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The bacterium Xylella fastidiosa (Xf) is a plant pathogen that can block the flow of water and nutrients through the xylem. Xf symptoms may be confounded with generic water stress responses. Here, we assessed changes in biochemical, biophysical and photosynthetic traits, inferred using biophysical models, in Xf-affected almond orchards under rainfed and irrigated conditions on the Island of Majorca (Balearic Islands, Spain). Recent research has demonstrated the early detection of Xf-infections by monitoring spectral changes associated with pigments, canopy structural traits, fluorescence emission and transpiration. Nevertheless, there is still a need to make further progress in monitoring physiological processes (e.g., photosynthesis rate) to be able to efficiently detect when Xf-infection causes subtle spectral changes in photosynthesis. This paper explores the ability of parsimonious machine learning (ML) algorithms to detect Xf-infected trees operationally, when considering a proxy of photosynthetic capacity, namely the maximum carboxylation rate (Vcmax), along with carbon-based constituents (CBC, including lignin), and leaf biochemical traits and tree-crown temperature (Tc) as an indicator of transpiration rates. The ML framework proposed here reduced the uncertainties associated with the extraction of reflectance spectra and temperature from individual tree crowns using high-resolution hyperspectral and thermal images. We showed that the relative importance of Vcmax and leaf biochemical constituents (e.g., CBC) in the ML model for the detection of Xf at early stages of development were intrinsically associated with the water and nutritional conditions of almond trees. Overall, the functional traits that were most consistently altered by Xf-infection were Vcmax, pigments, CBC, and Tc, and, particularly in rainfed-trees, anthocyanins, and Tc. The parsimonious ML model for Xf detection yielded accuracies exceeding 90% (kappa = 0.80). This study brings progress in the development of an operational ML framework for the detection of Xf outbreaks based on plant traits related to photosynthetic capacity, plant biochemistry and structural decay parameters., This research was supported by grant: ITS2017-095: Design and Implementation of control strategies for Xylella fastidiosa, Project 5. Government of the Balearic Islands, Spain. Data collection was partially supported by the European Union's Horizon 2020 research and innovation program through gran agreement XF-ACTORS (727987).

Trabajo presentado en el Simposium Científico Técnico Expoliva, celebrado online del 22 al 25 de septiembre de 2021., Los patógenos vasculares son los agentes causales de las principales enfermedades que amenazan la salud y la producción olivarera en todo el mundo. El uso de microorganismos endofíticos representa una estrategia desafiante y prometedora para el manejo de las enfermedades vasculares en el olivo. Aunque la investigación actual se ha centrado en analizar la estructura y diversidad de las comunidades microbianas que habitan el xilema del olivo, la caracterización de este nicho ecológico ha sido pasada por alto hasta la fecha. En este trabajo, se seleccionaron plantones jóvenes y olivos adultos de los genotipos 'Picual' y 'Arbequina' para caracterizar la composición química de la savia del xilema, así como su diversidad microbiana con el objetivo de mejorar la comprensión de los requisitos nutricionales de los microorganismos que habitan el xilema, incluyendo sus patógenos vasculares, para permitir el diseño de medios de crecimiento artificiales que ayuden a mejorar el cultivo del microbioma del olivo.

Trabajo presentado en la 3rd European Conference on Xylella fastidiosa (Building knowledge, protecting plant health), celebrada online el 29 y 30 de abril de 2021., Remote sensing efforts made as part of European initiatives via POnTE, XF-ACTORS and the JRC, as well as through regional programs, have focused, among others, on the development of algorithms for the early detection of Xylella fastidiosa (Xf)-induced symptoms. Airborne campaigns carried out between 2016 and 2019 collected high-resolution hyperspectral and thermal images from infected areas in the Apulia region (Italy), in the province of Alicante and on the island of Mallorca (Spain). The remote sensing imagery collections were performed alongside field surveys and laboratory analyses to assess the presence of Xf, and the severity and incidence of disease in olive and almond trees. Radiative transfer models and machine learning algorithms were used to quantify spectral plant traits for each individual infected tree, assessing their importance as pre visual indicators of Xf-induced stress. These studies conducted across species have demonstrated that specific spectral plant traits successfully revealed Xf induced symptoms at early stages, i.e., before visual symptoms appear. The results show that spectral plant traits contribute differently to symptom detection across host species (olive vs. almond), and that abiotic-induced stress affects the performance of the algorithms used for detecting infected trees. Together, the different European initiatives studying the use of remote sensing to support the monitoring of landscapes for Xylella fastidiosa detection lead us to conclude that the early detection of Xf-induced symptoms is feasible when high-resolution hyperspectral imagery and physically-based plant trait retrievals are used, obtaining accuracies exceeding 92% (kappa>0.8). These results are essential to enable the implementation of effective control and management of plant diseases using airborne- droneand satellite-based remote sensing technologies. Moreover, these large-scale hyperspectral and thermal imaging methods greatly contribute to the future operational monitoring of infected areas at large scales, well beyond what is possible from field surveys and laboratory analyses alone.

Trabajo presentado en la 3rd European Conference on Xylella fastidiosa (Building knowledge, protecting plant health), celebrada online el 29 y 30 de abril de 2021., Xylella fastidiosa (Xf) causes plant diseases that lead to massive economic losses in agricultural crops, making it one of the pathogens of greatest concern to agriculture nowadays. Detecting Xf at early stages of infection is crucial to prevent and manage outbreaks of this vector-borne bacterium. Recent remote sensing (RS) studies at different scales have shown that Xf-infected olive trees have distinct spectral features in the visible and infrared regions (VNIR). However, RS-based forecasting of Xf outbreaks requires tools that account for their spatiotemporal dynamics. Here, we show how coupling a spatial Xf-spread model with the probability of Xf-infection predicted by an RS-driven modeling algorithm based on a Support Vector Machine (RS-SVM) helps detecting the spatial Xf distribution in a landscape. To optimize such model, we investigated which RS plant traits (i.e., pigments, structural or leaf protein content) derived from high-resolution hyperspectral imagery and biophysical modelling are most responsive to Xf infection and damage. For that, we combined a field campaign in almond orchards in Alicante province (Spain) affected by Xf (n=1,426 trees), with an airborne campaign over the same area to acquire high-resolution thermal and hyperspectral images in the visible-near-infrared (400-850 nm) and short-wave infrared regions (SWIR, 950-1700 nm). We found that coupling the epidemic spread model and the RS-based model increased accuracy by around 5% (OA = 80%, kappa = 0.48 and AUC = 0.81); compared to the best performing RS-SVM model (OA = 75%; kappa = 0.50) that included as predictors leaf protein content, nitrogen indices (NIs), fluorescence and a thermal indicator, alongside pigments and structural parameters. The parameters with the greatest explanatory power of the RS model were leaf protein content together with NI (28%), followed by chlorophyll (22%), structural parameters (LAI and LIDFa), and chlorophyll indicators of photosynthetic efficiency. In the subset of almond trees where the presence of Xf was tested by qPCR (n=318 tress), the combined RS-spread model yielded the best performance (OA of 71% and kappa = 0.33). Conversely, the best-performing RS-SVM model and visual inspections produced OA and kappa values of 65% and 0.31, respectively. This study shows for the first time the potential of combining spatial epidemiological models and remote sensing to monitor Xf-disease distribution in almond trees.

Trabajo presentado en la 2nd European conference on Xylella fastidiosa (how research can support solutions), celebrada en Ajaccio el 29 y 30 de octubre de 2019., An outbreak of Xylella fastidiosa (Xf) subsp. multiplex ST6 was first identified in June 2017 in Alicante province (Valencian Community, Spain) affecting mainly almond trees, but also other species including ornamentals, cultivated Prunus spp. and landscape plants. The current demarcated area (DA) in Alicante covers 134,581 hectares, and affects more than 70 municipalities. Genetic analysis may provide evidence of biological, ecological and host-range diversity among Xf strains of the same subspecies and STs and help to elucidate the pathway of introduction and spread in an affected area. Pairwise comparisons of the chromosomal genomes of two ST6-sequenced strains isolated from almond trees in Alicante, ESVL and IVIA5901, showed an average nucleotide identity higher than 99.9%. Interestingly, the two strains differ for the presence of the plasmids pXF64-Hb_ESVL and pUCLA-ESVL detected only in the ESVL strain. The aim of this study was to determine the incidence and distribution of plasmid-bearing strains of Xf subsp. multiplex ST6 in Alicante. PCR tests were performed on 20 strains isolated from different hosts, and on approximately 100 DNA samples from infected almond trees collected in eight municipalities within the DA of Alicante. PCR results on the cultured isolates showed the occurrence of ST6-strains harbouring the two plasmids, or only the plasmid pUCLA-ESVL or none of them. Interestingly, in some of the DNA samples from infected almonds, only the plasmid pXF64-Hb_ESVL could be detected. More specifically, 4% of ST6-infected samples harboured the plasmid pXF64-Hb_ESVL, 12% only pUCLA-ESVL, 16% both plasmids, and 44% neither of the two plasmids. In five of the eight municipalities, we found samples harbouring the two plasmids, and only in the samples from the municipality of Benifato the PCR tests failed to amplify the targeted plasmids. Future work will increase the number of samples to cover the entire DA to better understand and relate the presence of plasmids to the epidemiology of this disease in combination with the use of multilocus sequence typing and genome sequencing of more Xf subsp. multiplex ST6 strains., The present work was presented in the framework of the Joint Annual Meeting of the EU Horizon 2020 Projects POnTE ‘Pest Organisms Threatening Europe' (GA 635646) and XF-ACTORS ‘Xylella fastidiosa Active Containment Through a multidisciplinary-Oriented Research Strategy' (GA 727987).

Trabajo presentado en la 2nd European conference on Xylella fastidiosa (how research can support solutions), celebrada en Ajaccio el 29 y 30 de octubre de 2019., A first case of infection caused by Xylella fastidiosasubsp. multiplexST6 was detected in an olive tree in the municipality of Villarejo de Salvanés, in the southeast of the Community of Madrid (Spain) in April 2018. This constituted the first detection of X. fastidiosain this crop on the Iberian Peninsula, and the second concerning other crops after the detection of the pathogen in almondtrees in Alicante, in the Valencian Community (Spain), where X. fastidiosasubsp. multiplexST6 was also identified, although until now it has not been detected causing infection in olive trees in that region.The Community of Madrid accounts for 27,000 hectares of olive trees, most of them in Villarejo de Salvanés. This area has a continental climate, with below zero minimum winter temperatures. After the official declaration of the outbreak, the actions established in the European Decision 2017/2352, theContingency Plan against X. fastidiosaof the Spanish Ministry of Agriculture, Fisheries and Food, and that of the Community of Madrid were immediately applied to eradicate the bacterium and prevent its spread.Among these measures, around 2,000 samples from olive trees and other host plants of the demarcated area (DA), 300 insect vectors, and olive trees from other municipalities were analysed. All of them gave a negative result by real-time PCR official protocols. As a direct consequence of the application of the eradication measures, the results obtained so far would indicate that this detection was a unique event and the bacterium would be neither disseminated nor established in the EU, although surveillance measures within the DA must continue as established in the Action Plan of the Community of Madrid.

Trabajo presentado en la European Conference on Xylella 2017 (Finding answers to a global problem), celebrada en Palma de Mallorca del 13 al 15 de noviembre de 2017., With the goal of understanding the nutritional requirements of xylem-limited microbiome, the chemical composition of xylem sap extracted from two Olea europaea var. europaea cultivars (Picual and Arbequina) was evaluated. Selected plants were established on their own feet or grafted onto rootstocks of Olea europaea var. sylvestris. Xylem sap of olives branches was collected using a external 45-cm external Scholander Super Chamber. Metabolome analysis was performed by proton nuclear magnetic resonance (NMR) spectroscopy-based study and the ionome analysis was performed by inductively coupled plasma with optical emission spectroscopy (ICP-OES). In total 26 metabolites were identified by NMR spectroscopy, including amino acids (alanine, arginine, aspartate, glutamate, glutamine, isoleucine, leucine, methionine, proline, threonine, tyrosine, and valine), sugars (glucose, fructose, mannitol, myo-inositol, and sucrose), organic acids (formic, fumaric and succinic acid), alcohols (ethanol and methanol), and other molecules. On the other hand ICP-OES allowed the quantification of 14 elements and five inorganic anions. Xylem sap analysis will lead to a better understanding of the biology and complex nutritional requirements of olive xylem-inhabiting microorganisms, including Xylella fastidiosa, and to help designing artificial growing media to improve culturing of the olive microbiome., Study supported by Projects 727987 XF-ACTORS (EU-H2020) & AGL2016-75606-R (MEIC Spain and FEDER-EU).

Xylella fastidiosa es una bacteria de cuarentena en la Unión Europea (bacteria incluida en la Lista A1 EPPO desde 1981), ya que se considera una grave amenaza para la UE y los países de la cuenca del Mediterráneo por la gran variedad de plantas huéspedes que infecta, así como por su forma de transmisión a través de insectos vectores de diversa naturaleza, y por la existencia en Europa de condiciones climáticas que favorecerían su desarrollo epidémico. Además, la bacteria posee un enorme potencial patogénico ya que infecta y causa enfermedad severa en diferentes cultivos agrícolas de gran importancia económica, incluyendo el almendro, cítricos, melocotón, olivo y vid, así como una gran variedad de árboles y plantas no cultivadas como arces, adelfas, y diferentes especies silvestres y forestales típicas del bosque mediterráneo como Quercus, olmos, jara, y retama, entre otros. Esto hace que a esta bacteria se la considere uno de los organismos fitopatógenos de más riesgo para la Unión Europea.

Downy mildew (DM) of opium poppy (Papaver somniferum) caused by Peronospora somniferi is one of the most destructive diseases of this crop due to the systemic nature of infection as compared with local infections caused by Peronospora meconopsidis, the other downy mildew pathogen of this crop. We developed an inoculation method using Peronospora somniferi sporangia as inoculum and demonstrated for the first time that local infection of leaves by sporangia give rise to systemic infections in the plant as well as of seeds. Our results also showed that this inoculation protocol was very effective in reproducing disease symptoms and assessing the resistance response to DM in opium poppy genotypes under field conditions. More interestingly, results indicate that up to 100% of seed samples from some genotypes showing a complete (symptomless) resistant phenotype were infected by the pathogen when seeds were analyzed by a P. somniferi-specific nested-PCR protocol. This latter aspect deserves further attention while breeding opium poppy for resistance to P. somniferi., Regional Government of Andalusia (project P10-AGR-6497) to MMB; European Union through the European Regional Development Fund (ERDF).

Trabajo presentado en la European Conference on Xylella 2017 (Finding answers to a global problem), celebrada en Palma de Mallorca del 13 al 15 de noviembre de 2017., A remote sensing campaign carried out in Puglia in summer 2016 collected hyperspectral and thermal images of ca 200,000 olive trees at sub-meter resolution. In the 1200 ha study area within the X. fastidiosa infected zone, 3500 trees were simultaneously evaluated in the field for severity of X. fastidiosa symptoms. The hyperspectral sensor used in this experiment acquired data in the visible and near-infrared spectral region (400-885 nm) with 260 bands of 6.5 nm FWHM at 1.85 nm/pixel and 12-bit radiometric resolution. The sensor was radiometrically calibrated in the laboratory, and images atmospherically corrected to obtain surface reflectance using total incoming irradiance measured in the field. The high resolution hyperspectral and thermal imagery acquired over the orchards allowed the delineation of individual trees using object-based crown detection algorithms. Crown temperature and hyperspectral indices were calculated for each tree to classify disease severity levels using different machine learning algorithms, including linear discriminant analysis, support vector machines and neural networks. The success and applicability of these early detection methods to other areas will be discussed in the context of a new airborne campaign planned in July 2017 over X. fastidiosa infected zones in Mallorca. In this new airborne campaign the impact of X. fastidiosa symptoms on crown reflectance will also be evaluated in the 800-1700 nm spectral region., This work was supported by JRC and has received funding from the European Union’s Horizon 2020 research and innovation programme under grant agreement N. 635646 “Pest Organisms Threatening Europe POnTE” and grant agreement N. 727987 “Xylella fastidiosa Active Containment Through a multidisciplinary-Oriented Research Strategy XF-ACTORS”.

Trabajo presentado en el XVIII Congreso de la Sociedad Española de Fitopotología, celebrado en Palencia del 20 al 23 de septiembre de 2016., Xylella fastidiosa (Xf) es una bacteria Gram-negativa de cuarentena en la Unión Europea, que crece restringida en el xilema de las plantas y presenta a menudo variantes patogénicas específicas (subspecies) de ciertos huéspedes. Xf causa importantes enfermedades y pérdidas de rendimientos en diferentes cultivos y plantas forestales y ornamentales, llegando a infectar más de 340 especies vegetales. La asignación genotípica de Xf a nivel de subespecie es necesaria para permitir realizar inferencias acerca de la biología de los aislados y/o su procedencia en el momento que se realiza alguna interceptación de esta bacteria en un país donde no está presente. En este estudio se ha desarrollado un nuevo protocolo basado en la minisecuención, que consiste en una reacción de extensión de un cebador en un único nucleótido (SNuPE), utilizando cuatro cebadores simultáneamente, y que permite la identificación de todas las subespecies de Xf (subsp. fastidiosa, multiples, sandyi y pauca) y de tres genotipos de Xf subsp. pauca, incluyendo la cepa CoDiRO causante de la enfermedad de decaimiento rápido del olivo en el sur de Italia. El protocolo resultó robusto para la predicción de todas las subespecies y genotipos de Xf en pruebas ciegas realizadas utilizando ADN extraído de: i) cultivos puros de Xf, ii) diferentes plantas infectadas por Xf de forma artificial o natural, y iii) insectos vectores portadores de Xf. Este protocolo SNuPE múltiple puede ser muy útil como una primera metodología de análisis rápida y robusta para la asignación taxonómica de cepas de Xf a nivel de subespecie o genotipo (Xf subsp. pauca), en estudios epidemiológicos de campo o para fines de cuarentena cuando se intercepte material infectado por la bacteria., Investigación financiada por el Proyecto POnTE H2020-SFS-2014-2015 Sustainable Food Security call (Topic SFS-03a-2014: Native and alien pests in agriculture and forestry).

Trabajo presentado en el 32nd Symposium European Society of Nematologists, celebrado en Braga del 28 de agosto al 1 de septiembre de 2016., This work has studied the diversity of plant-parasitic nematodes infecting olive in a wide-region in southern Spain (Andalusia). Our data indicate that there is a high diversity of plant-parasitic nematodes (including root-knot = Meloidogyne spp., root-lesion = Pratylenchus spp., cystforming = Heterodera mediterranea, stubby- root = Trichodorus spp., reniform = Rotylenchulus sp., dagger and needle = Longidorus spp., Xiphinema spp., ring = Criconemoides spp., Ogmo spp., and other migratory ectoparasitic nematodes viz. Amplimerlinius spp., Helicotylenchus spp., Neodolichorhynchus spp., Paratrophurus spp. Paratylenchus spp., Rotylenchus spp., Trophurus sp., Tylenchorhynchus spp.) associated with olive in Southern Spain that can exert different damage to olive roots depending on the olive variety and their abundance. The highest number of species was detected in Xiphinema and Longidorus, with description of eight and seven new species, respectively, apart from several new records for Spain. In addition, seven Meloidogyne species were detected in wi ld and cultivated olive, from which one is being described as a new species. Multivariate analyses allowed determining the agronomic and environmental factors that drive the structure and diversity of plant-parasitic nematodes infecting olive in Southern-Spain, that included: soil physicochemicol factors (texture, pH, sand and doy, and extractable K), climatic variables (mínimum and maximum temperatures) and the olive cultivar. Nematode abundance and diversity indexes (Richness, Shannon, Evenness and plant-parasitic index) were influenced by olive cultivar, and orchard and soil management predices. The plant-parasitic nematode abundance was significantly higher in orchards managed organically compared to those managed conventionally. These findings could be the result of the influence of the herbaceous plants that compase the soil cover, which may affect plant-parasit ic nematodes at the species level.

Trabajo presentado en la 3rd International Conference on Microbial Diversity (MD2015, "The Challenge of Complexity"), celebrada en Perugia del 27 al 29 de octubre de 2015., The biochemistry and microbial ecology of PDO Aloreña de Málaga table olive fermentations were investigated in two different industries (1 and 2) of province of Málaga (Spain). Monitoring of the physicochemical parameters such as pH, acidity, salt, color and texture of fruits was carried out during the entire process. No differences were detected in physicochemical arameters, which were characteristics for this type of elaboration. The microbial ecology was investigated using both culture-dependent and culture-independent methods in fruit and brine. The initial yeast population was different between both industries. While industry (1) showed counts around 4 - 5 log10 cfu/mL during the first month of fermentation, industry 2 presented lower oncentrations (1 – 3 log 10 cfu/mL), although, both locations reached the same yeast populations at the end of storage period (>5 log 10 cfu/mL). Bar - coded pyrosequencing analysis of the ITS rDNA region identified 109 and 118 operational taxonomic units (OTUs) in fruits and brines, espectively, and revealed the presence of yeast genera previously identified in table olives such as Pichia, Candida, Debaryomyces and Sacharomyces sp., but also, this is the first time that other predominant genera such as Zygostorulaspora, Penicillium and Aspergillus have been identified in this type of table olives. Small differences were found between most abundant yeasts in fruits and brines, during the fermentation process and between both industries.

Two haplotypes of Xylella fastidiosa subsp. pauca (Xfp) that correlated with their host of origin were identified in a collection of 90 isolates infecting citrus and coffee plants in Brazil, based on a single-nucleotide polymorphism in the gyrB sequence. A new single-nucleotide primer extension (SNuPE) protocol was designed for rapid identification of Xfp according to the host source. The protocol proved to be robust for the prediction of the Xfp host source in blind tests using DNA from cultures of the bacterium, infected plants, and insect vectors allowed to feed on Xfp- infected citrus plants. AMOVA and STRUCTURE analyses of microsatellite data separated most Xfp populations on the basis of their host source, indicating that they were genetically distinct. The combined use of the SNaPshot protocol and three previously developed multilocus SSR markers showed that two haplotypes and distinct isolates of Xfp infect citrus and coffee in Brazil and that multiple, genetically different isolates can be present in a single orchard or infect a single tree. This combined approach will be very useful in studies of the epidemiology of Xfp- induced diseases, host specificity of bacterial genotypes, the occurrence of Xfp host jumping, vector feeding habits, etc., in economically important cultivated plants or weed host reservoirs of Xfp in Brazil and elsewhere [Int Microbiol 2015; 18(1):13-24]., We acknowledge financial support from the EU grant ICA4-CT-2001-10005 and an ‘Intramural Project’ to B. B. Landa from the Spanish National Research Council (CSIC), as well as CNPq for a scholarship to J. R. S. Lopes in Brazil.

Trabajo presentado en la 3rd International Conference on Microbial Diversity (MD2015, "The Challenge of Complexity"), celebrada en Perugia del 27 al 29 de octubre de 2015., The objective of this research was to characterize the biochemistry and microbiology of spontaneous PDO Aloreña de Málaga table olive fermentations and define the bacterial biodiversity during the process. The study was carried out in two different locations in Malaga province (Spain) following the fermentation during 4 months. Biochemical analysis of olives showed a progressive increase in free acidity as consequent of a spontaneous fermentation, with pH and salt values characteristic of this type of product. Both locations showed a similar trend in microbiology enumeration by plating in selective media. Yeasts were the most abundant microorganisms during the time, while lactic acid bacteria (LAB) cell count revealed low concentration or even were not detected. The novel of this research is the study of the microbial ecology of olives fermentation using bar-coded pyrosequencing of 16S rDNA to characterize bacterial diversity. The bacterial α- and β- diversity differed between fruits and brines being higher for fruits especially at the beginning of the fermentation process and the opposite for the brines. Bacterial diversity changed during fermentation time, and a total of 323 and 150 operational taxonomic units (OTUs) were identified in fruits and brines, respectively. The predominant genera were Celerinantantimonas, Pseudomonas, Propionibacterium, and diverse Acetobacteraceae. Some differences in the bacterial diversity were found between industries, special at the beginning of the process, whereas no differences were found for the rest of physicochemical parameters such as color and texture in the final product.

Trabajo presentado en el Rhizosphere 4: Stretching the interface of life, celebrado en Maastricht del 21 al 25 de junio de 2015., Plant disease epidemics result from interactions of a susceptible host, a virulent pathogen and conducive environment. Shifts in any component can change disease expression to still unknown directions. Verticillium wilt (VW) of olive caused by the soilborne fungus Verticillium dahliae (Vd) is of major concern for olive industry in the Mediterranean basin. We carried out experiments using olive cvs. Picual and Arbequina, which grew in soil infested by the defoliating (D) or non-defoliating (ND) Vd pathotypes at 20, 24 and 28ºC and CO2 levels of 386, 550 and 750 ppm, representative of current and future SRES-IPCC A2 and B2 scenarios for southern Spain. Surface response models quantified the combined effects of temperature and CO2 on VW intensity and plant growth, demonstrating a differential effect of biotic and abiotic factors. Optimum VW development occurred at 20-24°C and current CO2, being faster and severe in 'Picual'/D. Raising CO2 delayed VW and Vd infection. The transcriptomic profile of olive cultivars in response to Vd infection and the three climate SRES scenarios was assessed using a 12-plex microarray of 37,449 olive unigenes (OLEAGEN project). Main differences in the expression profiles were due to olive genotype, followed by the climatic conditions at the three SRES scenarios and in a lesser extent by Vd infection. Thus, plants growing at current climate showed higher number of differentially expressed genes compared to plants growing at SRES-IPCC A2 and B2 scenarios with a general trend to decrease the number of significantly transcribed genes as temperature and CO2 increased. Furthermore, a higher number of genes (up to 10x) were expressed differentially in 'Arbequina' than in 'Picual' in response to environmental changes as well as to infection by the D pathotype with a low number of genes being common for both cultivars.

Durante los últimos años, el Grupo AGR136 Sanidad Vegetal del IAS-CSIC y la UCO ha centrado sus proyectos de investigación de financiación privada o pública en el desarrollo de conocimeintos y tecnologías sobre medidas de lucha aplicables para la gestión integrada de verticilosis del olivo. En este artículo presentamos sucintamente algunos de los objetivos perseguidos en dichos proyectos y los resultados más destacables alcanzados en el desarrollo de los mismos., Las investigaciones de los autores referidas en este artículo han sido financiadas por los proyectos AGL2008-00344 (Ministerio de Ciencia e Innovación), AGL2011-24935 y AGL2012-37521 (Ministerio de Economía y Competitividad), P08-AGR 3528, P10-AGR-5908, P10-AGR 6082, y P12- AGR 1486 (Consejería de Economía, Innovación y Ciencia, Junta de Andalucía), cofinanciados por fondos FEDER; y en parte por la Organización Interprofesional del Aceite de Oliva Español-CITOLIVA'.

Nowadays, there is a tendency in olive production systems to reduce tillage or keep a vegetative cover to reduce soil erosion and degradation. However, there is scarce information on the effects of different soil management systems (SMS) in soil bacterial community composition of olive groves. In this study, we have evaluated the effects of soil type and different SMS implemented to control weeds in the structure and diversity of bacterial communities of 58 soils in the two geographic areas that best represent the organic olive production systems in Spain. Bacterial community composition assessed by frequency and intensity of occurrence of terminal restriction profiles (TRFs) derived from terminal restriction fragment length polymorphism (T-RFLP) analysis of amplified 16S ribosomal deoxyribonucleic acid were strongly correlated with soil type/field site (Eutric/Calcaric) that differed mainly in soil particle size distribution and soil pH, followed by a strong effect of SMS, in that order. Canonical discriminant (CD) analysis of TRFs properly classified all of the olive orchard soils as belonging to their respective soil type or SMS. Furthermore, only a small set of TRFs were enough to clearly and significantly differentiate soil samples according to soil type or SMS. Those specific TRFs could be used as bioindicators to assess the effect of changes in SMS aimed to enhance soil quality in olive production systems. © 2014 Society for Applied Microbiology and John Wiley & Sons Ltd., Financial support for this research was provided by Projects AGL2008-00344 and AGL2009–12936-C03-01 from ‘Ministerio de Ciencia e Innovación’, AGL-2012–37521 and ‘Ministerio de Economia y Competitividad’ of Spain and Projects P08-AGR-03528, P08-AGR-03643 and P10-AGR-5908 from ‘Consejería de Economía, Innovación y Ciencia’ of Junta de Andalucía and FEDER financial support from the European Union. S. Aranda was recipient of a PhD. grant from ‘Consejo Nacional de Ciencia y Tecnología’ (CONACYT) México.

Trabajo presentado en el XVII Congreso de la Sociedad Española de Fitopatología, celebrado en Lleida del 7 al 10 de octubre de 2014., El objetivo de este estudio ha sido comparar la estructura y diversidad bacteriana de la rizosfera de olivar en árboles con síntomas de Verticilosis y en árboles asintomáticos no infectados, para identificar bacterias que potencialmente supriman la actividad de supervivencia y patogénica de Verticillium dahliae. Para ello, raíces y ramas jóvenes de olivos de 4 árboles con síntomas de Verticilosis y 4 árboles asintomáticos en cada una de tres fincas en Mengibar-Jaén fueron muestreadas en otoño (2012) y primavera (2013). En ambos periodos se aisló V. dahliae en el 80% de árboles con síntomas de Verticilosis, confirmándose su pertenencia al patotipo D, no aislándose de árboles asintomáticos. Para cada suelo, se caracterizaron sus propiedades fisicoquímicas, y se determinó la densidad de población de bacterias cultivables y la diversidad funcional de la rizosfera de cada árbol (ECOPLATE®). Adicionalmente, se caracterizó la estructura y diversidad de las comunidades bacterianas totales mediante análisis de polimorfismos de los fragmentos terminales de restricción (T-RFLP) y de pirosecuenciación de fragmentos amplificados del ARNr 16S. Las secuencias de pirosecuenciación obtenidas se agruparon en OTUs, y cada OTU fue alineado con la base de datos de Greengenes utilizando UCLUST para su identificación taxonómica, y a partir de las frecuencias normalizadas de OTUs se calcularon índices de diversidad α y β, y curvas de rarefacción. En el análisis de pirosecuenciación se obtuvieron un total de 100.769 secuencias de calidad con una media de 4.200 por árbol, y un total de 8.044 OTUs (especies). Los diferentes análisis multivariantes llevados a cabo (NMDS, PCA, CDA y cluster), indicaron que existe una tendencia de las muestras rizosféricas de cada árbol a agruparse de acuerdo a la finca de procedencia, así como al desarrollo o no de Verticilosis dentro de ésta. Los índices de diversidad α y β indicaron que la rizosfera de olivo sustenta una diversidad bacteriana muy elevada y en general altamente estable con una clara prevalencia de Alphaproteobacteria (Rhizobacteria) seguido de Actinobacteria (Actinomycetales). Sin embargo, el análisis discriminante y canónico nos permitió clasificar en el 100% de los casos los árboles con Verticilosis y no infectados en base a las frecuencias diferenciales de 16 OTUs, con 4 de ellos pertenecientes a 4 clases bacterianas presentando diferencias significativas., Financiado por Proyecto AGL2012-37521, fondos CICE-Junta de Andalucía (ayudas Grupo PAIDI AGR-136, P. Excelencia P10-AGR-5908) y cofinanciado por fondos FEDER (EU).

Trabajo presentado en el XVII Congreso de la Sociedad Española de Fitopatología, celebrado en Lleida del 7 al 10 de octubre de 2014., La temperatura y la concentración de dióxido de carbono (CO2) atmosférico son las variables ambientales asociadas al Cambio Climático que mayor impacto pueden ejercer sobre los patosistemas agrícolas, modificando hacia direcciones aún desconocidas la virulencia de los patógenos, así como el nivel de resistencia de las plantas a éstos. En este trabajo se ha realizado un estudio transcriptómico utilizando un microarray 12-plex 135K de 37.449 unigenes de olivo (Proyecto OLEAGEN) para determinar el efecto de la temperatura y CO2 ambiental sobre el desarrollo de la Verticilosis, y su interacción con el nivel de virulencia de patotipos de Verticllium dahliae (VD) y la susceptibilidad del genotipo de olivo en momentos previos a la aparición de síntomas. Para ello plantones de olivo de los cvs. Picual y Arbequina, crecieron en suelo infestado por cada uno de los patotipos defoliante (D) y no defoliante (ND) de VD, con condiciones de temperatura y CO2 representativas de los niveles actuales ambientales de ambos factores (20ºC, 386 ppm) así como sus estimaciones por modelos globales de simulación de clima para los escenarios AR4 del ICPP para el sur de España B2 (24ºC, 550 ppm) y A2 (28ºC, 750 ppm). Los resultados mostraron que, en las condiciones evaluadas, las principales diferencias en los perfiles de expresión son debidas al genotipo de olivo, seguidas de las condiciones ambientales y la infección por VD. Así, en plantas control (no inoculadas) se observaron grandes diferencias (P, Financiado por Proyecto AGL2012-37521, fondos CICE-Junta de Andalucía (ayudas Grupo PAIDI AGR-136, P. Excelencia P08-AGR-03528) y cofinanciado por fondos FEDER (EU).

Trabajo presentado en el XVII Congreso de la Sociedad Española de Fitopatología, celebrado en Lleida del 7 al 10 de octubre de 2014., Las plantas han desarrollado estrategias de selección y sustento en su rizosfera de grupos específicos de microorganismos antagonistas como medida de defensa frente a patógenos de suelo, fenómeno que ocurre en suelos supresivos a enfermedades causadas por patógenos de suelo . Por ello, el conocer los microrganismos responsables de este fenómeno puede contribuir al control de estas enfermedades mediante dos vías: i) Favoreciendo el desarrollo de supresividad de forma artificial; ii) Identificando y aislando los microorganismos responsables para su posterior aplicación como agentes de biocontrol de dichos patógenos. En las dos últimas décadas el estatus fitosanitario de los olivares se ha visto amenazado por la Verticilosis; la enfermedad más limitante para este cultivo en la cuenca Mediterránea . Nuestros estudios indican que al menos el 25% de suelos representativos de olivar de Andalucía presentan un alto nivel de supresividad a la Verticilosis . Por ello, estamos determinado los factores asociados con este fenómeno utilizando tanto enfoques “Clásicos” como “Moleculares de Segunda Generación (NGS)”. Para identificar los microorganismos que pueden determinar la supresividad natural de los suelos se han utilizado técnicas clásicas que demuestran: i) la transferencia de la supresividad de suelos altamente supresivos a conducivos mediante mezclas de éstos; ii) la pérdida de parte de la supresividad de los suelos cuando éstos son tratados con calor; y iii) las bacterias aisladas de suelos supresivos poseen capacidad antagonista frente a Verticillium dahliae in vitro y suprimen el desarrollo de la Verticilosis in planta . En un segundo paso mediante NGS se han caracterizado las comunidades bacterianas asociadas a la rizosfera de plantas crecidas en suelos supresivos y conducivos en presencia o no de V. dahliae y en experimentos de transferencia de suelos . Para ello se han secuenciado más de 252 muestras rizosféricas independientes y se han obtenido más de 750 .000 secuencias de calidad que se han podido agrupar en >15 .000 OTUs . Posteriormente, a partir de las frecuencias de OTUs se han calculado diversos índices de diversidad α y β para determinar la estructura de las poblaciones bacterianas asociadas al fenómeno de supresividad, y mediante diversos análisis multivariantes (Canónico Discriminante, Árboles de Clasificación) se han determinado OTUs específicos presentes en mayor frecuencia en suelos supresivos, así como OTUs que se transfieren de suelos supresivos a conducivos, que podrían estar asociados con la transferencia de la supresividad entre suelos., Financiado por fondos CICE-Junta de Andalucía (ayudas Grupo PAIDI AGR-136, P . Excelencia P10-AGR-5908) . Interprofesional del Aceite de Oliva Español-Citoliva y cofinanciado por fondos FEDER (EU).

Trabajo presentado en el XVII Congreso de la Sociedad Española de Fitopatología, celebrado en Lleida del 7 al 10 de octubre de 2014., La temperatura y la concentración de dióxido de carbono (CO2) atmosférico son las variables ambientales asociadas al Cambio Climático que mayor impacto pueden ejercer sobre los patosistemas agrícolas, modificando hacia direcciones aún desconocidas la virulencia de los patógenos, así como el nivel de resistencia de las plantas a éstos . En este trabajo se ha realizado un estudio transcriptómico utilizando un microarray 12-plex 135K de 37 .449 unigenes de olivo (Proyecto OLEAGEN) para determinar el efecto de la temperatura y CO2 ambiental sobre el desarrollo de la Verticilosis, y su interacción con el nivel de virulencia de patotipos de Verticllium dahliae (VD) y la susceptibilidad del genotipo de olivo en momentos previos a la aparición de síntomas . Para ello plantones de olivo de los cvs . Picual y Arbequina, crecieron en suelo infestado por cada uno de los patotipos defoliante (D) y no defoliante (ND) de VD, con condiciones de temperatura y CO2 representativas de los niveles actuales ambientales de ambos factores (20ºC, 386 ppm) así como sus estimaciones por modelos globales de simulación de clima para los escenarios AR4 del ICPP para el sur de España B2 (24ºC, 550 ppm) y A2 (28ºC, 750 ppm) . Los resultados mostraron que, en las condiciones evaluadas, las principales diferencias en los perfiles de expresión son debidas al genotipo de olivo, seguidas de las condiciones ambientales y la infección por VD . Así, en plantas control (no inoculadas) se observaron grandes diferencias (P, Financiado por Proyecto AGL2012-37521, fondos CICE-Junta de Andalucía (ayudas Grupo PAIDI AGR-136, P. Excelencia P08-AGR-03528) y cofinanciado por fondos FEDER (EU).

Trabajo presentado en la 5th OLIVEBIOTEQ Conference (International Conference for OliveTrees and Olive Products), celebrada en Ammam (Jordania) del 3 al 6 de noviemnre de 2014.-- Mateille, T. et al.., PESTOLIVE (Contribution of olive history for the management of soil-borne parasites in the Mediterranean Basin) is a project funded by ARIMNet, an ERANET action supported by the 7th European Framework Programme and by non-European Mediterranean countries. PESTOLIVE (www1.montpellier.inra.fr/CBGP/pestolive) aims at producing knowledge and tools for a new and efficient management of plant-parasitic nematodes (PPN) and plant-pathogenic fungi (PPF) in olive (Olea europaea L.) cropping systems and nurseries, while reducing the use of pesticides. Because of the anthropic continuum from Olea post-glacial refuges to oleasters (domestication) and then to olive-trees (breeding and cropping), the fragmentation of the PPN and PPF communities and of their natural enemies could explain the scattered diversity of the control techniques (especially resistance rootstocks, biocontrol, cropping strategies) developed and applied all around the Mediterranean Basin. The novelty of PESTOLIVE is based on i) the analysis and the management of the parasite diversity (ecology of communities) instead of controlling emblematic species (population approach) and ii) the involvement of knowledge about the historical co-adaptation of soil-borne parasite and natural enemies communities to olive-tree domestication (origins and past assemblages) and breeding that follows the history of O. europaea around the Mediterranean Basin., ARIMNet is coordinated by Dr. F. Jacquet and M. Ollagnon (INRA, France) and PESTOLIVE is co-funded by ANR (France), GDAR (Turkey), INIA (Spain), IRESA (Tunisia), MENESF CRS (Morocco) and NAGREF (Greece).

Ponencia presentada en el 11th International Verticillium Symposium, celebrado en Göttingen (Alemania) del 5 al 8 de mayo de 2013., Plants have evolved strategies of stimulating and supporting specific groups of antagonistic microorganisms in the rhizosphere as a defense against diseases caused by soilborne plant pathogens. Disease suppressive soils provide the best examples of this strategy. Suppressive soils have been described worldwide for many different pathogens, however for the fungal pathogen Verticillum dahliae the information is very scarce. Olive (Olea europaea L.) is one of the most important crops in Spain with > 2.4 million ha. During the last two decades the phytosanitary status of olive orchards is being threatened mainly due to Verticillium wilt. In previous studies a collection of rhizosphere soils from 90 olive orchards under different management systems and three rhizosphere soils from wild olive havens in Andalusia were characterized by their level of suppressiveness to Verticillium wilt. Results indicated that at least 25% of soils showed a high level of suppressiveness to Verticillium wilt. The objective of this study was to unravel the biotic and abiotic factors that may be associated with this phenomenon. For that purpose we selected a set of suppressive and conducive soils and have performed some “Classical” approaches to identify the biotic factors (microorganisms) involved in this specific suppression that included: 1) Transferring suppressiveness by adding small amounts of suppressive soil to conducive soil which have confirmed its biological nature. 2) Treating suppressive soil with heat to eliminate specific groups of microorganism which have demonstrated loss of suppressiveness at a certain level. 3) Isolating different microbial groups and correlating their presence with suppressiveness. 4) Screening representatives of microbial groups for in vitro and biocontrol activity against the target pathogen. In a second step using molecular approaches such as bar-coded pyrosequencing we have characterized all bacterial communities associated to the rhizosphere of plants growing in those selected soils. Sequences generated from pyrosequencing of rRNA gene amplicons with the GS Junior system (Roche) were processed using the Quantitative Insights Into Microbial Ecology (QIIME 1.6.0) pipeline. Flowgrams were clustered into OTUs at 97% pairwise identity using the seed-based UCLUST algorithm, and representative sequences from each OTU were aligned to the Greengenes bacteria database using PyNAST. In addition, α diversity and β diversity metrics together with rarefaction plots were also calculated to determine the bacterial population structure in those soils. Different multivariate analyses have allowed identifying some climatic parameters and physicochemical soil characteristics that are differentially associated to the level of suppressiveness of those soils as well as to determine which OTUs are specially enriched in the suppressive soils. Furthermore, specific OTUs have been identified as being transferred from the suppressive into the conducive soils in the ‘transferability experiment’ which might be associated as the OTUs being responsible of this transferable suppressive effect. Finally, experiments are being conducted to determine the influence of the indigenous microbiota on inducing plant defense mechanisms in olive plants grown in those suppressive soils. For that purpose both microarray and metagenomic sequencing of total DNA will be used to unravel genes differentially expressed or present under the suppressive conditions.

The phylum Nematoda includes the genus Longidorus, a remarkable group of invertebrates that are polyphagous root-ectoparasites of many plants including various agricultural crops and trees. Damage is caused by direct feeding on root cells as well as by transmitting nepoviruses. Species discrimination in Longidorus is complicated by phenotypic plasticity (intraspecific variability and minor interspecific differences) leading to potential misidentification. We conducted nematode surveys in cultivated and natural environments in southern Spain that detected 11 species of Longidorus. We developed a comparative study amongst these related species by considering morphological and morphometric features together with molecular data from nuclear ribosomal RNA genes [D2-D3 expansion segments of large ribosomal subunit (28S), internal transcribed spacer 1 (ITS1), and partial small ribosomal subunit (18S)]. The results of our molecular and phylogenetic analyses confirmed the morphological hypotheses and allowed the delimitation and discrimination of three new species of the genus, described herein as Longidorus baeticus sp. nov., Longidorus oleae sp. nov., and Longidorus andalusicus sp. nov., and eight known species (Longidorus alvegus, Longidorus crataegi, Longidorus fasciatus, Longidorus intermedius, Longidorus iuglandis, Longidorus magnus, Longidorus rubi, and Longidorus vineacola). Phylogenetic analyses of Longidorus spp. based on the three molecular markers resulted in a general consensus of these species grouping, as lineages were maintained for the majority of species (i.e. species with a conoid-rounded lip region, amphidial fovea asymmetrically bilobed, female tail bluntly rounded), but not in some others (i.e. positions of L. crataegi, L. intermedius, and L. rubi were quite variable). To date, this is the most complete phylogenetic analysis for Longidorus and Paralongidorus species, with the highest number of species included. No correspondence between phylogenetic trees and morphological characters was found for ribosomal markers, with the exception of amphidial shape. Thus, polyphasic identification, based on integration of molecular analysis with morphology, is a tool beyond doubt in Longidorus identification., This research was supported by grant AGL2009- 06955 from the ‘Ministerio de Ciencia e Innovación’ of Spain, grant 219262 FP7-ERANET ARIMNET (PESTOLIVE) from Instituto Nacional de Investi- gación y Tecnología Agraria y Alimentaria (INIA), grant AGR-136 from the ‘Consejería de Economía, Innvovación y Ciencia’ from Junta de Andalucía, and the European Social Fund.

Comunicación presentada en el 6th meeting of the IOBC-WPRS Working Group Multitrophic Interactions in Soil, celebrado en Córdoba (España) del 4 al 7 de abril de 2011.

Ponencia presentada en el 6th meeting of the IOBC-WPRS Working Group Multitrophic Interactions in Soil, celebrado en Córdoba (España) del 4 al 7 de abril de 2011.

Trabajo presentado en el 6th meeting of the IOBC-WPRS Working Group Multitrophic Interactions in Soil, celebrado en Córdoba (España) del 4 al 7 de abril de 2011.

Póster presentado en el 6th meeting of the IOBC-WPRS Working Group Multitrophic Interactions in Soil, celebrado en Córdoba (España) del 4 al 7 de abril de 2011., Olive (Olea europaea L. subs. europaea) is one of the most important crops in Spain with > 2.4 million ha. During the last two decades phytosanitary status of olive orchards is being threatened due to some diseases caused by soilborne pathogens, mainly Verticillium wilt (VW) caused by Verticillium dahliae (VD) which is steadily increasing both in severity and extension. The main objective of this study was to determine the effect of olive management system on the natural suppressiveness of olive orchards soils to VD and how this may be related with biological and functional indicators of soil quality. An in planta bioassay was developed for assessing the natural s uppressiveness to VD of a collection of 93 soils using highly cond ucive conditions for VW development. Results showed a wide range of variation among the differen t soils in their ability to suppress VD, with 25.8, 49.5 and 24.7% of soils showing low, moderate and h igh suppressiveness according to the VW severity developed in the test plants growing in those soils compared to that developed in plants in sterile and pasteurized artificial soils. Canonical multivariat e discriminant analysis is being used to identify physicochemical, functional or biological indicator s that may be involved in the natural suppression to VD of those soils., This research was supported by grants AGL2008-00344 and HA2008-0014 from ‘Ministerio de Ciencia e Innovación’ of Spain and the European Soc ial Fund.

Olive is a key crop, both economically and sociall y in southern Spain. Although the interaction of plant-parasitic nematodes with free- living nematodes could play an important role in their control, it is not known how the olive genoty pes may influence the rhizosphere environment and consequently the balance between plant-parasitic an d free-living nematodes. The aim of this work was to study the nematode communities associated with r hizospheric soil and roots of a collection of 16 olive cultivars from a world olive germplasm bank a t southern Spain and assess whether olive genotype determine differences in the nematofauna p resent in their rhizosphere. Classical nemato- logical analysis, including soil nematode extractio n, species counting and morphological identificatio n was performed. In addition, total soil DNA was extr acted from the same samples and used for analysis of genetic diversity of total nematode populations using Terminal Restriction Fragments Length Polymorphism (T-RFLPs) analysis of the 18S rDNA gen e. Results showed that 24 taxa belonging to 8 genera (including Criconemoides, Ditylenchus, Filenchus, Helicotylenc hus, Merlinius, Paratylenchus, Tylenchus, and Xiphinema ) and 8 families (including Aphelenchidae, Belonola imidae, Criconema- toidae, Hoplolaimidae, Longidoridae, Sphaerularioid ea, Tylenchidae and Tylenchulidae) of plant- parasitic nematodes were present, with one species ( Helicotylenchus digonicus ) being prevalent in all samples. There was no clear relationship between in cidence or population density of each nematode species with the olive genotype. General nematode m aturity index (MI) values were inconsistent between olive varieties and did not therefore disti nguish differences in the levels of soil disturbanc e. However, plant-parasitic nematode index (PPI) was s ignificantly modified by olive variety within the single field. In addition, T-RFLP analysis showed d ifferences between olive cultivars, with samples associated with specific varieties showing higher d iversity of nematode taxa., Authors gratefully acknowledge MICINN Projects AGL2 008-00344/AGR and HA2008-0014 and FEDER financial support from the European Union.

[EN] The invention relates to bacterial strains of Duganella spp., obtained from the rhizosphere of wild and cultivated olive trees. Preferably, the strains are CECT 7779, CECT 7780 and CECT 7781, and more preferable still, the bacterial strain is CECT 7780. In addition, the invention relates to combinations thereof with other microorganisms and to compositions comprising the aforementioned pro ducts , as well as to a method for the production of violacein and to the violacein produced for bioteclmological and agronomic use. [ES] La presente invención se refiere a unas cepas bacterianas de Duganella spp. obtenidas de la rizosfera de olivos silvestres y cultivados. Preferentemente las cepas son CECT 7779, CECT 7780 y CECT 7781 y más preferentemente la cepa bacteriana es CECT 7780. Además, la presente invención se refiere a sus combinaciones con otros microorganismos y a las composiciones que comprenden los productos anteriores, así como a un procedimiento para la producción de violaceína y a la violaceína producida para su aplicación biotecnológica y agronómica. Peer reviewed Consejo Superior de Investigaciones Científicas A1 Solicitud de patente con informe sobre el estado de la técnica

Las enfermedades son un componente de significación en la producción agrícola actual porque pueden reducir el rendimiento potencial o causar devastación en los cultivos de plantas; y la extensión y severidad de sus ataques va unida a los cambios que se están produciendo en los sistemas de manejo que dan lugar a mejoras en el rendimiento o modificaciones en los ambientes de producción. El control eficiente de las enfermedades requiere necesariamente la determinación exacta, rápida e informativa de su etiología, incluyendo la evaluación de la historia y potencial evolutivo en las poblaciones del agente causal, así como de la composición y diversidad de la microbiota beneficiosa asociada con el crecimiento vegetal, que hagan posible la aplicación eficiente de las acciones de prevención en que se fundamenta el manejo sostenible de dichas enfermedades. Las tecnologías de base molecular y biotecnológica ofrecen una excelente oportunidad para mejorar nuestras capacidades para satisfacer dichos requisitos. En este trabajo se ilustran tales oportunidades utilizando como ejemplo los resultados alcanzados por el Grupo AGR136 ‘Sanidad Vegetal’ durante el desarrollo de programa de investigación llevados a cabo sobre enfermedades importantes en la agricultura andaluza., Las investigaciones del Grupo AGR136 referidas en este trabajo fueron financiadas por proyectos de: Fundación Areces; Ministerio de Educación y Ciencia (AGR89-0533-CO2-01, AGF92-0910-CO2-01, AGF93-0740-CO2-01, AGF95-0985-CO2-01, OLI96-2131, AGF97-14791, FD97-0763-CO3-01, 1FD97-1322-CO4-02, AGL2000-1444-CO2-01, AGL 2003-00640, AGL 2004-01231, HF2004-0096, y AGL2005-00751); Unión Europea (AGRE-CT90-0051, ERB CT96-2015, y QLK5-CT1999-01523); y Junta de Andalucía (Grupo PAI AGR-136).

Método para la detección, identificación y cuantificación de Peronospora arborescens por PCR cuantitativa en tiempo real. El método para la cuantificación de Peronospora arborescens por PCR cuantitativa (qPCR) en una muestra biológica, comprende extraer el ADN contenido en dicha muestra biológica y amplificarlo mediante qPCR. De aplicación en la cuantificación de P. arborescens., Consejo Superior de Investigaciones Científicas (España), ALCALIBER SA, Universidad de Córdoba, A1 Solicitud de patentes con informe sobre el estado de la técnica

Prioridad: 22.07.2009. - Titulares: Universidad de Cordoba. - ALCALIBER S.A. - Consejos Superior de Investigaciones Científicas (CSIC), [ES] El método para la cuantificación de peronospora arborescens por PCR cuantitativa (qPCR) en una muestra biológica comprende extraer el ADN contenido en dicha muestra biológica y amplificarlo mediante qPCR. De aplicación en la cuantificación de P. arborescens, [EN]The method for quantifYing Peronospora arborescens by quantitative PCR (qPCR) in a biological sample comprises extracting the DNA contained in said biological sample and amplifying that DNA by means of qPCR. The invention is ofuse in the quantification of P. arborescens., Universidad de Cordoba, ALCALIBER SA, Consejo Superior de Investigaciones Científicas (España), A2 Solicitud de patentes sin informe sobre el estado de la técnica

Fecha de solicitud: 10-07-2008.- Titular: Consejo Superior de Investigaciones Científicas (CSIC), The method for detecting and identifying Peronospora arborescens is based on amplification of specific regions of said pathogen. To do this, a number of oligonucleotide primers specific for P. arborescens were designed, by means of enzymatic amplification reaction, for identifying and detecting said pathogen. This method is useful in agriculture and provides an early diagnosis of the disease (mildew) during asymptomatic states of plants, which facilitates efficient control thereof, and sanitary certification of the seed thereby preventing distribution of the pathogen in growing areas where it is not present., El método para detectar y identificar arborescens de los Peronospora es basado en la amplificación de las regiones específicas de patógeno dicho. Para hacer esto, un número de oligonucleótidos cebadores específicos para el P. los arborescens eran diseñados, por medio de la reacción de amplificación enzimática, porque de identificar y de detectar el patógeno dicho. Este método es útil en agricultura y proporciona un diagnóstico temprano de la enfermedad (moho) durante el estado asintomático de plantas, que facilita el control eficaz de eso, y la certificación sanitaria de la semilla de tal modo que evita la distribución del patógeno en áreas del crecimiento donde no está presente.