Discerning the relationship between geminiviral infection and vesicle trafficking using virus induced gene silencing

Tomato yellow leaf curl disease is one of the most important threats to tomato crops worldwide. One of its causal agents, Tomato yellow leaf curl Sardinian virus (TYLCSV) is a monopartite member of the genus Begomovirus from the family Geminiviridae. Due to the few proteins encoded by their viral genome, geminiviruses rely heavily on host cellular machineries and interact with a wide range of plant proteins to complete all processes required for infection, such as viral replication, movement and suppression or evasion of plant defence mechanisms. The identification of the host proteins involved in viral infection will be an important step towards the understanding of the mechanisms underlying this process. In our laboratory, transgenic Nicotiana benthamiana plants containing a green fluorescent protein (GFP) expression cassette flanked by two direct repeats of the intergenic region of TYLCSV have been constructed (2IR plants). When these plants are infected with TYLCSV, an overexpression of the reporter gene is observed in those cells where the virus is actively replicating. These plants have been used together with virus induced gene silencing (VIGS) in an effort to identify host genes involved in the infection process using a reverse genetics approach. Using this combined technique our group has identified two genes δ-COP and ARF 1, involved in retrograde vesicle trafficking, which are essential for the infectious process. We are currently assaying genes codifying proteins involved in different pathways of the vesicle trafficking system: Sar1b, γ subunit of AP1, Sec24, SYT1 and two that encode the heavy chain of triskelion proteins. Their effect over viral infection will be presented and discussedUniversidad de Málaga. Campues de Excelencia Internacional Andalucía Tech

Identifying the function of vesicle trafficking in geminiviral infection using virus induced gene silencing

Tomato yellow leaf curl Sardinian virus (TYLCSV) is one of the causal agent of the tomato yel-low leaf curl disease, one of the most important threats to tomato crops worldwide. TYLCSV is a monopartite member of the genus Begomovirus from the family Geminiviridae. To carry out a full infection, geminiviruses need to move inside the infected cell and from one cell to an-other for which they depend on diverse cellular factors. While cell-to-cell movement has been described to occur through plasmodesmata, the way in which geminiviruses move inside the host cells is yet unknown. The identification of the host proteins involved in viral infection will be an important step to-wards the understanding of the mechanisms underlying this process. In our laboratory, trans-genic Nicotianabenthamiana plants containing a green fluorescent protein (GFP) expression cassette flanked by two direct repeats of the intergenic region of TYLCSV have been construct-ed (2IR plants). When these plants are infected with TYLCSV, an overexpression of the reporter gene is observed in those cells where the virus replicates. These plants have been used to-gether with virus induced gene silencing (VIGS) in an effort to identify host genes involved in the infection process using a reverse genetics approach. Using this combined technique our group has identified two genes δ-COP and ARF 1, involved in retrograde vesicle trafficking, which are essential for the infectious process. We are current-ly assaying genes codifying proteins involved in different pathways of the vesicle trafficking system: Sar1b, γ subunit of AP1, Sec24, SYT1 and two that encode the heavy chain of triskelion proteins. Their effect over virus infection will be presented and discussed.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

Reconocimiento y evaluación para riegos de los suelos de la comarca de Andujar (Jaén) (Escala 1:50.000)

67 páginas.-- 14 perfiles de suelos.-- Memoria presenta en el V Curso Internacional de Edafología y Biología VegetalEste estudio resume las características morfológicas y analíticas de los suelos que constituyen el área del proyecto, principalmente para evaluar su actitud parar riegos. Se ha indicado también el método de trabajo seguido para llegar a dicha evaluación. Anteriormente se ha expresado que el proyecto tiene una aptitud mediana para regadío. Debe hacerse notar que dicha aptitud obedece sólo a las posibilidades del suelo natural o en estado actual para su utilización directa, sin considerar los efectos de mejora que puedan introducirse por medio de trabajos, cuyo fin es el de disminuir o eliminar alguno de los factores limitantes. Algunos de ellos, como la textura por ejemplo, no puede ser modificados en absoluto o son de difícil corrección. Otros como el drenaje, con la realización de ciertas obras, se vería reducidos al mínimo. Dejando aparte el factor climático, para el adecuado manejo y transformación de los suelos que integras el proyecto, sería de gran utilidad el mapa de suelos, de posiciones fisiográficas y clases para riegos que acompañar al estudio.Patrocinado por la UNESCO, OEA, Instituto de Cultura Hispánica, Consejo Superior de Investigaciones Científicas y Universidad de Sevilla y GranadaPeer reviewe

Very high-energy γ-ray observations of the Crab nebula and other potential sources with the GRAAL experiment

The “γ-ray astronomy at Almeria” (GRAAL) experiment uses 63 heliostat-mirrors with a total mirror area of ≈2500 m2 from the CESA-1 field at the “Plataforma Solar de Almeria” to collect Cherenkov light from air showers. The detector is located in a central solar tower and detects photon-induced showers with an energy threshold of 250±110 GeV and an asymptotic effective detection area of about 15 000 m2. A comparison between the results of detailed Monte-Carlo simulations and data is presented. Data sets taken in the period September 1999–September 2000 in the direction of the Crab pulsar, the active galaxy 3C 454.3, the unidentified γ-ray source 3EG J1835+59 and a “pseudosource” were analyzed for high energy γ-ray emission. Evidence for a γ-ray flux from the Crab pulsar with an integral flux of 2.2±0.4 above threshold and a significance of 4.5σ in a total measuring time of 7 h and 10 min on source was found. No evidence for emission from the other sources was found. Some difficulties with the use of heliostat fields for γ-ray astronomy are pointed out. In particular the effect of field-of-view restricted to the central part of a detected air shower on the lateral distribution and timing properties of Cherenkov light are discussed. Upon restriction the spread of the timing front of proton-induced showers sharply decreases and the reconstructed direction becomes biased towards the pointing direction. This is shown to make efficient γ-hadron separation difficult

Integrating The Five Practices of Exemplary Leadership Model into Entrepreneurship Education

Entrepreneurship educators can assess their students by focusing on leadership self-efficacy dimensions that align with desirable entrepreneurship behaviors. To support this claim, we used the Student Leadership Practices Inventory (S-LPI) to survey a group of 46 undergraduate students in Mexico and 49 undergraduate students in Spain that were involved in entrepreneurship education programs. Independent samples t-tests show statistically significant differences between the two groups. We also compared the whole sample in terms of gender and found no differences. We propose that educators integrate Five Practices of Exemplary Leadership model into de design and assessment of their curricular and co-curricular entrepreneurship development programs. Specifically, the model serves educators from different countries, in this case Mexico and Spain, by identifying the leadership behaviors that their students enact. By integrating the five practices of exemplary leadership model, educators can account for variables like nation of origin and gender, and identify differences between groups

The GRAAL Project

26th International Cosmic Ray Conference Salt Lake City, Utah August 17-25,1999The GRAAL Project (Gamma Ray Astronomy at ALmeria) makes use of the CESA-1 heliostat field at the “Plataforma Solar de Almeria” (Spain) as a gamma-ray telescope with an energy threshold of about 100 GeV. Cherenkov light generated by EAS is reflected by the heliostats and collected into photomultipliers (PMTs) with nonimaging secondary optics. Each PMT collects the light reflected by 13 - 18 heliostats of 40 m2 using a Winston cone. After successful tests with two collecting cones, a more advanced setup on a dedicated platform, using four collectors and 63 heliostats (total reflecting area of about 2500 m2 ) is being installed. A description of this setup together with Monte Carlo results about its excellent capabilities in the precise determination of pulse arrival times are presented

Importance of vesicle trafficking in the establichsment of a geminiviral infection

Importance of vesicle trafficking in the establishment of a geminiviral infection P. CANA-QUIJADA1, T. ROSAS-DÍAZ2, LOZANO-DURÁN R. 2 AND E.R. BEJARANO1 1Dpto. Biología Celular, Genética y Fisiología. Área de Genética Instituto de Hortofruticultura Subtropical y Mediterránea “La Mayora” (IHSM-UMA-CSIC), Universidad de Málaga, Málaga, Spain E-mail: [email protected] 2. Shanghai Center for Plant Stress Biology (PSC), Shanghai Institutes of Biological Sciences, Chinese Academy of Sciences, Shanghai 201602, China. INTRODUCTION Geminiviruses produces some of the most devastating diseases for agriculture worldwide. Geminiviral genomes encode only 5 to 7 proteins, forcing them to rely heavily on host cellular machineries and to interact with a high amount of host proteins in order to complete a full infection. The identification of the host proteins involved in viral infection will be an important step towards the understanding of the mechanisms underlying this process and develop new strategies to generate new sources of resistance. Previous efforts from our group have identified several genes involved in vesicle trafficking. OBJECTIVES The main aim of this work is to elucidate the role of the plant cell’s vesicle trafficking in a geminiviral infection. MATHERIALS AND METHODS Transgenic Nicotiana benthamiana plants containing a green fluorescent protein (GFP) expression cassette flanked by two direct repeats of the intergenic region of TYLCSV have been constructed (2IR plants). When these plants are infected with TYLCSV, an overexpression of the reporter gene is observed in those cells where the virus is actively replicating. 2IR plants were used in combination with virus induced gene silencing (VIGS), to identify vesicle trafficking genes involved in the infectious process. Viral replication was monitorized by GFP expression. Viral accumulation was determined using qPCR. RESULTS When silenced, four of the assessed genes reduced dramatically the viral amounts or completely abolished the infection. On the other hand, three of them had no significant effect over the infection and one of them seems to cause a slight increase in viral accumulation. CONCLUSIONS The vesicle trafficking machinery plays an essential role in geminivirus infection. Assays to determine whether the described effect is due to a lack of replication or movement of the virus inside the plant cells are in progress.Universidad de Málaga. Campues de Excelencia Internacional Andalucía Tech

Geminivirus replication protein decreases PCNA sumoylation at two acceptor sites

Geminiviruses are plant viruses with circular, single-stranded DNA (ssDNA) genomes that infect a broad range of plants causing substantial crop diseases worldwide. They replicate in nuclei of infected cells by using host DNA replication machinery and an essential protein encoded in their genome designated Rep (replication-associated protein). This multifunctional protein induces the accumulation of the host factors involved in replication and it is capable of interacting with a lot of plant proteins including PCNA (Proliferating Cell Nuclear Antigen), a processivity factor that coordinates a wide range of processes involved in maintenance, duplication and transmission of the genome, and the sumoylation enzyme that conjugates SUMO to target proteins (SUMO-conjugating enzyme- SCE). PCNA modification by SUMO, and also ubiquitin, has long been known to be of key importance for determining how DNA damage is processed by the replisome and for maintenance of overall genome integrity. In yeast, PCNA sumoylation has been associated to DNA repair involving homologous recombination (HR). Previously, we reported that Rep ectopic expression does not result in broad changes in the sumoylation pattern of plant cells, but it modifies the sumoylation state of selected host proteins. In this work, we show, using a reconstituted sumoylation system in Escherichia coli, that tomato PCNA is sumoylated at two residues, K254 and K164, and that co-expression of the Rep protein suppresses PCNA sumoylation at these lysines. Finally, we confirm that PCNA is sumoylated and that Rep also interferes with PCNA sumoylation in planta.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tec

Immune Evasion by Herpes Simplex Viruses

Infection with herpes simplex viruses type 1 (HSV-1) and type 2 (HSV-2) is extremely frequent in the human population, as well as recurrent reactivations due to lifelong infection. Infection and persistence of HSVs within healthy individuals likely results as a consequence of numerous molecular determinants evolved by these pathogens to escape both immediate and long-term host antiviral mechanisms. Indeed, HSVs harbor an arsenal of proteins that confer them stealth by negatively modulating immune function. Consequently, these viruses perpetuate within the host, altogether silently shedding onto other individuals. In this chapter, we discuss HSV determinants that interfere with cellular antiviral factors, as well as viral determinants that hamper innate and adaptive immune components intended to control such microbes. The identification of HSV evasion molecules that modulate the immune system, as well as the understanding of their mechanisms of action, should facilitate the design of novel prophylactic and therapeutic strategies to overcome infection and disease elicited by these viruses. This chapter is intended to provide an overview of the evasion mechanisms evolved by herpes simplex viruses to escape numerous host antiviral mediators

Justicia transicional: el camino para pacificar el conflicto armado en chiapas*

Han pasado 28 años del levantamientopolítico-militar encabezado por el EjércitoZapatista de Liberación Nacional (EZLN),el cual viene exigiendo al Estado mexicanoy sus instituciones la realización de losderechos de los que han sido desposeídos lospueblos originarios, en especial a la libertad,democracia, justicia, igualdad y paz. Paraalcanzar estas aspiraciones, se requiere laaplicación de medidas judiciales y políticassociales encaminadas a reparar violacionesmasivas de derechos humanos, utilizandoprocesos de justicia transicional, que ya se hanpuesto en práctica en otros países -procesosde paz o misiones de paz- para superar suscontextos de conflicto, represión e impunidad,o para tratar el posconflicto