Biology and Life Sciences

Abstract: The domestication of aromatic plants and the valorization of distillation by-products offer promising strategies for the development of sustainable botanical pesticides. This study evaluated the chemical composition and biocidal activities of essential oils (EOs) and hydrolate-derived extracts obtained from mother (CMSAMO1) and domesticated (SAMO1) populations of Satureja montana selected from Spanish germplasm. Chemical characterization by GC-MS revealed carvacrol and thymol as the dominant constituents in EOs and dichloromethane-extracted hydrolate organic fractions (WRD), while an activated-charcoal hydrolate extract was enriched in thymoquinone (WRC). Biological activities were assessed against the root-knot nematode Meloidogyne javanica, the tick Hyalomma lusitanicum, and the insect pests Spodoptera littoralis and Myzus persicae. All extracts showed strong nematicidal activity, with LD₅₀ values ranging from 0.079 to 0.323 μg μL⁻¹, and significantly reduced nematode reproduction in tomato plants under greenhouse conditions. Hydrolate-derived fractions displayed sustained inhibition of egg hatching compared with EOs. Against H. lusitanicum, extracts from the domesticated population were more active than those from the mother plant, with the WRD fraction exhibiting the highest ixodicidal activity and persistent repellency. Antifeedant assays revealed stronger effects against M. persicae than S. littoralis, particularly for hydrolate-derived fractions. Carvacrol, thymol, and thymoquinone were identified as the principal bioactive metabolites, with principal component analysis confirming their association with biological activity. Cytotoxicity assays demonstrated a favorable selectivity of carvacrol and thymol toward tick cells. These findings demonstrate that S. montana domestication and hydrolate valorization enhance the production of chemically stable, bioactive extracts suitable for sustainable biopesticide development within circular-economy frameworks.

Abstract: Ribonucleic acid (RNA) foundation models (FMs) are reshaping computational biology, using self-supervised pretraining on massive transcriptomic corpora to acquire generalisable molecular representations. Developed initially for sequence annotation and expression prediction, they are increasingly applied to plant science, supporting tasks from gene function inference to regulatory element discovery and stress-response modelling in both model and orphan crop species. This review surveys RNA FMs with a focus on plant-relevant applications. We catalogue the major model families, including DNABERT-derived RNA adaptations, PlantRNA-FM, and Nucleotide Transformer variants, comparing their pretraining objectives, input modalities, and evaluation protocols. We examine how these models are deployed across plant genomics, transcriptomics, and functional annotation pipelines, highlighting the opportunities and the constraints arising from species-specific data gaps, limited experimental validation, and interpretability challenges. Across applications, the decisive constraint is no longer architecture but data: corpus breadth, condition-resolved benchmarks, and experimentally anchored validation now dominate the marginal gains available from model design. We close with the key open questions and a roadmap for integrating RNA FMs into the plant systems biology toolkit, emphasising benchmark design, cross-species transfer, and experimentally grounded validation. Throughout, every claim about what a model can do is held to three questions of evidence: what data the model was trained on, which benchmark supports the claim, and whether experimental validation in a plant system exists.

Abstract: Public policies such as the EU Common Agricultural Policy (CAP) support organic conversion to advance agri-food security, market-price competitiveness and trade positioning, yet cultivar-matched fruit-quality evidence for high-value niche crops remains scarce. This pilot case study compared biometric, physicochemical, biochemical, and mineral fruit traits of three organically grown sea buckthorn (Hippophae rhamnoides L.) cultivars ('Mara', 'Dora', 'Clara') and 'Clara' under conventional management from a Romanian orchard, linked to CAP support payments (€620/€442/ha/year) and a farm-gate revenue scenario using export price benchmarks (€4.0/kg). Conventionally grown 'Clara' produced larger, heavier fruit (549.3 ± 26.5 mg vs. 218.7–282.0 mg; p < 0.001) and higher polyphenol content (1446.0 mg GAE/kg) than organic cultivars, but lower DPPH antioxidant activity (57.6%) and lower soluble solids/sugar content (p < 0.01). Organic price premiums (15–40%) plus the CAP maintenance payment closed 18–45% of the estimated revenue shortfall relative to conventional cultivation. This trade-off between fruit size/polyphenol content and sugar/antioxidant quality only partly aligns with CAP payments, indicating that market-price premiums and trade positioning, not subsidies alone, are needed to make organic conversion of niche high-value crops viable and support agri-food diversification and rural resilience.

Abstract: Red clover is a major and highly important forage legume in Northern Europe. It is a perennial crop commonly grown together with grasses and harvested multiple times per season. With decreasing phenotypic stability of red clover due to various factors, mainly caused by climate change, breeders face increasing challenges when improving dry matter yield and forage quality. Marker-assisted selection using genetic markers can aid breeders in making quick decisions with higher accuracy. Candidate markers can be found through genome-wide association studies (GWAS). This study looked at the performance of longitudinal multi-trait (MT) and single-trait (ST) models for GWAS. A collection of 532 red clover accessions, both diploid and tetraploid, was phenotyped at multiple locations and genotyped using pool-seq. The target traits, dry matter yield, neutral detergent fiber (NDF), net energy for lactation (NEL) and crude protein content are highly quantitative traits that can be affected by the plant’s growth that are affected by plant development stages and growth conditions. Using a longitudinal MT model, markers associated with traits over time were identified. Genomic regions that appear to contain genes involved in the regulation of both dry matter yield and forage quality traits were also identified. Furthermore, the observed effects of significantly associated single nucleotide polymorphisms (SNPs) showed that there were considerable non-additive genetic effects in some traits. This study sets the foundation for longitudinal MT GWAS for red clover and suggests ways for further improvement to gain a deeper understanding of the genetic mechanisms underlying dry matter yield and forage quality.

Abstract: Low-coverage (low-pass) whole-genome sequencing (lcWGS) followed by imputation is a cost-effective alternative to array genotyping, but its accuracy depends on how well the target genotypes are represented in the reference panel. We assessed lcWGS imputation quality for three cattle breeds differing in their representation in a multi-breed panel: Holstein, Aberdeen-Angus and the native Russian Kalmyk breed. Imputed genotypes of 86 Kalmyk, 75 Holstein, and 72 Aberdeen-Angus animals were compared with BovineSNP50 array genotypes at 46,715 shared SNPs. Concordance and dosage R² were 99.3% and 0.98 for Holstein and 99.2% and 0.98 for Aberdeen-Angus, and 94.4% and 0.86 for Kalmyk, for which R² exceeded 0.8 at MAF > 5%, an acceptable level for common-variant analyses. PCA and ADMIXTURE placed Kalmyk cattle closest to Central- and East-Asian breeds (Kazakh, Mongolian, Hanwoo) that are poorly represented in the panel. Kalmyk cattle also showed the fastest LD decay (r² ≈ 0.04 vs. ≈ 0.10 in Holstein at 450 kb). Both the absence of the breed from the panel and the fast LD decay are expected to reduce imputation accuracy, and their contributions cannot be separated in a study of three breeds. Imputation nearly doubled the coverage of known functional markers from OMIA and other sources (from 70 to 134 positions), although casein variants absent from the panel were not recovered. Imputed lcWGS data for Kalmyk cattle are suitable for GWAS on common variants and population-structure analysis, whereas rare-variant genotyping requires a breed-specific reference panel.

Abstract: The escalating global crisis of antimicrobial resistance (AMR), often characterized as a "silent pandemic," necessitates a paradigm shift in antibiotic discovery. Traditional cheminformatics pipelines, reliant on hand-crafted molecular descriptors and low-throughput screening, are increasingly insufficient to address the evolving threats of multidrug-resistant pathogens. This Opinion article explores the transformative integration of Large Language Models (LLMs) and foundational Transformer architectures into bacterial cheminformatics. We argue that the field is transitioning from simple discriminative modeling toward a generative and agentic era. By leveraging advanced molecular representations that treat chemical structures as a linguistic medium, LLMs enable the exploration of vast, structurally novel chemotypes previously inaccessible. Furthermore, LLMs are revolutionizing microbial genome mining and the identification of cryptic biosynthetic gene clusters through the decoding of "genomic language." We highlight recent breakthroughs, including foundational discrete diffusion models and autonomous discovery agents, while addressing the critical needs for data quality and rigorous biological validation. The future of antimicrobial research lies in multi-modal, foundational systems capable of bridging the gap between genomic potential and therapeutic reality.

Abstract: Orchard intensification has transformed fruit production through the combined adoption of higher planting densities, dwarfing rootstocks, localized irrigation/fertigation, and increasingly mechanized orchard management. While aboveground outcomes are well documented, belowground responses remain fragmented across species, orchard contexts, and measurement approaches. This systematic review synthesizes field-based evidence on how orchard intensification and its main operational drivers are associated with root system architecture (RSA) and root plasticity in fruit trees. Following systematic searching and study selection, we performed a narrative synthesis because heterogeneity in species, orchard age, intensification metrics, root assessment methods, and reported traits precluded robust quantitative meta-analysis. The reviewed evidence was more frequently structured around co-occurring drivers—particularly irrigation geometry, rootstock vigour, and soil physical constraints—than around planting density manipulations alone. Root distributions were often reported as predominantly shallow and spatially concentrated within managed zones, especially along tree rows under localized irrigation, with shifts in vertical and lateral allocation that appeared context-dependent. These responses varied with species, genotype, and pedoclimatic conditions, suggesting that intensification does not impose a single deterministic RSA outcome, but rather constrains the potential root zone volume, defined here as the soil volume theoretically available for root growth, within which root distribution may be plastically modulated. Overall, we propose an interpretative framework that may help organize the available evidence by distinguishing (i) the structural definition of an effective rooting domain under intensive management and (ii) plastic modulation of root distribution within that domain. The review also highlights a major evidence gap: the scarcity of factorial field experiments that isolate planting density from irrigation design, genotype, and soil physical constraints.

Abstract: Instead of constructing conventional greenhouses, existing fixed-tilt agrivoltaic racking can be converted to passive greenhouses with transparent plastic covering. The potential of this approach has not been tested experimentally, and there are two competing processes (soil temperatures beneath agrivoltaics are 2.6°C lower than control crops because of shading), but cold-frame and greenhouse temperature are higher than ambient because of the greenhouse effect, which enables extension of the growing season in northern latitudes. To fill this knowledge gap, this study explores the adaptation of an agrivoltaics rack into a plastic-covered greenhouse. Four thermal environments were monitored from late October through December in Ontario’s agricultural belt: air temperature under plastic-covered racks and control as well as root zone temperature (RZT) under plastic-covered racks and uncovered controls. The results showed that converting agrivoltaic racks into passive greenhouses increased average RZT relative to uncovered control during late autumn and early winter in Canada. From October to December, the under-rack RZT was 3.4°C warmer than the control, while the under-rack air was 5.4°C warmer. The highest temperature difference was observed in October and November, when the under-rack RZT was ~5.7°C and 4.2°C warmer than the control, respectively. Passive plastic enclosures alone are insufficient for deep-winter agriculture but can still contribute to crop season extension (16 days observed). These results suggest that agrivoltaic racks can be adapted into protected cultivation structures for northern-latitude agriculture, reducing the cost between 30-429% % compared to commercial hoop houses.

Abstract: Plant-pathogenic bacteria such as Pseudomonas syringae pv. syringae, P. syringae pv. actinidiae, and Xanthomonas campestris pv. campestris cause major economic losses in fruit and vegetable crops, creating demand for sustainable antibacterial alternatives. In this study, an ethyl acetate extract from Cladobotryum virescens G.R.W. Arnold (C10/110) was evaluated its in vitro antibacterial activity against these three phytopathogens and chemically characterized by UHPLC-ESI-HRMS/MS. Disk diffusion assays revealed clear dose-dependent antibacterial activity, with larger inhibition halos at higher extract concentrations. X. campestris pv. campestris showed the highest sensitivity, exhibiting larger and more uniform inhibition zones and the lowest EC₅₀ value (102 mg mL⁻¹), whereas both Pseudomonas pathovars showed lower susceptibility. Dose–response modeling confirmed a concentration-dependent inhibitory effect, particularly against X. campestris pv. campestris. Metabolomic profiling allowed the identification of 21 secondary metabolites, including phenolic acids, diketopiperazines, flavonoids, fatty acids and several destruxins, most of them reported for the first time in the Cladobotryum genus. These findings highlight C. virescens as a promising source of antibacterial metabolites with potential application in sustainable plant disease management.

Abstract: Agricultural intensification and increasing climatic variability can substantially affect ar-thropod communities and the ecological functions they provide in agroecosystems. This study evaluated coleopteran diversity, abundance, and community structure in wheat, maize, and sunflower under chemically treated (V1) and untreated low-input (V2) man-agement regimes during two climatically contrasting growing seasons (2024–2025). Col-eopterans were sampled using pitfall traps, sweep netting, and manual collection, and community responses were characterized using species richness, total abundance, Shan-non–Wiener diversity, Pielou’s evenness, and Bray–Curtis-based non-metric multidimen-sional scaling (NMDS). Untreated plots consistently supported greater species richness, total abundance, and Shannon diversity across all crop × year combinations, although the magnitude of these differences varied among crops and growing seasons. Evenness showed a less consistent response, reflecting variation in species dominance among as-semblages. The warmer and drier conditions recorded in 2024 contrasted with the gener-ally cooler and wetter conditions of 2025, while interannual changes in coleopteran diver-sity were crop dependent. NMDS ordination (stress = 0.112) revealed differentiation among crop × management × year assemblages, indicating that community composition varied in relation to crop type, management regime, and growing season rather than showing a simple separation attributable to management alone. Overall, the results high-light the importance of considering biodiversity conservation, interannual climatic varia-bility, and crop-specific responses when developing adaptive integrated pest management strategies.

Abstract: Plants have developed mechanisms to regulate carbon metabolism during growth and development under both optimal and stressful conditions. In the biotrophic interaction between Coffea spp. and Hemileia vastatrix, the plant restricts pathogen access to nutrients, whereas the fungus attempts to manipulate host metabolism. Here, we elucidate the metabolic mechanisms underlying post-haustorial resistance of the Kawisari coffee hybrid to H. vastatrix by integrating metabolomic, starch, enzymatic, and gene expression analyses to distinguish between resistant (R) and susceptible (S) responses. Polar untargeted metabolomics (GC-TOF-MS) revealed distinct modulation of sugars, amino acids, polyamines, and phenolics between R and S responses. Combining these metabolomic profiles with starch quantification, semi-high-throughput enzyme activity assays, and RT-qPCR gene expression analysis revealed a strategic reallocation of carbon in the resistant response. Specifically, resistance was associated with the activation of the pentose phosphate pathway, providing reducing power and precursors for secondary metabolism (phenolic defenses), coordinated with a tight control of starch utilization and nitrogen remobilization (namely through asparagine and ornithine) and polyamine signaling. These findings underscore the pivotal role of primary metabolic networks in supporting resistance to H. vastatrix and identify promising biochemical targets for breeding coffee cultivars with enhanced resistance to rust under climate change scenarios.

Abstract: Comparative visualization of bacterial genome structure is an important component of microbial genomics, enabling the inspection of genome conservation, rearrangements, inversions, insertions and deletions, and large-scale differences in gene organization. I present bactiviz, a lightweight command-line tool for generating linear collinearity maps of bacterial genomes. Implemented as a single-binary application in Rust, bactiviz uses MUMmer for nucleotide-level pairwise alignment and produces publication-ready, self-contained SVG figures. The tool is designed specifically for multi-replicon bacterial assemblies and supports chromosomes, plasmids, and other sequence records within GenBank and FASTA inputs. bactiviz provides optional origin normalization of circular replicons using a cascading gene-identification strategy based on gene symbols, locus tags, and product annotations, thereby accommodating heterogeneous real-world GenBank annotations. Conserved regions between adjacent genomes are represented as identity-scaled ribbons, including strand-aware visualization of inversions, while annotated genomic features are rendered as directional gene arrows. An automated label-placement procedure prioritizes user-specified genes and places additional labels where sufficient space is available, reducing overlap while retaining informative gene-level annotation at different output scales. A redundancy-resolution procedure further reduces visually cluttered alignments caused by repetitive sequences and duplicated genomic regions. The resulting SVG output is resolution-independent and can be directly incorporated into manuscripts or edited using standard vector-graphics software. bactiviz is intended for analyses in which a defined ordering of genomes is to be visualized rather than for all-against-all or multiple-genome alignment. By combining MUMmer-based alignment with native multi-replicon handling, genome-orientation normalization, exhaustive feature rendering, and automated annotation placement, bactiviz provides a lightweight and reproducible workflow for producing interpretable linear comparative-genomics figures.

Abstract: Citrus fruits are considered the most consumed fruits in the world; however, their production is becoming less efficient and their cost has increased, mainly due to the rapid spread of Huanglongbing (HLB), a disease whose vector is an insect called Diaphorina citri. Controlling this vector is essential to controlling the disease, which is complicated by the lack of knowledge about the population dynamics of D. citri. Mexico is the fourth largest citrus producer internationally. Therefore, the objective of this study was to evaluate the popula-tion dynamics and aggregation indices of D. citri in “Valencia Late” orange crops in three locations in northern Veracruz State, Mexico. To achieve this, minimum and maximum temperatures, precipitation, and relative humidity were recorded. Linear regression mod-els were developed between these environmental parameters and the number of adult psyllids and nymphs. The results showed a positive association between the development of new outbreaks and the presence of population peaks of nymphs and adults in Febru-ary-March and July-August. Therefore, it is suggested that broad-spectrum insecticides and mineral oils be applied during this period. Rainfall reduced the presence of nymphs at the three locations. The calculated aggregation indices showed that the distribution of nymphs was aggregated, and that of adults tended to be so, but during periods of low density, their distribution was random.

Abstract: Internal greening of the artichoke (Cynara scolymus L.) floral receptacle — commonly termed the “heart” — is an unresolved postharvest quality defect that reduces marketability, particularly for export. No published study to date has directly scored this specific defect against controlled nitrogen or biostimulant treatments in artichoke. Here we synthesize established plant-physiology principles (nitrogen's role in chlorophyll biosynthesis, the carbon-to-nitrogen ratio's influence on the vegetative-to-reproductive transition, and biostimulant-driven cell division) with crop-specific evidence from globe artichoke field trials that measured chlorophyll content and receptacle biochemistry — though not visible internal greening itself — under varying nitrogen and biostimulant regimes. We propose that late-stage nitrogen and biostimulant application during head formation is the most physiologically plausible driver of internal greening, and we explain why the defect becomes visually apparent only after boiling (chlorophyll unmasking following cell-wall breakdown, rather than heat-induced pigment formation). We present this as a testable hypothesis, not a proven causal mechanism, and outline the controlled field trial — scoring visible internal greening directly — needed to confirm it.

Abstract: Medicinal quality of Scutellaria baicalensis Georgi is decided by baicalin and wogonoside in root. In arid and semi-arid regions this crop is grown, salinized soils constrain establishment but reshape specialized metabolism. We used field density trials, salt-stress experiments, UPLC-MS/MS and time-series RNA-seq to ask whether agronomic practices and salt-induced transcriptional changes alter root flavone content. Planting density showed a clear yield-quality trade-off: higher density increased belowground biomass per unit area but reduced individual root growth and flavone content, with 100,000 plants mu-1 providing a practical balance between population yield and root quality under the tested Xinjiang field conditions. In established seedlings, 0.5% NaCl caused no visible damage over 24 h and specifically raised baicalin and baicalein at 12 h, while wogonin-type flavones remained nearly unchanged. Time-series RNA-seq identified thousands of differentially expressed genes between 2 and 24 h. WGCNA identified a turquoise module that was overrepresented with genes for carbohydrate metabolism and cytoskeletal/microtubule processes. Salt-responsive NAC, MYB and WRKY transcription factors co-expressed with core structural genes (PAL, 4CL, CHS, CHI); one candidate, SbNAC076, was up-regulated mainly at 24 h after the metabolite peak, consistent with a role in sustaining rather than triggering accumulation. These findings outline the temporal dynamics of salt-induced branch-selective flavone accumulation and suggest candidate regulators for functional validation, with implications for quality-focused cultivation on saline marginal lands.

Abstract: Cross-referencing an Arabidopsis thaliana locus against UniProt accessions, Gene Ontology (GO) terms, TAIR10/Araport11 genomic coordinates, InterPro domain annotations, Plant Ontology terms, and PubMed literature currently requires the integration of several independently formatted flat files. In many laboratories, these tasks are performed using ad hoc scripts that are developed for individual analyses and are rarely parallelized. Here, I present araseq (https://codeberg.org/gsablok/araseq), a single Rust binary that implements thirteen cross-referencing operations as clap subcommands and, optionally, exposes the same functionality through an axum-based web server. All operations execute within a user-configurable Rayon thread pool and follow a common read-index-filter paradigm: identifier lists are normalized where appropriate, reference files are streamed into in-memory hash maps, and the resulting maps are filtered according to the requested identifiers. I describe the implementation and independently reproduce two representative algorithms in Python using the TAIR10/Araport11 sample files distributed with the repository to assess algorithmic correctness. The evaluation confirms the expected behavior of the normalized matching workflow while also identifying an inconsistency in identifier normalization between modules. These findings indicate that araseq provides a compact and reusable framework for routine cross-referencing of A. thaliana annotation resources.

Abstract: Endophytic fungi are an important source of bioactive compounds with potential applications in sustainable pest management. In this study, we isolated an Epicoccum species from Bethencourtia palmensis, an endemic plant from the Canary Islands, and performed bioactivity-guided fractionation of its ethyl acetate extract. The extract exhibited significant bioactivity against several agricultural pests, including Myzus persicae, Spodoptera littoralis, Botrytis cinerea, Dickeya dadantii and Pseudomonas syringae, and against the tick Rhipicephalus bursa. After bioguided fractionation, the major constituent of the extract and responsible for most of its bioactivity was identified as 2,3-dihydro-2-hydroxy-2,4-dimethyl-5-trans-propenylfuran-3-one (BPF3-1). Its structure was elucidated by 1D and 2D NMR experiments and high-resolution electrospray ionization mass spectrometry (HRESIMS) data analysis, and its absolute configuration was established by comparison of experimental and calculated electronic circular dichroism (ECD) spectra. This is the first report of BPF3-1 production by an Epicoccum species and the first documentation of its insect antifeedant and acaricidal activities. The culture medium was subsequently optimized to enhance BPF3-1 production. Maximum production was achieved using a medium supplemented with fructose and mycological peptone with a 487% production over control, and this formulation was selected for an initial fermentation scale-up in a bench-top bioreactor. Under these conditions, the maximum BPF3-1 concentration was reached three days earlier than in flask cultures, demonstrating the feasibility of process intensification and representing a first step toward industrial-scale production.

Abstract: Graphene oxide (GO) is a widely used biocompatible nanomaterial, whose unique carbon-based structure bears an abundance of oxygen-containing functional groups. An increasing number of investigations have been implemented on the effects of GO on plants due to its hydrophilicity and biocompatibility. Employing the GO (6 mg/L) synthesized by our group it was found that GO promoted the growth rate of poplar roots tissue culture seedlings. The intuitive TEM images of these root cell samples depicted that high concentrations of GO ( > 6 mg/L) can lead to two physiological changes (plasmolysis and an increased number of starch grains). From these images, it can be seen that GO adhered to the cell membrane on the inner side of the cell wall. In order to gain insight about GO’s oxidation on the adventitious roots, antioxidant enzymes’activities, such as polyphenol oxidase (POD), catalase (CAT) and superoxide dismutase (SOD) were measured.The data showed that SOD、POD and CAT were increased significantly compared with the control group, the range of the increase in multiples is between 2.8 and 4.5. Transcriptome gene testing data indicated that related gene expressions were enhanced at low concentration of GO, but inhibited at high concentration. This work confirms that GO of appropriate size and concentration can promote the growth of poplar tissue culture seedlings.

Abstract: This study evaluated the effects of supplemental LED lighting on the photomorphogenesis and phytochemical characteristics of flowers of Chrysanthemum indicum L. Four lighting treatments were applied, including three red and blue LED spectral ratios (30:70, 55:45, and 78:22) under 6 h of daily supplemental lighting, and a natural sunlight control. Supplemental photosynthetic photon flux density was maintained at 100 µmol·m⁻²·s⁻¹. The highest average flower number (196.2 flowers·plant⁻¹) was obtained under the control treatment (F1), whereas the greatest average fresh flower biomass (55.5 g·plant⁻¹), dry flower biomass (7.4 g·plant⁻¹), essential oil content (0.695%, DW), and essential oil yield (10.01 kg·ha⁻¹) were achieved under F4 (R:B = 78:22). These parameters gradually declined with decreasing red light proportions. Gas chromatography revealed that the flower essential oil was dominated by 2,2,6-trimethyl-3-keto-6-vinyl tetrahydropyran, with the highest relative abundance under F3 (R:B = 55:45; 39.9%) and the lowest under F2 (32.5%). No significant differences were observed in the antimicrobial activity of the essential oils, whereas the strongest anti-inflammatory activity was detected under F3. These findings provide a scientific basis for optimizing supplemental LED spectra to maximize flower productivity, essential oil quality, and pharmaceutical value of C. indicum in cultivation systems.

Abstract: The European Union (EU) recently adopted Regulation (EU) 2026/1388 for plants developed with new genomic techniques (NGTs). Under this Regulation, Category 1 NGT plants are exempt from most EU requirements applicable to genetically modified organisms, provided that they meet, among other things, the criteria of equivalence defined in Annex I. Although these criteria consist of simplified molecular characteristics and numerical thresholds, their molecular, analytical, and regulatory verification is more demanding than the legislative wording may suggest at a first glance. Demonstrating compliance requires comprehensive identification of relevant genetic modifications, alignment with genome annotations, evaluation of the gene pool for conventional breeding, assessment of polyploid genomes, and attribution of genetic changes to the use of NGTs. Accordingly, enforcement of Annex I will require robust technical guidance defining sequencing standards, bioinformatic tasks, annotation requirements, attribution specifications, and harmonized counting methodologies. To this end, we propose a seven-step workflow from data-generation to regulatory evaluation in order to ensure a reliable and standardized verification process for Category 1 NGT categorization.