Unravelling Micromonospora interactions with its host plant and the associated microbioma

[ES] La Unión Europea depende en gran medida de las importaciones de soja (> 70%) como fuente de proteínas, ya que la producción local apenas cubre el 5% de la demanda interna. Por ello, es necesario explorar fuentes alternativas para reducir esta dependencia. Entre las leguminosas, Lupinus angustifolius es una opción dado su alto valor proteico y su uso para la alimentación animal y humana. Esta leguminosa es una planta autóctona del continente europeo, que está bien adaptada a las condiciones climáticas de otras regiones como puede ser Australia o América. También crece de forma silvestre en suelos pobres gracias a su capacidad para fijar nitrógeno en simbiosis con bacterias. La adaptación de dicha planta puede deberse en parte a los microorganismos asociados a sus raíces, que le proporcionan estabilidad y resistencia, además de moléculas promotoras del crecimiento vegetal y nutrientes. Las comunidades microbianas asociadas a las plantas se ven influenciadas por diversos factores como son el genotipo/especie del huésped, el tipo de suelo, compartimento de la planta y estación climática, entre otros. Separar estos factores para saber cuáles son los que más influyen en la asociación de microorganismos a las plantas es una tarea muy complicada puesto que ninguno se da de forma independiente. En el primer capítulo de esta tesis doctoral, se abordó esta temática estudiando las variaciones estacionales y geográficas de la microbiota del suelo, y caracterizando el microbioma asociado a la planta Lupinus angustifolius en diferentes condiciones de cultivo mediante técnicas independientes de cultivo. En el segundo capítulo, el objetivo fue el aislamiento e identificación molecular de la comunidad bacteriana presente en los distintos tejidos de la planta y la generación de una colección de cepas asociada al microbioma de L. angustifolius. Con los resultados obtenidos en los dos primeros capítulos, se describió por primera vez el microbioma core de la planta L. angustifolius. En el tercer y último capítulo de esta tesis doctoral se trató de descifrar las interacciones de Micromonospora con su planta huésped y el microbioma asociado, empleando para tal fin la información obtenida en los capítulos anteriores. Se desarrollaron siete comunidades sintéticas que se inocularon en experimentos in planta, en condiciones de invernadero en un suelo con su comunidad natural, y en un sistema gnotobiótico con un sustrato estéril. Posteriormente se evaluó mediante técnicas independientes de cultivo cómo se ensamblaban los microorganismos a la raíz y cuál era el efecto de las distintas SynComs en la planta huésped y el microbioma circundante. [EN] The European Union highly depends on soy imports (> 70%) as a protein source since local production barely covers 5% of its internal demand. Thus, it is necessary to explore alternative sources to reduce this dependency. Among legumes, Lupinus angustifolius is an important alternative given its high protein value and use for animal and human nutrition. This legume is a native plant of Europe, well adapted to the climatic conditions of many countries. It also thrives in poor soils due to its capacity to fix nitrogen. Plant adaptation may be partly due to the microorganisms associated with its roots, providing stability and resilience, in addition to plant growth promoting molecules and nutrients. Plant-associated microbial communities are influenced by several factors such as host genotype/species, soil type, plant compartment and climatic season, among others. Separating these factors to understand which are the most influential in the association of microorganisms to plants is a very complex task as they do not occur independently. In the first chapter of this doctoral thesis, this topic was addressed by studying seasonal and geographical variations in the soil microbiota, and characterizing the microbiome associated with the plant Lupinus angustifolius under different cultivation conditions using an independent culture methodology. The results of the soil samples analysed suggest that the difference in the microbial community composition observed between the two sampling locations, Cabrerizos and Salamanca, was partly due to differences in soil conditions. None of the communities analysed (bacterial and fungal) showed differences in alpha diversity (Shannon index) between the climatic seasons in which the samples were collected. Beta diversity (Bray-Curtis-based principal coordinate analysis) for both microbial communities separated the samples into two groups according to soil type. In the case of bacteria, it was observed that, in addition, subgroups were formed according to the climatic seasons for the Salamanca soil. Interestingly, this also occurred with the fungal communities, where the samples were separated by season in both soil types. These results suggest that the main difference in soil microbial communities is due to edaphic properties, although environmental factors such as temperature, humidity or rainfall also influence the diversity of soil microbial communities. In addition, the microbiome associated with the legume Lupinus angustifolius cultivated under natural and greenhouse conditions was also characterized. For this purpose, wild and greenhouse-grown plants were collected from the same locations and analysed by 16S rRNA gene and ITS-2 gene profiling. Bacterial communities were characterized in the different plant compartments (rhizosphere, roots, nodules and leaves) while ITS profiles were restricted to the soil and rhizosphere. As previously reported for other plants, the highest richness was found in the rhizosphere, followed by the roots, leaves, and nodules. Within the rhizosphere, the bacterial richness in the in Salamanca plants was lower, especially for the field samples, probably affected by a pH below 7 and high amounts of P and K. In general, the compartments from the plants grown under greenhouse conditions showed a slightly higher bacterial diversity when compared to the wild plants. Within the fungal communities, the Shannon index was significantly higher in soil than rhizosphere samples (P1% and designed several isolation protocols. A total of 722 bacterial strains were isolated. As expected, the highest number of isolates was obtained in the rhizosphere compartment and a similar pattern was observed with a decreasing diversity gradient starting from the rhizosphere followed by the roots, leaves and nodules. In total, 87 different genera were identified, of which 19 had more than 10 isolates. The most abundant strains were identified in the genera Pseudomonas, Streptomyces, Agrobacterium, Bacillus and Pseudoclavibacter. In this work, 51.9% of the searched genera were isolated, and 74.7% of the isolated genera were identified by metagenomics, but 19.6% could not be detected in any plant compartment by metagenomics. Plant pathogenicity assays showed that 29% of the L. angustifolius isolates were potentially pathogenic for Arabidopsis thaliana Col-0. In turn, 394 strains (55%) were found to be non-pathogenic and 116 (16%) promoted the growth of A. thaliana. Analysis of metagenomics and culturomics results identified a core microbiome of the host plant L. angustifolius that included Acidovorax, Bradyrhizobium, Caulobacter, Chitinophaga, Flavobacterium, Kribella, Massilia, Pseudomonas, Pseudoxanthomonas, Rhizobium, Sphingomonas, Streptomyces and Variovorax. The composition and diversity of the identified host plant-associated bacteriome varied slightly between sampling locations and growing conditions. The genera identified as the core microbiome were present in more than 80% of the samples analysed. In chapter three of this work, the aim was to decipher the interactions of Micromonospora with its host plant and the associated microbiome, using the information obtained in the previous two chapters. Seven different synthetic communities (SynComs) were designed using bacterial strains isolated from the rhizosphere and roots of L. angustifolius to study their effect on the root and rhizosphere of the plant. In addition, we wanted to learn if the selected strains had any effect on the host plant and the natural bacterial communities present in the cultivation soils. After obtaining the genomes of the bacterial strains included in the different SynComs, a comparative genomic analysis was carried out, confirming that all the selected strains had genes with functions related to plant association and growth promotion. Plants were grown for 8 weeks in unsterilised soil under greenhouse conditions, and several plant parameters were measured and compared against the control plants (uninoculated). The plants inoculated with SynCom_7 showed the best growth and development. Furthermore, 16S rRNA gene profiling showed that the soil samples were the most diverse, followed by rhizosphere and roots (alpha diversity) (Figs. 54 and 55). Beta diversity grouped the samples into three clusters according to compartments: soil, rhizosphere and roots. In addition, a clustering pattern was observed for the SynComs inoculated in the root samples. All consortia that contained the nitrogen fixer, Bradyrhizobium sp. in the synthetic community formed one cluster, while the rest of the SynComs were recovered in a second cluster. The analysis of the bacterial composition of the bulk soil samples confirmed that the synthetic communities did not affect the composition of the soil where the plant was growing. However, when we studied the bacterial composition in the rhizosphere, a slight variation was observed, and the bacterial community of root samples was greatly influenced by the inoculated SynComs. The second part of this chapter consisted in the evaluation of the different SynComs on L. angustifolius plants grown in sterile soil under a gnotobiotic system. As in the first experiment, several growth parameters were registered, observing that plants inoculated with SynCom_7 showed the highest growths, again. Pseudomonas sp. Strain CRA141 showed the closest association with the roots. This result is not unexpected as it is well known that many Pseudomonas strains associate to plant roots. In addition, it was found that Micromonospora sp. Lupac 08 was detected in the rhizosphere and roots, and while this actinobacterium is not part of the core microbiome, it could be considered a "satellite" microorganism with important beneficial functions for the plant. Plant gene expression was related to the effect of the SynComs inoculated. When inoculated consortia included the Bradyrhizobium strain, very little differences were found when compared to the control plants, however, when only the Micromonospora strain and/or the other members of the SynComs were added, the differential gene expression increased threefold (Fig. 62). Gene ontology enrichment analyses revealed that those functions that were enriched by inoculating the different SynComs were clearly related to plant-microbe interaction functions. The same was observed for the enriched metabolic pathways when KEGG analysis was performed

Pharmaceutical Pollution in Aquatic Environments: A Concise Review of Environmental Impacts and Bioremediation Systems

MO received a Ph.D. grant from the Junta de Castilla y Leon (Spain). Open Access Funding was provided by the University of Helsinki.The presence of emerging contaminants in the environment, such as pharmaceuticals, is a growing global concern. The excessive use of medication globally, together with the recalcitrance of pharmaceuticals in traditional wastewater treatment systems, has caused these compounds to present a severe environmental problem. In recent years, the increase in their availability, access and use of drugs has caused concentrations in water bodies to rise substantially. Considered as emerging contaminants, pharmaceuticals represent a challenge in the field of environmental remediation; therefore, alternative add-on systems for traditional wastewater treatment plants are continuously being developed to mitigate their impact and reduce their effects on the environment and human health. In this review, we describe the current status and impact of pharmaceutical compounds as emerging contaminants, focusing on their presence in water bodies, and analyzing the development of bioremediation systems, especially mycoremediation, for the removal of these pharmaceutical compounds with a special focus on fungal technologies.Junta de Castilla y LeonUniversity of Helsink

Pharmaceutical Pollution in Aquatic Environments: A Concise Review of Environmental Impacts and Bioremediation Systems

The presence of emerging contaminants in the environment, such as pharmaceuticals, is a growing global concern. The excessive use of medication globally, together with the recalcitrance of pharmaceuticals in traditional wastewater treatment systems, has caused these compounds to present a severe environmental problem. In recent years, the increase in their availability, access and use of drugs has caused concentrations in water bodies to rise substantially. Considered as emerging contaminants, pharmaceuticals represent a challenge in the field of environmental remediation; therefore, alternative add-on systems for traditional wastewater treatment plants are continuously being developed to mitigate their impact and reduce their effects on the environment and human health. In this review, we describe the current status and impact of pharmaceutical compounds as emerging contaminants, focusing on their presence in water bodies, and analyzing the development of bioremediation systems, especially mycoremediation, for the removal of these pharmaceutical compounds with a special focus on fungal technologies.Peer reviewe

Metabolic Capability of Penicillium oxalicum to Transform High Concentrations of Anti-Inflammatory and Analgesic Drugs

Olicón-Hernández would like to thank Consejo Nacional de Ciencia y Tecnología (CONACyT) for the postdoc fellowships (231581/454815). E. A. would like to thank the Ministry of Economy and Competitiveness (MINECO) and European Regional Development Fund (ERDF) funds [RYC-2013-12481] and [CTM- 2017-84332, MINECO/AEI/FEDER, UE].Non-steroidal anti-inflammatory drugs (NSAIDs) and analgesics are two of the most employed drug groups around the world due to their use in the treatment of edema and pain. However, they also present an ecological challenge because they are considered as potential water pollutants. In this work, the biodegradation of four NSAIDs (diclofenac, ibuprofen, naproxen and ketoprofen) and one analgesic (acetaminophen) at 50 µM (initial concentration) by Penicillium oxalicum, at both flask and bioreactor bench scales, was evaluated. An important co-metabolic mechanism as part of the global bioremediation process for the elimination of these drugs was observed, as in some cases it was necessary to supplement glucose to achieve a 100% removal rate: both individually and as a complex mixture. Identical behavior in the implementation of a fluidized bench-scale batch bioreactor, inoculated with pellets of this fungus and the complex mix of the drugs, was observed. The role of the cytochrome P450 enzymes (CYP) in the biodegradation of the drugs mix were evidenced by the observation of hydroxylated by-products. The results on the reduction of toxicity (micro and phyto) were not conclusive; however, a reduction in phytotoxicity was detected.MINECO-ERDF funds [MINECO/AEI/ERDF, UE] RYC-2013-12481 CTM-2017-84332Consejo Nacional de Ciencia y Tecnologia (CONACyT) 231581/45481

Contextos educativos : revista de educación

Resumen basado en el de la publicaciónTítulo, resumen, palabras clave en español e inglesSe centra en el diseño de un Proyecto de intervención en educación del ocio para la infancia en Honduras desde un enfoque humanista. Para ello se describe el contexto de aplicación del proyecto atendiendo al marco legal educativo vigente en Honduras; la descripción del Centro de Innovación Educativa en el que se contextualiza el proyecto de intervención, perteneciente a la Universidad Pedagógica Nacional Francisco Morazán; y los programas y actividades que dan contenido a la intervención diseñada.ES

Six novel Micromonospora species associated with the phyllosphere and roots of leguminous plants: Micromonospora alfalfae sp. nov., Micromonospora cabrerizensis sp. nov., Micromonospora foliorum sp. nov., Micromonospora hortensis sp. nov., Micromonospora salmantinae sp. nov., and Micromonospora trifolii sp. nov

13 páginas, 4 figuras, 2 tablasSix actinobacterial strains isolated from diverse legume tissues collected in various locations in Spain were characterized to determine their taxonomic status. Using 16S rRNA gene sequencing, the strains were primarily identified as members of the genus Micromonospora with more than 99 % similarity. Digital DNA-DNA hybridization values and average nucleotide identities between the six strains and the nearest type strains confirmed that each strain represented a novel species. Genome sequences were analysed to infer their metabolic profiles, their potential to produce secondary metabolites and plant growth promoting features. Chemotaxonomic and physiological studies were carried out to complete the phenotypic characterization and to distinguish the new Micromonospora species. The genomic and phenotypic characterization of the Micromonospora strains strongly support their classification as representatives of new species with the following names: Micromonospora alfalfae sp. nov., Micromonospora cabrerizensis sp. nov., Micromonospora foliorum sp. nov., Micromonospora hortensis sp. nov., Micromonospora salmantinae sp. nov. and Micromonospora trifolii sp. nov., with the type strains MED01T, LAH09T, PSH25T, NIE111T, PSH03T and NIE79T, respectively.This research was supported by the Spanish Ministry of Sciences, Innovation and Universities (MICINN) under project PGC2018-096185-B-I00 to M.E.T.Peer reviewe