Agrobiotechnology Institute

Orf virus (ORFV) is the etiological agent of Contagious Ecthyma, a global disease that mainly affects sheep, goats, wild ruminants, and humans. Here, we determined the complete genome sequence of two ORFV strains from Spain (NAV and ARA) and two from Argentina (HRE and CHB), representing the first report from the Iberian Peninsula and from South America. The assembled genomes of the ARA, CHB, HRE, and NAV strains of ORFV were found to be 137 891, 137 160, 137 340, and 137 214bp long, respectively, each contained 132 genes, and all showed high amino-acid identity and similar lengths compared to the reference strain NZ2. We performed a microsatellite analysis to identify molecular signatures associated with host species (sheep or goat). In addition, the analysis of 32 selected genes showed that the median nucleotide substitution rate for the worldwide cluster 3 that includes the four samples described in this study, was 2.6 x10-5 subs/site/year (9.2x10-8-6.7x10-5-95% HPD) placing the TMRCA (median divergence time) around 1730. Genetic characterization of ORFV strains not only allows epidemiological studies but also represents the first step towards developing molecular tools for diagnostics and vaccines.

Abstract Pattern-recognition receptors (PRRs) initiate plant pattern-triggered immunity (PTI), encompassing receptor-like kinases (RLKs) and receptor-like proteins (RLPs). Eucalyptus grandis , an economically important species worldwide, is a long-lived organism that faces multiple disease pressures. The deployment of PRR-based breeding tools offers a route to broad, non-race-specific resistance that can remain effective across outbreaks. The objective of this work was to identify and characterise the PRR repertoire of E. grandis using public multi-omics data. Two state-of-the-art predictors were compared with a multi-species curation and a meta-analysis was conducted compiling transcriptomic data under biotic stress. A set of 730 PRR candidates were consistently identified across three sources (~ 1.6% of protein-coding genes), of which 283 were differentially expressed (PRR-DEGs). A PFAM domain-based scheme was applied to standardise classification across tools. RLKs (TM-K-LRR) predominated over RLPs (TM-LRR), while many carried malectin or malectin-like ectodomains. PRR genes were unevenly distributed across the genome: chromosome 6 had the highest count and the densest clustering. Gene-family expansion appeared mainly driven by duplication, with extensive tandem arrays supported by segmental, proximal and dispersed events. Gene Ontology and cis-elements annotations in PRR/PRR-DEGs showed significant enrichment in terms related to cell cycle development, hormonal regulation and stress response. We proposed a catalogue of 16 PRR which resulted in DEGs against at least three pathogens, suggesting their broad spectrum and robustness. Most of them presented orthologues with cross-taxon evidence of defence roles. This study delineates novel multi-pathogen candidate PRR genes, providing valuable information to assist Eucalyptus breeding programs.

Potassium (K) is an essential macronutrient that plays a central role in grapevine physiology and fruit quality. This study aimed to evaluate the effects of different K concentrations in the nutrient solution on berry composition, color development, and mineral concentration in leaves, petioles, and berries of fruit-bearing cuttings of Vitis vinifera L. cv. Tempranillo. Fruit-bearing cuttings were grown under controlled greenhouse conditions and irrigated with modified half-strength Hoagland solutions containing 0%, 25%, 50%, 75%, or 100% K and a control nutrient solution. Berry quality parameters, including total soluble solids, acidity, anthocyanins, and phenolic maturity, were significantly influenced by K availability. Moderate K treatments (50–75%) produced the most favorable outcomes in terms of sugar accumulation, anthocyanin concentration, and nutrient balance. The results suggest that optimized K fertilization is critical for improving grape quality while avoiding potential negative effects associated with deficiency or excess. These findings provide valuable insights for K management in viticulture.

ABSTRACT In Arabidopsis, exposure to growth-promoting microbial volatile compounds (VCs) enhances CLPC2 levels. This chaperone forms part of the CLP protease complex, which ensures the correct functioning of essential processes in plastids. Previous studies indicated considerable functional redundancy of CLPC2 with its dominant paralogue CLPC1. However, the function and action mechanism of CLPC2 still remain unknown. Here we found that CLPC2-lacking clpc2-2 mutants were unresponsive to microbial VCs, whereas clpc1-1 knockout mutants exhibited a WT-like response to VCs when grown on sucrose-containing medium. Unlike clpc1-1 , clpc2-2 plants presented a fully functional photosystem II and lower than WT stomatal conductance. Furthermore, clpc2-2 plants, but not clpc1-1 plants, produced wrinkled seeds with delayed embryonic development and reduced postgerminative establishment rates that resembled those of mutants lacking P and R components of the CLP proteolytic core. Proteomic analyses revealed that knocking out of CLPC2 enhanced the levels of chloroplastic proteins that are essential for growth, embryo development and seedling establishment. These changes differed from those promoted by the lack of CLPC1, but partially resembled those promoted by CLPPR core inactivation. Nearly 40% of the proteins differentially accumulated by the lack of CLPC2 were VC-responsive. Notably, 35S promoter-driven CLPC2 expression promoted changes in the proteome similar to those promoted by the lack of CLPC1. Collectively, our findings highlighted contrasting functional and molecular specificities for CLPC1 and CLPC2, and provided strong evidence that CLPC2 plays specific roles in CLP complex-mediated regulation of plant growth, photosynthesis, embryogenesis, postgerminative seedling establishment and microbial VC responsiveness.

Agriculture significantly contributes to greenhouse gas (GHG) emissions, yet fluxes from irrigated semi-arid systems remain poorly quantified. This study investigates CO2, CH4, N2O, and NH3 fluxes in a short-term lettuce experiment under semi-arid conditions. The objective was to quantify flux variability and identify key environmental and management drivers. High-frequency soil gas flux measurements were conducted under three treatments: irrigated soil (I), irrigated soil with plants (IP), and irrigated soil with plants plus NH4NO3 fertilizer (IPF). Environmental factors, including solar radiation, soil temperature, water-filled pore space, and relative projected leaf area, were monitored. A Random Forest model identified main flux determinants. Fluxes varied with plant function, growth, and fertilization. IP exhibited net CO2 uptake through photosynthesis, whereas I and IPF showed net CO2 emissions from soil respiration and fertilizer-induced disruption of plant function, respectively. CH4 uptake occurred across treatments but decreased with plant presence. Fertilization in IPF triggered episodic N2O (EF = 0.1%) and NH3 emissions (EF = 0.97%) linked to nitrogen input. Vegetated semi-arid soils can act as CO2 sinks when nitrogen is optimally managed. Excess or poorly timed nitrogen delays CO2 uptake and increases reactive nitrogen losses. Methanotrophic activity drives CH4 dynamics and is influenced by plants and fertilization. Maintaining crop vigor and applying precision nitrogen management are essential to optimize productivity while mitigating GHG and NH3 emissions in semi-arid lettuce cultivation.

Protein ubiquitination regulates cell biology through diverse avenues, from quality control-linked protein degradation to signaling functions such as modulating protein-protein interactions and enzyme activation. Mass spectrometry-based proteomics has allowed proteome-scale quantification of hundreds of thousands of ubiquitination sites (ubi-sites), however the functional importance and regulatory roles of most ubi-sites remain undefined. Here, we assembled a human reference ubiquitinome of 108,341 ubi-sites by harmonizing public proteomics data. We identified a core subset of ubi-sites under evolutionary constraint through alignment of ubiquitin proteomics data from six non-human species, and determined ultra-conserved ubi-sites recurring at regulatory hotspots within protein domains. Perturbation proteomics revealed that these highly conserved ubi-sites are more likely to regulate signaling functions rather than proteasomal degradation. To further prioritize functional ubi-sites with roles in cellular signaling, we constructed a functional score for more than 100,000 ubi-sites by integrating evolutionary, proteomic, and structural features using machine learning. Our score identifies ubi-sites regulating diverse protein functions and rationalizes mechanisms of genetic disease. Finally, we employed chemical genomics to validate the functional relevance of high-scoring ubi-sites and leveraged genetic code expansion to demonstrate that ubiquitination of K320 in the RNA-regulator ELAVL1 disrupts RNA binding. Our work reveals systems-level principles of the ubiquitinome and provides a powerful resource for studying protein ubiquitination.

The increasing of industrial activities and urbanization have led to the accumulation of heavy metals, which pose a high risk to human health and ecosystems. Adsorption using biochar from bamboo waste is an alternative solution as an adsorbent. This study evaluated the performance of biochar from pyrolysis of bamboo waste at various temperatures (300–600°C) for adsorption of lead (Pb²⁺) ions. The pyrolysis was conducted under a nitrogen gas atmosphere to prevent oxidation during thermal decomposition. Fourier Transform Infrared (FTIR) analysis revealed an increase in the stability of carbonyl (C=O) and carbon–nitrogen (C–N) functional groups with increasing temperature, which contributed to a stronger affinity for metal ions. Brunauer–Emmett–Teller (BET) analysis showed that biochar produced at 400 °C exhibited the most favorable surface characteristics, with a surface area of 178.56 m²/g, a pore volume of 0.091 cm³/g, and an average pore diameter of 2.05 nm. This sample also demonstrated the highest Pb²⁺ adsorption capacity of 52.4 mg/g. Despite higher porosity, the biochar produced at 600 °C showed diminished adsorption efficiency due to pore structure collapse and decreased carbon content. The findings suggest that biochar synthesized at 400 °C has strong potential for use in environmental remediation applications, particularly in mitigating lead contamination in water systems.

Climate change is impacting the performance of plants worldwide. However, the impact on ferns, the second-most diverse lineage of vascular plants, has received little attention. Here, we investigated the effects of one of the most claimed scenarios of the climatic change: drought (D), high temperature (HT) and high CO2 concentration (HCO2) on a fern (Nephrolepis exaltata) and a commonly studied angiosperm (Brassica oleracea) at photosynthetic, anatomical, and metabolic levels. Leaf anatomy was slightly affected by stress conditions in both species. Multivariate analysis demonstrated that B. oleracea's physiological responses to HCO2 were greater than N. exaltata's. Lipids and primary metabolites levels differed in response to stress in B. oleracea. Notably, the combination of D, HT, and HCO2 exacerbated the changes in primary metabolites, reducing amino and organic acids levels. Interestingly, phosphatidylcholine and phosphatidylethanolamine levels showed varied responses, increasing under HT and decreasing under HCO2 or combined stress in B. oleracea. In contrast, the fern was mostly unresponsive to D, HT, HCO2, and the combination among them at the metabolic level. Beyond providing important information concerning the trade-off between carbon uptake and stress acclimation mechanisms, our study indicates minor fern responses to D, HT, HCO2, suggesting differential impacts of climate change on ferns and angiosperms.

The response of plants to elevated atmospheric [CO2] is highly dynamic and influenced by developmental stage, yet its role in photosynthetic acclimation remains underexplored. This study examines the physiological and molecular responses of wheat (Triticum durum, var. Amilcar) to elevated [CO2] (700 ppm vs. 400 ppm) at two distinct developmental stages: the vegetative stage at the end of the elongation stage and the reproductive stage at the beginning of ear emergence (Z39 and Z51, respectively). Wheat plants at the developmental stage Z39, cultivated under elevated [CO2], maintained photosynthetic rates despite a carbohydrate build-up. However, at Z51, photosynthetic acclimation became more evident as the decline in Rubisco carboxylation capacity (Vcmax) persisted, but also stomatal conductance and diffusion were decreased. This was accompanied by the up-regulation of the CA1 and CA2 genes, likely as a compensatory mechanism to maintain CO2 supply. Additionally, hormonal adjustments under elevated [CO2], including increased auxin and bioactive cytokinins (zeatin and isopentenyl adenine), may have contributed to delayed senescence and nitrogen remobilization, sustaining carbon assimilation despite biochemical constraints. These findings highlight the developmental regulation of photosynthetic acclimation, emphasizing the need for the stage-specific assessments of crop responses to future atmospheric conditions.

The article examines the specific features of implementing a quality management system in accordance with the requirements of DSTU ISO 9001:2015 in the operations of a young meat processing enterprise specializing in the production of high-protein snacks. The relevance of applying international standards in the food industry is substantiated, considering the need to harmonize national practices with global market requirements and to strengthen consumer trust. An analysis of the enterprise’s quality management system revealed several problematic aspects, including the absence of a quality policy, insufficient process formalization, lack of systematic risk monitoring, and no practice of internal audits. Based on this analysis, proposals were developed for the integration of quality management system elements, including a quality policy, a process interaction scheme, and an internal audit procedure. The proposed approaches are aimed at strengthening production task discipline, increasing process transparency, and fostering a culture of continuous improvement. The expected results of implementing these measures include reduced production costs, stronger competitive positions, enhanced corporate image, and the creation of prerequisites for entering international markets. It is concluded that systematic implementation of ISO 9001 in the meat processing sector should be considered not only as a means of confirming product quality but also as a strategic tool for improving operational efficiency, oriented toward sustainable development and long-term market resilience.