Stratified application of seed treatment components reduces neonicotinoid phytotoxicity and maintains physiological quality of soybean seeds during storage

ABSTRACT

Industrial seed treatment protects soybean seeds against pests and diseases, but prolonged storage causes progressive physiological quality loss driven by phytotoxicity from neonicotinoid insecticides. To our knowledge, no prior research has evaluated whether stratified application, by applying components in ordered layers, reduces phytotoxicity while maintaining seed quality during storage. We evaluated 10 treatment processes combining fungicide, insecticides, polymer, and finishing powder in stratified or simultaneous sequences on soybean seed physiological quality, stored for 135 days at 20°C. A completely randomized 10 × 4 factorial design (10 processes × four storage periods: 0, 45, 90, and 135 days) with four replications was used. Physiological quality was assessed through germination in paper roll plus vermiculite, seedling emergence, accelerated aging, and modified accelerated aging in substrate. A phytotoxicity index was calculated for each test and analyzed via multivariate analysis and k-means clustering. Simultaneous application of all components (Mix) showed the highest phytotoxicity, reducing germination by 4 percentage points (pp) and vigor by up to 6 pp compared to stratified treatments. Treatments with fungicide in the first stage formed a low-phytotoxicity cluster, while Mix occupied an isolated high-phytotoxicity cluster. Seeds maintained viability for up to 90 days and vigor for up to 45 days under controlled storage. Stratified application, particularly with fungicide and polymer applied first, significantly reduces neonicotinoid phytotoxicity and represents a practical strategy to extend seed safety margins in the soybean seed production chain.

Key words
application technology; Glycine max L.; seed quality; seed safety; vigor

INTRODUCTION

Industrial seed treatment (IST) protects seeds against pests and diseases through application of phytosanitary products (Reis et al. 2023, Reis et al. 2026). However, the increased duration of contact between these products and seeds during storage, driven by logistical requirements in seed commercialization, presents a critical challenge: many active ingredients (AIs), particularly neonicotinoid insecticides, induce phytotoxicity that intensifies during storage, progressively reducing seed quality (Carvalho et al. 2020; Pereira et al. 2021).

In the Brazilian seed industry, the average time between industrial treatment and planting is approximately three months. However, depending on the region and weather-related planting delays, some seed lots can be stored for up to six months. Therefore, evaluating extended storage limits covering this timeframe is crucial.

Seed deterioration during storage is accelerated by chemical-induced phytotoxicity, elevated temperature and humidity, and high slurry volumes (Santos et al. 2018, Nardelli et al. 2025). Previous studies have documented that simultaneous application of multiple components increases chemical exposure intensity and phytotoxic effects (Rocha et al. 2025). However, the impact of sequence and stratification of application, applying treatment components sequentially rather than simultaneously, has shown superiority in corn but remains unexplored in soybeans (Medeiros et al. 2026).

Despite extensive research on formulation chemistry and AI interactions in IST, the role of application sequence and physical layering of components on seed surface dynamics and physiological quality during storage remains unexplored in soybean. To our knowledge, no prior study has investigated whether stratified application, that is, applying treatment components in ordered, sequential layers on the seed surface, can attenuate phytotoxic effects while maintaining protective efficacy throughout storage.

The logical basis for this approach rests on preventing the insecticide from coming into direct contact with the seed coat during its peak permeability window. We hypothesize that by applying a preliminary layer (e.g., polymer or fungicide), the resulting physical barrier significantly reduces phytotoxicity (Reis et al. 2026). Therefore, the objective of this study was to evaluate the effects of sequential and simultaneous application processes of fungicide, insecticides, polymer, and finishing powder on the physiological quality and phytotoxicity of soybean seeds during controlled storage.

MATERIALS AND METHODS

Soybean seeds of cultivar Monsoy 6410 from the 2023/24 crop season were used. The experiment was conducted in a completely randomized design with a 10 × 4 factorial arrangement and four replications, comprising 10 IST processes and four storage periods after IST (initial evaluation period, 45, 90, and 135 days). Seeds were weighed in 1-kg portions and subjected to treatment using a Momesso Arktos Laboratório L5K machine equipped with 15 Hz frequency and operated for up to 30 seconds. The treatment involved sequential stratification of components across up to three stages, each lasting 10 seconds.

The 10 treatment processes combined five commercial components. The fungicide consisted of metalaxyl-M (20 g.L^-1), thiabendazole (150 g.L^-1), and fludioxonil (25 g.L^-1), applied at 100 mL.100 kg^-1 of seeds. The first insecticide was cyantraniliprole (600 g.L^-1), applied at 60 mL.100 kg^-1, while the second insecticide was thiamethoxam (350 g.L^-1), applied at 200 mL.100 kg^-1 of seeds. The polymer Biocroma red (density 1.10 g.cm^-3) was applied at 100 mL.100 kg^-1, and the finishing powder Biogloss biogrow was applied at 100 g.100 kg^-1. In the control treatment, consisting only of fungicide and polymer, distilled water was used to equalize the final volume of the slurry, which was standardized for all treatments at 460 mL.100 kg^-1. Table 1 presents the stratification sequence of components for each treatment. Whenever insecticide is listed as a single stage component, both insecticide products were combined and applied simultaneously within that stage.

Table 1
Sequential stratification in the application of components and commercial product dosage for soybean seed treatment.

ID	Seed treatment process			F	I1	I2	P	FP	W	FSV
	Stage 1	Stage 2	Stage 3	(mL)	(mL)	(mL)	(mL)	(g)	(mL)	(mL)
Control	F+P	-	-	100	-	-	100	-	260	460
F/I+P	F	I+P	-	100	60	200	100	-	-	460
F/I+P/FP	F	I+P	FP	100	-	-	100	100	-	460
F/P/I	F	P	I	100	60	200	100	-	-	460
F+P/I	F+P	I	-	100	60	200	100	-	-	460
F+P/I/FP	F+P	I	FP	100	60	200	100	100	-	460
F+P/FP/I	F+P	FP	I	100	60	200	100	100	-	460
I/P/F	I	P	F	100	60	200	100	-	-	460
Mix	F+P+I	-	-	100	60	200	100	-	-	460
Mix/FP	F+P+I	FP	-	100	60	200	100	100	-	460

• ID: identification code; F: fungicide; I¹: insecticide (Cyantraniliprole); I²: insecticide (Thiamethoxam); I: simultaneous application of I¹ and I² at the same stage; P: polymer; FP: finishing powder; W: water; FSV: final slurry volume; product dosage: mL or g·100 kg^-1 of seeds; stage 1, 2 and 3: application sequence stratification, each applied for 10 seconds using a laboratory seed treating machine; –: not applied, only mixture.

Following treatment, seeds were stored in a biochemical oxygen demand (BOD) incubator at 20 ± 1°C in multifoil paper bags (approximately 250 g each, randomly selected and identified for each storage period). Physiological quality evaluations were performed at initial evaluation period (72 hours after treatment), 45, 90, and 135 days of storage, using the following tests:

• Germination in paper towel roll plus vermiculite (RP+V): Four replicates of 50 seeds per treatment were placed between two sheets of paper towel moistened with distilled water in an amount equivalent to three times the paper weight, plus commercial medium vermiculite moistened (100 mL per roll, 1:1 vermiculite:water ratio) distributed in a uniform layer over the paper. Rolls were maintained in a Mangelsdorf-type germinator at 25 ± 2°C, with evaluation of normal seedlings at five (first count, FC_RP+V) and eight days after sowing (Carvalho et al. 2024, MAPA 2025);

• Seedling emergence in controlled conditions (SE): Four replicates of 50 seeds per treatment were placed in a substrate composed of a mixture of sand and soil (2:1 volume ratio) arranged in plastic trays irrigated at 60% of water-holding capacity at sowing and maintained in a greenhouse at 25 ± 2°C under alternating light and dark regimes (12 h each). Evaluation was performed five days after sowing, recording the percentage of emerged seedlings (Krzyzanowski et al. 2020);

• Accelerated aging in paper (AA): The test was conducted in plastic boxes adapted with suspended aluminum mesh, containing 40 mL of distilled water. A single layer of seeds was uniformly distributed on the mesh, boxes were sealed with plastic film, and maintained in a BOD chamber at 41 ± 1°C for 48 hours (Marcos-Filho 2020). Afterwards, four replicates of 50 seeds each were sown between two overlapping sheets of paper towel moistened with distilled water in an amount equivalent to 2.5 times the paper weight. Papers were rolled and maintained in a germinator at 25 ± 2°C, with evaluation of normal seedlings at 5 days after sowing (MAPA 2025).

• Modified accelerated aging in substrate (AAS): Like the AA test, but after 48 h at 41 ± 1°C, seeds were sown in a substrate composed of sand and soil (2:1 volume ratio) placed in a plastic tray, moistened at 60% of water-holding capacity. Trays were maintained in a growth chamber at 25 ± 2°C under alternating light and dark regimes (12 h each), with evaluation at eight days after sowing, recording the percentage of emerged seedlings (Rocha et al. 2025).

Statistical analysis

Data were first evaluated for compliance with analysis of variance (ANOVA) assumptions. Analysis of variance was then performed in the RStudio environment, and when the F-test was significant at the 5% level, treatment means were compared using the Scott–Knott’s test at 5% significance (R Core Team 2024). Polynomial regressions were fitted to the data from each physiological test to evaluate the behavior of variables across storage periods.

For each analyzed variable, a phytotoxicity index (iX) was calculated according to Eq. 1. In this index, the control treatment in each storage period is used as the reference (C), and only reductions in physiological quality relative to the control generate positive values, which are interpreted as phytotoxicity; improvements relative to the control are not the focus of this index. Negative iX values (i.e., treatments performing better than the control) were set to 0, so that the index exclusively represents phytotoxic effects. The index is dimensionless, but expressed as a percentage, and it was calculated for each physiological test and each process of ST.

(1) $\mathrm{iX}\left(\mathrm{\%}\right)=\frac{(\bar{\mathrm{C}}-\mathrm{T})}{\bar{\mathrm{C}}}\times 100$

where: iX: Phytotoxicity percentage index for the specific physiological quality test; C: Mean value of the analyzed variable for the control treatment in the same storage period; T: Value of one of the replicates of the analyzed variable for the ST processes.

These phytotoxicity indices (iFC_RP+V, iRP+V, iSE, iAA, iAAS) were then used as input variables for principal component analysis (PCA) to reduce data dimensionality and identify the main patterns of variation among treatments (Jolliffe 2002). The first two principal components were retained for subsequent analyses. The k-means algorithm was applied to the scores of these two components to cluster treatments, with the number of clusters fixed at k = 3 based on exploratory analysis. The algorithm was run 25-folds with different random initializations, and the partition with the lowest intra-cluster variance was selected (Hartigan and Wong 1979). Results were visualized in a biplot showing treatment scores, loading vectors of the original variables, and confidence ellipses for each cluster. PCA was performed using the FactoMineR package, and k-means clustering using the stats package (Lê et al. 2008).

RESULTS AND DISCUSSION

All ANOVA assumptions were met. In the variables first count of germination on paper towel roll plus vermiculite (FC_RP+V), standard germination (RP+V), seedling emergence under controlled conditions (SE), and accelerated aging on paper (AA), no significant interaction between factors was observed (p > 0.05). For accelerated aging in substrate (AAS), a significant interaction was detected.

In first count germination testing, the control, F+P/I, and F+P/FP/I treatments yielded germination values 4 percentage points (pp) higher than the Mix process (p ≤ 0.05; Fig. 1a). During 135-day storage, all treatments maintained 94% normal seedlings (Fig. 1b). In germination testing, the treatment with simultaneous application of all components was inferior to the other processes, showing a 4 pp reduction in normal seedlings in the Mix treatment relative to the control (Fig. 2a). Although no significant difference among storage periods was detected up to 90 days, polynomial regression indicated a marginal downward trend (Fig. 2b), with significant decline observed only at 135 days.

Figure 1
Normal seedlings (%) of soybean obtained from the first count of germination by the rolled paper plus vermiculite as affected by (a) seed treatment processes (n = 16) and (b) storage periods (n = 40). Coefficient variation = 3.16%*.

Figure 2
Normal seedlings (%) of soybean obtained from germination by the rolled paper plus vermiculite as affected by (a) seed treatment processes (n = 16) and (b) storage periods (n = 40). Coefficient variation = 2.58%***.

These findings align with corn ST research. Medeiros et al. (2026) documented identical patterns in maize, in which stratified neonicotinoid applications demonstrated superior physiological quality compared to simultaneous mixing. The consistency across both crops indicates that application order fundamentally influences physiological quality.

The overall phytotoxicity reduction and maintenance of germination values observed in the best-performing treatments can be attributed to the technological benefits of the stratified application. By creating a physical and spatial separation of the active ingredients on the seed coat with the initial applications, the stratification prevents the direct and immediate contact of highly concentrated phytotoxic products with the seed tissue, ensuring a safer and more localized chemical exposure. Furthermore, stratification does not increase the total processing time nor subject the seed to detrimental mechanical overexposure; it simply introduces a slight sequential delay in activating specific application nozzles to ensure successive layering, maintaining the operational efficiency of the machinery.

In seedling emergence testing (SE), treatments with insecticide applied first (I/P/F), Mix, and Mix+FP were inferior to the other processes, but they did not differ from F+P/I/FP and F+P/FP/I (Fig. 3a). Within this lower-performing group, the I/P/F treatment, for example, exhibited 6 pp lower emergence vigor compared to the control. Through 90 days of storage, no significant differences were observed among treatments, but regression analysis indicated a linear deterioration over time, with a decline of 1.2 pp per month, culminating at 135 days with emergence close to 88% (Fig. 3b).

Figure 3
Emerged seedlings (%) of soybean obtained from emergence in controlled conditions as affected by (a) seed treatment processes (n = 16) and (b) storage periods (n = 40). Coefficient variation = 3.81%***.

Medeiros et al. (2026) observed that insecticide-first treatments in corn exhibited deterioration rates of 0.85–0.94 pp per month, compared to only 0.38–0.53 pp per month when fungicide plus polymer preceded insecticide application. While this pattern across crops highlights a shared susceptibility to neonicotinoids, physiological sensitivity to these treatments is highly genotype-dependent. The intensity of phytotoxicity can vary significantly according to the specific genetics and inherent seed coat characteristics of each soybean cultivar rather than representing an absolute universal principle.

The differential performance derives from the differential imbibition window. At initial application, the seed coat is physiologically dry and highly permeable, generating a steep water potential gradient that strongly favors absorption of water and solutes (McDonald 1999). Products applied first penetrate deeply into seed tissues. During subsequent rapid imbibition, the seed coat progressively hydrates, and its pores become occupied by initially-deposited components, progressively reducing permeability and limiting absorption of subsequent applications to more superficial layers (Salanenka and Taylor 2011, Yang et al. 2018).

Therefore, insecticide-first treatments such as I/P/F likely underperform because insecticide, applied first when seed coat permeability is maximum, penetrates deeply and maximizes phytotoxic tissue contact (Reis et al. 2026). Conversely, treatments with fungicide applied first (e.g., F+P/I), which statistically grouped with the control in the highest performance category (Fig. 3a), succeeded because fungicide and polymer likely established an initial protective layer. When neonicotinoid was subsequently applied to an increasingly impermeable seed coat, it may have remained more superficial, minimizing phytotoxic exposure. The simultaneous Mix treatment represents the worst scenario: all components compete for absorption at maximum permeability, resulting in intense penetration of all compounds. These mechanisms are proposed based on observed physiological patterns and require further experimental validation to confirm the role of sequential application in modulating chemical penetration and phytotoxicity.

Through 90 days of storage at 20°C, physiological quality remained stable with no significant differences between initial evaluation period and 90 days, both for germination (Fig. 2b) and emergence (Fig. 3b). This 90-day inflection point represents the temporal threshold in which oxidative stress and metabolic changes accumulate sufficiently to compromise seed quality.

Seeds treated with phytosanitary products experience chronic metabolic stress as cellular machinery attempts to detoxify and compartmentalize absorbed chemicals. During the initial 90 days, antioxidant defense systems effectively neutralize reactive oxygen species (ROS) generated by chemical detoxification (Carvalho et al. 2022). However, sustained chemical exposure progressively depletes antioxidant reserves. Beyond 90 days, antioxidant capacity becomes limiting, permitting ROS accumulation and uncontrolled oxidative damage to cellular membranes and proteins (Pereira et al. 2024, Li et al. 2025).

Rocha et al. (2025) documented that neonicotinoid-treated soybean maintained stable germination through 60 days, under uncontrolled storage conditions, with pronounced deterioration beginning beyond 90. Medeiros et al. (2026) confirmed that corn seeds treated with F+P/I sequences maintained vigor for up to 12 months, but neonicotinoid-treated lots showed accelerated deterioration rates of 0.85–0.94 pp per month beyond the initial stability phase. The 90-day threshold in soybean represents an earlier deterioration inflection, possibly reflecting greater metabolic sensitivity of this crop to chemical stress, under controlled storage conditions at 20°C.

In the AA, the Mix treatment exhibited pronounced vigor loss, achieving only 68% germination, 6 pp below stratified treatments, indicating that treatment with Mix components may accentuate quality loss regarding stratified treatments (Fig. 4a). During extended storage, this deterioration intensified markedly, declining at a rate of approximately 12 pp per month (Fig. 4b), far exceeding the 1–2 pp per month observed under non-stressed conditions, according to quality tests previously presented.

Figure 4
Normal seedlings (%) of soybean obtained from germination after accelerated aging as affected by (a) seed treatment processes (n = 16) and (b) storage periods (n = 40). Coefficient variation = 5.47%***.

In accelerated aging in substrate (AAS), all treatments maintained similar vigor through 45 days, except for the I/P/F, Mix, Mix+FP, and control treatments (Fig. 5). In this storage period, most underperforming treatments had insecticide present in the first application layer. However, beyond this threshold, treatment responses diverged dramatically, revealing complex interactions between finishing powder addition and substrate-mediated stress, particularly with advancing storage beyond 90 days.

Figure 5
Emerged seedlings (%) of soybean obtained from emergence in controlled conditions, after modified accelerated aging in substrate, as affected by seed treatment processes and storage periods (n = 4). Coefficient variation = 7.72%***.

Treatments receiving finishing powder demonstrated notably reduced vigor at 135 days. Simultaneously, Mix and I/P/F treatments lacking finishing powder declined to below 25% emerged seedlings by 135 days, representing 17 pp per month deterioration in the Mix process, reinforcing the relationship between insecticide in the first application layer and intensified phytotoxic effects.

Suzukawa et al. (2018) concluded that IST soybean seeds exhibit more intense physiological loss across storage compared to untreated controls, with accelerated aging amplifying these effects substantially. The authors attributed this pattern to seed-treatment-imposed metabolic costs that manifest progressively and are dramatically magnified under stress conditions. Furthermore, Reis et al. (2025) verified that neonicotinoids are the most aggressive components, reducing germination potential even without storage.

PCA of five phytotoxicity indices, for each response variable, revealed that the first two components explained 90.3% of total variation (PC1 = 76.1%, PC2 = 14.2%; Fig. 6). PC1 represented the general level of phytotoxicity, while PC2 contrasted distinct response patterns to phytotoxic stress.

Figure 6
Biplot of principal component analysis showing the grouping of seed treatment processes based on phytotoxicity indices across all storage periods. Points represent seed treatment process colored by the first stage of application. Ellipses delimit the three identified clusters. Vectors indicate the five evaluated variables.

Among the indices, the first count phytotoxicity index on germination (iFC_RP+V) with loading 0.956 and the emergence phytotoxicity index after accelerated aging in substrate (iAAS) with loading 0.921 made the largest contributions to PC1, followed by accelerated aging on paper (iAA, loading 0.862) and standard germination on germination (iRP+V, loading 0.857). The emergence index under controlled conditions (iSE) predominated in PC2 with loading 0.643.

K-means clustering identified three biologically distinct treatment phenotypes. Cluster 1 aggregated five low-phytotoxicity process treatments (control, F/I+P, F+P/I, F/I+P/FP, F/P/I) on the negative PC1 axis, all of these treatments featuring fungicide in the first application layer. Cluster 2 isolated the Mix treatment on the positive PC1 axis (PC1 = 4.32), indicating maximum phytotoxicity and biologically distinct behavior, reinforcing the potential of correctly designed stratification processes and their effects in attenuating phytotoxicity in treated seeds. Cluster 3 congregated four intermediate-phytotoxicity treatments (F+P/FP/I, F+P/I/FP, I/P/F, Mix/FP) in intermediate position. The separation between clusters accounted for 84.3% of total variation.

Carvalho et al. (2020) and Rocha et al. (2025) demonstrated that phytotoxicity manifests across multiple physiological dimensions simultaneously, germination, emergence, stress resilience, and membrane integrity, rather than as a single unidimensional trait. Our multivariate analysis confirms this multidimensionality: the high loadings of distinct indices on PC1 indicate that treatments simultaneously affecting multiple physiological pathways achieve the greatest overall phytotoxic impact.

The polymer’s function transcends simple adhesion. Applied in initial positions, the polymer creates a hydrophobic surface layer that acts as a diffusion barrier, reducing direct contact between subsequent applications and seed tissues (Avelar et al. 2012, Medeiros et al. 2026, Reis et al. 2026). When the fungicide and polymer are applied first, the polymer coating physically blocks subsequent insecticide penetration into deep seed tissues. The neonicotinoid remains largely external to the seed or penetrates only the outermost layers. This spatial separation, achieved through application sequence, dramatically reduces phytotoxic exposure and has a direct impact on physiological quality.

In contrast, when the insecticide is applied first (I/P/F), it penetrates deeply before any protective layer can form. The subsequent polymer application can only protect against further external exposure, but it cannot reverse the internal penetration already achieved.

Stratified treatment processes achieved substantially greater maintenance of seed viability and vigor over extended storage by reducing cumulative physiological costs imposed by industrial treatment. Combined with controlled-temperature storage (20°C), stratified treatments maintained high seed quality for up to 90 days post-treatment, with 45 days for vigor through the most aggressive test (AAS) and 90 days for viability, representing a critical seed safety margin and logistical flexibility gain.

The fundamental principle emerging is that AI arrangement during treatment significantly determines physiological quality maintenance during storage. Application of fungicide or fungicide plus polymer in initial stages provides superior protection against neonicotinoid phytotoxic effects compared to immediate insecticide application. This likely operates through multiple synchronized mechanisms: reduced slurry volume minimizes absorption intensity, initial polymer application physically restricts subsequent insecticide penetration, and sequential application allows partial drying between stages, reducing total metabolic stress.

Further research is necessary to evaluate stratification efficacy across different soybean cultivars and industry formulations. Critical investigations should include field validation of agronomic efficacy, particularly regarding whether intermediate polymer positioning affects insecticide persistence and pest control, as well as experiments with alternative cultivars to determine whether stratification benefits generalize across soybean genetic backgrounds (Reis et al. 2026).

CONCLUSION

The properly designed sequential stratification of seed treatment components reduces phytotoxic effects compared to simultaneous application of all components, thus presenting potential for use in the soybean seed production chain.

Under controlled baseline storage conditions at 20°C, seeds maintained viability for up to 90 days and vigor for up to 45 days post-treatment. It is important to note that these specific safety margins cannot be directly extrapolated to non-climate-controlled storage environments subject to thermal fluctuations. However, under these baseline conditions, stratified application processes, particularly those with fungicide and polymer in the first stage, tended to extend these safety margins, as evidenced most clearly by accelerated aging tests.

ACKNOWLEDGMENTS

The authors would like to acknowledge the Seedcare Institute Syngenta for their technical support, seed treatment resources provided during the experimental phase, and financial assistance. We also thank the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), the Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG), and the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) for their financial support, including scholarships and a research productivity grant (CNPq).

• How to cite:
  Gusmão, L. V., Barros, E. S., Reis, V. U. V., Maciel, D. C., Tavares, G. I. S., Fernandes, K. A. C., Alberti, I. A., Cruz, T. A., Caetano, C. C., Santos, H. O. and Carvalho, E. R. (2026). Stratified application of seed treatment components reduces neonicotinoid phytotoxicity and maintains physiological quality of soybean seeds during storage. Bragantia, 85, e20260072. https://doi.org/10.1590/1678-4499.20260072
• FUNDING
  Coordenação de Aperfeiçoamento de Pessoal de Nível Superior
  Finance code: 001
  Conselho Nacional de Desenvolvimento Científico e Tecnológico
  Grant No.: 307200/2025-6
• DECLARATION OF USE OF ARTIFICIAL INTELLIGENCE TOOLS
  The authors declare that artificial intelligence tools (Perplexity) were used solely to check and improve English grammar and language fluency. The authors take full responsibility for the scientific content, original ideas, and integrity of this manuscript.

DATA AVAILABILITY STATEMENT

The datasets generated and analyzed during this study are available from the corresponding author upon reasonable request.

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Edited by

• Section Editor:
  Hector Valenzuela https://orcid.org/0000-0001-8415-5453

Publication Dates

• Publication in this collection
  10 Aug 2026
• Date of issue
  2026

History

• Received
  23 Mar 2026
• Accepted
  27 May 2026