Biotecnología agrícola

The domestication of plants can be considered as the first genetic manipulation of living organisms and it is associated with the origin of agriculture, and represents the major contribution of biotechnology to the development of mankind. The challenges the agricultural sector faces in the coming decades are population growth and the increasing demand for raw materials for food, feed, as well as the consumer preferences, environmental and legal aspects, and the overall globalisation of the economy. The agricultural biotechnology applications will undoubtedly play a role in meeting these challenges. Spain has competent scientific groups devoted to basic and applied research, with a wide knowledge on agricultural biotechnology. The transfer of the business sector needs to the scientific field, coordinating the R&D companies and research groups from universities and public research organisms is an objective to be attained in the future.La domesticación de plantas, primera manipulación genética de los organismos vivos y origen de la Agricultura, representa la mayor contribución de la biotecnología al desarrollo de la humanidad. Los retos a superar en las próximas décadas para el sector agrario están determinados por el aumento demográfico y la demanda de materias primas para alimentación humana y animal, por las preferencias en el consumo, por aspectos medioambientales y legales, y por la globalización de la economía. Las aplicaciones de la biotecnología agrícola van a jugar un papel incuestionable en el logro de estos retos. Nuestro país posee unos buenos grupos en investigación básica y aplicada, con amplios conocimientos de la agrobiotecnología. El traslado de las necesidades del sector empresarial a la investigación, coordinando los departamentos de I+D+I de las empresas y los grupos de investigación de las universidades y organismos públicos de investigación es una finalidad a conseguir en el futuro

Impact of Postharvest Handling on Preharvest Latent Infections Caused by Monilinia spp. in Nectarines

Latent infections caused by Monilinia spp. in nectarines cause great economic losses since they are not detected and rejected at harvest and can appear at any time post-harvest, even at the consumer’s home. The effect of a pre-cooling chamber, water dump operation, and cold-storage chamber on the activation and/or development of preharvest latent infections caused by Monilinia spp. on nectarines were studied under different postharvest conditions: (a) cold storage for 0, 1, or 3 d at 4 °C at either 75% relative humidity (RH) or 100% RH before water dumping, (b) water dumping for 10 minutes at 15 °C, and (c) cold storage for 0, 3, or 10 d at 4 °C at either 75% RH or 100% RH after water dumping. These storage conditions were transformed to fungal physiological time. For visualization of the latent infections caused by Monilinia spp., the nectarines were placed in sterile paper bags and frozen at −20 °C for 48 h in order to damage the epidermis. To compare different handling scenarios, the incidence of latent infection was modelled for physiological time description by a modified Gompertz model. The activation and/or development of preharvest natural latent infections caused by Monilinia spp. at postharvest was mainly related to temperature and incubation time at postharvest. Storing nectarines with any postharvest handling less than 11 days at 4 °C avoids brown rot symptoms and reduced the activation and/or development of pre-harvest latent infections caused by Monilinia spp., while more cold days caused the exponential phase of latent infection activation and/or development. The Gompertz model employed could be used for predicting the activation and/or development of latent infection caused by Monilinia spp. at postharvest conditions and looks at the postharvest life. To our knowledge, this is the first time that the effects of post-harvest handling on latent infections in fruit have been studied.info:eu-repo/semantics/publishedVersio

Development of brown rot epidemics in Spanish peach orchards

A new approach to modelling epidemics of brown rot caused by Monilinia spp. in Ebro Valley peach orchards has been developed. This compartmental model was subdivided according to the phenological stages in which the disease can develop (blossom, immature fruit, and ripe fruit). Information host susceptibility, primary and secondary inoculum sources and latent infections in immature fruit was taken into account. The compartmental model is described by a system of differential equations, and is simple enough to allow an analytical study of the main epidemiological factors that determine the rate of disease progress during a single growing season. The proposed model fits well to the epidemic pattern of brown rot observed in northeastern Spain. The transmission of the disease as a nonlinear term implied that small changes in the infection rate had a large effect on the development of the disease. The model has confirmed the usefulness of removing mummies (infected fruit that remains in the crop during winter) from the field to reduce the final incidence of the disease. In addition, all control measures that reduce the rate of secondary infection in ripe fruit, either through the use of more resistant varieties or the use of fungicides, are effective in reducing brown rot incidence. The proposed epidemic model is flexible and allows to add complexities to the system and evaluate the effectiveness of different control strategies.info:eu-repo/semantics/publishedVersio

Transformation of Penicillium rubens 212 and Expression of GFP and DsRED Coding Genes for Visualization of Plant-Biocontrol Agent Interaction

Strain 212 of Penicillium rubens (PO212) is an effective fungal biological control agent against a broad spectrum of diseases of horticultural plants. A pyrimidine auxotrophic isolate of PO212, PO212_18.2, carrying an inactive pyrG gene, has been used as host for transformation by positive selection of vectors containing the gene complementing the pyrG1 mutation. Both integrative and autonomously replicating plasmids transformed PO212_18.2 with high efficiency. Novel PO212-derived strains expressed green (sGFP) and red (Ds-Red Express) fluorescent reporter proteins, driven by the A. nidulans gpdA promoter. Fluorescence microscopy revealed constitutive expression of the sGFP and Ds-Red Express proteins, homogenously distributed across fungal cells. Transformation with either type of plasmid, did not affect the growth and morphological culture characteristics, and the biocontrol efficacy of either transformed strains compared to the wild-type, PO212. Fluorescent transformants pointed the capacity of PO212 to colonize tomato roots without invading plant root tissues. This work demonstrates susceptibility of the biocontrol agent PO212 to be transformed, showing that the use of GFP and DsRed as markers for PO212 is a useful, fast, reliable and effective approach for studying plant–fungus interactions and tomato root colonization

Labeling of Monilinia fructicola with GFP and Its Validation for Studies on Host-Pathogen Interactions in Stone and Pome Fruit

To compare in vivo the infection process of Monilinia fructicola on nectarines and apples using confocal microscopy it is necessary to transform a pathogenic strain with a construct expressing a fluorescent chromophore such as GFP. Thus, germinated conidia of the pathogen were transformed with Agrobacterium tumefaciens carrying the plasmid pPK2-hphgfp that allowed the expression of a fluorescent Hph-GFP chimera. The transformants were selected according to their resistance to hygromycin B, provided by the constitutive expression of the hph-gfp gene driven by the glyceraldehyde 3P dehydrogenase promoter of Aspergillus nidulans. The presence of T-DNA construct in the genomic DNA was confirmed by PCR using a range of specific primers. Subsequent PCR-mediated analyses proved integration of the transgene at a different genomic location in each transformant and the existence of structural reorganizations at these insertion points. The expression of Hph-GFP in three independent M. fructicola transformants was monitored by immunodetection and epifluorescence and confocal microscopy. The Atd9-M. fructicola transformant displayed no morphological defects and showed growth and pathogenic characteristics similar to the wild type. Microscopy analysis of the Atd9 transformant evidenced that nectarine infection by M. fructicola was at least three times faster than on applesinfo:eu-repo/semantics/publishedVersio

Occupation and mobility in high-mountain agropastoral societies: a proposal for an ethnoarchaeological study in the Jbel Sirwa (Anti-Atlas, Morocco)

Desde los orígenes de la domesticación, las sociedades agropastoriles de alta montaña han sido un ejemplo excepcional de adaptación y resiliencia a territorios inhóspitos, ya que requieren de un delicado equilibrio entre las actividades de trashumancia y cultivo para poder asegurar la continuidad del grupo. Los trabajos arqueológicos sobre estas sociedades han dejado tras de sí numerosas preguntas sobre la relación entre los humanos y el medio de montaña. Todos estos interrogantes nos han llevado a estudiar la materialidad de la vida cotidiana de las poblaciones agropastoriles del Jbel Sirwa. Así pues, desde el proyecto ARCHEOMOBAS, ofrecemos esta propuesta metodológica interdisciplinar para llevar a cabo estudios etnoarqueológicos de este tipo de comunidades en territorios montañosos.Since the first days of domestication, high mountain agropastoral societies have provided an exceptional example of adaptation and resilience to inhospitable territories, as the maintenance of population groups requires a delicate balance between transhumance and cultivation activities. Archaeological research into these societies has left numerous questions about the relationship between humans and the mountain environment unresolved. These questions have motivated the authors to explore the materiality of the daily life of the agropastoral populations of the Jbel Sirwa. Thus, with this proposal, the ARCHEOMOBAS project outlines an interdisciplinary methodological approach intended to carry out ethnoarchaeological studies of this kind of community in mountainous territories

Supervivencia de la terapia biológica en pacientes con artritis reumatoide de origen paraguayo

Introduction: The survival of biological therapies (BT) is considered an indirect measure of their efficacy and tolerability, which is why there is a growing interest in studying it. Currently, information on the survival of BT in Paraguayan patients with rheumatoid arthritis (RA) is limited. The development of biological therapies has allowed a better control of the inflammatory activity in rheumatoid arthritis (RA) and a greater number of patients achieve conremission. There is an increasing interest in studying the survival and efficacy of these agents in real life clinical practice. Objective: To analyze the survival of biological therapies administered to patients with RA from the Rheumatology Service of the Hospital Central del Instituto de Previsión Social (H.C.I.P.S). Methodology: Observational study of patients with RA who received biological therapies with follow-up at the Rheumatology Service of the H.C.I.P.S from January 2005 to December 2015. Survival was determined using the Kaplan Meier estimator. To analyze the effect of certain covariates, we used the Cox regression model. Results: 229 patients were included, and 258 TB were identified (adalimumab 58.1%, tocilizumab 20, 9% etanercept 18.9% and rituximab 1.9%). The mean survival of adalimumab, etanercept and tocilizumab was 246, 276 and 246 weeks respectively. In the analysis, the number of previous biological therapies was significantly associated with an increased risk of withdrawal. The number of previous biological therapies and treatment with tocilizumab were associated with an increased risk of withdrawal due to an adverse event. When analyzing the group of treatments withdrawn due to inefficacy, the number of previous biological therapies and adalimumab treatment were associated with an increased risk of discontinuation for this reason. Conclusion: In this series of Paraguayan patients with RA, the survival was greater for treatment with etanercept and variables associated with a lower survival vary according to the reason for withdrawal of treatment.Introducción: La supervivencia de las terapias biológicas (TB) es considerada una medida indirecta de su eficacia y tolerabilidad, por lo cual existe un interés creciente en estudiarla. Actualmente la información sobre la supervivencia de las TB en pacientes paraguayos con artritis reumatoide (AR) es limitada. Objetivo: Analizar la supervivencia de las TB administradas a pacientes con AR del Servicio de Reumatología del Hospital Central del Instituto de Previsión Social (IPS). Metodología: Estudio observacional, de una serie de pacientes con diagnóstico de AR que recibieron TB en el Servicio de Reumatología del Hospital Central del IPS durante el periodo comprendido entre enero del 2005 a diciembre del 2015. La supervivencia fue determinada con el estimador de Kaplan Meier. Para analizar el efecto de las covariables se utilizó el modelo de regresión de Cox. Resultados: Se incluyó a 229 pacientes, en los que se identificaron a 258 TB (adalimumab 58,1%, tocilizumab 20, 9% etanercept 18,9% y rituximab1,9%). La supervivencia media del adalimumab, etanercept y tocilizumab fue de 246, 276 y 246 semanas respectivamente. En el análisis global, el número de terapias biológicas previas (NT B) se asoció de forma significativa a un mayor riesgo de retirada. El NTB y el tratamiento con tocilizumab se asociaron a un mayor riesgo de retirada por un evento adverso. Al analizar el grupo de tratamientos retirados por ineficacia, el NTB y el tratamiento con adalimumab se asociaron a un mayor riesgo de discontinuación por este motivo. Conclusión: En esta serie de pacientes con AR de origen paraguayo, se objetivó que la supervivencia media fue mayor para el tratamiento con etanercept y que las variables asociadas a una menor supervivencia varían según el motivo de retirada del tratamiento

Proficiency of real-time PCR detection of latent Monilinia spp. infection in nectarine flowers and fruit

Rapid and reliable detection of Monilinia latent infections is needed to prevent and control dispersion of Monilinia spp. in infected localities and non-infected countries. A fast multiplex quantitative real-time PCR method (qPCR) for the detection and identification of Monilinia spp. latent infections in blossoms and fruit of nectarine trees (Prunus persica var. nucipersica) was tested in an inter-laboratory trial. The test performance study involving five laboratories was conducted to validate the sensitivity and specificity of several real-time PCR platforms for the detection of low amounts of Monilinia DNA (latent infections), using a common protocol, and to identify possible difficulties when these tests were implemented by diagnostic laboratories or national reference centres. The method has two hydrolysis probes distinguishing between Monilinia fructicola and M. fructigena/M. laxa. Validation included test performance accuracy, analytical specificity and sensitivity, repeatability, and reproducibility, as defined by standard PM7/98 of the European Plant Protection Organization (EPPO). All qPCR platforms detected Monilinia latent infections and mycelium samples with both hydrolysis probes, and healthy flowers and fruit samples gave negative results. The method specificity was consistent between different laboratories, despite different equipment used, and there were no laboratories with z-scores in the unacceptable region. Monilinia fructicola latent infection samples were correctly detected by all laboratories, but some M. laxa samples were cross-detected as if they were M. fructicola. Monilinia laxa cross-detection could be compensated by including the allelic discrimination step in qPCR runs, which permitted differentiating between M. fructicola and M. laxa samples. The inter-laboratory comparison demonstrated the robustness of the developed method and confirmed in-house validation data. This method could be used to detect latent infections of Monilinia in asymptomatic nectarine fruit and flowers

Epidemiological Studies of Brown Rot in Spanish Cherry Orchards in the Jerte Valley

Departamento de ​Protección Vegetal (INIA)Cherry brown rot caused by Monilinialaxa was observed and estimated in organic cherry orchard located in the Jerte Valley between 2013 and 2018 (Cáceres, Spain). Climatic variables were collected from this orchard and also from a nearby weather station. The primary inoculum of the pathogen recorded in March was detected in overwintered mummified fruits, ground mummies, and necrotic twigs and was a function of the average temperature of the previous three months (December, January, and February). The first symptoms of brown rot could be observed on flowers until fruit set in April. The months of March and April were identified as the critical period for cherry brown-rot development. A significant positive correlation was identified between brown rot observed at harvest and the mean number of consecutive days in each fortnight of March and April when the percent relative humidity was above 80%. Brown-rot incidence observed over the 6 years ranged from 0 to 38%. More than 11 days with relative humidity >80% in each fortnight of critical period would mean 100% of cherry brown rot at harvest. A forecasting model could be used to predict brown rot infection in Jerte Valley cherries.This work was supported by national project CC13-047 from INIA and CICYTEX, AGL2017- 84389-C2-2-R from the Ministry of Science, Innovation and Universities (MCIU, Spain), and by supra-autonomic operational group FITOSCEREZO funded by Horizon 2020 in the framework of the national program for the Rural Development 2014–2020 by the European Agricultural Fund for Rural Development (EAFRD) and the Spanish Ministry of Agriculture, Fisheries, and Food with the Agricultural European Innovation Partnership (EIP-AGRI).Peer reviewed12 Págs. (This article belongs to the Special Issue Control of Fungal Diseases in Crops