Regeneration of plants from somatic embryos of Verticillium dahliae-resistant wild olive genotypes

Regeneration capacity, via somatic embryogenesis, of several wild olive genotypes differing in their response to Verticillium wilt (resistant genotypes Stop Vert, Out Vert, Ac4 and Ac 18 and the susceptible Ac 15) has been evaluated. To induce somatic embryogenesis, methodologies previously used in cultivated (high ratio cytokinin/auxin) or wild olive (low ratio cytokinin/auxin) were used. Obtained results revealed the importance of genotype, explant type, mineral formulation and hormonal balance in the induction process, ca. use of apical buds obtained from micropropagated shoots following the methodology of Mazri et al. (2013) in cultivated olive (4 days in liquid medium MS ½, 30 µM TDZ – 0.54 µM ANA, 8 weeks in basal medium MS ½, followed by subculturing in ECO basal medium supplemented with 0.5 µM 2iP, 0.44 µM BA and 0.25 µM IBA) was adequate to obtain somatic embryos in 2 genotypes, Stop Vert and Ac18, but no embryogenic response was observed in the other three. An analysis of genetic stability on Stop Vert, using SSR and RAPDs markers, was carried out in embryogenic callus, plants regenerated form this callus and micropropagated shoots in comparison with the mother plant. Polymorphism was only observed in the banding pattern generated by RAPDs in one of the 10 callus samples evaluated, resulting in a variation rate of 0.07%. This is the first time in which plants have been regenerated via somatic embryogenesis in wild olive.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech. Research project: Junta de Andalucía P11-AGR799

Somatic embryogenesis in explants of adult wild olive trees

In this investigation, somatic embryogenesis in explants of adult wild olive trees, using the protocol developed by Mazri et al. (2013, Sci. Hort. 159: 88-95), was induced. Four genotypes differring in the level of resistance to the fungal pathogen Verticillium dahliae were used: Ac18, StopVert and OutVert (symptomless resistant genotypes, Jiménez-Fernández et al. 2015, Plant Pathology, in press) and Ac15 (susceptible genotype, Jiménez-Díaz, IAS-CSIC, Córdoba, personal communication). Three types of explants from actively growing cultures were used: first pair of leaves, petioles and shoot apex. Firstly, all explants were cultured on a liquid induction medium with MS mineral elements at 0.5X and 30 μM TDZ-0.5 μM NAA, for 4 days at 80rpm. Afterwards, explants were transferred to basal MS with 0.5X macroelements, for 8 weeks. Finally, calli were cultured on expression ECO medium supplemented with 0.25 μM IBA, 0.5 μM 2ip and 0.44 μM BA, for several subcultures. Cultures were incubated under darkness at 25 degrees. Embryogenic calli were observed on shoot apex (StopVert) or leaf primordia (Ac18) explants.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

Packaging Process Optimization in MultiheadWeighers with Double-Layered Upright and Diagonal Systems

[EN] In multihead weighers, packaging processes seek to find the best combination of passage hoppers whose product content provides a total package weight as close as possible to its (nominal) label weight. The weighing hoppers arranged in these machines dispense the product quantity that each package contains through computer algorithms designed and executed for this purpose. For its part, in the packaging process for double-layered multihead weighers, all hoppers are arranged in two levels. The first layer comprises a group of weighing hoppers, and the second comprises a set of booster hoppers placed uprightly or diagonally to each weighing hopper based on design of the machine. In both processes, the initial machine configuration is the same; however, the hopper selection algorithm works differently. This paper proposes a new packaging process optimization algorithm for double-layer upright and diagonal machines, wherein the hopper subset combined has previously been defined, and the packaging weight is expressed as actual values. As part of its validation, product filling strategies were implemented for weighing hoppers to assess the algorithm in different scenarios. Results from the process performance metrics prove that the new algorithm improves processes by reducing variability. In addition, results reveal that some machine configurations were also able to improve their operation.We express our gratitude for the support from Universidad Simon Bolivar, and Universitat Politecnica de Valencia.Garcia-Jimenez, R.; García-Díaz, JC.; Pulido-Rojano, ADJ. (2021). Packaging Process Optimization in MultiheadWeighers with Double-Layered Upright and Diagonal Systems. Mathematics. 9(9):1-20. https://doi.org/10.3390/math9091039S1209

Regeneración de plantas, vía embriogénesis somática, a partir de material adulto de olivo silvestre

La embriogénesis somática es una herramienta poderosa para la clonación de genotipos de interés. En algunas especies como el olivo, esta técnica se ve limitada por las dificultades que presenta la regeneración a partir de material adulto. En este trabajo, se ha inducido embriogénesis somática a partir de material adulto de Olea europaea var. sylvestris siguiendo el protocolo desarrollado por Mazri et al. (Scient. Hort. 159: 88-95, 2013) para el cultivar de olivo Dahbia. Se emplearon 4 genotipos con distinto nivel de resistencia al patógeno fúngico Verticillium dahliae: AC18, StopVert and Outvert (genotipos resistentes) y AC15 (genotipo susceptible) (Jiménez-Díaz, IAS-CSIC, Córdoba, comunicación personal). Inicialmente, los explantos se cultivaron en oscuridad en medio líquido de inducción conteniendo la formulación mineral MS con los macroelementos a la mitad, y un suplemento de 30 µM TDZ-0.5 µM NAA durante 4 días, a 80 rpm; posteriormente, fueron transferidos al mismo medio sin reguladores del crecimiento. A las 8 semanas, el callo que proliferó se cultivó en medio de expresión ECO suplementado con 0.25 µM IBA, 0.5 µM 2iP y 0.44 µM BA, durante varios subcultivos. Sólo los explantos de yema apical del genotipo Stopvert formaron callo embriogénico con una frecuencia del 5%. Tras la multiplicación de este callo, una parte del mismo se transfirió a medio de maduración ECO con membranas de acetato de celulosa para la maduración de embriones. Los embriones maduros mostraron un porcentaje de germinación del 35%, lo que ha permitido la recuperación de plantas. La estabilidad genética del material obtenido se analizó mediante marcadores microsatélites. Se compararon plantas regeneradas a partir de embriones somáticos con el callo embriogénico del que procedían, plantas iniciadas a partir de yemas laterales y mantenidas in vitro mediante proliferación de axilares y la planta donante.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tec

Generation of organotypic multicellular spheres by magnetic levitation : model for the study of human hematopoietic stem cells microenvironment

Q4Q3Background and Objective: The characteristics of human hematopoietic stem cells are conditioned by the microenvironment of the bone marrow, where they interact with other cell populations, such as mesenchymal stem cells and endothelial cells; however, the study of this microenvironment is complex. The objective of this work was to develop a 3D culture system by magnetic levitation that imitates the microenvironment of human HSC. Methods and Results: Human bone marrow-mesenchymal stem cells, umbilical cord blood-hematopoietic stem cells and a non-tumoral endothelial cell line (CC2811, LonzaⓇ) were used to develop organotypic multicellular spheres by the magnetic levitation method. We obtained viable structures with an average sphericity index greater than 0.6, an average volume of 0.5 mm3 and a percentage of aggregation greater than 70%. Histological studies of the organotypic multicellular spheres used hematoxylin and eosin stains, and an evaluation of vimentin expression by means of immunohistochemistry demonstrated an organized internal structure without picnotic cells and a high expression of vimentin. The functional capacity of human hematopoietic stem cells after organotypic multicellular spheres culture was evaluated by multipotency tests, and it was demonstrated that 3D structures without exogenous Flt3L are autonomous in the maintenance of multipotency of human hematopoietic stem cells. Conclusions: We developed organotypic multicellular spheres from normal human cells that mimic the microenvironment of the human hematopoietic stem cells. These structures are the prototype for the development of complex organoids that allow the further study of the biology of normal human stem cells and their potential in regenerative medicine.https://orcid.org/0000-0002-9152-5552https://orcid.org/0000-0003-3075-9854https://orcid.org/0000-0002-0084-0339https://orcid.org/0000-0003-1881-9367N/

The AOLI low-order non-linear curvature wavefront sensor: a method for high sensitivity wavefront reconstruction

The Adaptive Optics Lucky Imager (AOLI) is a new instrument under development to demonstrate near diffraction limited imaging in the visible on large ground-based telescopes. We present the adaptive optics system being designed for the instrument comprising a large stroke deformable mirror, fixed component non-linear curvature wavefront sensor and photon-counting EMCCD detectors. We describe the optical design of the wavefront sensor where two photoncounting CCDs provide a total of four reference images. Simulations of the optical characteristics of the system are discussed, with their relevance to low and high order AO systems. The development and optimisation of high-speed wavefront reconstruction algorithms are presented. Finally we discuss the results of simulations to demonstrate the sensitivity of the system.Comment: 10 pages. To be published in Proc SPIE 8447: Adaptive Optics Systems II

Transformación de olivo con el gen AtNPR1 para inducir tolerancia a patógenos fúngicos.

El gen NPR1 codifica un componente esencial de la respuesta SAR mediada por ácido salicílico (AS). Tras la infección por el patógeno, la acumulación de AS libera los monómeros NPR1 en el citoplasma, los cuáles son translocados al núcleo activando la expresión de genes relacionados con la patogénesis (PR). La sobreexpresión del gen NPR1 de Arabidopsis thaliana ha incrementado la resistencia a hongos, bacterias y virus, en distintas especies. El objetivo de esta investigación fue sobreexpresar este gen en olivo con objeto de evaluar su efecto en la tolerancia a dos hongos de suelo, el hemibiotrofo Verticillium dahliae (Vd), una de las mayores amenazas del cultivo y el necrotrofo Rosellinia necatrix, un patógeno emergente en nuevas plantaciones. Se obtuvieron 3 líneas transgénicas, a partir de una línea embriogénica derivada de semilla del cv. Picual. Las líneas mostraron diferencias en el nivel de expresión del transgen en hoja, aunque estas diferencias no afectaron a los niveles de actividad endoquitinasa basal, similar a la de plantas control. La respuesta a Vd varió con el patotipo; así, todas las plantas murieron 50 días tras su inoculación con la cepa defoliante (D) V-138. Por otra parte, la respuesta a patotipos no defoliantes (ND) también fue variable, en función de la raza; tras la inoculación con la cepa V1242 (ND, raza 2), los síntomas aparecieron transcurridos 44-55 días, siendo la línea NPR1-780, con mayor expresión del transgen, la que mostró menor índice de severidad de la enfermedad. Esta línea también mostró un comportamiento superior al control tras la inoculación con la cepa V1558 (ND, raza 1), aunque las diferencias no fueron tan acusadas. En la respuesta a R. necatrix, las líneas transgénicas mostraron un ligero retraso en el desarrollo de la enfermedad con valores AUDPC entre 7-15% inferiores al control.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

CD69 expression on regulatory T cells protects from immune damage after myocardial infarction.

Increasing evidences advocate for an important function of T cells in controlling immune homeostasis and pathogenesis after myocardial infarction (MI), although the underlying molecular mechanisms remain elusive. In this study, a broad analysis of immune markers in 283 patients revealed a significant CD69 overexpression on Treg cells after MI. Our results in mice showed that CD69 expression on Treg cells increased survival after left-anterior-descending coronary artery (LAD)-ligation. Cd69-/- mice developed strong IL-17+ γδT cell responses after ischemia that increased myocardial inflammation and, consequently, worsened cardiac function. CD69+ Treg cells, by induction of AhR-dependent CD39 ectonucleotidase activity, induced apoptosis and decreased IL-17A production in γδT cells. Adoptive transfer of CD69+ Treg cells to Cd69-/- mice after LAD-ligation reduced IL-17+ γδT cell recruitment, thus increasing survival. Consistently, clinical data from two independent cohorts of patients indicated that increased CD69 expression in peripheral blood cells after acute MI was associated with a lower risk of re-hospitalization for heart failure (HF) after 2.5 years of follow-up. This result remained significant after adjustment for age, sex and traditional cardiac damage biomarkers. Our data highlight CD69 expression on Treg cells as a potential prognostic factor and a therapeutic option to prevent HF after MI.This study was supported by competitive grants from the Ministerio de Ciencia e Innovación (MCIN), through the Carlos III Institute of Health (ISCIII)-Fondo de Investigación Sanitaria (PI22/01759) to P.M.; RTI2018-094727-B-100 to J. M-G; Comunidad de Madrid grants S2017/BMD-3671-INFLAMUNE-CM to P.M. and FSM.; Fundació La Marató TV3 (20152330 31) to J.M-G and F.S-M.; Ministerio de Ciencia e Innovación (MCIN) RTI2018-099357-B-I00, and CIBERFES (CB16/10/00282), Human Frontier Science Program (grant RGP0016/2018), and Leducq Transatlantic Networks (17CVD04) to JAE. AC is supported by Marie Skłodowska- Curie grant (agreement No. 713673). R.B-D. is supported by Formación de Profesorado Universitario (FPU16/02780) program from the Spanish Ministry of Education, Culture and Sports. The CNIC is supported by the ISCIII, the MCIN and the Pro CNIC Foundation, and is a Severo Ochoa Center of Excellence (SEV-2015-0505).S

The Fourteenth Data Release of the Sloan Digital Sky Survey: First Spectroscopic Data from the extended Baryon Oscillation Spectroscopic Survey and from the second phase of the Apache Point Observatory Galactic Evolution Experiment

The fourth generation of the Sloan Digital Sky Survey (SDSS-IV) has been in operation since July 2014. This paper describes the second data release from this phase, and the fourteenth from SDSS overall (making this, Data Release Fourteen or DR14). This release makes public data taken by SDSS-IV in its first two years of operation (July 2014-2016). Like all previous SDSS releases, DR14 is cumulative, including the most recent reductions and calibrations of all data taken by SDSS since the first phase began operations in 2000. New in DR14 is the first public release of data from the extended Baryon Oscillation Spectroscopic Survey (eBOSS); the first data from the second phase of the Apache Point Observatory (APO) Galactic Evolution Experiment (APOGEE-2), including stellar parameter estimates from an innovative data driven machine learning algorithm known as "The Cannon"; and almost twice as many data cubes from the Mapping Nearby Galaxies at APO (MaNGA) survey as were in the previous release (N = 2812 in total). This paper describes the location and format of the publicly available data from SDSS-IV surveys. We provide references to the important technical papers describing how these data have been taken (both targeting and observation details) and processed for scientific use. The SDSS website (www.sdss.org) has been updated for this release, and provides links to data downloads, as well as tutorials and examples of data use. SDSS-IV is planning to continue to collect astronomical data until 2020, and will be followed by SDSS-V.Comment: SDSS-IV collaboration alphabetical author data release paper. DR14 happened on 31st July 2017. 19 pages, 5 figures. Accepted by ApJS on 28th Nov 2017 (this is the "post-print" and "post-proofs" version; minor corrections only from v1, and most of errors found in proofs corrected