Agave sisalana, widely cultivated in northeastern Brazil, holds great socioeconomic importance, although its leaves remain underutilized. This study evaluated their potential as feedstock for integrated first- (1G) and second-generation (2G) ethanol production. In the 1G route, leaf juice underwent acid hydrolysis and fermentation, yielding 40.70 g/L ethanol with 74.00% efficiency using Ethanol Red®. In the 2G route, a fractional factorial design optimized bagasse pretreatment; the best condition (30 min, 180°C, 15% solids, 0.5% acid) achieved 60.31% cellulose and 80.32% hemicellulose conversion. Although inhibitors were generated, detoxification enabled efficient fermentation, resulting in 27.44 g/L ethanol with an 83.59% yield using the GMO strain Celere2L. Overall, 80.90 L of ethanol per ton of leaves was obtained, demonstrating A. sisalana's potential as a sustainable feedstock for integrated 1G–2G ethanol production and reinforcing its relevance for biofuel development and energy diversification in semi-arid regions.

Sugarcane is a globally important crop, widely used in both the food industry and biofuel production. Weed infestations can significantly reduce its productivity, and to address this challenge, herbicides such as glyphosate are commonly applied. Glyphosate acts as a structural analog of phosphoenolpyruvate (PEP), inhibiting the enzyme 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS), thereby disrupting the shikimate pathway and impairing the biosynthesis of essential aromatic amino acids required for plant growth and development. However, the limited tolerance of sugarcane to glyphosate restricts herbicide application to doses that are often insufficient for effective weed control. Previous studies have shown that amino acid substitutions in the EPSPS enzyme, particularly the T102I/P106A (TIPA) mutations, can confer glyphosate resistance in plants. In this study, we evaluated whether a specific mutation in the endogenous ScEPSPS gene could enhance glyphosate tolerance in sugarcane without compromising morphological, physiological, and biometric parameters evaluated under greenhouse conditions. The mutated gene was constitutively overexpressed in transgenic sugarcane plants, which survived glyphosate concentrations as high as 5% (v/v of the commercial formulation, 106.5 mM of glyphosate acid equivalent). Molecular analyses revealed approximately 25- to 80-fold overexpression of the transgene in resistant lines compared with non-transgenic controls. High-performance liquid chromatography (HPLC) quantification showed elevated levels of phenylalanine, tyrosine, and tryptophan 14 days after herbicide application in plants overexpressing the mutated ScEPSPS. Although the mutation affected certain histological and physiological parameters, biometric assessments indicated that the overall phenotype remained largely unchanged. The successful overexpression of the mutated endogenous ScEPSPS gene represents a promising biotechnological strategy to enhance glyphosate tolerance in sugarcane without compromising plant growth and development under greenhouse conditions.

The expansion of arid and semi-arid regions, consequent to the intensification of desertification processes attributable to global warming, exerts a deleterious effect on the agricultural production of energy crops, with current estimates indicating that a further 23% of global agricultural areas will suffer from desertification by 2100, precipitating crises in these sectors. Agave species have the capacity to thrive in these marginal environments characterized by aridity and elevated temperatures. These plants can serve as a source of biomass for the production of biofuels, a process that mitigates the environmental impacts of the transport sector while promoting the utilization of drylands, thereby eliminating competition with food crops. Given the paucity of knowledge regarding the soil microbiota and rhizosphere in minimal technological input Agave plantations, the objective of this study was to evaluate the microbiological and chemical soil properties of Agave sisalana and Agave hybrids (H11648 and H400f) farming systems. The analyses, which were carried out using microbial quantification, enzyme stoichiometry and enzymatic vector calculations, demonstrated that the microbiome of these plants is active and well-structured in terms of nutritional acquisition. It was observed that the Agave fields’ microbiome is very similar to that of the native vegetation. This finding suggests that the soil and rhizosphere microbiota are healthy and stable in the Agave fields evaluated, even with the implementation of agronomic exploitation models, as the chemical analysis of the soil reveals that all measured parameters are consistent with those of soils suitable for crop production. These observations persist even in long-established Agave plantations of varying ages that have never received any type of implement or soil correction. Thus, the integration of the chemical and biological data through principal component analysis, redundancy analysis and Permutational Multivariate Analysis of Variance (PERMANOVA) enabled the differentiation of the soil among the three Agave species, which shows the influence of the plant genotype on its microbiota.

Biochar has emerged as a scalable negative emission technology with promising agronomic and environmental benefits. In this context, arid and semi-arid regions represent promising zones for carbon removal through soil biochar application, particularly when exploring new non-food biomass sources with high energy potential, such Agave wercklei. This study aims to evaluate the properties of biochars produced from Agave wercklei and investigate the role of pyrolysis atmosphere by comparing nitrogen (N2) and recirculated pyrolysis gas (PyGR) across four temperatures (350, 425, 500, and 575 degrees C). PyGR-based biochars exhibited higher carbon content (up to 71.36 %), energy density, and enriched surface chemistry with increased aliphatic functional groups, retention of dissolved organic carbon, while N2-based biochars showed greater aromaticity, higher thermal stability, and lower content of volatile compounds. Differences were also observed in handling and interfacial properties, with PyGR-based biochars presenting superior flowability and higher hydrophobicity, while N2-based biochars exhibited greater water retention. Multivariate analysis revealed that the pyrolysis atmosphere plays a crucial rule in key indicators, including atomic ratios (H/C, O/C), aromatic indices, and thermal degradation profiles. Thus, the use of inert gas in bench-scale studies may thus misrepresent the carbon permanence and agronomic behavior of biochars produced under realistic industrial conditions. Furthermore, Agave wercklei demonstrated favorable biochar properties, positioning it as a valuable biomass for producing biochar in semi-arid regions.

Drylands cover 41% of Earth's surface and are expanding due to climate change, requiring innovative agricultural strategies for resource efficiency. Agave, a non-conventional drought-tolerant monocot cultivated in Mexico and Brazil, is a sustainable crop with high biomass productivity and low water requirements, supporting industries like biofuels, textiles, and beverages. With high biomass productivity and extremely low water requirements, Agave species are increasingly recognized as one of the most sustainable crops for semi-arid and marginal regions. Despite its environmental resilience, agave cultivation is significantly constrained by insect pests that reduce yield and economic returns. While Bacillus thuringiensis (Bt) Cry proteins have revolutionized pest management in other crops, their application in Agave spp. remains unexplored due to technical barriers in genetic transformation. Here, we argue that the integration of Bt technology into Agave species is both technically feasible and strategically necessary. We discuss recent advances in plant transformation and regeneration that lower historical barriers in monocots and outline how Cry-based resistance could be incorporated into Agave spp. within integrated pest management frameworks. By synthesizing current knowledge and identifying key research priorities, we suggest that Bt-engineered Agave spp. could represent a realistic and timely strategy to enhance pest control, sustainability, and resilience in dryland agricultural systems. © 2026 The Author(s). Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

In humans and laboratory species, specialized pro-resolving mediators (SPM), a class of cell-signaling molecules originating from the metabolism of PUFA, emerged as mediators of inflammation resolution, but their role in other species remains almost unknown. In dairy cows, postpartum uterine involution involves a physiological transient inflammation, which may evolve into a pathologic inflammation (endometritis). This observational retrospective cohort study characterized the SPM profiles of plasma (2, 10, and 21 d postpartum; DPP) and uterine fluid (21 DPP) of healthy cows (n = 19), and their deviations in cows with endometritis (n = 12). Compared with healthy cows, at 21 DPP, cows with endometritis showed greater uterine cytology proportion of neutrophils, macrophages, M2 phenotype macrophages, and uterine fluid concentrations of the inflammatory mediators prostaglandin E2 (7,360.7 vs. 2,647.6 pg/mL), LTB4 (36.5 vs. 16.4 pg/mL), and chemerin (1.6 vs. 1.3 pg/mL). This indicates that in healthy cows the endometrial inflammatory stimuli already declined by 21 DPP, whereas in cows with endometritis, inflammation was active. Compared with healthy cows, at 21 DPP, cows with endometritis showed greater uterine fluid concentrations of the SPM 18-HEPE (42.2 vs. 19.8 pg/mL), resolvin E2 (RvE2; 2.7 vs. 1.1 pg/mL), and RvE4 (1.5 vs. 0.5 pg/mL). This may indicate that in healthy cows the pro-resolving mechanisms were already declining, as inflammation is controlled, whereas in cows with endometritis the pro-resolving mechanisms were still underway in an attempt to control inflammation. From the SPM, only RvE1 was detected in plasma, whereas MaR2, RvE1, RvE2, and RvE4 were detected in the uterine fluid, indicating their local action at low concentrations. In conclusion, this foundational study is consistent with a role of SPM in postpartum uterine inflammation resolution mechanisms.

The macauba palm (Acrocomia aculeata) is an emerging oilseed species with promising applications in biodiesel production, as well as in food and cosmetic industries. Native to the Neotropics, it is in the early stages of domestication and distributed across diverse environments and edaphoclimatic conditions. However, genomic studies of macauba are limited due to the scarcity of publicly available sequence data, as it is considered a nonmodel plant. In this study, we present an exploratory analysis of a transcriptome dataset comprising seven different organs (roots, bulbs, male and female flowers, leaves, leaf sheath, and fruits). A total of 22,703 transcripts were assembled into a single reference dataset. Of these, 9729 transcripts (42.85 %) were annotated using KEGG orthology. Gene expression profiling revealed 306, 32, 41, 67, 92, 158 and 916 organ-specific transcripts in leaves, leaf sheaths, bulbs, female flower, male flower, fruit and root, respectively. Comparative analysis with oil palm (Elaeis guineensis) and date palm (Phoenix dactylifera) revealed 55 gene families exclusive to macauba palm. In addition, 221 transcripts related to drought stress were identified through functional annotation and grouped into 112 gene families. Root libraries revealed 7091 fungal transcripts - approximately 3.9 % of all reads - mainly derived from arbuscular mycorrhizal fungi (AMF) Rhizophagus spp. These findings highlight the central role of signal transduction pathways in response to environmental stresses in macauba palm. The transcriptome dataset generated in this study provides a valuable genomic resource for future genotype-phenotype investigations in macauba palm. Furthermore, the presence of AMF-associated transcripts suggests a potentially important role for these symbiotic fungi in macauba palm growth and development.

Xylose metabolism in Saccharomyces cerevisiae remains a significant bottleneck due to the difficulty in identifying functional and efficient xylose isomerases (XI). In the present study, publicly available metagenomic and metatranscriptomic datasets of rumen microbiota from different herbivorous mammals were used to prospect novel XIs sequences. Seven putative XIs from moose, camel, cow, and sheep were cloned into a strain modified for xylose metabolism. Out of those, five XIs demonstrated activity and efficiently converted xylose into xylulose, resulting in ethanol as the final product. A XI from camel rumen microbiota exhibited a KM of 16.25 mM, indicating high substrate affinity. The strains expressing enzymes XI11 and XI12, obtained from sheep rumen microbiota, were able to deplete 40 g/L of xylose within 72 and 96 h, achieving theoretical ethanol yields of 90% and 88%, respectively. These results are comparable to those obtained with Orpinomyces sp. ukk1 XI, a benchmark enzyme previously reported as highly efficient in S. cerevisiae. This study also provides the first report on the successful expression of XIs mined from the ruminal microbiotas of sheep and camels in S. cerevisiae, expanding the perspectives for the optimization of fermentation processes and the production of lignocellulosic biofuels from xylose.

Some consumers are replacing cow milk by plant-based milk alternatives (PBMA); however, the current knowledge regarding the mineral profile of PBMA is limited. This study aimed to characterize the mineral profile of commercial milk (n = 80) and PBMA types (n = 60; soya, rice, oat, almond, coconut, and hazelnut) by inductively coupled plasma-optical emission spectroscopy, along with a modification of the Sandell-Kolthoff reaction (iodine determination). A single fixed-effect statistical model considering the type of beverage as an independent variable was applied. Soya PBMA presented higher contents of Ca, Mg, Cu, and Mg than commercial milk, and similar contents of K and P. In contrast, commercial milk had higher contents of S, Zn, and Se, with the latter being below the limit of quantification (10 mu g/ kg) in all PBMA types. Both almond and hazelnut PBMA displayed I contents similar to those of commercial milk. In terms of mineral ratios, PBMA types presented a higher Ca/P compared with commercial milk, being aligned with dietary guidelines, whereas commercial milk and soya PBMA showing lower Na/K values, which are beneficial for cardiovascular health. Due to their variability, it is difficult to say with certainty that PBMA can reliably substitute milk as a source of minerals.

Efficient utilization of complex biomass-derived sugars and tolerance to inhibitors are key requirements for the viability of lignocellulosic-based biorefineries. In this study, a two-stage evolution of an industrial yeast strain engineered with a xylose isomerase pathway yielded strain AceY.14, which exhibited improved fermentative performance and increased tolerance to acetic acid. Whole-genome sequencing of the evolved strain identified SNPs in ZWF1, a component of the pentose phosphate pathway (PPP), and in the G1 cyclin gene CLN3, both of which were functionally validated through CRISPR and reverse engineering. The zwf1E191D mutation reduced xylitol accumulation, alleviating inhibition of xylose isomerase and enhancing flux through the non-oxidative branch of the PPP, while the frameshift cln3T556fs mutation unexpectedly improved acetic acid tolerance and xylose consumption in the evolved strain, also affecting cell size and growth. Genome sequencing of AceY.14 also revealed a significant reduction in the xylA gene copy number, likely decreasing the metabolic burden associated with high xylose isomerase expression. A synergistic effect was observed in the isu1Δ/zwf1Δ double mutant, further boosting xylose consumption rates. A diploid derivative (AceY-2n) demonstrated high productivity and robustness in fermentations using hydrolysates from various lignocellulosic feedstocks, highlighting the strain's potential for industrial-scale applications. These findings reveal novel metabolic targets for strain optimization and offer valuable insights for the rational engineering of yeast platforms for sustainable biofuel and bioproduct production.

Blue light (BL) plays an important role in stomatal opening, finely tuning plant responses to environmental conditions. While the BL signaling pathway is well understood in C3 and C4 plants, its role in crassulacean acid metabolism (CAM) plants remains uncertain. Traditionally, stomata in CAM plants were considered insensitive to BL stimulation, and as a result, research on such interaction was overlooked for a long time. Only recently, studies have found that the BL signaling cascade is active in CAM plants. Here, we investigated the effects of BL irradiance on stomatal responses in Agave, a highly productive CAM plant, by stimulating Agave leaves with BL and taking measurements of leaf gas exchange during the morning (closed stomata) and in the afternoon (open stomata). Our findings revealed that BL had no significant effect on stomatal opening during the morning period. However, BL increased stomatal conductance (gs) by 67.3

Small and medium-scale biorefineries are processing facilities designed to produce a portfolio of value-added products with applications in different productive sectors. The Territory of the Southern Coast (TSC) of Bahia in Brazil has a high potential to provide agricultural and forest biomass for these bioindustries. This study focuses on quantifying the biomass of key agricultural crops in the TSC from 1999 to 2019, utilizing data from the Municipal Agricultural Production (MAP) survey conducted by the Brazilian Institute of Geography and Statistics (IBGE). The biomass is subsequently categorized based on its applications in pharmaceutical and other industries, employing the Classification and Regression Tree (CART) algorithm, specifically using the Gini Index. Thus, our results reveal a noteworthy trend over 20 years through a time series analysis. For example, banana production (1.3 Mt) surpassed cocoa production (1.1 Mt) despite cocoa cultivation covering a vast 5.8 million hectares expanse compared to the smaller banana footprint. Cocoa and cassava are significant contributors, representing 41.95

The use of plants from the Agave genus as biomass for biofuel presents great potential for energy transition due to its physiological traits and non-competitiveness with food crops. Brazil, the largest producer of sisal, presents a promising opportunity to use agave residues as substrate for anaerobic digestion. However, few studies address the potential for conversion of raw juice extracted from these plants into methane. This study evaluated the physicochemical and composition characteristics of raw agave juice, as well as its impacts on anaerobic digestion for biogas production and energy recovery. Biochemical Methane Potential assays were conducted with agave juice at concentrations of 0.5, 2.5, and 5 gVS/L, both with and without nutritional supplementation. The substrate concentration and nutritional supplementation increased methane production by 3.77- and 2-fold, respectively, showing that increasing substrate concentrations at constant substrate/inoculum ratio and nutritional supplementation significantly boosts methane production from agave. This optimization yielded methane yields of up to 626.97 +/- 33.20 NmL CH4/gVS, along with 49.81 kWh of electric energy and 332.98 MJ of thermal energy for each ton of processed agave leaves. These findings enhance our understanding of optimizing biogas production from agave juice, supporting the transition to sustainable energy.

Plant growth and development are regulated processes influenced by molecular, cellular, physiological mechanisms, and environmental factors. Additionally, meristems play a crucial role in regulating the development of lateral buds (axillary buds), thereby modifying the overall architecture of the plant. In plants of the Saccharum spp. complex, buds play a significant role in the formation of new plants. The establishment of a new field for these plants is achieved through the distribution of stem cuttings containing 3 to 4 buds per segment, pre- sprouted seedlings (PSS) utilizing a single axillary bud, or plants derived from in vitro meristem cultivation. The objective of this study was to identify key genes involved in the pre-emergence regulation of buds by comparing a commercial sugarcane hybrid, characterized by low sprouting speed, with an energy cane cultivar, which exhibits high sprouting speed. To elucidate the regulation of gene expression in buds during the pre- emergence phase, axillary buds were collected during the first 48 h post-planting for transcriptional profiling using RNA-Seq analysis. The results reveal a distinct difference in expression profiles at 0, 24, and 48 h after planting. Notable differences were observed in the expression patterns of genes associated with hormone biosynthesis and degradation, axillary bud dormancy release, cellulose deposition, cell growth, division, expansion, and sugar metabolism. Furthermore, genes related to lactate metabolism were identified as potential negative markers for bud sprouting in Saccharum spp. hybrids. These findings are unprecedented, offering novel insights into the gene regulation mechanisms of axillary buds in sugarcane and energy cane. This contribution holds significant implications for both the scientific community and the agro-industrial sector.

The Agave genus is recognized for its diversity, economic importance, and adaptability to arid and semi-desert climates. Among its species, Agave sisalana stands out for the production of sisal, a resilient natural fiber with various industrial applications and potential to be used as a raw material in biofuel production, due to the accumulation of fermentable sugars in its biomass. Although this species presents significant agronomic potential, challenges in conventional breeding hinder its cultivation. A viable alternative for genetic improvement is the development of transgenic plants that incorporate desirable agronomic traits through the controlled insertion of genes of interest into their genome. The objective of study was to develop a protocol for obtaining transgenic plants of Agave sisalana 'RLV19,' a species widely cultivated in the semi-arid region of Bahia-BA, Brazil, using the cp4-epsps gene aimed at constitutive gene expression. Two transgenic plants of Agave sisalana 'RLV19' were regenerated via organogenesis. PCR analyses and CP4-EPSP protein expression by RT-PCR confirmed the presence and expression of the transgene in these plants. This is the first report of A. sisalana transgenic plants expressing the cp4-epsps gene, and to the best of our knowledge, there have been no prior reports on protocols for the production of transgenic plants of this species.

AIMS:The impact of promoter selection on the overexpression of the XKS1 gene in Saccharomyces cerevisiae is investigated with a focus on optimizing xylose metabolism for second-generation ethanol production. The goal was to identify how different promoters affect the fermentation performance of laboratory and industrial yeast strains under various media conditions. METHODS AND RESULTS:Four constitutive promoters-TEF1p, ADH1p, PGK1p, and TDH3p-were tested to overexpress XKS1 in two strains of S. cerevisiae, one laboratory strain (BY4742) and one industrial strain (PE-2B), both engineered with a heterologous xylose isomerase pathway. The strains were evaluated in defined (YNB) and complex (YPDX) media, as well as a synthetic sugarcane hydrolysate, over a 144-h fermentation period. Promoter choice significantly influenced cell growth, xylose consumption, and ethanol production. In the laboratory strain, TEF1p yielded the highest ethanol production in YPDX, while TDH3p promoted higher biomass formation. In the industrial strain, ADH1p, TEF1p, and PGK1p led to high ethanol yields in YPDX, with ADH1p showing superior performance in the synthetic hydrolysate. RT-qPCR reveals lower XKS1 expression levels render a better trait for BY4742, while the opposite is observed for PE-2B. CONCLUSIONS:It is demonstrated that promoter selection is crucial for optimizing XKS1 expression and xylose metabolism in S. cerevisiae. Promoters must be carefully tailored to the yeast strain and fermentation conditions to maximize ethanol production, providing strategic insights for enhancing the industrial fermentation of lignocellulosic biomass.

The agave plant’s economic chain generates a significant waste, which has potential for sustainable bioenergy through pyrolysis. However, the diversity and heterogeneity of chemical composition of agave species may pose challenges. This study investigates the impact of species heterogeneity on the pyrolysis of three different agave species (Agave sisalana, Agave tequilana, and Agave wercklei), aiming to correlate pyrolysis and biomass properties. Solid characterization and Py-GC/MS were used to understand agave physicochemical characteristics and organic group distribution in volatiles, respectively.A multi-step model and advanced numerical methods were employed for the kinetic study. The physicochemical characteristics showed similar values but a distinct distribution of inorganic compounds, predominantly composed of alkaline metals (6-11%w.b.), potentially influencing the organic groups’ distribution in the volatiles. High relative areas of aliphatic components (13-28% at 773K and 16-36% at 873K) and low quantity of acidic groups (<2%) could be attributed to the catalytic deoxygenation promoted by alkaline metals. These findings are significant for future application of agave in bio-oil production by pyrolysis, as commercial biomasses often yield a high content of oxygenated and acid groups. For the kinetic study, six decomposition profiles were identified in the pyrolysis, encompassing the decomposition of extractives, saponins, lignocellulose, and oxalate salts. The similarity in profiles resulted in approximately equivalent kinetic parameter values and mechanisms among the species. The average values of Ea ranged from 71-324 kJ mol-1, k0 values varied between 107-1022, and the reaction mechanisms included n-order and Avrami-Erofeyev types. The validity of the parameters was verified through curve reconstruction. The inorganic composition was chosen as the parameter related to pyrolysis characteristics, as it was the only parameter that differed significantly among the species. Based on the data, normalization and the proposed model demonstrated satisfactory values of R² (>0.9251), QOF (>94%), and MSE (<2.73×10-3). This underscores the model's potential to describe decomposition profiles solely based on knowledge of inorganic composition, regardless of agave species.

Reducing feeding costs without compromising rabbit health, productivity, and meat quality is essential to increase the competitiveness of rabbit meat. This study tested the hypothesis that supplementing rabbits' diet with oat hay and whole carrots, either singly or in combination, does not affect the composition and nutritional quality of the meat's lipid fraction. Eighty rabbits were divided into four groups: control (CC, fed concentrate feed exclusively), CT (whole carrot supplementation), OH (oat hay supplementation), and CO (whole carrot and oat hay). In the CC group, concentrate feed supplied 66.0 g/rabbit of total fat, decreasing by 22.8 % and 22.6 % with carrot or oat hay supplementation alone, and by 39.2 % when both were combined. Conversely, supplementation added 6.14, 18.96, and 25.1 g/rabbit of total fat in the CT, OH, and CO groups, respectively. The longissimus thoracis et lumborum (LTL) muscles used in this trial were sampled following complete processing of the rabbits, and their fatty acid profile remained mostly unchanged. Oat hay supplementation increased the proportion of odd-numbered and branched-chain fatty acids plus the C18 biohydrogenation intermediates (P = 0.024) and reduced the contents of total cholesterol by 26.5 % (P = 0.003) and total vitamin E by 56.3 % (P < 0.001). Additionally, CO supplementation improved the n-3 PUFA percentage and decreased the n-6/n-3 ratio in the LTL muscles. In conclusion, oat hay and carrot supplementation induced minor lipid profile changes but showed potential nutritional benefits, such as lower cholesterol and improved fatty acid balance. However, the supplementation strategies tested reduced total vitamin E contents in rabbit meat, potentially reducing oxidative stability.

Strobilurin fungicides inhibit mitochondrial respiration, leading to ATP depletion and oxidative stress. Although widely used in agriculture, these chemicals are ineffective against the cacao pathogen Moniliophthora perniciosa. Here, we show that M. perniciosa tolerates high concentrations of the commercial strobilurin azoxystrobin in vitro. Transcriptomic analysis revealed that short-term exposure triggers upregulation of genes related to catabolic pathways, including the glyoxylate cycle and fatty acid degradation, alongside repression of genes involved in anabolic processes, such as cell division and ribosome biogenesis. Simultaneously, genes associated with cellular detoxification and oxidative stress responses were strongly induced. These alterations suggest that M. perniciosa remodels its metabolism to counteract fungicide toxicity. Remarkably, long-term exposure to azoxystrobin led to the emergence of a resistant mutant harboring mutations in two putative growth and transcriptional regulators. This work provides new insights into the molecular basis of strobilurin resistance and informs strategies for more effective fungicide deployment in agriculture.