Response to fungal exudates of the rhizosphere isolate Pseudomonas sp. UMAF110 involves a GGDEF/EAL domain-containing protein

Pseudomonas sp. UMAF110, isolated from rhizosphere soil in Spain, display in vitro antagonism towards the pythopathogenic fungus Rosellinia necatrix and is able grow in fungal exudates (BM-RE medium). A transposon mutant library of this strain was constructed and several mutants were selected by their reduced competitiveness in BM-RE medium. Pseudomonas sp. UMAF110-G3, which contains the transposon into a gene encoding a putative REC/PAS/GGDEF/EAL protein, was selected for further characterization. Blastn searches using the sequence of the gene interrupted by the transposon in UMAF110-G3, here called cmpA (c-di-GMP Metabolizing Protein), yielded a single positive hit (98% cover, 78% identity) with a gene from a terpene-degrading Pseudomonas sp. strain isolated from soil. Context analysis of the cmpA gene in Pseudomonas sp. UMAF110 showed that this gene is located downstream from several genes involved in flagellar motility/chemotaxis. RT-PCR experiments further confirmed that cmpA form a transcriptional unit with the che gene cluster. Expression analysis of cmpA by qRT-PCR clearly showed upregulation of this gene after transfer of Pseudomonas sp. UMAF110 cells to BM-RE medium, suggesting a role for this operon in response to fungal exudates. Deletion of cmpA in Pseudomonas sp. UMAF110 did not affect the ability of the strain to form biofilms under the conditions tested. However, overexpression of wild type CmpA in Pseudomonas putida KT2440 negatively regulated biofilm formation in this strain. Together, these results suggest that CmpA could be involved in signal transduction pathways regulating flagellar motility/chemotaxis in response to fungal exudates.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

Transformación vía Agrobacterium tumefaciens para inducir tolerancia a la podredumbre blanca del aguacate

Comunicación presentada en el VII World Avocado Congress, celebrado en Cairns (Australia) del 5 al 9 de septiembre de 2011.[EN] One of the most important limiting factors for avocado production in Spain is the disease caused by the fungus Rosellinia necatrix . Genetic manipulation could be useful for the introduction of fungal resistance traits into this crop. A n efficient Agrobacterium - mediated transformation protocol for avocado using AGL1 Agrobacterium strain and somatic embryos as the target material has been established by our group, although embryo conversion rate into plants needs to be improved. For that reason, we are using the strawberry, another Rosellinia necatrix ́s host, as model species to test the effect of several transgenes (two of fungal origin, chit 42 chitinase and β - 1,3 - glucanase from Trichoderma harzianum , and one of plant origin, At NPR1), on inducing tolerance to this fungus. Strawberry transformation with the β - 1,3 - glucanase gene has allowed the selection of two lines, β6 and β10, with enhanced tolerance to R. necatrix while no positive results were obtained following transformation with the chit - 42 gene. In relation to the At NPR1 gene more than 30 independent transgenic lines have been obtained whose tolerance to R. necatrix is currently under evaluation. Concerning avocado transformation, more than 10 independent transgenic lines (derive d from an embryogenic line of an immature Duke7 zygotic embryo) have been obtained with At NPR1 gene. Plants have been recovered from one line and efforts are underway to recover plants from other lines following micrografting of the transgenic sprouted sho ots onto in vitro germinated seedlings.[ES] Uno de los factores limitantes de la producción de aguacate en España es la enfermedad causada por el hongo R. necatrix. La manipulación genética podría ser de utilidad para introducir caracteres de resistencia en este cultivo. Se ha establecido un sistema eficiente de transformación en aguacate usando la cepa de Agrobacterium AGL1 y células embriogénicas como diana, sin embargo, la conversión en plantas de los embriones transgénicos necesita ser mejorada. Por esta razón, estamos utilizando la fresa, otro huésped de R. necatrix, como especie modelo para testar el efecto de varios transgenes (2 de origen fúngico, la quitinasa chit-42 y la β-1,3-glucanasa de Trichoderma harzianum, y uno derivado de plantas, AtNPR1), en la inducción de tolerancia a este patógeno tras la transformación de esta especie. La transformación de fresa con el gen de β -1,3-glucanasa ha permitido la selección de dos líneas, β6 y β10, con mayor tolerancia a R. necatrix, mientras que no se han obtenido resultados positivos con el gen chit-42. En relación con el gen AtNPR1, se han obtenido más de 30 líneas transgénicas independientes, cuya tolerancia frente a R. necatrix se está evaluando en la actualidad. En relación con la transformación de aguacate, más de 10 líneas transgénicas independientes (derivadas de una línea embriogénica obtenida a partir de un embrión zigótico inmaduro del cv. Duke 7) se han obtenido con el gen AtNPR1. Se han recuperado plantas de una línea y actualmente se está intentando recuperar plantas de otras líneas mediante microinjerto de los embriones transgénicos germinados.Este trabajo se ha realizado en el marco del proyecto AGL2008 - 05453 - C02 - 01/AGR.Peer Reviewe

Two-step persulfate and Fenton oxidation of naphthenic acids in water

WILEY: "This is the peer reviewed version of the following article: Journal of Chemical Technology and Biotechnology 93 (2018): 2262-2270, which has been published in final form at http://doi.org/10.1002/jctb.5569. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Use of Self-Archived Versions."BACKGROUD: In the current study, two-step persulfate and Fenton oxidation has been investigated for the mineralization of naphthenic acids at 80 °C and initial pH ≈ 8. This pH evolves during the persulfate oxidation step towards the optimum for Fenton oxidation (≈ 3). The effects of persulfate and H2O2doses, iron concentration, duration of the persulfate oxidation step and operating temperature have been assessed. RESULTS: The combined treatment allowed up to ≈ 80% mineralization of cyclohexanoic acid using fairly low relative amounts of reagents (20 and 30% of the stoichiometric for persulfate and H2O2, respectively). For mineralization of cyclohexanoic acid, 115 and 87 kJ mol-1were obtained as representative values of the apparent activation energy for the persulfate and Fenton oxidation steps, respectively. The system was also successfully tested with other naphthenic acids, including cyclohexanebutyric acid, 2-naphthoic acid and 1,2,3,4-tetrahydro-2-naphthoic acid. Treatment of the naphthenic acids tested by this system gave rise to easily biodegradable effluents consisting mainly of short-chain organic acids. The biodegradability was confirmed by the BOD5/COD ratio and respirometric tests. CONCLUSION: The results show the potential application of this approach as a promising cost-effective solution for the treatment of naphthenic acids-bearing aqueous wastes. This approach has significant advantage compared with the single thermally-activated persulfate or Fenton oxidation, since it allows a high mineralization at reduced reagent cost upon replacing part of the persulfate by less expensive H2O2.We are grateful to the Chinese Scholarship Council (CSC) for supporting the Ph.D. program of Xiyan Xu (CSC, File No. 201308410047). Spanish MINECO is also gratefully acknowledged for the financial support through the project CTQ2013-41963-

Application of intensified Fenton oxidation to the treatment of hospital wastewater: Kinetics, ecotoxicity and disinfection

Fenton oxidation has proved to be effective for the treatment of a real hospital wastewater. The intensification of the process viz. the improvement on the efficiency of H2O2consumption, by increasing the temperature, is necessary to operate in a single step and avoid high reaction times and the need of large H2O2 and iron concentrations. This approach would allow taking advantage of the heat energy contained in the laundry stream since the washing process is performed at high temperature to assure disinfectionof the hospital textiles. In this work, temperatures within the range of 50-90 °C have been tested, which allowed an effective oxidation of the real wastewater ([COD]0 = 365 mg L-1, [Phenols]0 = 8.4 mg L-1, [total coliforms]0 = 4.16 × 106 MPN/100 mL, 5 toxicity units) with the stoichiometric amount of H2O2 (1000 mg L-1) and a relatively low iron concentration (25 mg L-1 Fe3+). Operating at 90 °C, complete disappearance of phenolic compounds and 70% and 50% reduction of COD and TOC were achieved in 1 h reaction time. Time evolution of those global parameters were appropriately fitted to pseudo-first and second order kinetic equations, providing a useful approach for design purposes. The main by-products were short-chain organic acids (oxalic and formic), being the final effluents non-toxic. Disinfection of the hospital wastewater was also achieved at all the operating temperatures tested, as confirmed by the absence of coliformsThis research has been supported by the CM through the project S2013/MAE-2716 and by the Spanish MINECO through the project CTQ2013-41963-R. M. Munoz thanks the Spanish MINECO for a Juan de la Cierva-Incorporación postdoctoral contract (IJCI-2014-19427

Exploring the Use of Fruit Callus Culture as a Model System to Study Color Development and Cell Wall Remodeling during Strawberry Fruit Ripening

Cell cultures derived from strawberry fruit at different developmental stages have been obtained to evaluate their potential use to study different aspects of strawberry ripening. Callus from leaf and cortical tissue of unripe-green, white, and mature-red strawberry fruits were induced in a medium supplemented with 11.3 µM 2,4-dichlorophenoxyacetic acid (2,4-D) under darkness. The transfer of the established callus from darkness to light induced the production of anthocyanin. The replacement of 2,4-D by abscisic acid (ABA) noticeably increased anthocyanin accumulation in green-fruit callus. Cell walls were isolated from the different fruit cell lines and from fruit receptacles at equivalent developmental stages and sequentially fractionated to obtain fractions enriched in soluble pectins, ester bound pectins, xyloglucans (XG), and matrix glycans tightly associated with cellulose microfibrils. These fractions were analyzed by cell wall carbohydrate microarrays. In fruit receptacle samples, pectins were abundant in all fractions, including those enriched in matrix glycans. The amount of pectin increased from green to white stage, and later these carbohydrates were solubilized in red fruit. Apparently, XG content was similar in white and red fruit, but the proportion of galactosylated XG increased in red fruit. Cell wall fractions from callus cultures were enriched in extensin and displayed a minor amount of pectins. Stronger signals of extensin Abs were detected in sodium carbonate fraction, suggesting that these proteins could be linked to pectins. Overall, the results obtained suggest that fruit cell lines could be used to analyze hormonal regulation of color development in strawberry but that the cell wall remodeling process associated with fruit softening might be masked by the high presence of extensin in callus cultures

Variación intra-anual de la clorofila-a y nutrientes en un sistema río-llanura de inundación hidráulicamente perturbado en la cuenca del río Grijalva

Antecedentes: La interconexión lateral en el sistema rio-llanura de inundación Maluco fue reducida parcialmente por una carretera construida sobre una barrera natural. Objetivo: Determinar si las concentraciones de clorofila-a (Cla-a) y nutrientes contrastan en diferentes niveles de agua (NA) durante un ano hidrológico en el humedal fluvial. Métodos: El NA, Cla-a y nutrientes (NO2-, NO3-, NH4+ y PT) se midieron simultáneamente seis veces en dos estaciones durante dos años. Las diferencias esperadas de las concentraciones de Cla-a y nutrientes, relacionadas a la fluctuación intra-anual del NA, fueron analizadas por medio de técnicas multivariadas. La hipótesis sostiene que no se detectaran diferencias intra anuales de Cla-a y nutrientes debido a la reducción de la interconexión lateral por el efecto físico del terraplen. Resultados: Ninguna variable fue descartada con el ACP. La distribución intra-anual de las muestras resulto similar según la rutina de perfiles de similitud. Coincidentemente: 1) más del 54% de los valores de Cla-a y PT indicaron condiciones hipereutroficas durante todo el año y 2) ninguno de los valores de WL sobrepaso la altura del terraplen (4.5 msnm). Además, las concentraciones máximas de Cla-a se registraron en los WL mínimos. Conclusiones: La similitud intra-anual de la Cla-a y los nutrientes inorgánicos se puede vincular a que la llanura de inundación estuvo desconectada del rio Grijalva, ya que la inundación no sobrepaso la altura del terraplén durante todo el tiempo de muestreo. La tendencia intra-anual independiente al tiempo de la persistencia de las altas concentraciones de Cla-a y PT en el sistema rio llanura de inundación abre la oportunidad de estimar el caudal de entrada mínimo requerido para restaurar los procesos biogeoquímicos en un estudio a largo plazo.Background: The lateral connectivity in the Maluco river-floodplain system was partially reduced by the highway built on a natural barrier. Goal: To determine if the concentrations of chlorophyll-a (Chl-a) and nutrients contrast in different water levels (WL) during a hydrologic year in the Maluco river-floodplain system. Methods: The WL, Chl-a, and nutrients (NO2-, NO3-, NH4+, and TP) were simultaneously measured six times in two sites during two years. The expected differences of Chl-a and nutrients concentrations, related to the intra-annual fluctuation of the WL, were tested through multivariate techniques. The hypothesis holds that there will be no intra-annual differences of Chl-a and nutrients due to the reduction of lateral interconnection by the physical effect of the embankment. Results: No variable was discarded during PCA. The intra-annual distribution of samples was similar according to the similarity profile routine. Furthermore: 1) more than 54% of the Chl-a and TP values indicated hypereutrophic conditions throughout the year, and 2) none of the values of WL surpassed the height of the bankfull (4.5 masl). The maximum concentrations of Chl-a were registered at the minimum WL values. Conclusions: The intra-annual similarity of Chl-a and inorganic nutrients can be linked to the disconnection between the floodplain and the Grijalva river since the bankfull was not overflowed throughout the sampling time. The intra-annual trend independent at the time of persistence of the high Chl-a and TP concentrations in the river-floodplain system opens up the opportunity to estimate the minimum input flow required to biogeochemical processes in a long-term study

Optimización de la transfección de protoplastos para la edición génica en fresa

Esta investigación ha sido financiada por los fondos FEDER EU, el Ministerio de Economía y Competitividad de España (AGL2017-86531-C2-1-R), y el contrato FPI PRE2018-085509.La fresa es un fruto blando de gran importancia económica, particularmente en Andalucía. La mejora de las cualidades organolépticas del fruto y la disminución del reblandecimiento para alargar la vida postcosecha del fruto, son unos de los principales objetivos de los programas de mejora en este cultivo. El reblandecimiento del fruto es consecuencia del desmantelamiento de la pared celular, la disolución de la lámina media y la pérdida de turgencia. En fresa, el silenciamiento mediante la transformación en antisentido de genes que codifican poligalacturonasas (PG) aumenta la firmeza del fruto y la vida postcosecha (Paniagua et al., 2020). Por tanto, estos genes son excelentes dianas para la edición génica con el fin de mejorar la calidad del fruto de la fresa. La transfección de protoplastos con complejos preensamblados Cas9-sgRNA permite la producción de plantas editadas vía CRISPR/Cas9 libres de ADN foráneo, que podrían ser consideradas como no transgénicas. En esta investigación, se ha optimizado un protocolo para la transfección de protoplastos de fresa, con el objetivo final de producir plantas no transgénicas con el gen de poligalacturonasa FaPG1 mutado. Como fuente de material vegetal se utilizaron hojas de plantas de Fragaria x ananassa, cv. ‘Chandler’, micropropagadas en medio Murashige y Skoog (MS) suplementado con 2 mg/L de BA. Para la extracción de protoplastos se utilizó el protocolo descrito por Barceló et al. (2019). A las 24 h del aislamiento, los protoplastos fueron transfectados con el plásmido pHBT-sGFP(S65T)-NOS que contiene el gen marcador GFP, mediante un tratamiento con polietilenglicol (PEG), como se describe en Yoo et al. (2007). Se evaluaron, entre otras variables, el efecto de la concentración y tiempo de incubación en PEG y la concentración de ADN. Los valores más altos de protoplastos con actividad GFP a las 48 h de la transfección, entre el 15-18%, se obtuvieron tras la incubación en 20% de PEG en presencia de 5 µg de ADN.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

Track trajectories with model uncertainty using a robust differentiator

[EN] In this article, we present the Levant’s Robust Differentiator applied to robot manipulators whose objective is to follow a desired trajectory. The robots’ dynamic model is unknown. The velocity obtained using the Robust Differentiator is applied in the control structure in order to fulfill the tracking task. A comparative study is made between the Levant’s Robust Differentiator and the most-used techniques to calculate the velocity. Experimental results are presented.[ES] En este artículo se presenta el uso de un diferenciador robusto de Levant aplicado a robots manipuladores cuyo objetivo es realizar el seguimiento de una trayectoria deseada. El modelo dinámico de los robots es desconocido. La velocidad obtenida empleando el diferenciador robusto se aplica en la estructura de control con la finalidad de cumplir con la tarea de seguimiento. Se realiza un estudio comparativo entre el diferenciador robusto de Levant y las técnicas más usadas para calcular la velocidad. Son presentados resultados experimentales.Los autores agradecen a PRODEP (PROMEP) con el número de Folio BUAP–811 y los proyectos PAPIIT 116314 y 114617 por el apoyo recibido, a CONACYT por la Cátedra CONACYT–CICESE 2017 y al Laboratorio de Robótica de la Facultad de Ciencias de la Electrónica de la Benemérita Universidad Autónoma de Puebla.Sánchez-Sánchez, P.; Gutiérrez–giles, A.; Pliego–jiménez, J.; Arteaga–pérez, M. (2019). Seguimiento de trayectorias con incertidumbre del modelo usando un diferenciador robusto. Revista Iberoamericana de Automática e Informática. 16(4):423-434. https://doi.org/10.4995/riai.2019.10265SWORD423434164Alcocer, A. and Robertsson, A. and Valera, A. and Johansson, R., 2003. Force Estimation and Control in Robot Manipulators. Proceedings of 7th Symposium on Robot Control (SYROCO'03) 55-60. Wroclaw, Poland https://doi.org/10.1016/S1474-6670(17)33369-4Atassi, A. N. and Khalil, H. K., 2000. Separation results for the stabilization of nonlinear systems using di_erent high-gain observer designs. Systems and Control Letters, 39(15), 183-191. https://doi.org/10.1016/S0167-6911(99)00085-7Berghuis, H. and Nijmeijer, H., 1994. Robust control of robots via linear estimated state feedback. IEEE Transactions on Automatic Control, 39(10), 2159-2162.https://doi.org/10.1109/9.328807Calafiore, G. and Indri, M. and Bona, B., 1997. Robot dynamic calibration: Optimal excitation trajectories and experimental parameter estimation. Journal of Robotic Systems 18(2), 55-68. https://doi.org/10.1002/1097-4563(200102)18:23.3.CO;2-FCruz-Zavala, E. and Moreno, J. A. and Fridman, L. M., 2010. Diferenciador Robusto Exacto y Uniforme. Proceedings of AMCA 2010, 1-6.Dabroom, A. M. and Khalil, H. K., 1994. Numerical differentiation using high gain observers. Proceedings of of the IEEE Conference on Decision and Control, 4790-4795. https://doi.org/10.1109/CDC.1997.649776Diop, S. and Grizzle, J. and Moraal, P. and Stefanopoulou, A., 1994. Interpolation and numerical differentiation for observer design. Proceedings of the American Control Conference, 1329-1333. https://doi.org/10.1109/ACC.1994.752275Hacksel, P. J. and Salcudean, S. E., 1994. Estimation of Environment Forces and Rigid-Body Velocities using Observers. Proc. IEEE International Conference on Robotics and Automation, 931-936. San Diego, CA, USA https://doi.org/10.1109/ROBOT.1994.351233Kelly, R. and Ortega, R. and Ailon, A. and Loria, A., 1994. Global regulation of flexible joint robots using approximate differentiation. IEEE Transactions on Automatic Control 39(6), 1222-1224. https://doi.org/10.1109/9.293181Kelly, R. and Santibáñez, V., 2003. Control de Movimiento de Robots Manipuladores. Prentice-HallKhalil, H. K., 1996. Nonlinear Systems. Prentice-HallKhatib, O., 1987. A Unified Approach for Motion and Force Control of Robot Manipulators: The Operational Space Formulation. IEEE Journal of Robotics and Automation 3(1), 43-53. https://doi.org/10.1109/JRA.1987.1087068Kumar, B. and Dutta-Roy, S. C., 1988. Design of digital differentiators for low frequencies. Proceedings of the IEEE, 76(3), 287-289. https://doi.org/10.1109/5.4408Levant, A., 1998. Sliding order and sliding accuracy in sliding mode control. International Journal of Control, 58(6), 1247-1263. https://doi.org/10.1080/00207179308923053Levant, A., 1998. Robust exact diferentiation via sliding mode technique. Automatica, 34(3), 379-384. https://doi.org/10.1016/S0005-1098(97)00209-4Levant, A., 2003. Higher-order sliding modes, diferentiation and output-feedback control. International Journal of Control, 76(9), 924-941. https://doi.org/10.1080/0020717031000099029Loria, A., 2016. Observers are Unnecessary for Output-Feedback Control of Lagrangian Systems. 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La construcción de la noción de Cosmovisión Maya en Guatemala

Con el presente escrito deseamos contribuir a la reflexión del proceso de conformación histórica, política y cultural de la noción Panmaya de Cosmovisión Maya, la cual ha sido propuesta, difundida y practicada por intelectuales y activistas de los pueblos de origen maya asentados en territorio guatemalteco. Proponemos  que este proceso resulta un ejemplo de apropiación, adopción y reinterpretación de un término académico migrado a contextos comunitarios y políticos. Esbozaremos el contexto en el que surge esta noción desde donde se enuncia la Cosmovisión Maya como concepto aglutinador de los procesos de fortalecimiento, visibilización, reivindicación y revitalización de la cultura maya en Guatemala en los años posteriores al conflicto armado interno

Synthesis of Nylon 6/Modified Carbon Black Nanocomposites for Application in Uric Acid Adsorption

High uric acid levels cause different clinic conditions. One of them is hyperuricemia, which leads to kidney damage. A solution for eliminating uric acid in the blood is by hemodialysis, which is performed using nanocomposite membranes. In this work, Nylon 6 nanocomposites were synthesized with modified carbon black (MCB), which were considered candidate materials for hemodialysis membranes. The modification of carbon black was made with citric acid using the variable-frequency ultrasound method. The new MCB was characterized by Fourier transform infrared spectroscopy (FTIR), thermogravimetric analysis (TGA), X-ray diffraction (XRD), transmission electron microscopy (TEM), and dispersion tests. Nylon 6/MCB nanocomposites were processed using the ultrasound-assisted melt-extrusion method to improve the dispersion procedure of the nanoparticles. The Nylon 6/MCB nanocomposites were characterized by FTIR, TGA, and differential scanning calorimetry (DSC). These were assessed for the absorption of toxins and hemocompatibility. MBC and nanocomposites showed excellent uric acid removal (78–82%) and hemocompatibility (1.6–1.8%). These results suggest that Nylon 6/MCB nanocomposites with low loading percentages can be used on a large scale without compatibility problems with blood