as collimators in compact range facilities, needs to be large in terms of wavelength. Their analysis is very inefficient, when it is carried out with fullwave/MoM techniques, and it is not very appropriate for designing with PO techniques. Also, fast GO formulations do not offer enough accuracy to reach performance results. The proposed algorithm is based on a GO-PWS hybrid scheme, using analytical as well as non-analytical formulations. On one side, an analytical treatment of the polygonal rim reflectors is carried out. On the other side, non-analytical calculi are based on efficient operations, such as M2 order 2-dimensional FFT. A combination of these two techniques in the algorithm ensures real ad-hoc design capabilities, reached through analysis speedup. The purpose of the algorithm is to obtain an optimal conformal serrated-edge reflector design through the analysis of the field quality within the quiet zone that it is able to generate in its forward half space.

Two way clustering of Microarray Data using a Hybrid Approach

The Microarray technique is rather powerful, as it allows to test up thousands of genes at a time, but this produces an overwhelming set of data files containing huge amounts of data, which is quite difficult to pre-process, separate, classify and correlate for interesting conclusions to be extracted. Modern machine learning, data mining and clustering techniques based on information theory, are needed to read and interpret the information contents buried in those large data sets. Independent Component Analysis method can be used to correct the data affected by corruption processes or to filter the uncorrectable one and then clustering methods can group similar genes or classify samples. In this paper a hybrid approach is used to obtain a two way unsupervised clustering for a corrected microarray data.

Experience with PV-diesel hybrid village power systems in southern Morocco

In October 2002, under the auspices of Spanish Cooperation, a pilot electrification project put into operation two centralised PV-diesel hybrid systems in two different Moroccan villages. These systems currently provide a full-time energy service and supply electricity to more than a hundred of families, six community buildings, street lighting and one running water system. The appearance of the electricity service is very similar to an urban one: one phase AC supply (230V/50Hz) distributed up to each dwelling using a low-voltage mini-grid, which has been designed to be fully compatible with a future arrival of the utility grid. The management of this electricity service is based on a “fee-for-service” scheme agreed between a local NGO, partner of the project, and electricity associations created in each village, which are in charge of, among other tasks, recording the daily energy production of systems and the monthly energy consumption of each house. This register of data allows a systematic evaluation of both the system performance and the energy consumption of users. Now, after four years of operation, this paper presents the experience of this pilot electrification project and draws lessons that can be useful for designing, managing and sizing this type of small village PV-hybrid system

A semantic scheduler architecture for federated hybrid clouds

Cloud computing is one the most relevant computing paradigms available nowadays. Its adoption has increased during last years due to the large investment and research from business enterprises and academia institutions. Among all the services cloud providers usually offer, Infrastructure as a Service has reached its momentum for solving HPC problems in a more dynamic way without the need of expensive investments. The integration of a large number of providers is a major goal as it enables the improvement of the quality of the selected resources in terms of pricing, speed, redundancy, etc. In this paper, we propose a system architecture, based on semantic solutions, to build an interoperable scheduler for federated clouds that works with several IaaS (Infrastructure as a Service) providers in a uniform way. Based on this architecture we implement a proof-of-concept prototype and test it with two different cloud solutions to provide some experimental results about the viability of our approach.

Social and content hybrid image recommender system for mobile social networks

One of the advantages of social networks is the possibility to socialize and personalize the content created or shared by the users. In mobile social networks, where the devices have limited capabilities in terms of screen size and computing power, Multimedia Recommender Systems help to present the most relevant content to the users, depending on their tastes, relationships and profile. Previous recommender systems are not able to cope with the uncertainty of automated tagging and are knowledge domain dependant. In addition, the instantiation of a recommender in this domain should cope with problems arising from the collaborative filtering inherent nature (cold start, banana problem, large number of users to run, etc.). The solution presented in this paper addresses the abovementioned problems by proposing a hybrid image recommender system, which combines collaborative filtering (social techniques) with content-based techniques, leaving the user the liberty to give these processes a personal weight. It takes into account aesthetics and the formal characteristics of the images to overcome the problems of current techniques, improving the performance of existing systems to create a mobile social networks recommender with a high degree of adaptation to any kind of user.

Epitaxial self-alignment: A new route to hybrid active plasmonic nanostructures

Concepts of lateral ordering of epitaxial semiconductor quantum dots (QDs) are for the first time transferred to hybrid nanostructures for active plasmonics. We review our recent research on the self-alignment of epitaxial nanocrystals of In and Ag on ordered one-dimensional In(Ga)As QD arrays and isolated QDs by molecular beam epitaxy. By changing the growth conditions the size and density of the metal nanocrystals are easily controlled and the surface plasmon resonance wavelength is tuned over a wide range in order to match the emission wavelength of the QDs. Photoluminescence measurements reveal large enhancement of the emitted light intensity due to plasmon enhanced emission and absorption down to the single QD level.

CPV hybrid system in ISFOC building, first results

PV Off-Grid systems have demonstrated to be a good solution for the electrification of remote areas [1]. A hybrid system is one kind of these systems. The principal characteristic is that it uses PV as the main generator and has a backup power supply, like a diesel generator, for instance, that is used when the CPV generation is not enough to meet demand. To study the use of CPV in these systems, ISFOC has installed a demonstration hybrid system at its headquarters. This hybrid system uses CPV technology as main generator and the utility grid as the backup generator. A group of batteries have been mounted as well to store the remaining energy from the CPV generator when nedeed. The energy flows are managed by a SMA system based on Sunny Island inverters and a Multicluster-Box (figure 1). The Load is the air-conditioning system of the building, as it has a consumption profile higher than the CPV generator and can be controlled by software [2]. The first results of this system, as well as the first chances of improvement, as the need of a bigger CPV generator and a better management of the energy stored in the batteries, are presented in this paper.

Hybrid behavioral-analytical loss model for a high frequency and low load DC/DC buck converter

This work presents a behavioral-analytical hybrid loss model for a buck converter. The model has been designed for a wide operating frequency range up to 4MHz and a low power range (below 20W). It is focused on the switching losses obtained in the power MOSFETs. Main advantages of the model are the fast calculation time (below 8.5 seconds) and a good accuracy, which makes this model suitable for the optimization process of the losses in the design of a converter. It has been validated by simulation and experimentally with one GaN power transistor and three Si MOSFETs. Results show good agreement between measurements and the model

Envelope Amplifier Based on a Hybrid Series Converter With The Slow-envelope Technique

High frequency dc-dc switching converters are used as envelope amplifiers in RF transmitters. The dc-dc converter should operate at very high frequency to track an envelope in the MHz range to supply the power amplifier. One of the circuits suitable for this application is a hybrid topology composed of a switched converter and a linear regulator in series that work together to adjust the output voltage to track the envelope with accuracy. This topology can take advantage of the reduced slew-rate technique where switching dc-dc converter provides the RF envelope with limited slew rate in order to avoid high switching frequency and high power losses, while the linear regulator performs fine adjustment in order to obtain the exact replica of the RF envelope. The combination of this control technique with this topology is proposed in this paper. Envelopes with different bandwidth will be considered to optimize the efficiency of the dc-dc converter. The calculations and experiments have been done to track a 2MHz envelope in the range 0-12V for an EER RF transmitter.

Hybrid incremental modeling based on least squares and fuzzy K-NN for monitoring tool wear in turning processes

There is now an emerging need for an efficient modeling strategy to develop a new generation of monitoring systems. One method of approaching the modeling of complex processes is to obtain a global model. It should be able to capture the basic or general behavior of the system, by means of a linear or quadratic regression, and then superimpose a local model on it that can capture the localized nonlinearities of the system. In this paper, a novel method based on a hybrid incremental modeling approach is designed and applied for tool wear detection in turning processes. It involves a two-step iterative process that combines a global model with a local model to take advantage of their underlying, complementary capacities. Thus, the first step constructs a global model using a least squares regression. A local model using the fuzzy k-nearest-neighbors smoothing algorithm is obtained in the second step. A comparative study then demonstrates that the hybrid incremental model provides better error-based performance indices for detecting tool wear than a transductive neurofuzzy model and an inductive neurofuzzy model.