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Publication Open Access Comparative genomics of native plasmids from plant pathogenic Gammaproteobacteria(Oxford University Press, 2025-04-01) Urriza Leoz, Miriam; Dimaria, Giulio; Oliveira, Luiz Orlando de; Catara, Vittoria; Murillo Martínez, Jesús; Agronomía, Biotecnología y Alimentación; Agronomia, Bioteknologia eta Elikadura; Institute for Multidisciplinary Research in Applied Biology - IMAB; Universidad Pública de Navarra / Nafarroako Unibertsitate PublikoaPlasmids are key in the evolution and adaptation of plant pathogenic Gammaproteobacteria (PPG), yet their diversity and functional contributions remain underexplored. Here, comparative genomics revealed extensive variation in plasmid size, replicon types, mobility, and genetic content across PPG. Most plasmids are small (< 200 kb), except in Pantoea, exhibiting high coding densities (76% to 78%). Five ancestral replicon types were identifed across multiple orders, indicating vertical descent yet effcient horizontal transfer across taxa, although with limited genetic conservation. Virulence plasmids are widespread (56% to 68%) but differ in virulence gene content across orders: type III effector (T3E) genes are common in Pseudomonas and Xanthomonas, but rare in Enterobacterales and Xylella, aligning with their smaller effector repertoires. Plasmids frequently carry regulatory genes, highlighting their role in bacterial phenotype modulation. Distinct patterns were observed among orders: Enterobacterales plasmids often harbor thiamin biosynthesis operons and transcriptional regulators but lack post-transcriptional regulators, while most Pseudomonas and Xanthomonas plasmids are mobile, enriched in T3E genes, and exhibit high insertion sequence densities, fostering DNA mobility. Resistance to ultraviolet light is common, but not to antimicrobial compounds. These fndings highlight the dynamic role of plasmids in spreading adaptive traits, shaping virulence, and driving the evolution of plant pathogenic bacteria.Publication Open Access Drought stress modifies the source-sink dynamics of nitrogen-fixing soybean plants prioritizing roots and nodules(Wiley, 2025-05-26) Rubia Galiano, María Isabel; Larrainzar Rodríguez, Estíbaliz; Arrese-Igor Sánchez, César; Ciencias; Zientziak; Institute for Multidisciplinary Research in Applied Biology - IMAB; Universidad Pública de Navarra / Nafarroako Unibertsitate PublikoaSoybean plants are one of the most cultivated legume crops worldwide. Their ability to establish nitrogen-fixing symbiosis with rhizobium bacteria allows the reduction of molecular nitrogen to ammonium, contributing to a reduction in the dependence on nitrogen fertilizers. However, nitrogen fixation is highly sensitive to environmental stresses, such as water deficit, and the regulatory mechanisms underlying this inhibition remain debatable. In the current study, we analyzed carbon (C) allocation dynamics in drought-stressed soybean plants following the application of [U-C-13]-sucrose to source leaves. Three sets of plants were analyzed: well-watered plants, mild drought, and severe drought-stressed plants. C-13 distribution was monitored for up to 6 h post-application. Under optimal water conditions, C-13 was mainly allocated to young (sink) leaves. During drought stress, transport trends changed, prioritizing C allocation primarily to the roots and nodules to a lesser extent. Metabolite profiling identified drought- and tissue-specific variations in the levels of the major C and N compounds.Publication Open Access Drought limits tree growth more than greenness and reproduction: insights from five case studies in Spain(KeAi Communications, 2025-08-01) Camarero, Jesús Julio; Rubio-Cuadrado, Álvaro; González de Andrés, Ester; Valeriano, Cristina; Pizarro, Manuel; Imbert Rodríguez, Bosco; Lo, Yueh-Hsin; Blanco Vaca, Juan Antonio; Ciencias; Zientziak; Institute for Multidisciplinary Research in Applied Biology - IMABDroughts impact forests by influencing various processes such as canopy greenness, tree growth, and reproduction, but most studies have only examined a few of these processes. More comprehensive assessments of forest responses to climate variability and water shortages are needed to improve forecasts of post-drought dynamics. Iberian forests are well-suited for evaluating these effects because they experience diverse climatic conditions and are dominated by various conifer and broadleaf species, many of which exhibit masting. We assessed how greenness, evaluated using the normalized difference vegetation index (NDVI), tree radial growth, and seed or cone production responded to drought in five tree species (three conifers: silver fir (Abies alba), Scots pine (Pinus sylvestris), and stone pine (Pinus pinea); two broadleaves: European beech (Fagus sylvatica) and holm oak (Quercus ilex) inhabiting sites with different aridity. We correlated these data with the standardized precipitation evapotranspiration index (SPEI) using the climate window analysis (climwin) package, which identifies the most relevant climate window. Drought constrained growth more than greenness and seed or cone production. Dry conditions led to high seed or cone production in species found in cool, moist sites (silver fir, beech, and Scots pine). We also found negative associations of cone production with summer SPEI in the drought-tolerant stone pine, which showed lagged growth−cone negative correlations. However, in the seasonally dry holm oak forests, severe droughts constrained both growth and acorn production, leading to a positive correlation between these variables. Drought impacts on greenness, growth, seed, and cone production depended on species phenology and site aridity. A negative correlation between growth and reproduction does not necessarily indicate trade-offs, as both may be influenced by similar climatic factors.Publication Open Access Integrated biowaste management by composting at a university campus: process monitoring and quality assessment(MDPI, 2025-03-07) Álvarez-Alonso, Cristina; Pérez-Murcia, María Dolores; Martínez-Sabater, Encarnación; Irigoyen Iriarte, Ignacio; Sánchez Arizmendiarrieta, Joseba; Plana, Ramón; López, Marga; Nogués, Isabel; Bustamante, María Ángeles; Agronomía, Biotecnología y Alimentación; Agronomia, Bioteknologia eta Elikadura; Institute for Multidisciplinary Research in Applied Biology - IMABThe sustainable management of biowaste, mainly food and pruning waste, is currently a challenge due to the increase in its production. The CaMPuSTAJE program, which has been implemented on the campus of the Public University of Navarre (UPNA) since 2019, is an excellent example of how the institution is addressing its strategic interests in sustainable waste management. The principal aim of this program is to manage the biowastes generated by the campus canteens through a simple community composting facility, involving UPNA students and graduates. This program aims to promote experiential learning and applied research in sustainability and circular economy, managing their own waste in a circular and local way. Thus, four composting sets of the CaMPuSTAJE program were evaluated by monitoring the process and the main chemical properties of the composting samples. Also, final composts were fully characterized to ensure the process reproducibility and efficiency and the absence of any hazard in the end-products. The final composts showed a significant agronomic quality, had low content of potentially toxic elements, and were free from phytotoxicity, thus being able to be reintroduced as an organic amendment at the university campus itself.Publication Open Access Baculovirus genetic diversity and population structure(MDPI, 2025-05-07) López Ferber, Miguel; Caballero Murillo, Primitivo; Williams, Trevor; Agronomía, Biotecnología y Alimentación; Agronomia, Bioteknologia eta Elikadura; Institute for Multidisciplinary Research in Applied Biology - IMABBaculoviruses can naturally regulate lepidopteran populations and are used as biological insecticides. The genetic diversity of these viruses affects their survival and efficacy in pest control. For nucleopolyhedroviruses, occlusion-derived virions and the occlusion body facilitate the transmission of groups of genomes, whereas this is not the case for granuloviruses. We review the evidence for baculovirus genetic diversity in the environment, in the host insect, and in occlusion bodies and virions. Coinfection allows defective genotypes to persist through complementation and results in the pseudotyping of virus progeny that can influence their transmissibility and insecticidal properties. Genetic diversity has marked implications for the development of pest resistance to virus insecticides. We conclude that future research is warranted on the physical segregation of genomes during virus replication and on the independent action of virions during infection. We also identify opportunities for studies on the transmission of genetic diversity and host resistance to viruses.Publication Open Access Broccoli (Brassica oleracea var. italica) biomass as a resource for obtaining glucosinolate extracts to control postharvest fungal diseases(Springer, 2025-05-27) Eugui Arrizabalaga, Daniel; Fernández San Millán, Alicia; Velasco, Pablo; Veramendi Charola, Jon; Rodríguez, Víctor Manuel; Poveda Arias, Jorge; Agronomía, Biotecnología y Alimentación; Agronomia, Bioteknologia eta Elikadura; Institute for Multidisciplinary Research in Applied Biology - IMABBroccoli (Brassica oleracea var. italica) is a crop of great agronomic and economic importance worldwide. Because its edible parts are the inflorescences, large quantities of non-commercial biomass are produced each year in the field and in the food industry. In order to develop a circular economy around the broccoli crop, the present work develops glucosinolates (GSL) extracts with antimicrobial capacity for postharvest use in tomato, apple and table white grape against fungal diseases produced by the pathogens Botrytis cinerea, Alternaria alternata and Penicillium expansum. GSL extracts from organic crop management reported a higher content of GSLs than conventional management. These extracts are not effective in the control of A. alternata and P. expansum, possibly due to the absence of sinigrin. Furthermore, the extracts were ineffective in the control of B. cinerea on table white grapes, possibly due to the non-climacteric fruit condition and an absence in the induction of ethylene-mediated plant defenses. However, intact GSL extracts were effective in controlling B. cinerea on apple, while the addition of myrosinase enzyme caused effectiveness also on tomato and apple. Therefore, obtaining GSL extracts with biopesticidal capacity against B. cinerea in postharvest could be a circular economy strategy for broccoli agriculture and industry.Publication Open Access Comparative enzymatic browning transcriptome analysis of three apple cultivars unravels a conserved regulatory network related to stress responses(BMC, 2025-04-12) Bielsa González, Francisco Javier; Grimplet, Jérôme; Irisarri, Patricia; Miranda Jiménez, Carlos; Errea, Pilar; Pina, Ana; Agronomía, Biotecnología y Alimentación; Agronomia, Bioteknologia eta Elikadura; Institute for Multidisciplinary Research in Applied Biology - IMABEnzymatic browning (EB) endangers the adaptation of apple fruit cultivars to new markets, affecting organoleptic properties and producing economic losses. Polyphenol oxidases and polyphenol compounds play a key role in EB development in apple. However, the regulation of apple response to EB remains to be uncovered. In this study, three apple cultivars with different EB phenotypes ranging from low to high browning in apple pulp were used to study transcriptomic changes over time after fresh cutting (0, 30 and 60 min). This study allowed the identification of 1448 differentially expressed genes (DEGs), revealing both shared and genotype-specific responses, particularly in the affected metabolic pathways associated with EB. At 60 min (T60 vsT0), 77 DEGs were shared by all genotypes, suggesting a conserved regulatory network. This network included genes encoding for protein families such as calcium-binding proteins, heat-shock proteins, redox-responsive transcription factors, WRKY family transcription factors, zinc finger family proteins and disease resistance proteins among others. A co-expressed gene cluster, identified through Weighed Gene Co-Expression Network Analysis (WGCNA), was found to correlate with EB and included 323 genes enriched in several biological terms according to Gene Ontology analysis. Moreover, a more detailed analysis of identified WGCNA gene cluster regulatory sequences allowed the detection of cis-regulatory elements belonging to CAMTA, WRKY and WUSCHEL transcription factor families. The identification of these sequences alongside with an abundant and diverse amount of overexpressed transcription factors from various families (WRKY, ERF, GRAS, GATA, etc.) point out to a highly regulated stress-response that is strictly connected to innate plant immunity. These findings provide valuable insights into the molecular mechanism involved in apple fresh-cut browning and offer new potential targets for EB regulation.Publication Open Access Alfalfa yield estimation using the combination of Sentinel-2 and meteorological data(Elsevier, 2025-03-19) Gámez Guzmán, Angie Lorena; Segarra, Joel; Vatter, Thomas; Santesteban García, Gonzaga; Araus, José Luis; Aranjuelo Michelena, Iker; Agronomía, Biotecnología y Alimentación; Agronomia, Bioteknologia eta Elikadura; Institute for Multidisciplinary Research in Applied Biology - IMAB; Gobierno de Navarra / Nafarroako GobernuaContext: Alfalfa (Medicago sativa L.) is one of the world's most important forages for livestock feeding. Timely yield estimates could provide information to guide management decisions to improve production. Since alfalfa crops typically undergo multiple harvests in a year and demonstrate rapid regrowth, satellite remote sensing techniques present a promising solution for alfalfa monitoring. Objective: To generate alfalfa yield estimation models at three phenological stages (early vegetative, late vegetative, and budding stages) using vegetation indices (VIs) derived from satellite Sentinel-2 images and their combination with meteorological data. Methods: We analyzed fields located in Navarre (northern Spain) over two consecutive seasons (2020 and 2021). To generate the yield estimation models, we applied a conventional multilinear regression and two machine learning algorithms (Least Absolute Shrinkage and Selection Operator - LASSO and Random Forest - RF). Results: Regardless of the statistical approach, the three phenological stages were not optimal when either VIs or meteorological data were used singularly as the predictor. However, the combination of VIs and meteorological data significantly improved the yield estimations, and in the case of LASSO model reached percentages of variance explained (R2) and normalized root mean square error (nRMSE) of R2= 0.61, nRMSE= 0.16 at the budding stage, but RF reached a R2= 0.44, nRMSE= 0.22 at the late vegetative stage, and R2= 0.36, nRMSE= 0.24 at the early vegetative stage. The most suitable variables identified were the minimum temperature, accumulated precipitation, the renormalized difference vegetation index (RDVI) and the normalized difference water index (NDWI). The RF model achieved more accurate yield estimations in early and late vegetative stages, but LASSO at bud stage. Conclusion: These models could be used for alfalfa yield estimations at the three phenological stages prior to harvest. The results provide an approach to remotely monitor alfalfa fields and can guide effective management strategies from the early development stages.Publication Open Access Cubierta vegetal bajo las cepas: una alternativa al control de las malas hierbas en los viñedos(INTIA (Tecnologías e Infraestructuras Agroalimentarias), 2023) Abad Zamora, Francisco Javier; Cibriain Sabalza, Félix; Sagüés Sarasa, Ana; Santesteban García, Gonzaga; Lezáun San Martín, Juan Antonio; Fabo Boneta, Jesús María; Virto Quecedo, Íñigo; Imbert Rodríguez, Bosco; Marín Arroyo, Remedios; Garbisu Crespo, Carlos; Ciencias; Zientziak; Agronomía, Biotecnología y Alimentación; Agronomia, Bioteknologia eta Elikadura; Institute for Multidisciplinary Research in Applied Biology - IMAB; Institute on Innovation and Sustainable Development in Food Chain - ISFOODEn este artículo se presentan los resultados obtenidos con una cubierta vegetal de trébol sembrada bajo las cepas para competir con las malas hierbas, de manera que no sea necesario recurrir al empleo de herbicidas o laboreos intercepas.Publication Open Access A novel Enterococcus-based nanofertilizer promotes seedling growth and vigor in wheat (Triticum aestivum L.)(MDPI, 2024-10-14) Batool, Salma; Safdar, Maryam; Naseem, Saira; Sami, Abdul; Saleem, Rahman Shah Zaib; Larrainzar Rodríguez, Estíbaliz; Shahid, Izzah; Ciencias; Zientziak; Institute for Multidisciplinary Research in Applied Biology - IMABExcessive use of chemical fertilizers poses significant environmental and health concerns. Microbial-based biofertilizers are increasingly being promoted as safe alternatives. However, they have limitations such as gaining farmers¿ trust, the need for technical expertise, and the variable performance of microbes in the field. The development of nanobiofertilizers as agro-stimulants and agro-protective agents for climate-smart and sustainable agriculture could overcome these limitations. In the present study, auxin-producing Enterococcus sp. SR9, based on its plant growth-promoting traits, was selected for the microbe-assisted synthesis of silver nanoparticles (AgNPs). These microbial-nanoparticles SR9AgNPs were characterized using UV/Vis spectrophotometry, scanning electron microscopy, and a size analyzer. To test the efficacy of SR9AgNPs compared to treatment with the SR9 isolate alone, the germination rates of cucumber (Cucumis sativus), tomato (Solanum lycopersicum), and wheat (Triticum aestivum L.) seeds were analyzed. The data revealed that seeds simultaneously treated with SR9AgNPs and SR9 showed better germination rates than untreated control plants. In the case of vigor, wheat showed the most positive response to the nanoparticle treatment, with a higher vigor index than the other crops analyzed. The toxicity assessment of SR9AgNPs demonstrated no apparent toxicity at a concentration of 100 ppm, resulting in the highest germination and biomass gain in wheat seedlings. This work represents the first step in the characterization of microbial-assisted SR9AgNPs and encourages future studies to extend these conclusions to other relevant crops under field conditions.Publication Open Access Unlocking Spanish pear genetic diversity: strategies for construction of a national core collection(Springer Nature, 2024-11-04) Irisarri, Patricia; Urrestarazu Vidart, Jorge; Ramos-Cabrer, Ana; Pereira-Lorenzo, Santiago; Velázquez-Barrera, María Encarnación; Díaz-Hernández, María Belén; Dapena, Enrique; Urbina, Valero; Dalmases, Josep; Ríos-Mesa, Domingo; Crespo Martínez, Sara; Loidi Erviti, Maite; Santesteban García, Gonzaga; Ascasíbar-Errasti, Javier; Errea, Pilar; Miranda Jiménez, Carlos; Pina, Ana; Agronomía, Biotecnología y Alimentación; Agronomia, Bioteknologia eta Elikadura; Institute for Multidisciplinary Research in Applied Biology - IMABSpanish pear germplasm collections are crucial for preservation, research, and breeding efforts. However, genetic diversity and structure is unknown at national level. A coordinated national project analyzed 1251 accessions from 7 Spanish pear collections using an internationally recognized set of 14 SSRs to enhance the utilization of these collections. Key findings included the identification of 760 unique genotypes (490 diploids and 270 triploids). Notably, genotypes represented by a single accession accounted for 49% of the total, indicating high vulnerability of this material. Using a Bayesian clustering method revealed two main genetic groups, G1 containing most foreign cultivars and G2 retaining local Spanish cultivars, which were further divided into two other subgroups using a nested approach, revealing moderate but significant differentiation among them. The populations were renamed according to the origin of the reference samples assigned to each group as 'South' (G1.1), 'Western Europe-1' (G1.2), 'Western Europe-2' (G2.1) and 'No-Pyrus communis' (G2.2). The results led to the creation of a 'generalist' collection, aiming to maximize genetic diversity representativeness, starting with 68 genotypes but expanding to 111 to achieve better allele recovery. This core collection is a valuable resource for genetic studies and conservation, enhancing efforts to preserve pear biodiversity.Publication Open Access Randomized clinical trial: bone bioactive liquid improves implant stability and osseointegration(MDPI, 2024-10-01) Al Madhoun, Ashraf; Meshal, Khaled; Carrió, Neus; Ferrés‐Amat, Eduard; Ferrés-Amat, Elvira; Barajas Vélez, Miguel Ángel; Jiménez-Escobar, Ana Leticia; Al-Madhoun, Areej Said; Saber, Alaa; Abou Alsamen, Yazan; Marti, Carles; Atari, Maher; Ciencias de la Salud; Osasun Zientziak; Institute for Multidisciplinary Research in Applied Biology - IMAB; Universidad Pública de Navarra / Nafarroako Unibertsitate PublikoaImplant stability can be compromised by factors such as inadequate bone quality and infection, leading to potential implant failure. Ensuring implant stability and longevity is crucial for patient satisfaction and quality of life. In this multicenter, randomized, double-blind clinical trial, we assessed the impact of a bone bioactive liquid (BBL) on the Galaxy TS implant's performance, stability, and osseointegration. We evaluated the impact stability, osseointegration, and pain levels using initial stability quotient (ISQ) measurements, CBCT scans, and pain assessment post-surgery. Surface analysis was performed using scanning electron microscopy (SEM) and atomic force microscopy (AFM). In vitro studies examined the BBL's effects on dental pulp pluripotent stem cells' (DPPSCs') osteogenesis and inflammation modulation in human macrophages. All implants successfully osseointegrated, as demonstrated by the results of our clinical and histological studies. The BBL-treated implants showed significantly lower pain scores by day 7 (p < 0.00001) and improved stability by day 30 (ISQ > 62.00 ± 0.59, p < 8 × 10-7). By day 60, CBCT scans revealed an increased bone area ratio in BBL-treated implants. AFM images demonstrated the BBL's softening and wettability effect on implant surfaces. Furthermore, the BBL promoted DPPSCs' osteogenesis and modulated inflammatory markers in human primary macrophages. This study presents compelling clinical and biological evidence that BBL treatment improves Galaxy TS implant stability, reduces pain, and enhances bone formation, possibly through surface tension modulation and immunomodulatory effects. This advancement holds promise for enhancing patient outcomes and implant longevity.Publication Open Access Converging pathways: new approaches to integrate vocational education training and higher education(Routledge, 2024-11-14) Farrán Blanch, Inmaculada; Núñez Aldaz, Imanol; Institute for Advanced Research in Business and Economics - INARBE; Institute for Multidisciplinary Research in Applied Biology - IMABThe evolving professional landscape necessitates educational innovation to prepare students for emerging challenges. This paper explores the integration of Vocational Education and Training (VET) and Higher Education (HE) through double-degree programs, challenging the conventional perception of these pathways as independent. Recognising the transformative potential in synergising competencies from VET and HE, we advocate for enhancing students' employability through collaborative curriculum design and shared learning objectives. Addressing limitations in the current VET-to-HE transition, we identify four criteria for successful double programs (i) coherent competency map, (ii) unified methodology, (iii) distinctive profile, (iv) own signalling. The paper concludes with a case study of an emerging dual VET/HE program in Navarra (Spain) built on the criteria outlined above.Publication Open Access New insights on Arabidopsis thaliana root adaption to ammonium nutrition by the use of a quantitative proteomic approach(MDPI, 2019-02-14) Coleto, Inmaculada; Vega-Mas, Izargi; Glauser, Gaëtan; González Moro, María Begoña; Marino, Daniel; Ariz Arnedo, Idoia; Ciencias; Zientziak; Institute for Multidisciplinary Research in Applied Biology - IMABNitrogen is an essential element for plant nutrition. Nitrate and ammonium are the two major inorganic nitrogen forms available for plant growth. Plant preference for one or the other form depends on the interplay between plant genetic background and environmental variables. Ammonium-based fertilization has been shown less environmentally harmful compared to nitrate fertilization, because of reducing, among others, nitrate leaching and nitrous oxide emissions. However, ammonium nutrition may become a stressful situation for a wide range of plant species when the ion is present at high concentrations. Although studied for long time, there is still an important lack of knowledge to explain plant tolerance or sensitivity towards ammonium nutrition. In this context, we performed a comparative proteomic study in roots of Arabidopsis thaliana plants grown under exclusive ammonium or nitrate supply. We identified and quantified 68 proteins with differential abundance between both conditions. These proteins revealed new potential important players on root response to ammonium nutrition, such as H+-consuming metabolic pathways to regulate pH homeostasis and specific secondary metabolic pathways like brassinosteroid and glucosinolate biosynthetic pathways.Publication Embargo Upgrading and validating a soil water balance model to predict stem water potential in vineyards(Elsevier, 2024-12-15) Mirás-Ávalos, José M.; Escalona, José M.; Pérez-Álvarez, Eva Pilar; Romero Azorín, Pascual; Botia, Pablo; Navarro, Josefa; Torres Molina, Nazareth; Santesteban García, Gonzaga; Uriarte, David; Intrigliolo, Diego S.; Buesa, Ignacio; Agronomía, Biotecnología y Alimentación; Agronomia, Bioteknologia eta Elikadura; Institute for Multidisciplinary Research in Applied Biology - IMABEfficient water management is pivotal for viticulture sustainability. Decision support tools can advise on how to optimize irrigation or on the feasibility of growing grapes in rainfed conditions, but reliable algorithms for assessing vine water status are required. In this context, the aim of the current study was to upgrade a soil water balance model specific for vineyards by incorporating meteorological, soil and vine vigor in equations that transform the fraction of transpirable soil water into midday stem water potential (Ψstem). The model's sensitivity to variations in the magnitude of input parameters was analyzed. Furthermore, the model was tested in a broad scope of Spanish vineyards with different grapevine cultivars (both red and white), rootstocks, plant age, soil and climatic conditions, and water regimes, totaling 129 scenarios. The model was only slightly sensitive to variations in the magnitude of most inputs, except for the fraction of transpirable water at which leaf stomatal conductance begin to decline. Moreover, the model satisfactorily reproduced the evolution of Ψstem over the growing season, although it slightly overestimated the measured ¿stem values, as the slopes of the fitted regression lines were lesser than 1 on most occasions, 76 out of 129. Nonetheless, the coefficients of determination for these relationships were greater than 0.9, except for 21 datasets. Mean errors averaged 0.024 ± 0.015 MPa, while root mean square errors averaged 0.27 ± 0.01 MPa. The index of agreement was greater than 0.75 in 51 datasets, with only three datasets showing an index of agreement lower than 0.5. Nevertheless, the deviations between observed and simulated Ψstem values did not alter the classification of the water stress undergone by grapevines. This upgraded model could constitute the core of a decision support system for water management in vineyards, applicable to both rainfed and irrigated conditions.Publication Open Access The effectiveness of polyhydroxyalkanoate (PHA) extraction methods in gram-negative pseudomonas putida U(MDPI, 2025-01-09) Getino, Luis; García, Irene; Cornejo Ibergallartu, Alfonso; Mateos, Raúl; Ariza-Carmona, Luisa M.; Sánchez-Castro, Natalia; Morán Juez, José Fernando; Olivera, Elías R.; Chamizo Ampudia, Alejandro; Ciencias; Zientziak; Institute for Multidisciplinary Research in Applied Biology - IMAB; Institute for Advanced Materials and Mathematics - INAMAT2Bioplastics are emerging as a promising solution to reduce pollution caused by petroleum-based plastics. Among them, polyhydroxyalkanoates (PHAs) stand out as viable biotechnological alternatives, though their commercialization is limited by expensive downstream processes. Traditional PHA extraction methods often involve toxic solvents and high energy consumption, underscoring the need for more sustainable approaches. This study evaluated physical and chemical methods to extract PHAs from Pseudomonas putida U, a bacterium known to produce poly-3-hydroxyoctanoate P(3HO). Lyophilized cells underwent six extraction methods, including the use of the following: boiling, sonication, sodium hypochlorite (NaClO), sodium dodecyl sulfate (SDS), sodium hydroxide (NaOH), and chloroform. Physical methods such as boiling and sonication achieved yields of 70% and 60%, respectively, but P(3HO) recovery remained low (30–40%). NaClO extraction provided higher yields (80%) but resulted in significant impurities (70%). NaOH methods offered moderate yields (50–80%), with P(3HO) purities between 50% and 70%, depending on the conditions. Spectroscopic and analytical techniques (FTIR, TGA, NMR, GPC) identified 0.05 M NaOH at 60 °C as the optimal extraction condition, delivering high P(3HO) purity while minimizing environmental impact. This positions NaOH as a sustainable alternative to traditional halogenated solvents, paving the way for more eco-friendly PHA production processes.Publication Open Access Mixtures of insect-pathogenic viruses in a single virion: towards the development of custom-designed insecticides(American Society for Microbiology, 2021) López Ferber, Miguel; Lent, Jan W. M. van; Beperet Arive, Inés; Simón de Goñi, Oihane; Williams, Trevor; Caballero Murillo, Primitivo; Agronomía, Biotecnología y Alimentación; Agronomia, Bioteknologia eta Elikadura; Institute for Multidisciplinary Research in Applied Biology - IMABAlphabaculoviruses (Baculoviridae) are pathogenic DNA viruses of Lepidoptera that have applications as the basis for biological insecticides and expression vectors in biotechnological processes. These viruses have a characteristic physical structure that facilitates the transmission of groups of genomes. We demonstrate that coinfection of a susceptible insect by two different alphabacu-lovirus species results in the production of mixed-virus occlusion bodies containing the parental viruses. This occurred between closely related and phylogeneti-cally more distant alphabaculoviruses. Approximately half the virions present in proteinaceous viral occlusion bodies produced following coinfection of insects with a mixture of two alphabaculoviruses contained both viruses, indicating that the viruses coinfected and replicated in a single cell and were coenveloped within the same virion. This observation was confirmed by endpoint dilution assay. Moreover, both viruses persisted in the mixed-virus population by coinfection of insects during several rounds of insect-to-insect transmission. Coinfection by viruses that differed in genome size had unexpected results on the length of viral nucleocapsids, which differed from those of both parental viruses. These results have unique implications for the development of alphabaculoviruses as biological control agents of insect pests. IMPORTANCE Alphabaculoviruses are used as biological insecticides and expression vectors in biotechnology and medical applications. We demonstrate that in caterpillars infected with particular mixtures of viruses, the genomes of different baculovirus species can be enveloped together within individual virions and occluded within proteinaceous occlusion bodies. This results in the transmission of mixed-virus populations to the caterpillar stages of moth species. Once established, mixed-virus populations persist by coinfection of insect cells during several rounds of insect-to-insect transmission. Mixed-virus production technology opens the way to the development of custom-designed insecticides for control of different combinations of caterpillar pest species.Publication Open Access Use of biocides to minimize microbial contamination in Spodoptera exigua multiple nucleopolyhedrovirus preparations(Elsevier, 2020-08-13) Presa-Parra, Ehdibaldo; Lasa, Rodrigo; Reverchon, Frédérique; Simón de Goñi, Oihane; Williams, Trevor; Institute for Multidisciplinary Research in Applied Biology - IMABThe presence of contaminant microbes in baculovirus-based insecticides is regulated by phytosanitary product registration authorities. We aimed to determine whether the abundance of microbes in suspensions of Spodoptera exigua multiple nucleopolyhedrovirus occlusion bodies (OBs) could be reduced by treatment with a range of biocidal compounds. The diversity of contaminant bacteria was determined by next-generation sequencing of the 16S rRNA gene. Overall, 97.9% of sequences detected were Gammaproteobacteria (mostly Pseudomonas spp. and Enterobacteriaceae) and 2.1% were Firmicutes (mostly Enterococcus spp.). Colloidal silver, benzalkonium chloride and chlorhexidine digluconate were identified as highly effective biocides. Incubation of OB suspensions with high concentrations of colloidal silver (450 mg/l) or benzalkonium chloride (6000 mg/l) resulted in marked reductions in colony forming unit counts over a 180 day period at 4° or 25 °C. Benzalkonium chloride and colloidal silver treatments, at either 4 or 25 °C, did not affect the insecticidal activity of OBs over an 80 day period. However, OB activity decreased following 180 days of treatment by benzalkonium chloride at either 4 or 25 °C, or by colloidal silver at 25 °C, but not at 4 °C. Counts of OBs revealed a significant decrease in OB numbers in benzalkonium chloride-treated suspensions after 180 days at both temperatures, whereas colloidal silver-treated OBs were not affected. Benzalkonium chloride also caused aggregation of OBs at the concentration tested. We conclude that biocidal compounds can markedly reduce the abundance of contaminant microorganisms in OB suspensions, and can be accompanied by reductions in OB infectivity and OB numbers in some circumstances. Future studies should focus on lower concentrations of biocides that do not affect OBs in long-term storage.Publication Open Access Rational application of treated sewage sludge with urea increases GHG mitigation opportunities in Mediterranean soils(Elsevier, 2017-02-01) Calleja Cervantes, María Eréndira; Aparicio Tejo, Pedro María; Villadas Latorre, Pablo José; Irigoyen Iriarte, Ignacio; Irañeta, Iosu; Fernández-González, A.J.; Fernández-López, M.; Menéndez, S.; Producción Agraria; Nekazaritza Ekoizpena; Ciencias del Medio Natural; Natura Ingurunearen Zientziak; Institute for Multidisciplinary Research in Applied Biology - IMABPublication Open Access High irradiance increases NH4+ tolerance in Pisum sativum: higher carbon and energy availability improve ion balance but not N assimilation(Elsevier, 2011-03-02) Ariz Arnedo, Idoia; Artola Rezola, Ekhiñe; Asensio, Aarón C.; Cruchaga Moso, Saioa; Aparicio Tejo, Pedro María; Morán Juez, José Fernando; Ciencias del Medio Natural; Natura Ingurunearen Zientziak; IdAB. Instituto de Agrobiotecnología / Agrobioteknologiako Institutua; Institute for Multidisciplinary Research in Applied Biology - IMABThe widespread use of NO3− fertilization has had a major ecological impact. NH4+ nutrition may help to reduce this impact, although high NH4+ concentrations are toxic for most plants. The underlying tolerance mechanisms are not yet fully understood, although they are thought to include the limitation of C, the disruption of ion homeostasis, and a wasteful NH4+ influx/efflux cycle that carries an extra energetic cost for root cells. In this study, high irradiance (HI) was found to induce a notable tolerance to NH4+ in the range 2.5–10 mM in pea plants by inducing higher C availability, as shown by carbohydrate content. This capacity was accompanied by a general lower relative N content, indicating that tolerance is not achieved through higher net N assimilation on C-skeletons, and it was also not attributable to increased GS content or activity in roots or leaves. Moreover, HI plants showed higher ATP content and respiration rates. This extra energy availability is related to the internal NH4+ content regulation (probably NH4+ influx/efflux) and to an improvement of the cell ionic balance. The limited C availability at lower irradiance (LI) and high NH4+ resulted in a series of metabolic imbalances, as reflected in a much higher organic acid content, thereby suggesting that the origin of the toxicity in plants cultured at high NH4+ and LI is related to their inability to avoid large-scale accumulation of the NH4+ ion.