Búsqueda de dianas en Podosphaera Xanthii para el desarrollo de nuevas fitoterapias antifúngicas

La resistencia a fungicidas en uno de los principales problemas de la agricultura y esto es especialmente patente en el caso de los oídios. En España y en las principales áreas productoras del mundo, el oídio de las cucurbitáceas es una amenaza muy grave, y Podosphaera xanthii es considerado como el principal agente causante de la enfermedad. Hasta la fecha, la aplicación de fungicidas y el uso de variedades resistentes son las principales herramientas para el control de la enfermedad. En cualquier caso, el oídio sigue imponiendo serias limitaciones en la producción agrícola, siendo necesario el desarrollo de nuevas estrategias de control. En este estudio se pretende proporcionar información sobre las bases moleculares de P. xanthii que pueda ser de utilidad para el desarrollo de nuevas herramientas de fitoprotección. Para lograr este objetivo, estamos centrando nuestra atención en un conjunto de proteínas fúngicas carentes de función, determinado en un estudio anterior. Ante la falta de homología con proteínas funcionalmente anotadas, para conocer la posible función de dichas proteínas, en primer lugar, llevamos a cabo un análisis in silico detallado de las proteínas que incluye modelado 3D, predicción de posibles ligandos e identificación de dominios funcionales. En segundo lugar, para la identificación de proteínas clave para la patogénesis de P. xanthii, silenciamos proteínas del hongo con función predicha bioinformáticamente mediante silenciamiento génico inducido por hospedador (HIGS) empleando Agrobacterium tumefaciens como vector para la expresión transitoria de construcciones de silenciamiento en células de melón, y desde ellas, al hongo diana. Finalmente, las proteínas con un fenotipo claro de silenciamiento serán seleccionadas para la caracterización de su actividad biológica. En este congreso se mostrarán los resultados más relevantes obtenidos hasta la fecha, relativos a la asimilación de azufre.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech. Este trabajo ha sido financiado por una ayuda del Programa Estatal de I+D+i Orientada a los Retos de la Sociedad (AGL2016-76216-C2-1-R), cofinanciada con fondos FEDER (UE). Los autores agredecen además ayudas del Plan Propio de Investigación de la Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

Growth performance, in vitro antioxidant properties and chemical composition of the halophyte Limonium algarvense Erben are strongly influenced by the irrigation salinity

Limonium algarvense Erben (sea lavender) is a halophyte species with potential to provide natural ingredients with in vitro antioxidant, anti-inflammatory, neuroprotective and antidiabetic properties. This study reports for the first time the 1) cultivation of sea lavender in greenhouse conditions under irrigation with freshwater (approx. 0 mM NaCl) and saline aquaculture wastewater (300 and 600 mM NaCl), and 2) the influence of the irrigation salinity on the plant performance (e.g growth, number of produced leaves and flowers), in vitro antioxidant properties [radical scavenging activity (DPPH and ABTS), ferric reducing antioxidant power (FRAP), metal chelating properties on copper (CCA) and iron (ICA)], toxicity (in vitro on three mammalian cell lines) and chemical composition (determined by LC-ESI-HRMS/MS). The freshwater-irrigated plants had better growth performance than those irrigated with saltwater. Extracts from wild plants, had the highest antioxidant activity, but those from cultivated ones kept high in vitro antioxidant properties and interesting chemical profile. The flowers' extracts of plants irrigated with 300 mM NaCl had the highest antioxidant activities against DPPH, whereas those from freshwater-irrigated plants were more active on ABTS, CCA and FRAP. Most of the extracts showed nil toxicity. The flowers' extracts displayed the highest diversity of compounds, mainly quercetin, apigenin, luteolin, naringenin and their glycoside derivatives. Moreover, their abundance varied with the irrigation salinity. These data indicate that sea lavender plants can be successfully cultivated in greenhouse conditions under fresh- and saltwater irrigation, maintaining interesting biological and chemical properties.Funding Agency Portuguese Foundation for Science and Technology Portuguese National Budget CCMAR/Multi/04326/2019 GreenVet project ALG-01-0145-FEDER-028876 XtrerneAquaCrops FA-05-2017-028 Lisboa-01-0145-FEDER-022125-RNEM-IST ID/QUI/00100/201 Portuguese Foundation for Science and Technology SFRH/BD/116604/2016 CEECIND/00425/2017info:eu-repo/semantics/publishedVersio

A crowdsourcing database for the copy-number variation of the spanish population

Background: Despite being a very common type of genetic variation, the distribution of copy-number variations (CNVs) in the population is still poorly understood. The knowledge of the genetic variability, especially at the level of the local population, is a critical factor for distinguishing pathogenic from non-pathogenic variation in the discovery of new disease variants. Results: Here, we present the SPAnish Copy Number Alterations Collaborative Server (SPACNACS), which currently contains copy number variation profiles obtained from more than 400 genomes and exomes of unrelated Spanish individuals. By means of a collaborative crowdsourcing effort whole genome and whole exome sequencing data, produced by local genomic projects and for other purposes, is continuously collected. Once checked both, the Spanish ancestry and the lack of kinship with other individuals in the SPACNACS, the CNVs are inferred for these sequences and they are used to populate the database. A web interface allows querying the database with different filters that include ICD10 upper categories. This allows discarding samples from the disease under study and obtaining pseudo-control CNV profiles from the local population. We also show here additional studies on the local impact of CNVs in some phenotypes and on pharmacogenomic variants. SPACNACS can be accessed at: http://csvs.clinbioinfosspa.es/spacnacs/. Conclusion: SPACNACS facilitates disease gene discovery by providing detailed information of the local variability of the population and exemplifies how to reuse genomic data produced for other purposes to build a local reference database.This work is supported by Grants PID2020-117979RB-I00 from the Spanish Ministry of Science and Innovation; by the Institute of Health Carlos III (project IMPaCT-Data, exp. IMP/00019, IMP/00009 and PI20/01305), co-funded by the European Union, European Regional Development Fund (ERDF, “A way to make Europe”)

Does co-infection with vector-borne pathogens play a role in clinical canine leishmaniosis?

The severity of canine leishmaniosis (CanL) due to Leishmania infantum might be affected by other vector-borne organisms that mimic its clinical signs and clinicopathological abnormalities. The aim of this study was to determine co-infections with other vector-borne pathogens based on serological and molecular techniques in dogs with clinical leishmaniosis living in Spain and to associate them with clinical signs and clinicopathological abnormalities as well as disease severity. Sixty-one dogs with clinical leishmaniosis and 16 apparently healthy dogs were tested for Rickettsia conorii, Ehrlichia canis, Anaplasma phagocytophilum and Bartonella henselae antigens by the immunofluorescence antibody test (IFAT) and for E. canis, Anaplasma spp., Hepatozoon spp., Babesia spp. and filarioid DNA by polymerase chain reaction (PCR). Among the dogs examined by IFAT, the seroprevalences were: 69% for R. conorii, 57% for E. canis, 44% for A. phagocytophilum and 37% for B. henselae ; while the prevalences found by PCR were: 8% for Ehrlichia / Anaplasma, 3% for Anaplasma platys and 1% for H. canis. No other pathogen DNA was detected. Statistical association was found between dogs with clinical leishmaniosis and seroreactivity to R. conorii antigen (Fisher's exact test: P = 0.025, OR = 4.1, 95% CI = 1-17) and A. phagocytophilum antigen (Fisher's exact test: P = 0.002, OR = 14.3, 95% CI = 2-626) and being positive to more than one serological or molecular tests (co-infections) (Mann-Whitney test: U = 243, Z = -2.6, n = 14, n = 61, P = 0.01) when compared with healthy dogs. Interestingly, a statistical association was found between the presence of R. conorii, E. canis, A. phagocytophilum and B. henselae antibodies in sick dogs and some clinicopathological abnormalities such as albumin and albumin/globulin ratio decrease and increase in serum globulins. Furthermore, seroreactivity with A. phagocytophilum antigens was statistically associated with CanL clinical stages III and IV. This study demonstrates that dogs with clinical leishmaniosis from Catalonia (Spain) have a higher rate of co-infections with other vector-borne pathogens when compared with healthy controls. Furthermore, positivity to some vector-borne pathogens was associated with more marked clinicopathological abnormalities as well as disease severity with CanL

Detection of white root rot in avocado trees by remote sensing

White root rot, caused by the soilborne fungus Rosellinia necatrix, is an important constraint to production for a wide range of woody crop plants such as avocado trees. The current methods of detection of white root rot are based on microbial and molecular techniques, and their application at orchard scale is limited. In this study, physiological parameters provided by imaging techniques were analyzed by machine learning methods. Normalized difference vegetation index (NDVI) and normalized canopy temperature (canopy temperature - air temperature) were tested as predictors of disease by several algorithms. Among them, logistic regression analysis (LRA) trained on NDVI data showed the highest sensitivity and lowest rate of false negatives. This algorithm based on NDVI could be a quick and feasible method to detect trees potentially affected by white root rot in avocado orchards.This work was supported by a grant from Consejería de Economía, Innovación, Ciencia y Empleo, Junta de Andalucía (P12-AGR-0370) to M. Barón, Ministerio de Economía, Industria y Competitividad Consejo Superior de Investigaciones Científicas and European Regional Development Fund (RECUPERA 2020/20134R060) to M. Barón, and Agencia de Innovación y Desarrollo de Andalucía (AGL14-52518-C2-1-R). C. Vida was recipient of a Ph.D. fellowship from the FPI program (MINECO)

Comparative histochemical analyses of oxidative burst and cell wall reinforcement in compatible and incompatible melon-powdery mildew (Podosphaera fusca) interactions.

The spatial–temporal expression patterns of oxidative burst and cell wall reinforcement were analyzed in leaves of resistant and susceptible melon (Cucumis melo L.) cultivars in response to Podosphaera fusca (Fr.) Braun & Shishkoff, the main causal agent of powdery mildew in cucurbits. Extensive development of powdery mildew mycelia and a progressive increase in haustorial count were recorded in the susceptible cultivar after 4 d, while in the resistant cultivar powdery mildew failed to grow and small brownish and necrotic leaf areas were frequently observed. Rapid generation of the reactive oxygen intermediates hydrogen peroxide and superoxide radicals 4 h after pathogen challenge, but before the fungal haustoria formation, stood upstream in the cascade of events induced during these interactions. This oxidative burst was followed by the accumulation of strengthening polymers of callose and lignin at the cell wall of attacked resistant plant cells. Interestingly, the transcriptional levels of phenylalanine ammonia-lyase (PAL), an important enzyme for phenylpropanoid metabolism, did not significantly change throughout the experiments. Although these physiological changes were observed in both cultivars, their faster kinetics and amplitude in the resistant line compared to the susceptible cultivar governed the differential visual response of these cultivars against P. fusca. These findings, along with data obtained in previous studies, have provided the bases for an integrated model in which the spatial–temporal response patterns of these resistance mechanisms have been arranged, which may ultimately lead to successful protection of melon plants against P. fusca.This study was supported by grants from Plan Nacional de Recursos y Tecnologías Agroalimentarias from Ministerio de Educación y Ciencia (AGL2001-1837-CO2-01; AGL2004-06056-CO2-01). DR was supported by a grant from the former Ministerio de Ciencia y Tecnología.Peer reviewe