Entomopathogenic microbial potential in the management of Fall Armyworm, Spodoptera frugiperda (J.E. Smith) in Maize Production

Entomopathogenic microbial potential in the management of Fall Armyworm, Spodoptera frugiperda (J.E. Smith) in Maize Production | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Entomopathogenic microbial potential in the management of Fall Armyworm, Spodoptera frugiperda (J.E. Smith) in Maize Production Iwebaffa Amos Edet, Oluwafolake Adenike AKINBODE,, Iwebafa George Oluwadamilare, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7022858/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Fall armyworm ( Spodoptera frugiperda ) poses a significant threat to maize production in sub-Saharan Africa, particularly in Nigeria, where infestation levels continue to disrupt food security. This study investigates the efficacy of entomopathogenic microbial inoculants, delivered through biochar-based formulations, as a sustainable strategy for managing S. frugiperda in maize cultivation. Field and screenhouse trials were conducted to evaluate the effects of various treatment combinations involving Bacillus thuringiensis, Trichoderma spp., and synthetic insecticide(Ampligo) under sprayed and non-sprayed conditions. Agronomic traits disease incidence, and yield parameterswere assessed. Results revealed that the combination of NPK (50 kg/ha), Mycorrhiza, and biochar (T2) significantly improved plant growth, reduced armyworm damage, and enhanced yield performance, closely rivaling chemical control method. Principal Component Analysis (PCA) confirmed that T2 contributed to superior vegetative vigour (PC1) and physiological stability (PC2). These findings supported the integration of microbial inoculants and biochar as part of an environmentally friendly and scalable Integrated Pest Management (IPM) approach. Spodoptera frugiperda maize biochar microbial inoculants Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Figure 8 Figure 9 Figure 10 INTRODUCTION Maize ( Zea mays L. ) remains one of the most important cereal crops globally, serving as a staple food for both humans and livestock and as a key raw material in various industrial applications, including biofuel production (Togola et al ., 2025,FAO, 2023, Swati et al., 2024 ). However, maize cultivation is increasingly threatened by biotic stresses, particularly from insect pests such as the fall armyworm ( Spodoptera frugiperda ), which has emerged as a major invasive species in tropical and subtropical agroecosystems (Paude et al ., 2022 Mlambo et al., 2024 ; Kasoma et al., 2021 , Zanzana, et al., 2024 ). Originally native to the Americas, S. frugiperda was first detected in Africa in 2016 and has since spread rapidly across the continent, with severe infestations recorded in West Africa, including Nigeria’s key maize-growing regions (Makgoba et al ., 2020; Tambo et al., 2020 Anjorin et al., 2021 ). The larvae feed aggressively on maize leaves, tassels, and ears, leading to substantial reductions in photosynthetic capacity and yield. Unchecked infestations can result in yield losses ranging from 20–50% (, Anjorin et al., 2021 ; Matova et al., 2020 ; Kris 2024). Adding to the challenge, the pest's subterranean pupation—typically 3–5 cm below the soil surface—enables it to evade surface-applied insecticides (Thirawut et al., 2023 ). Climate change had further compounded the fall armyworm threat (Zanzana et al., 2024 ). The fluctuation in rainfall patterns and the rising temperatures have further expanded the pest’s survival window, reproductive capacity, and geographical range, thereby intensifying its impact on food security in vulnerable regions such as southwestern Nigeria (Yan et al., 2022 , Mir et al., 2024). Conventional pest management strategies rely heavily on synthetic insecticides. While these chemicals are effective, their overuse has led to pesticide resistance, environmental contamination, and its adverse effects on non-targeted organisms, which includes beneficial insects (Sangaraju et al., 2024 , Pereira et al., 2024 , Verma et al .,2024 ,Tyagi., et al., 2024, Yao et al ., 2023; Koskey et al., 2021 ). These have resulted into growing interest in sustainable alternatives, particularly biological control methods employing entomopathogenic and plant growth-promoting microorganisms. Microbial agents such as Bacillus thuringiensis (Bt), Pseudomonas putida , and Trichoderma spp. have shown promise for their dual roles in pest suppression and plant health promotion (Irsad et al ., 2023; Martínez-Salgado et al ., 2023). Bt, a spore-forming Gram-positive bacterium, produces insecticidal crystal proteins (Cry and Cyttoxins) that disrupts the midgut epithelium of larval insects, that ultimately results into mortality (Pradeep et al., 2021, Ma et al., 2023 ). Trichoderma spp., in addition acts as an alternative biopesticides, also enhances plant vigour by the inducement of systemic resistance that improves nutrient acquisition (Waleed et al., 2024 Sood et al., 2020 Lanzuise et al., 2022 , Bharti et al., 2024 ).As recent studies have highlighted the synergistic potentials of using biochar—a stable, carbon-rich soil amendment derived from pyrolyzed agricultural biomass—as a carrier for microbial inoculants. Biochar improves soil structure, water-holding capacity, and nutrient availability, thereby the enhancing the microbial survival and efficacy in the field (Luo et al.,2025, Ali et al.,2025 Jatuwong, et al 2025 , Shiv et al 2023 ). When combined with entomopathogenic microorganisms, biochar can support integrated pest and soil fertility management in a sustainable and scalable manner (Fritz, et al., 2022 , Xiang et al.,2022 ). This study aims to evaluate the efficacy of entomopathogenic microbial inoculants formulated with biochar-based carriers for the sustainable management of Spodoptera frugiperda in maize cultivation. The specific objectives are to: Assess the effectiveness of biochar as a carrier for microbial inoculants in enhancing soil fertility and pest suppression. Evaluate the biocontrol potential of Bacillus thuringiensis and microbial consortia against fall armyworm under both controlled and field conditions, and to investigate the impact of biochar-based biopesticide applications on maize productivity, soil health, and integrated pest management strategies in smallholder farming systems. MATERIALS AND METHODS Microbial Isolation and Characterization Soil samples were collected from maize rhizospheres at the Institute of Agricultural Research and Training (IAR&T), Ibadan, Nigeria. Serial dilutions up to 10⁷ CFU/mL were prepared and plated on nutrient agar and King’s B medium using the spread plate technique. Distinct colonies were isolated and subjected to Gram staining and a suite of biochemical assays, including catalase, oxidase, and indole-3-acetic acid (IAA) production. For precise identification, molecular characterization of the bacterial isolates was performed via amplification of the 16S rRNA gene using universal primers 27F (5'-AGAGTTTGATCMTGGCTCAG-3') and 1492R (5'-TACGGYTACCTTGTTACGACTT-3'). PCR products were purified and sequenced via Sanger sequencing. The sequences recorded were analyzed using BLAST against the NCBI GenBank database and aligned using MEGA 14 for phylogenetic tree construction. Key entomopathogenic strains—Bacillus thuringiensis, Pseudomonas putida, and Klebsiella variicola—were screened for biocontrol attributes such as siderophore production (CAS assay),(Chiho et al., 2021) phosphate solubilization (Pikovskaya’s medium),(Singh et al ., 2016) and chitinaseGomaa et al., 2012 and protease activity.(Bakkiet al., 2024 ) Experiment 1: In-Vitro Bioassay against Spodoptera frugiperda Pure cultures of B. thuringiensis were grown in Luria-Bertani (LB) broth at 30°C and harvested at 24, 48, 72, and 96 hours. Cultures were centrifuged at 12,000 rpm for 15 minutes to separate the supernatant and cell pellets. Bioassays were conducted by incorporating varying concentrations (1 × 10⁶ to 1 × 10⁹ CFU/mL) of Bt suspensions into an artificial diet fed to third-instar S. frugiperda larvae, reared under controlled laboratory conditions. Mortality was recorded at 12-hour intervals over 96 hours. Lethal concentration (LC₅₀) and lethal time (LT₅₀) were calculated using Probit analysis in R v4.2.0. Experiment 2: Screenhouse Trial (Artificial Infestation) A screenhouse experiment was conducted to evaluate the efficacy of microbial and biochar treatments under controlled pest pressure. Maize seeds (variety SUWAN 1 ) were sown in sterilized soil and grown to the V3 growth stage. Each plant was manually infested with one first-instar larva of S. frugiperda . Treatments included: Biochar + Bacillus thuringiensis Biochar + Trichoderma spp. Ampligo® (Chlorantraniliprole + Lambda-cyhalothrin) as chemical control Untreated control Each treatment was applied via foliar spray at 500 mL per plot using a hand-held atomizer. Damage severity was assessed using digital image analysis with Image J v1.45i, quantifying percentage leaf area loss. Larval mortality was monitored daily. Experiment 4: Field Trials under Natural Infestation Field trials were conducted at the IAR&T experimental station (7°26′N, 3°54′E), Ibadan, Nigeria, from April to August 2024. A split-plot design was used: Main plots: Sprayed vs. non-sprayed Plot Sub-plots: Seven treatment combinations: T1: Mycorhiza + Biochar T2: NPK Fertilizer (50 kg N/ha) + Mycorrhiza + Biochar T3: NPK Fertilizer (50 kg N/ha) T4: NPK Fertilizer (100 kg N/ha) T5: Untreated Control T6: Ampligo (Chemical insecticide) T7: Trichoderma spp. Biopesticides and insecticides were applied at 30 days after planting (DAP) and repeated at 10-day intervals for three cycles. Data Collection and Analysis Agronomic parameters assessed included: Growth metrics: Plant stand, plant height (PH_6wk, PH_8wk), stem girth (6wk and 8wk), ear height, and number of leaves (6wk and 8wk) Phenology: Days to tasseling, days to silking Structural and yield traits: Root lodging, stalk lodging, ear aspect, ear and plant at harvest, grain weight, and field weight Disease and pest scores: Foliar damage scores using Bajracharya’s 0–5 scale(Karki, et al 2023 ) Chlorophyll content and leaf moisture were also evaluated. Bajracharya (0–5) scale: Score Description 0 No visible damage 1 Window-paning on whorl leaves 2 Few small holes in upper leaves 3 Ragged holes and partial whorl damage 4 Extensive damage to whorl and upper leaves 5 Whorl destruction and drying of plant Pest infestation and disease severity were monitored weekly using the five-point sampling method. Populations of natural enemies (Coccinellidae and Chrysopidae) were also recorded to assess ecological balance. All data were analyzed using Analysis of Variance (ANOVA) in R Studio v4.2.0, and treatment means were separated using Duncan’s Multiple Range Test (DMRT) at p ≤ 0.05. Visualizations and summary statistics were generated in R Studio v4.2.0 RESULT The molecular and cultural characterization confirmed the identification of three entomopathogenic strains: Bacillus thuringiensis, Pseudomonas putida , and Klebsiella variicola . All isolates exhibited key biocontrol and plant growth-promoting traits. B. thuringiensis showed strong chitinase and protease enzyme activity. P. putida and K. variicola also demonstrated consistent phosphate solubilization and siderophore production. These results confirm the dual potential of the isolates for both pest suppression and nutrient mobilization. In-vitro bioassay of Bacillus thuringiensis against Spodoptera frugiperda , bioassay results (Fig. 1 ) showed a progressive increase in larval mortality with increased concentration and incubation duration of B. thuringiensis cultures. The LC₅₀ value decreased from 3.4 × 10⁷ CFU/mL at 24 hours to 2.6 × 10⁷ CFU/mL at 72 hours. The LT₅₀ for the 72-hour culture was recorded at 48.2 hours. The mortality data followed a sigmoid response curve, confirming concentration-dependent efficacy across all exposure intervals. Screenhouse Experiment: Artificial Infestation (Fig. 2 ) All bio-inoculated treatments significantly reduced foliar damage compared to the untreated control. The biochar + Bacillus thuringiensis treatment achieved the highest larval mortality (78%) and the lowest mean severity score (1.7 on a 0–5 scale), indicating a 65% reduction in leaf area loss relative to the control (4.3, p < 0.01). Trichoderma-treated plants recorded a 59% larval mortality rate, while the chemical control, Ampligo®, achieved 71% mortality. The agronomic performance of maize under seven different treatments (T1–T7) and sprayed and non-sprayed plot is summarized in Fig. 3 . The graph illustrates the mean values of multiple traits, including growth, structural integrity, and phenological development, enabling comparative evaluation across integrated nutrient and pest management strategies: Vegetative Growth and Canopy Development with Plant height measured at 6 and 8 weeks (PH_6wk, PH_8wk) was highest under Treatment T2 (50 kg N/ha + Mycorrhiza + Biochar), closely followed by T3 (Inorganic N only) and T7 (Trichoderma spp.). Stem Girth and Structural Integrity Stem girth at both 6 and 8 weeks followed a comparable trend, with T2 and T1 outperforming other treatments. The increase in girth of stem under biochar treatments was from enhanced root development and water retention, contributing to better nutrient translocation and structural stability. Treatments with synthetic insecticide alone (T6) or no input (T5) lagged in this regard, which suggested a lack of supportive soil structure or biological stimulation. Phenological Traits: Silking and Tasseling Days to silk and tasseling were shortest in T2, These indicated accelerated reproductive maturity. which suggested that microbial consortia, in combination with moderate nitrogen application, promoted early phenological progression—a trait beneficial in short-season agroecologies. In contrast, T1 and T5 exhibited the longest durations to silk and tassel, that reflected delayed development under reduced nutrient input or stress conditions. Lodging Resistance and Plant Quality Root and stalk lodging recorded remained generally low across the treatments, although slightly elevated in the untreated control (T5) and T6. The best structural resilience was however recorded in biochar-integrated plots (T2, T1),This supported the role of enhanced root architecture and mechanical strength via improved soil conditioning. Ear Development and Yield Proxy Traits Ear height, ear aspect, and plant-at-harvest ratings were consistently higher in T2 and T3, with T2 thus exhibiting superior uniformity in ear placement and harvest quality. These traits are positively correlated with grain yield and harvestability. However the lower scores recorded in T5 and T6 suggested underperformance under nutrient-deficient or solely chemical management systems. Chlorophyll Content and Leaf Moisture Response (Fig. 4 ) presented the chlorophyll concentration and leaf moisture status measured at 6 and 8 weeks after planting (WAP) across the seven treatment combinations, stratified by spray status. These parameters serve as vital physiological indicators of plant health and stress tolerance. Chlorophyll Content (Fig. 4 ) At 6 weeks (Chlor_6wk), the SPAD chlorophyll values ranged from 44.0 ± 1.9 in Treatment 4 (100 kg N/ha) to 52.7 ± 2.1 in Treatment 2 (50 kg N/ha + Mycorrhiza + Biochar), This highlighted the significant impact of integrated nutrient management on early-stage photosynthetic potential. Both T1 and T2 (biochar-based microbial treatments) consistently showed higher SPAD readings than the untreated control (T5). While sprayed plots generally recorded higher chlorophyll values than their non-sprayed counterparts (~ 1.5 SPAD units), this difference was not statistically significant (p > 0.05). By 8 weeks (Chlor_8wk), chlorophyll levels increased in all treatments, It thus peaked in the sprayed T6 (Ampligo; 65.0 ± 2.3 SPAD) and sprayed T7 (Trichoderma spp.; 63.1 ± 2.0). However, biochar-microbial treatments maintained competitive chlorophyll values with the avoidance of the trust on synthetic insecticides, which further underscores their ability to sustained photosynthetic efficiency under pest pressure. Leaf Moisture Content Leaf moisture percentages showed a slightly different trend (Fig. 4 ). Moisture content at harvest ranged from 19.4 ± 0.9% in non-sprayed T2 to a maximum of 25.2 ± 1.1% in sprayed T6. Although sprayed plots had ~ 1% more moisture on average, the difference was not significant across treatments. Treatments T1 and T2 maintained moderate moisture levels (22.3–23.1%) which suggested that biochar contributed to soil water retention and delayed desiccation, especially under non-sprayed conditions. Grain and Field Weight Response Across Treatments Figure 5 displays the mean values for 100-grain weight and field weight across the seven treatments under both sprayed and non-sprayed conditions. These indicators serve as critical yield components that reflected the combined effects of nutrient input and pest management strategy.Field Weight Field weight ( Fig. 5 ) was highest in Treatment 3 (T3: 50 kg N/ha only), this reached 1.10 ± 0.25 kg in non-sprayed plots and 0.80 ± 0.15 kg in sprayed plots. Treatment 2 (T2: 50 kg N/ha + Mycorrhiza + Biochar) followed with 0.68 ± 0.10 kg (No spray) and 0.58 ± 0.08 kg (Yes spray), This outperformed the untreated control (T5: 0.72 ± 0.12 kg vs. 0.67 ± 0.10 kg). The lowest values recorded were observed in T7 (Trichoderma alone: 0.12 ± 0.03 kg, No spray) and T6 (Ampligo: 0.22 ± 0.05 kg, No spray), although the sprayed raised T6’s yield to 0.55 ± 0.07 kg. Grain Weight(Fig. 5 ) The grain weight trends mirrored field weight. T3 again produced the highest yield, with 1.05 ± 0.30 kg (No spray) and 0.80 ± 0.18 kg (Yes spray). T2 was a close second, this achieved 0.95 ± 0.20 kg (No spray) and 0.75 ± 0.12 kg (Yes spray). T1 (Mycorrhiza + Biochar) followed, while T5 showed only moderate gains (0.38 ± 0.10 kg vs. 0.45 ± 0.12 kg). Notably, insecticide spray improved grain weight in poorly performed treatments (T4–T7) but reduced yield slightly in T2 and T3, likely due to phytotoxic stress or disruption of microbial-plant symbiosis. Pest Infestation and Damage Suppression(Fig. 6 ): illustrated the mean foliar damage caused by Spodoptera frugiperda at 6 and 8 weeks after planting across the seven treatments under both sprayed and non-sprayed regimes. Infestation levels varied significantly among treatments and over time. At 6 weeks, the highest infestation scores were recorded in the non-sprayed untreated control (T5: 3.2 ± 0.15) and chemical control (T6: 3.1 ± 0.12). In contrast, biochar–microbial treatments, particularly T1 (Mycorrhiza + Biochar: 2.8 ± 0.10) and T2 (Inorganic N + Mycorrhiza + Biochar: 2.4 ± 0.08), showed significantly reduced damage. The Foliar sprayed further enhanced control across treatments, with sprayed T2 recorded the lowest damage at 2.3 ± 0.09, with, outperformed even T6 (sprayed: 3.0 ± 0.13).and by 8 weeks, a general decline in infestation was observed across all plots. The non-sprayed T5 remained the most infested (2.5 ± 0.14), while non-sprayed T2 dropped to 2.3 ± 0.10. Notably, sprayed T2 (1.8 ± 0.07) and T1 (2.0 ± 0.08) exhibited the most substantial reduction,that outperformed even the sprayed chemical treatment T6 (2.2 ± 0.09).Hence, the T2 (Inorganic N + Mycorrhiza + Biochar) was the most resilient treatment against S. frugiperda, which maintained low infestation scores at both time points. Thus, It suggested that biological soil amendments not only promote plant health but also serve as a strategic tool for sustainable pest suppression, that instilled a viable alternative to exclusive chemical reliance in smallholder maize systems. Disease Severity Across Treatments (Fig. 7 ): displayed the mean disease severity scores (0–5 scale) for six foliar diseases—Blight, Curvularia, Ear Rot, Rust, Streak, and Stripe—across the seven treatment combinations (T1–T7) under both sprayed and non-sprayed conditions. Across all diseases, severity scores remained in the low range (1.0–3.5), with no treatment these completely eliminated the symptoms.Blight: Scores in non-sprayed plots ranged from 1.1 to 1.4, with the lowest in T5 (1.0 ± 0.12) and highest in T4 (1.4 ± 0.15). Sprayed plots consistently recorded higher values (up to 1.7 ± 0.18 in T1), suggesting a marginal spray-induced increase. Curvularia: Disease severity in non-sprayed plots was relatively stable (~ 1.25–1.35), with a minor dip in T3 (1.25 ± 0.10). Sprayed plots spiked early in T1 (1.9 ± 0.20) before settling across treatments (~ 1.3–1.6). Ear Rot: Highest in sprayed T7 (2.5 ± 0.18) and sprayed T2 (1.8 ± 0.14), while the non-sprayed T4 recorded the lowest severity (1.2 ± 0.13). A steady increase in sprayed plots indicated elevated vulnerability. Rust: Disease levels were minimal (1.0–1.5) across all treatments. T2 (sprayed: 1.5 ± 0.13; non-sprayed: 1.3 ± 0.11) showed slightly elevated rust compared to others. Streak: Fluctuated between 1.0 and 1.6. The highest was observed in non-sprayed T6 (1.6 ± 0.14) and sprayed T2 (1.5 ± 0.12). Stripe: Showed the most notable spray-related impact. While non-sprayed plots remained between 1.1 and 1.5, sprayed treatments surged to as high as 2.2 ± 0.18 (T3), with all others which exceeded 1.8. Principal Component Analysis (PCA) ( Fig. 8 and Fig. 9) : Trait Performance and Treatment Differentiation thus,Figs. 8 and 9 illustrated the observed physiological values of the on PC1 and PC2 scores across the seven treatment combinations (T1–T7), separated by foliar spray plot (Spray vs. Non-Spray). These principal components collectively explain the majority of variability among growth, physiological, and yield-related traits associated with fall armyworm management. PC1 (Fig. 8 ):PC1 scores captured the composite variance in traits linked to vegetative vigor and biomass productivity (e.g., plant height, leaf number, grain and field weight). The highest PC1 scores were recorded under Treatment T4 (100 kg N/ha) and T2 (50 kg N/ha + Mycorrhiza + Biochar), especially in sprayed plots, with mean scores exceeding 2.0 and 1.5, respectively. Non-sprayed plots, particularly T1 and T5 (Mycorrhiza + Biochar and untreated control), displayed negative PC1 scores, indicating limited contributions to the primary trait dimension under non-chemical regimes. PC2 (Fig. 9): PC2 scores varied within ± 1.5 and primarily captured physiological resilience, chlorophyll content, and structural integrity (e.g., lodging resistance). Non-sprayed T2 showed the highest negative score (− 1.1), suggesting its distinct influence on physiological traits. Biochar-microbial treatments (T1 and T2) maintained relatively stable PC2 scores regardless of spray status, while T6 (Ampligo) scored lower on PC2 despite high PC1 values, suggesting reduced physiological stability. Trait–Treatment Interaction ( Fig. 10): Correlation and Physiological Response to Management Strategies: The multivariate response of key agronomic traits to treatment and spray regimes (Fig. 10) illustrated the physiological and structural adjustments of maize under different pest and nutrient management strategies. Traits such as field weight, grain weight, plant height, stem girth, and leaf number were assessed across seven treatments (T1–T7), with clear delineation between sprayed and non-sprayed conditions with Growth Parameters and Structural Development Plant height (PH_6wk and PH_8wk) was highest under T2 (Inorganic N + Mycorrhiza + Biochar) and T3 (50 kg N only), with foliar spraying further enhancing early vegetative development, particularly under T2. Stem girth at both 6 and 8 weeks mirrored this trend, Photosynthetic Potential and Canopy Growth Leaf number at 6 and 8 weeks was also highest in T2 and T7 (Trichoderma). These treatments likely benefited from phytohormonal stimulation by Trichoderma spp. and improved root nutrient acquisition, respectively. Interestingly, leaf development was slightly suppressed in sprayed T6 (Ampligo), suggesting a possible trade-off due to insecticide-induced stress or negative impact on beneficial microbial populations. The yield Components (Fig. 10): Biomass Accumulation and Kernel Fill Grain and field weight responses aligned with vegetative vigor trends. T3 (50 kg N) recorded the highest field and grain weight under sprayed conditions, while T2 closely followed, demonstrating that microbial and biochar inputs can partially offset the need for synthetic inputs. The untreated control (T5) and chemical-only plot (T6) showed inconsistent performance, reinforcing the advantage of integrated biological systems. DISCUSSION This study confirms that combining moderate levels of inorganic nitrogen(NPK Fertilizer) (50 kg N/ha) with biochar and microbial Mycorrhiza (T2) significantly enhances maize growth, structure, and yield potential across both chemical and non-chemical pest management systems. Treatments T2 and T1 consistently outperformed chemical-only controls in both physiological stability and agronomic performance. Notably, PC2 and trait regression results revealed that biochar–microbial combinations support chlorophyll retention, stem integrity, and kernel development—traits essential for resilience under fall armyworm infestation. Microbial Isolation and Functional Characterization Molecular analysis confirmed the identity of three entomopathogenic strains: Bacillus thuringiensis, Pseudomonas putida, and Klebsiella variicola, each of the microbes identified gives a characteristics biocontrol traits. Although B. thuringiensis exhibited strong chitinase and protease activity, which aligned with its known larvicidal properties (Anuj Ranjan, et al.,2024, Martínez-Zavala et al.,2019, Franco-Rivera et al., 2004 ). Additionally, P. putida and K. variicola demonstrated phosphate solubilization and siderophore productionwhich indicated the potential for promoting nutrient availability and inducing systemic resistance (Al-Turki et al., 2023 Goswami et al.,2013). These attributed characteristics traits makes the isolated strains not only as biopesticides but also as Mycorrhiza with dual functionality in integrated pest and soil fertility management. Similar dual-action benefits have also been observed in prior work on rhizospheric bacteria by Kowalska et al. ( 2020 ), where microbial consortia enhanced maize growth while suppressing pests. The in-Vitro Bioassay of Bacillus thuringiensis Against Spodoptera frugiperda Larval mortality increased with both Bt concentration and incubation time. The LC₅₀ value declined markedly from 3.4 × 10⁷ CFU/mL at 24 hours to 2.6 × 10⁷ CFU/mL at 72 hours,which suggested an enhanced potency in older cultures, this is possibly due to the increased accumulation of Cry and Cyt toxins (Lianget al., 2025 ,Karshanal et al.,2023). The Larval response showed a typical sigmoid mortality curve, and LT₅₀ recorded indicated that the 72-hour formulations achieved rapid knockdown effects within 48.2 hours. These results are consistent with the recent findings by Karshanal et al. (2023), who reported that Bt-based products significantly outperformed synthetic alternatives in terms of larval suppression without harming non-target species. Hence,all bio-inoculated treatments significantly reduced foliar damage compared to the untreated control. The biochar + B. thuringiensis combination achieved the highest larval mortality (78%) and the lowest mean severity score (1.7 on a 0–5 scale),which represented a 65% reduction in leaf area loss compared to the control (4.3, p < 0.01). Trichoderma-treated plants showed moderate protection (59% mortality), while Ampligo®, the chemical control, reached 71% efficacy. The enhanced protection conferred by biochar-based microbial treatments likely stems from improved microbial colonization and persistence in the phyllosphere, as reported by Kaul et al., ( 2021 ). The Biochar provided a conducive microhabitat for microbial adhesion, while the microbial metabolites exert both direct toxicity and systemic induced resistance in the plant. In otherword,, the Ampligo treatment, though effective, did not outperform the Bt + biochar treatment and may pose long-term resistance risks and ecological drawbacks, as previously cautioned by Chen et al. ( 2023 ).In the field experiment the line graph indicated that T2 consistently outperformed other treatments in nearly all agronomic parameters, which emphasizes the value of integrated Mycorrhiza and biochar inputs. These enhancements in plant stature, leaf production, and reproductive maturity under T2 are indicative of improved nutrient uptake and systemic plant health. The consistent pattern across sprayed and non-sprayed plots further highlighted the robustness of biological amendments under varying pest pressures.T1 (Mycorrhiza + biochar without inorganic N) showed intermediate performance, affirming the growth-promoting role of microbial consortia even in the absence of synthetic nitrogen(NPK fertilizer). In contrast, T5 (untreated) and T6 (chemical control) showed limited enhancement of growth traits,which implicated the failure and the limitations of non-integrated or purely chemical approaches. These observations aligned with reports presented by Huey et al. ( 2020 ) and Sudip et al. (2023), which found that combining biochar with beneficial microbes significantly improves maize agronomy through better root colonization, stress tolerance, and nutrient mobilization. The progressive increase in chlorophyll from weeks 6 to 8 reflects sustained canopy development and continued nutrient availability. T2 outperformed the other treatments which may be due to enhanced nitrogen uptake and microbial nutrient mineralization, this also corroborated the findings by Mthiyane et al., ( 2024 ) and Liu, et al. ( 2024 ), who reported that biochar and microbial consortia enhance chlorophyll biosynthesis and delay senescence. Hence,the highest chlorophyll readings at 8 weeks occurred in sprayed T6 and T7 plots, which also suggested effective pest suppression via synthetic or microbial means and may reduce the defoliation and stress in the maize plant thereby preserving photosynthetic tissues (Zhao, et al., 2019 and Gekas et al., 2013, ). Nonetheless, T2’s non-sprayed performance was statistically comparable, this reinforced the robustness of biochar-based microbial treatments in the enhancement of physiological resilience even in the absence of foliar insecticides. The moisture data also aligned with these trends.Hence the biochar's porous structure contributed to improved water retention and buffering against mid-season droughts, as previously documented by Jiawei et al. (2025). While the chemical treatments achieved higher moisture content, this may reflect delayed maturity or water accumulation due to denser canopies rather than physiological efficiency.(Duan et al.,2024) The enhanced vegetative vigour in T2 reflected synergistic nutrient availability from both inorganic and biological sources. Similarly, the number of leaves at 6 and 8 weeks was highest in T2 and T7, confirming the promotive effects of microbial Mycorrhiza and biochar on canopy development. These findings are consistent with previous reports where biochar-amended rhizospheres improved leaf formation and photosynthetic efficiency (Jabborova et al., 2021). The chlorophyll and moisture results validated the physiological benefits of biochar-integrated microbial strategies. Treatments T1 and T2 consistently improved plant vitality indicators under both pest-suppressed and unsprayed conditions.(Liu et al.,2022) These findings support the incorporation of biochar-based microbial inputs into sustainable maize production systems to enhance crop resilience, reduce chemical dependency, and maintain agro-ecological balance (Luo et al.,2025) also from the results it can be confirmed that biochar-amended microbial inoculants (T1 and T2) effectively suppress fall armyworm infestation, rivaling or surpassing the efficacy of the synthetic insecticide (T6: Ampligo). The early damage reduction observed in T2 suggests that Mycorrhiza–biochar synergy disrupts the pest’s lifecycle, potentially by enhancing plant vigour and induced systemic resistance (Chen et al., 2023 Chisonga et al.,2023). Also at 8 weeks,the armyworm pest infestation remained significantly lower in T2 and T1 regardless of spray status,which indicated residual protective effects which may be due to persistent microbial colonization and biochar's soil amendment properties. Which is consistent with Kasoma, et al. ( 2023 ) discovery, who emphasized the role of entomopathogenic microbes in long-term pest suppression under field conditions. With the addition of foliar spray to T1 and T2 plot further provided additive benefits, that suggested the beneficial effect of integrated pest management (IPM) strategies combined with microbial bioinoculants with minimal chemical input can maintain efficacy with reduced ecological risk (Zhou et al., 2024 ). Moreover, lower pest pressure in biochar-amended plots can also enhance the effectiveness of natural enemies and reduce the likelihood of resistance buildup. The yield advantage of T3 (inorganic N) have emphasised the centrality of nitrogen in supporting kernel development, even under pest stress. However, T2's comparable performance indicated that biochar-borne microbial inoculants can substitute partially for synthetic N while sustaining productivity. This synergy is consistent with the findings of Kohira et al. ( 2025 ) and Beilei et al. (2023), who demonstrated that biochar improves nutrient and moisture retention critical to grain filling. The modest yield reductions in sprayed T2 and T3 treatments also suggested a potential trade-off between pest control and plant health, due to phytotoxicity or chemical interference with microbial colonization (Yu et al., 2023 ). Hence these findings emphasized the importance of balancing chemical inputs with biological amendments for optimized yield.(Rajiv et el.,2024) Moreover, the gains observed in lower-yielding treatments following spraying (T6 and T7) confirm that insecticide application remains effective where biological support is lacking. However, their limited ecological benefits and dependence on external inputs highlighted the superior agronomic efficiency of integrated approaches in T2.Treatment 2 (moderate N + biochar + Mycorrhiza) emerged as the most efficient and sustainable strategy, delivering yields comparable to full chemical regimes with fewer ecological trade-offs. While biopesticide and biochar-based treatments (T1, T2, T7) were effective in the reduction of insect pest pressure of Spodoptera frugiperda, their impact on disease suppression was less pronounced. Disease severity scores across most treatments and conditions remained relatively low, but a consistent pattern emerged—sprayed plots tended to exhibit slightly elevated disease severity, particularly for Stripe.(Rajiv et al., 2024) This disease increase may be attributed to foliar spray effects such as physical cuticle damage or increased leaf moisture retention, which promoted opportunistic fungal colonization (Abdelaaziz et al., 2025). The pronounced rise in Stripe under foliar sprays (+ 0.7 units on average) signals a potential trade-off in integrated pest management (IPM)—while insecticides effectively suppress pests, they may inadvertently create favorable microclimates for disease development.(Golan et al., 2023 ). Moreover, biochar–microbial amendments (T1, T2, T7) did not consistently suppress foliar diseases when compared to the untreated control (T5). This could suggest that under moderate pest pressure, the antagonistic activity of endophytes against foliar pathogens may be insufficient unless augmented by fungicides (Waqas et al., 2017 ). The analysis reveals that chemical insecticide application, while effective against pests, may unintentionally elevate foliar disease risk, particularly under humid conditions. This finding supports a refined IPM approach—integrating microbial Mycorrhiza with low-phytotoxicity or botanically derived insecticides, and pairing them with targeted fungicide applications when disease pressure is high(Cucu et al., 2025 ) Such a strategy could optimize both pest and disease management while minimizing environmental impacts in maize cultivation systems(Henri et al., 2025 ) Spray Interaction Effects The overall, sprayed plots exhibited elevated trait values, but the differential between sprayed and unsprayed conditions was smallest in T2 and T1. Thus suggesting that microbial inoculants and biochar build resilience even in the absence of synthetic chemical inputs. The stable performance of non-sprayed T2 plots hence supported the findings by Zheng et al. ( 2025 ), who noted that biochar–microbial systems sustained physiological function under biotic stress. Thus, reaffirming the synergistic effect of microbial Mycorrhiza and biochar on water and nutrient uptake (Li et al.,, 2023). The biochar matrix may have contributed to improved soil aeration and microbial colonization, which then resulted in superior physiological support(Khan, et al., 2024 ) The PCA effectively differentiated treatment impacts based on composite trait performance: PC1 Interpretation – Growth and Yield Efficiency shows that the treatments with integrated inputs—particularly T2 (50 kg N/ha + Mycorrhiza + Biochar)—demonstrated the most balanced trait enhancement, scoring high on PC1. This supports the previous findings that moderate inorganic fertilization supplemented with microbial consortia and biochar enhances nutrient use efficiency, early vigour, and yield accumulation (Faloye et al., 2024 ). In contrast, lower PC1 values in T5 and T7 shows the limitations of untreated or Trichoderma-only strategies in the absence of nutrient support.(Sanjay et al.,2018) PC2 Interpretation – Physiological Integrity and Disease Mitigation:shows the High PC2 scores in T1 and T2 which highlighted their role in sustaining physiological traits—such as chlorophyll content, moisture retention, and disease resistance—independent of yield-centric gains.(Lopes and Reynolds 2012 ) This also reinforces the benefits of microbe-driven resilience, as corroborated by Muaz et al. (2024) and Chaudhary et al. ( 2022 ), who reported that endophyte-rich systems mitigate stress effects and promote systemic resistance.(Muaz et al., Hassan 2025) The Trade-offs with Synthetic Control:show from the observation thatT6 (Ampligo), while enhancing yield metrics (PC1), consistently underperformed on PC2. This indicates potential disruption of beneficial plant–microbe interactions or physiological processes by chemical inputs—an ecological trade-off also noted by Burlakoti et al. ( 2024 ). Conclusion and recommendation Therefore future trials should focus on refining spray timing and exploring microbial consortia capable of dual pest and disease suppression to fully realize the potential of integrated pest management (IPM) systems in maize cultivation. The PCA evidently showed the multidimensional efficacy of integrated treatments like T2, which maximized both productivity (PC1) and physiological health (PC2). 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PMID: 37196943 Srivastava RK Sanju Purohit, Edris Alam, Md Kamrul Islam Advancements in soil management Optimizing crop production through interdisciplinary approaches,Journal of Agriculture and Food Research Volume 18,2024,101528,ISSN 2666 – 1543 https://doi.org/10.1016/j.jafr.2024.101528 ( https://www.sciencedirect.com/science/article/pii/S2666154324005659 ) Srivastava RK Sanju Purohit, Edris Alam, Md Kamrul Islam Advancements in soil management Optimizing crop production through interdisciplinary approaches,Journal of Agriculture and Food Research,Volume 18,2024,101528,ISSN 2666 – 1543, https://doi.org/10.1016/j.jafr.2024.101528.(https://www.sciencedirect.com/science/article/pii/S2666154324005659 ) Abdelaaziz Farhaoui M, Taoussi S-E, Laasli I, Legrifi NE, Mazouni A, Meddich M, Hijri R, Lahlali (2025) Arbuscular Mycorrhizal Fungi and Their Role in Plant Disease Control: A State-of-the-Art,The Microbe,2025,100438,ISSN 29501946, https://doi.org/10.1016/j.microb.2025.100438 ( https://www.sciencedirect.com/science/article/pii/S2950194625002067 ) Golan K, Kot I, Kmieć K, Górska-Drabik E (2023) Approaches to Integrated Pest Management in Orchards: Comstockaspis perniciosa (Comstock) Case Study. 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Agronomy 13(9):2203. https://doi.org/10.3390/agronomy13092203 Khan S, Irshad S, Mehmood K, Hasnain Z, Nawaz M, Rais A, Gul S, Wahid MA, Hashem A, Abd_Allah EF, Ibrar D (2024) Biochar Production and Characteristics, Its Impacts on Soil Health, Crop Production, and Yield Enhancement. Rev Plants 13(2):166. https://doi.org/10.3390/plants13020166 Faloye OT, Ajayi AE, Kamchoom V, Akintola OA, Oguntunde PG (2024) Evaluating Impacts of Biochar and Inorganic Fertilizer Applications on Soil Quality and Maize Yield Using Principal Component Analysis. Agronomy 14(8):1761. https://doi.org/10.3390/agronomy14081761 Mahato S, Bhuju S (2018) Jiban Shrestha Effect Of Trichoderma Viride As Biofertilizer On Growth And Yield Of Wheat. Malaysian Journal of Sustainable Agriculture (MJSA) 2(2) (2018) 01–05 http://doi.org/10.26480/mjsa.02.2018.01.05 Lopes M, Reynolds M (2012) Stay-green in spring wheat can be determined by spectral reflectance measurements (normalized difference vegetation index) independently from phenology. J Exp Bot 63:3789–3798. 10.1093/jxb/ers071 Muaz Ameen A, Mahmood A, Sahkoor MA, Zia Muhammad Saad Ullah,The role of endophytes to combat abiotic stress in plants,Plant Stress,12, 2024,100435,ISSN 2667-064X, https://doi.org/10.1016/j.stress.2024.100435 Chaudhary P, Agri U, Chaudhary A, Kumar A, Kumar G (2022) Endophytes and their potential in biotic stress management and crop production. Front Microbiol 13:933017. 10.3389/fmicb.2022.933017 Burlakoti S, Devkota AR, Poudyal S, Kaundal A (2024) Beneficial Plant–Microbe Interactions and Stress Tolerance in Maize. Appl Microbiol 4(3):1000–1015. https://doi.org/10.3390/applmicrobiol4030068 Hassan, Etesami The dual nature of plant growth-promoting bacteria: Benefits, risks, and pathways to sustainable deployment,Current Research in Microbial Sciences Volume 9,2025,100421,ISSN 2666–5174, https://doi.org/10.1016/j.crmicr.2025.100421 ( https://www.sciencedirect.com/science/article/pii/S2666517425000835 ) Additional Declarations No competing interests reported. Supplementary Files AMOSALONE.xlsx Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. 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version.\u003c/p\u003e","description":"","filename":"image10.png","url":"https://assets-eu.researchsquare.com/files/rs-7022858/v1/a82de90d665f117149989699.png"},{"id":87685713,"identity":"8242aaa9-12c0-4bdc-a9d5-05c18e749f26","added_by":"auto","created_at":"2025-07-28 02:31:39","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1839858,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7022858/v1/fb65751d-c1f2-4fd8-bd05-082dbd7a0c5c.pdf"},{"id":86171156,"identity":"bd86ad05-3c45-4458-bd6c-022ec55764b7","added_by":"auto","created_at":"2025-07-07 14:29:06","extension":"xlsx","order_by":0,"title":"","display":"","copyAsset":false,"role":"supplement","size":11096,"visible":true,"origin":"","legend":"","description":"","filename":"AMOSALONE.xlsx","url":"https://assets-eu.researchsquare.com/files/rs-7022858/v1/2bc0fb277e4ec82259553acd.xlsx"}],"financialInterests":"No competing interests reported.","formattedTitle":"\u003cp\u003eEntomopathogenic microbial potential in the management of Fall Armyworm, \u003cem\u003eSpodoptera frugiperda \u003c/em\u003e(J.E. Smith) in Maize Production\u003c/p\u003e","fulltext":[{"header":"INTRODUCTION","content":"\u003cp\u003eMaize (\u003cem\u003eZea mays L.\u003c/em\u003e) remains one of the most important cereal crops globally, serving as a staple food for both humans and livestock and as a key raw material in various industrial applications, including biofuel production (Togola et al ., 2025,FAO, 2023, Swati et al., \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). However, maize cultivation is increasingly threatened by biotic stresses, particularly from insect pests such as the fall armyworm (\u003cem\u003eSpodoptera frugiperda\u003c/em\u003e), which has emerged as a major invasive species in tropical and subtropical agroecosystems (Paude et al ., 2022 Mlambo et al., \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2024\u003c/span\u003e; Kasoma et al., \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2021\u003c/span\u003e, Zanzana, et al., \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2024\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eOriginally native to the Americas, \u003cem\u003eS. frugiperda\u003c/em\u003e was first detected in Africa in 2016 and has since spread rapidly across the continent, with severe infestations recorded in West Africa, including Nigeria\u0026rsquo;s key maize-growing regions (Makgoba \u003cem\u003eet al\u003c/em\u003e., 2020; Tambo et al., 2020 Anjorin et al., \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). The larvae feed aggressively on maize leaves, tassels, and ears, leading to substantial reductions in photosynthetic capacity and yield. Unchecked infestations can result in yield losses ranging from 20\u0026ndash;50% (, Anjorin et al., \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Matova et al., \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Kris 2024). Adding to the challenge, the pest's subterranean pupation\u0026mdash;typically 3\u0026ndash;5 cm below the soil surface\u0026mdash;enables it to evade surface-applied insecticides (Thirawut et al., \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2023\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eClimate change had further compounded the fall armyworm threat (Zanzana et al., \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). The fluctuation in rainfall patterns and the rising temperatures have further expanded the pest\u0026rsquo;s survival window, reproductive capacity, and geographical range, thereby intensifying its impact on food security in vulnerable regions such as southwestern Nigeria (Yan et al., \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2022\u003c/span\u003e, Mir et al., 2024).\u003c/p\u003e\u003cp\u003eConventional pest management strategies rely heavily on synthetic insecticides. While these chemicals are effective, their overuse has led to pesticide resistance, environmental contamination, and its adverse effects on non-targeted organisms, which includes beneficial insects (Sangaraju et al., \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2024\u003c/span\u003e, Pereira et al., \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2024\u003c/span\u003e, Verma et al .,2024 ,Tyagi., et al., 2024, Yao \u003cem\u003eet al\u003c/em\u003e., 2023; Koskey et al., \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). These have resulted into growing interest in sustainable alternatives, particularly biological control methods employing entomopathogenic and plant growth-promoting microorganisms.\u003c/p\u003e\u003cp\u003eMicrobial agents such as \u003cem\u003eBacillus thuringiensis\u003c/em\u003e (Bt), \u003cem\u003ePseudomonas putida\u003c/em\u003e, and \u003cem\u003eTrichoderma\u003c/em\u003e spp. have shown promise for their dual roles in pest suppression and plant health promotion (Irsad \u003cem\u003eet al\u003c/em\u003e., 2023; Mart\u0026iacute;nez-Salgado \u003cem\u003eet al\u003c/em\u003e., 2023). Bt, a spore-forming Gram-positive bacterium, produces insecticidal crystal proteins (Cry and Cyttoxins) that disrupts the midgut epithelium of larval insects, that ultimately results into mortality (Pradeep et al., 2021, Ma et al., \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). \u003cem\u003eTrichoderma\u003c/em\u003e spp., in addition acts as an alternative biopesticides, also enhances plant vigour by the inducement of systemic resistance that improves nutrient acquisition (Waleed et al., 2024 Sood et al., \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2020\u003c/span\u003e Lanzuise et al., \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e2022\u003c/span\u003e, Bharti et al., \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2024\u003c/span\u003e ).As recent studies have highlighted the synergistic potentials of using biochar\u0026mdash;a stable, carbon-rich soil amendment derived from pyrolyzed agricultural biomass\u0026mdash;as a carrier for microbial inoculants. Biochar improves soil structure, water-holding capacity, and nutrient availability, thereby the enhancing the microbial survival and efficacy in the field (Luo et al.,2025, Ali et al.,2025 Jatuwong, et al \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e2025\u003c/span\u003e, Shiv et al 2023 ). When combined with entomopathogenic microorganisms, biochar can support integrated pest and soil fertility management in a sustainable and scalable manner (Fritz, et al., \u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e2022\u003c/span\u003e, Xiang et al.,2022 ).\u003c/p\u003e\u003cp\u003eThis study aims to evaluate the efficacy of entomopathogenic microbial inoculants formulated with biochar-based carriers for the sustainable management of \u003cem\u003eSpodoptera frugiperda\u003c/em\u003e in maize cultivation.\u003c/p\u003e\u003cp\u003eThe specific objectives are to:\u003c/p\u003e\u003cp\u003eAssess the effectiveness of biochar as a carrier for microbial inoculants in enhancing soil fertility and pest suppression. Evaluate the biocontrol potential of \u003cem\u003eBacillus thuringiensis\u003c/em\u003e and microbial consortia against fall armyworm under both controlled and field conditions, and to investigate the impact of biochar-based biopesticide applications on maize productivity, soil health, and integrated pest management strategies in smallholder farming systems.\u003c/p\u003e"},{"header":"MATERIALS AND METHODS","content":"\u003cp\u003eMicrobial Isolation and Characterization\u003c/p\u003e\u003cp\u003eSoil samples were collected from maize rhizospheres at the Institute of Agricultural Research and Training (IAR\u0026amp;T), Ibadan, Nigeria. Serial dilutions up to 10⁷ CFU/mL were prepared and plated on nutrient agar and King\u0026rsquo;s B medium using the spread plate technique. Distinct colonies were isolated and subjected to Gram staining and a suite of biochemical assays, including catalase, oxidase, and indole-3-acetic acid (IAA) production.\u003c/p\u003e\u003cp\u003eFor precise identification, molecular characterization of the bacterial isolates was performed via amplification of the 16S rRNA gene using universal primers 27F (5'-AGAGTTTGATCMTGGCTCAG-3') and 1492R (5'-TACGGYTACCTTGTTACGACTT-3'). PCR products were purified and sequenced via Sanger sequencing. The sequences recorded were analyzed using BLAST against the NCBI GenBank database and aligned using MEGA 14 for phylogenetic tree construction. Key entomopathogenic strains\u0026mdash;Bacillus thuringiensis, Pseudomonas putida, and Klebsiella variicola\u0026mdash;were screened for biocontrol attributes such as siderophore production (CAS assay),(Chiho et al., 2021) phosphate solubilization (Pikovskaya\u0026rsquo;s medium),(Singh et al ., 2016) and chitinaseGomaa et al., 2012 and protease activity.(Bakkiet al., \u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e2024\u003c/span\u003e)\u003c/p\u003e\u003cp\u003eExperiment 1: \u003cem\u003eIn-Vitro\u003c/em\u003e Bioassay against \u003cem\u003eSpodoptera frugiperda\u003c/em\u003e\u003c/p\u003e\u003cp\u003ePure cultures of \u003cem\u003eB. thuringiensis\u003c/em\u003e were grown in Luria-Bertani (LB) broth at 30\u0026deg;C and harvested at 24, 48, 72, and 96 hours. Cultures were centrifuged at 12,000 rpm for 15 minutes to separate the supernatant and cell pellets. Bioassays were conducted by incorporating varying concentrations (1 \u0026times; 10⁶ to 1 \u0026times; 10⁹ CFU/mL) of Bt suspensions into an artificial diet fed to third-instar \u003cem\u003eS. frugiperda\u003c/em\u003e larvae, reared under controlled laboratory conditions. Mortality was recorded at 12-hour intervals over 96 hours. Lethal concentration (LC₅₀) and lethal time (LT₅₀) were calculated using Probit analysis in R v4.2.0.\u003c/p\u003e\u003cp\u003eExperiment 2: Screenhouse Trial (Artificial Infestation)\u003c/p\u003e\u003cp\u003eA screenhouse experiment was conducted to evaluate the efficacy of microbial and biochar treatments under controlled pest pressure. Maize seeds (variety SUWAN 1 ) were sown in sterilized soil and grown to the V3 growth stage. Each plant was manually infested with one first-instar larva of \u003cem\u003eS. frugiperda\u003c/em\u003e. Treatments included:\u003c/p\u003e\u003cp\u003eBiochar\u0026thinsp;+\u0026thinsp;\u003cem\u003eBacillus thuringiensis\u003c/em\u003e\u003c/p\u003e\u003cp\u003eBiochar\u0026thinsp;+\u0026thinsp;Trichoderma spp.\u003c/p\u003e\u003cp\u003eAmpligo\u0026reg; (Chlorantraniliprole\u0026thinsp;+\u0026thinsp;Lambda-cyhalothrin) as chemical control\u003c/p\u003e\u003cp\u003eUntreated control\u003c/p\u003e\u003cp\u003eEach treatment was applied via foliar spray at 500 mL per plot using a hand-held atomizer. Damage severity was assessed using digital image analysis with Image J v1.45i, quantifying percentage leaf area loss. Larval mortality was monitored daily.\u003c/p\u003e\u003cp\u003eExperiment 4: Field Trials under Natural Infestation\u003c/p\u003e\u003cp\u003eField trials were conducted at the IAR\u0026amp;T experimental station (7\u0026deg;26\u0026prime;N, 3\u0026deg;54\u0026prime;E), Ibadan, Nigeria, from April to August 2024. A split-plot design was used:\u003c/p\u003e\u003cp\u003eMain plots: Sprayed vs. non-sprayed Plot\u003c/p\u003e\u003cp\u003eSub-plots: Seven treatment combinations:\u003c/p\u003e\u003cp\u003eT1: Mycorhiza\u0026thinsp;+\u0026thinsp;Biochar\u003c/p\u003e\u003cp\u003eT2: NPK Fertilizer (50 kg N/ha)\u0026thinsp;+\u0026thinsp;Mycorrhiza\u0026thinsp;+\u0026thinsp;Biochar\u003c/p\u003e\u003cp\u003eT3: NPK Fertilizer (50 kg N/ha)\u003c/p\u003e\u003cp\u003eT4: NPK Fertilizer (100 kg N/ha)\u003c/p\u003e\u003cp\u003eT5: Untreated Control\u003c/p\u003e\u003cp\u003eT6: Ampligo (Chemical insecticide)\u003c/p\u003e\u003cp\u003eT7: \u003cem\u003eTrichoderma\u003c/em\u003e spp.\u003c/p\u003e\u003cp\u003eBiopesticides and insecticides were applied at 30 days after planting (DAP) and repeated at 10-day intervals for three cycles.\u003c/p\u003e\u003cp\u003eData Collection and Analysis\u003c/p\u003e\u003cp\u003eAgronomic parameters assessed included:\u003c/p\u003e\u003cp\u003eGrowth metrics: Plant stand, plant height (PH_6wk, PH_8wk), stem girth (6wk and 8wk), ear height, and number of leaves (6wk and 8wk)\u003c/p\u003e\u003cp\u003ePhenology: Days to tasseling, days to silking\u003c/p\u003e\u003cp\u003eStructural and yield traits: Root lodging, stalk lodging, ear aspect, ear and plant at harvest, grain weight, and field weight\u003c/p\u003e\u003cp\u003eDisease and pest scores: Foliar damage scores using Bajracharya\u0026rsquo;s 0\u0026ndash;5 scale(Karki, et al \u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e2023\u003c/span\u003e)\u003c/p\u003e\u003cp\u003eChlorophyll content and leaf moisture were also evaluated.\u003c/p\u003e\u003cp\u003eBajracharya (0\u0026ndash;5) scale:\u003c/p\u003e\u003cp\u003eScore Description\u003c/p\u003e\u003cp\u003e0 No visible damage\u003c/p\u003e\u003cp\u003e1 Window-paning on whorl leaves\u003c/p\u003e\u003cp\u003e2 Few small holes in upper leaves\u003c/p\u003e\u003cp\u003e3 Ragged holes and partial whorl damage\u003c/p\u003e\u003cp\u003e4 Extensive damage to whorl and upper leaves\u003c/p\u003e\u003cp\u003e5 Whorl destruction and drying of plant\u003c/p\u003e\u003cp\u003ePest infestation and disease severity were monitored weekly using the five-point sampling method. Populations of natural enemies (Coccinellidae and Chrysopidae) were also recorded to assess ecological balance.\u003c/p\u003e\u003cp\u003eAll data were analyzed using Analysis of Variance (ANOVA) in R Studio v4.2.0, and treatment means were separated using Duncan\u0026rsquo;s Multiple Range Test (DMRT) at p\u0026thinsp;\u0026le;\u0026thinsp;0.05. Visualizations and summary statistics were generated in R Studio v4.2.0\u003c/p\u003e"},{"header":"RESULT","content":"\u003cp\u003eThe molecular and cultural characterization confirmed the identification of three entomopathogenic strains: \u003cem\u003eBacillus thuringiensis, Pseudomonas putida\u003c/em\u003e, and \u003cem\u003eKlebsiella variicola\u003c/em\u003e. All isolates exhibited key biocontrol and plant growth-promoting traits. \u003cem\u003eB. thuringiensis\u003c/em\u003e showed strong chitinase and protease enzyme activity. \u003cem\u003eP. putida\u003c/em\u003e and \u003cem\u003eK. variicola\u003c/em\u003e also demonstrated consistent phosphate solubilization and siderophore production. These results confirm the dual potential of the isolates for both pest suppression and nutrient mobilization.\u003c/p\u003e\u003cp\u003e\u003cem\u003eIn-vitro\u003c/em\u003e bioassay of \u003cem\u003eBacillus thuringiensis\u003c/em\u003e against \u003cem\u003eSpodoptera frugiperda\u003c/em\u003e, bioassay results (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e) showed a progressive increase in larval mortality with increased concentration and incubation duration of \u003cem\u003eB. thuringiensis\u003c/em\u003e cultures. The LC₅₀ value decreased from 3.4 \u0026times; 10⁷ CFU/mL at 24 hours to 2.6 \u0026times; 10⁷ CFU/mL at 72 hours. The LT₅₀ for the 72-hour culture was recorded at 48.2 hours. The mortality data followed a sigmoid response curve, confirming concentration-dependent efficacy across all exposure intervals.\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003eScreenhouse Experiment: Artificial Infestation (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e)\u003c/p\u003e\u003cp\u003eAll bio-inoculated treatments significantly reduced foliar damage compared to the untreated control. The biochar\u0026thinsp;+\u0026thinsp;\u003cem\u003eBacillus thuringiensis\u003c/em\u003e treatment achieved the highest larval mortality (78%) and the lowest mean severity score (1.7 on a 0\u0026ndash;5 scale), indicating a 65% reduction in leaf area loss relative to the control (4.3, p\u0026thinsp;\u0026lt;\u0026thinsp;0.01). Trichoderma-treated plants recorded a 59% larval mortality rate, while the chemical control, Ampligo\u0026reg;, achieved 71% mortality.\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003eThe agronomic performance of maize under seven different treatments (T1\u0026ndash;T7) and sprayed and non-sprayed plot is summarized in Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e. The graph illustrates the mean values of multiple traits, including growth, structural integrity, and phenological development, enabling comparative evaluation across integrated nutrient and pest management strategies: Vegetative Growth and Canopy Development with Plant height measured at 6 and 8 weeks (PH_6wk, PH_8wk) was highest under Treatment T2 (50 kg N/ha\u0026thinsp;+\u0026thinsp;Mycorrhiza\u0026thinsp;+\u0026thinsp;Biochar), closely followed by T3 (Inorganic N only) and T7 (Trichoderma spp.). Stem Girth and Structural Integrity\u003c/p\u003e\u003cp\u003eStem girth at both 6 and 8 weeks followed a comparable trend, with T2 and T1 outperforming other treatments. The increase in girth of stem under biochar treatments was from enhanced root development and water retention, contributing to better nutrient translocation and structural stability. Treatments with synthetic insecticide alone (T6) or no input (T5) lagged in this regard, which suggested a lack of supportive soil structure or biological stimulation.\u003c/p\u003e\u003cp\u003ePhenological Traits: Silking and Tasseling\u003c/p\u003e\u003cp\u003eDays to silk and tasseling were shortest in T2, These indicated accelerated reproductive maturity. which suggested that microbial consortia, in combination with moderate nitrogen application, promoted early phenological progression\u0026mdash;a trait beneficial in short-season agroecologies. In contrast, T1 and T5 exhibited the longest durations to silk and tassel, that reflected delayed development under reduced nutrient input or stress conditions.\u003c/p\u003e\u003cp\u003eLodging Resistance and Plant Quality\u003c/p\u003e\u003cp\u003eRoot and stalk lodging recorded remained generally low across the treatments, although slightly elevated in the untreated control (T5) and T6. The best structural resilience was however recorded in biochar-integrated plots (T2, T1),This supported the role of enhanced root architecture and mechanical strength via improved soil conditioning.\u003c/p\u003e\u003cp\u003eEar Development and Yield Proxy Traits\u003c/p\u003e\u003cp\u003eEar height, ear aspect, and plant-at-harvest ratings were consistently higher in T2 and T3, with T2 thus exhibiting superior uniformity in ear placement and harvest quality. These traits are positively correlated with grain yield and harvestability. However the lower scores recorded in T5 and T6 suggested underperformance under nutrient-deficient or solely chemical management systems.\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003eChlorophyll Content and Leaf Moisture Response (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003e) presented the chlorophyll concentration and leaf moisture status measured at 6 and 8 weeks after planting (WAP) across the seven treatment combinations, stratified by spray status. These parameters serve as vital physiological indicators of plant health and stress tolerance.\u003c/p\u003e\u003cp\u003eChlorophyll Content (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003e)\u003c/p\u003e\u003cp\u003eAt 6 weeks (Chlor_6wk), the SPAD chlorophyll values ranged from 44.0\u0026thinsp;\u0026plusmn;\u0026thinsp;1.9 in Treatment 4 (100 kg N/ha) to 52.7\u0026thinsp;\u0026plusmn;\u0026thinsp;2.1 in Treatment 2 (50 kg N/ha\u0026thinsp;+\u0026thinsp;Mycorrhiza\u0026thinsp;+\u0026thinsp;Biochar), This highlighted the significant impact of integrated nutrient management on early-stage photosynthetic potential. Both T1 and T2 (biochar-based microbial treatments) consistently showed higher SPAD readings than the untreated control (T5). While sprayed plots generally recorded higher chlorophyll values than their non-sprayed counterparts (~\u0026thinsp;1.5 SPAD units), this difference was not statistically significant (p\u0026thinsp;\u0026gt;\u0026thinsp;0.05).\u003c/p\u003e\u003cp\u003eBy 8 weeks (Chlor_8wk), chlorophyll levels increased in all treatments, It thus peaked in the sprayed T6 (Ampligo; 65.0\u0026thinsp;\u0026plusmn;\u0026thinsp;2.3 SPAD) and sprayed T7 (Trichoderma spp.; 63.1\u0026thinsp;\u0026plusmn;\u0026thinsp;2.0). However, biochar-microbial treatments maintained competitive chlorophyll values with the avoidance of the trust on synthetic insecticides, which further underscores their ability to sustained photosynthetic efficiency under pest pressure.\u003c/p\u003e\u003cp\u003eLeaf Moisture Content\u003c/p\u003e\u003cp\u003eLeaf moisture percentages showed a slightly different trend (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003e). Moisture content at harvest ranged from 19.4\u0026thinsp;\u0026plusmn;\u0026thinsp;0.9% in non-sprayed T2 to a maximum of 25.2\u0026thinsp;\u0026plusmn;\u0026thinsp;1.1% in sprayed T6. Although sprayed plots had\u0026thinsp;~\u0026thinsp;1% more moisture on average, the difference was not significant across treatments. Treatments T1 and T2 maintained moderate moisture levels (22.3\u0026ndash;23.1%) which suggested that biochar contributed to soil water retention and delayed desiccation, especially under non-sprayed conditions.\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003eGrain and Field Weight Response Across Treatments\u003c/p\u003e\u003cp\u003eFigure \u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003e displays the mean values for 100-grain weight and field weight across the seven treatments under both sprayed and non-sprayed conditions. These indicators serve as critical yield components that reflected the combined effects of nutrient input and pest management strategy.Field Weight\u003c/p\u003e\u003cp\u003eField weight ( Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003e) was highest in Treatment 3 (T3: 50 kg N/ha only), this reached 1.10\u0026thinsp;\u0026plusmn;\u0026thinsp;0.25 kg in non-sprayed plots and 0.80\u0026thinsp;\u0026plusmn;\u0026thinsp;0.15 kg in sprayed plots. Treatment 2 (T2: 50 kg N/ha\u0026thinsp;+\u0026thinsp;Mycorrhiza\u0026thinsp;+\u0026thinsp;Biochar) followed with 0.68\u0026thinsp;\u0026plusmn;\u0026thinsp;0.10 kg (No spray) and 0.58\u0026thinsp;\u0026plusmn;\u0026thinsp;0.08 kg (Yes spray), This outperformed the untreated control (T5: 0.72\u0026thinsp;\u0026plusmn;\u0026thinsp;0.12 kg vs. 0.67\u0026thinsp;\u0026plusmn;\u0026thinsp;0.10 kg). The lowest values recorded were observed in T7 (Trichoderma alone: 0.12\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03 kg, No spray) and T6 (Ampligo: 0.22\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05 kg, No spray), although the sprayed raised T6\u0026rsquo;s yield to 0.55\u0026thinsp;\u0026plusmn;\u0026thinsp;0.07 kg.\u003c/p\u003e\u003cp\u003eGrain Weight(Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003e)\u003c/p\u003e\u003cp\u003eThe grain weight trends mirrored field weight. T3 again produced the highest yield, with 1.05\u0026thinsp;\u0026plusmn;\u0026thinsp;0.30 kg (No spray) and 0.80\u0026thinsp;\u0026plusmn;\u0026thinsp;0.18 kg (Yes spray). T2 was a close second, this achieved 0.95\u0026thinsp;\u0026plusmn;\u0026thinsp;0.20 kg (No spray) and 0.75\u0026thinsp;\u0026plusmn;\u0026thinsp;0.12 kg (Yes spray). T1 (Mycorrhiza\u0026thinsp;+\u0026thinsp;Biochar) followed, while T5 showed only moderate gains (0.38\u0026thinsp;\u0026plusmn;\u0026thinsp;0.10 kg vs. 0.45\u0026thinsp;\u0026plusmn;\u0026thinsp;0.12 kg). Notably, insecticide spray improved grain weight in poorly performed treatments (T4\u0026ndash;T7) but reduced yield slightly in T2 and T3, likely due to phytotoxic stress or disruption of microbial-plant symbiosis.\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003ePest Infestation and Damage Suppression(Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003e): illustrated the mean foliar damage caused by Spodoptera frugiperda at 6 and 8 weeks after planting across the seven treatments under both sprayed and non-sprayed regimes. Infestation levels varied significantly among treatments and over time.\u003c/p\u003e\u003cp\u003eAt 6 weeks, the highest infestation scores were recorded in the non-sprayed untreated control (T5: 3.2\u0026thinsp;\u0026plusmn;\u0026thinsp;0.15) and chemical control (T6: 3.1\u0026thinsp;\u0026plusmn;\u0026thinsp;0.12). In contrast, biochar\u0026ndash;microbial treatments, particularly T1 (Mycorrhiza\u0026thinsp;+\u0026thinsp;Biochar: 2.8\u0026thinsp;\u0026plusmn;\u0026thinsp;0.10) and T2 (Inorganic N\u0026thinsp;+\u0026thinsp;Mycorrhiza\u0026thinsp;+\u0026thinsp;Biochar: 2.4\u0026thinsp;\u0026plusmn;\u0026thinsp;0.08), showed significantly reduced damage. The Foliar sprayed further enhanced control across treatments, with sprayed T2 recorded the lowest damage at 2.3\u0026thinsp;\u0026plusmn;\u0026thinsp;0.09, with, outperformed even T6 (sprayed: 3.0\u0026thinsp;\u0026plusmn;\u0026thinsp;0.13).and by 8 weeks, a general decline in infestation was observed across all plots. The non-sprayed T5 remained the most infested (2.5\u0026thinsp;\u0026plusmn;\u0026thinsp;0.14), while non-sprayed T2 dropped to 2.3\u0026thinsp;\u0026plusmn;\u0026thinsp;0.10. Notably, sprayed T2 (1.8\u0026thinsp;\u0026plusmn;\u0026thinsp;0.07) and T1 (2.0\u0026thinsp;\u0026plusmn;\u0026thinsp;0.08) exhibited the most substantial reduction,that outperformed even the sprayed chemical treatment T6 (2.2\u0026thinsp;\u0026plusmn;\u0026thinsp;0.09).Hence, the T2 (Inorganic N\u0026thinsp;+\u0026thinsp;Mycorrhiza\u0026thinsp;+\u0026thinsp;Biochar) was the most resilient treatment against S. frugiperda, which maintained low infestation scores at both time points. Thus, It suggested that biological soil amendments not only promote plant health but also serve as a strategic tool for sustainable pest suppression, that instilled a viable alternative to exclusive chemical reliance in smallholder maize systems.\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003eDisease Severity Across Treatments (Fig.\u0026nbsp;\u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e7\u003c/span\u003e): displayed the mean disease severity scores (0\u0026ndash;5 scale) for six foliar diseases\u0026mdash;Blight, Curvularia, Ear Rot, Rust, Streak, and Stripe\u0026mdash;across the seven treatment combinations (T1\u0026ndash;T7) under both sprayed and non-sprayed conditions. Across all diseases, severity scores remained in the low range (1.0\u0026ndash;3.5), with no treatment these completely eliminated the symptoms.Blight: Scores in non-sprayed plots ranged from 1.1 to 1.4, with the lowest in T5 (1.0\u0026thinsp;\u0026plusmn;\u0026thinsp;0.12) and highest in T4 (1.4\u0026thinsp;\u0026plusmn;\u0026thinsp;0.15). Sprayed plots consistently recorded higher values (up to 1.7\u0026thinsp;\u0026plusmn;\u0026thinsp;0.18 in T1), suggesting a marginal spray-induced increase. Curvularia: Disease severity in non-sprayed plots was relatively stable (~\u0026thinsp;1.25\u0026ndash;1.35), with a minor dip in T3 (1.25\u0026thinsp;\u0026plusmn;\u0026thinsp;0.10). Sprayed plots spiked early in T1 (1.9\u0026thinsp;\u0026plusmn;\u0026thinsp;0.20) before settling across treatments (~\u0026thinsp;1.3\u0026ndash;1.6).\u003c/p\u003e\u003cp\u003eEar Rot: Highest in sprayed T7 (2.5\u0026thinsp;\u0026plusmn;\u0026thinsp;0.18) and sprayed T2 (1.8\u0026thinsp;\u0026plusmn;\u0026thinsp;0.14), while the non-sprayed T4 recorded the lowest severity (1.2\u0026thinsp;\u0026plusmn;\u0026thinsp;0.13). A steady increase in sprayed plots indicated elevated vulnerability.\u003c/p\u003e\u003cp\u003eRust: Disease levels were minimal (1.0\u0026ndash;1.5) across all treatments. T2 (sprayed: 1.5\u0026thinsp;\u0026plusmn;\u0026thinsp;0.13; non-sprayed: 1.3\u0026thinsp;\u0026plusmn;\u0026thinsp;0.11) showed slightly elevated rust compared to others.\u003c/p\u003e\u003cp\u003eStreak: Fluctuated between 1.0 and 1.6. The highest was observed in non-sprayed T6 (1.6\u0026thinsp;\u0026plusmn;\u0026thinsp;0.14) and sprayed T2 (1.5\u0026thinsp;\u0026plusmn;\u0026thinsp;0.12).\u003c/p\u003e\u003cp\u003eStripe: Showed the most notable spray-related impact. While non-sprayed plots remained between 1.1 and 1.5, sprayed treatments surged to as high as 2.2\u0026thinsp;\u0026plusmn;\u0026thinsp;0.18 (T3), with all others which exceeded 1.8.\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003ePrincipal Component Analysis (PCA) ( Fig.\u0026nbsp;\u003cspan refid=\"Fig8\" class=\"InternalRef\"\u003e8\u003c/span\u003e and Fig.\u0026nbsp;9) : Trait Performance and Treatment Differentiation thus,Figs.\u0026nbsp;\u003cspan refid=\"Fig8\" class=\"InternalRef\"\u003e8\u003c/span\u003e and 9 illustrated the observed physiological values of the on PC1 and PC2 scores across the seven treatment combinations (T1\u0026ndash;T7), separated by foliar spray plot (Spray vs. Non-Spray). These principal components collectively explain the majority of variability among growth, physiological, and yield-related traits associated with fall armyworm management.\u003c/p\u003e\u003cp\u003ePC1 (Fig.\u0026nbsp;\u003cspan refid=\"Fig8\" class=\"InternalRef\"\u003e8\u003c/span\u003e):PC1 scores captured the composite variance in traits linked to vegetative vigor and biomass productivity (e.g., plant height, leaf number, grain and field weight).\u003c/p\u003e\u003cp\u003eThe highest PC1 scores were recorded under Treatment T4 (100 kg N/ha) and T2 (50 kg N/ha\u0026thinsp;+\u0026thinsp;Mycorrhiza\u0026thinsp;+\u0026thinsp;Biochar), especially in sprayed plots, with mean scores exceeding 2.0 and 1.5, respectively.\u003c/p\u003e\u003cp\u003eNon-sprayed plots, particularly T1 and T5 (Mycorrhiza\u0026thinsp;+\u0026thinsp;Biochar and untreated control), displayed negative PC1 scores, indicating limited contributions to the primary trait dimension under non-chemical regimes.\u003c/p\u003e\u003cp\u003ePC2 (Fig.\u0026nbsp;9):\u003c/p\u003e\u003cp\u003ePC2 scores varied within \u0026plusmn;\u0026thinsp;1.5 and primarily captured physiological resilience, chlorophyll content, and structural integrity (e.g., lodging resistance).\u003c/p\u003e\u003cp\u003eNon-sprayed T2 showed the highest negative score (\u0026minus;\u0026thinsp;1.1), suggesting its distinct influence on physiological traits.\u003c/p\u003e\u003cp\u003eBiochar-microbial treatments (T1 and T2) maintained relatively stable PC2 scores regardless of spray status, while T6 (Ampligo) scored lower on PC2 despite high PC1 values, suggesting reduced physiological stability.\u003c/p\u003e\u003cp\u003eTrait\u0026ndash;Treatment Interaction ( Fig.\u0026nbsp;10): Correlation and Physiological Response to Management Strategies:\u003c/p\u003e\u003cp\u003eThe multivariate response of key agronomic traits to treatment and spray regimes (Fig.\u0026nbsp;10) illustrated the physiological and structural adjustments of maize under different pest and nutrient management strategies. Traits such as field weight, grain weight, plant height, stem girth, and leaf number were assessed across seven treatments (T1\u0026ndash;T7), with clear delineation between sprayed and non-sprayed conditions with\u003c/p\u003e\u003cp\u003eGrowth Parameters and Structural Development\u003c/p\u003e\u003cp\u003ePlant height (PH_6wk and PH_8wk) was highest under T2 (Inorganic N\u0026thinsp;+\u0026thinsp;Mycorrhiza\u0026thinsp;+\u0026thinsp;Biochar) and T3 (50 kg N only), with foliar spraying further enhancing early vegetative development, particularly under T2. Stem girth at both 6 and 8 weeks mirrored this trend,\u003c/p\u003e\u003cp\u003ePhotosynthetic Potential and Canopy Growth\u003c/p\u003e\u003cp\u003eLeaf number at 6 and 8 weeks was also highest in T2 and T7 (Trichoderma). These treatments likely benefited from phytohormonal stimulation by Trichoderma spp. and improved root nutrient acquisition, respectively. Interestingly, leaf development was slightly suppressed in sprayed T6 (Ampligo), suggesting a possible trade-off due to insecticide-induced stress or negative impact on beneficial microbial populations.\u003c/p\u003e\u003cp\u003eThe yield Components (Fig.\u0026nbsp;10): Biomass Accumulation and Kernel Fill\u003c/p\u003e\u003cp\u003eGrain and field weight responses aligned with vegetative vigor trends. T3 (50 kg N) recorded the highest field and grain weight under sprayed conditions, while T2 closely followed, demonstrating that microbial and biochar inputs can partially offset the need for synthetic inputs. The untreated control (T5) and chemical-only plot (T6) showed inconsistent performance, reinforcing the advantage of integrated biological systems.\u003c/p\u003e\u003cp\u003e\u003c/p\u003e"},{"header":"DISCUSSION","content":"\u003cp\u003eThis study confirms that combining moderate levels of inorganic nitrogen(NPK Fertilizer) (50 kg N/ha) with biochar and microbial Mycorrhiza (T2) significantly enhances maize growth, structure, and yield potential across both chemical and non-chemical pest management systems. Treatments T2 and T1 consistently outperformed chemical-only controls in both physiological stability and agronomic performance. Notably, PC2 and trait regression results revealed that biochar\u0026ndash;microbial combinations support chlorophyll retention, stem integrity, and kernel development\u0026mdash;traits essential for resilience under fall armyworm infestation.\u003c/p\u003e\u003cp\u003eMicrobial Isolation and Functional Characterization\u003c/p\u003e\u003cp\u003eMolecular analysis confirmed the identity of three entomopathogenic strains: Bacillus thuringiensis, Pseudomonas putida, and Klebsiella variicola, each of the microbes identified gives a characteristics biocontrol traits. Although B. thuringiensis exhibited strong chitinase and protease activity, which aligned with its known larvicidal properties (Anuj Ranjan, et al.,2024, Mart\u0026iacute;nez-Zavala et al.,2019, Franco-Rivera et al., \u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e2004\u003c/span\u003e). Additionally, P. putida and K. variicola demonstrated phosphate solubilization and siderophore productionwhich indicated the potential for promoting nutrient availability and inducing systemic resistance (Al-Turki et al., \u003cspan citationid=\"CR56\" class=\"CitationRef\"\u003e2023\u003c/span\u003e Goswami et al.,2013).\u003c/p\u003e\u003cp\u003eThese attributed characteristics traits makes the isolated strains not only as biopesticides but also as Mycorrhiza with dual functionality in integrated pest and soil fertility management. Similar dual-action benefits have also been observed in prior work on rhizospheric bacteria by Kowalska et al. (\u003cspan citationid=\"CR58\" class=\"CitationRef\"\u003e2020\u003c/span\u003e), where microbial consortia enhanced maize growth while suppressing pests.\u003c/p\u003e\u003cp\u003eThe in-Vitro Bioassay of Bacillus thuringiensis Against Spodoptera frugiperda\u003c/p\u003e\u003cp\u003eLarval mortality increased with both Bt concentration and incubation time. The LC₅₀ value declined markedly from 3.4 \u0026times; 10⁷ CFU/mL at 24 hours to 2.6 \u0026times; 10⁷ CFU/mL at 72 hours,which suggested an enhanced potency in older cultures, this is possibly due to the increased accumulation of Cry and Cyt toxins (Lianget al., \u003cspan citationid=\"CR59\" class=\"CitationRef\"\u003e2025\u003c/span\u003e,Karshanal et al.,2023).\u003c/p\u003e\u003cp\u003eThe Larval response showed a typical sigmoid mortality curve, and LT₅₀ recorded indicated that the 72-hour formulations achieved rapid knockdown effects within 48.2 hours. These results are consistent with the recent findings by Karshanal et al. (2023), who reported that Bt-based products significantly outperformed synthetic alternatives in terms of larval suppression without harming non-target species.\u003c/p\u003e\u003cp\u003eHence,all bio-inoculated treatments significantly reduced foliar damage compared to the untreated control. The biochar\u0026thinsp;+\u0026thinsp;B. thuringiensis combination achieved the highest larval mortality (78%) and the lowest mean severity score (1.7 on a 0\u0026ndash;5 scale),which represented a 65% reduction in leaf area loss compared to the control (4.3, p\u0026thinsp;\u0026lt;\u0026thinsp;0.01). Trichoderma-treated plants showed moderate protection (59% mortality), while Ampligo\u0026reg;, the chemical control, reached 71% efficacy.\u003c/p\u003e\u003cp\u003eThe enhanced protection conferred by biochar-based microbial treatments likely stems from improved microbial colonization and persistence in the phyllosphere, as reported by Kaul et al., (\u003cspan citationid=\"CR62\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). The Biochar provided a conducive microhabitat for microbial adhesion, while the microbial metabolites exert both direct toxicity and systemic induced resistance in the plant.\u003c/p\u003e\u003cp\u003eIn otherword,, the Ampligo treatment, though effective, did not outperform the Bt\u0026thinsp;+\u0026thinsp;biochar treatment and may pose long-term resistance risks and ecological drawbacks, as previously cautioned by Chen et al. (\u003cspan citationid=\"CR63\" class=\"CitationRef\"\u003e2023\u003c/span\u003e).In the field experiment the line graph indicated that T2 consistently outperformed other treatments in nearly all agronomic parameters, which emphasizes the value of integrated Mycorrhiza and biochar inputs. These enhancements in plant stature, leaf production, and reproductive maturity under T2 are indicative of improved nutrient uptake and systemic plant health. The consistent pattern across sprayed and non-sprayed plots further highlighted the robustness of biological amendments under varying pest pressures.T1 (Mycorrhiza\u0026thinsp;+\u0026thinsp;biochar without inorganic N) showed intermediate performance, affirming the growth-promoting role of microbial consortia even in the absence of synthetic nitrogen(NPK fertilizer). In contrast, T5 (untreated) and T6 (chemical control) showed limited enhancement of growth traits,which implicated the failure and the limitations of non-integrated or purely chemical approaches.\u003c/p\u003e\u003cp\u003eThese observations aligned with reports presented by Huey et al. (\u003cspan citationid=\"CR64\" class=\"CitationRef\"\u003e2020\u003c/span\u003e) and Sudip et al. (2023), which found that combining biochar with beneficial microbes significantly improves maize agronomy through better root colonization, stress tolerance, and nutrient mobilization.\u003c/p\u003e\u003cp\u003eThe progressive increase in chlorophyll from weeks 6 to 8 reflects sustained canopy development and continued nutrient availability. T2 outperformed the other treatments which may be due to enhanced nitrogen uptake and microbial nutrient mineralization, this also corroborated the findings by Mthiyane et al., (\u003cspan citationid=\"CR69\" class=\"CitationRef\"\u003e2024\u003c/span\u003e) and Liu, et al. (\u003cspan citationid=\"CR70\" class=\"CitationRef\"\u003e2024\u003c/span\u003e), who reported that biochar and microbial consortia enhance chlorophyll biosynthesis and delay senescence. Hence,the highest chlorophyll readings at 8 weeks occurred in sprayed T6 and T7 plots, which also suggested effective pest suppression via synthetic or microbial means and may reduce the defoliation and stress in the maize plant thereby preserving photosynthetic tissues (Zhao, et al., \u003cspan citationid=\"CR71\" class=\"CitationRef\"\u003e2019\u003c/span\u003e and Gekas et al., 2013, ). Nonetheless, T2\u0026rsquo;s non-sprayed performance was statistically comparable, this reinforced the robustness of biochar-based microbial treatments in the enhancement of physiological resilience even in the absence of foliar insecticides.\u003c/p\u003e\u003cp\u003eThe moisture data also aligned with these trends.Hence the biochar's porous structure contributed to improved water retention and buffering against mid-season droughts, as previously documented by Jiawei et al. (2025). While the chemical treatments achieved higher moisture content, this may reflect delayed maturity or water accumulation due to denser canopies rather than physiological efficiency.(Duan et al.,2024)\u003c/p\u003e\u003cp\u003eThe enhanced vegetative vigour in T2 reflected synergistic nutrient availability from both inorganic and biological sources. Similarly, the number of leaves at 6 and 8 weeks was highest in T2 and T7, confirming the promotive effects of microbial Mycorrhiza and biochar on canopy development. These findings are consistent with previous reports where biochar-amended rhizospheres improved leaf formation and photosynthetic efficiency (Jabborova et al., 2021).\u003c/p\u003e\u003cp\u003eThe chlorophyll and moisture results validated the physiological benefits of biochar-integrated microbial strategies. Treatments T1 and T2 consistently improved plant vitality indicators under both pest-suppressed and unsprayed conditions.(Liu et al.,2022) These findings support the incorporation of biochar-based microbial inputs into sustainable maize production systems to enhance crop resilience, reduce chemical dependency, and maintain agro-ecological balance (Luo et al.,2025) also from the results it can be confirmed that biochar-amended microbial inoculants (T1 and T2) effectively suppress fall armyworm infestation, rivaling or surpassing the efficacy of the synthetic insecticide (T6: Ampligo). The early damage reduction observed in T2 suggests that Mycorrhiza\u0026ndash;biochar synergy disrupts the pest\u0026rsquo;s lifecycle, potentially by enhancing plant vigour and induced systemic resistance (Chen et al., \u003cspan citationid=\"CR63\" class=\"CitationRef\"\u003e2023\u003c/span\u003e Chisonga et al.,2023).\u003c/p\u003e\u003cp\u003eAlso at 8 weeks,the armyworm pest infestation remained significantly lower in T2 and T1 regardless of spray status,which indicated residual protective effects which may be due to persistent microbial colonization and biochar's soil amendment properties. Which is consistent with Kasoma, et al. (\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e2023\u003c/span\u003e) discovery, who emphasized the role of entomopathogenic microbes in long-term pest suppression under field conditions.\u003c/p\u003e\u003cp\u003eWith the addition of foliar spray to T1 and T2 plot further provided additive benefits, that suggested the beneficial effect of integrated pest management (IPM) strategies combined with microbial bioinoculants with minimal chemical input can maintain efficacy with reduced ecological risk (Zhou et al., \u003cspan citationid=\"CR84\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). Moreover, lower pest pressure in biochar-amended plots can also enhance the effectiveness of natural enemies and reduce the likelihood of resistance buildup.\u003c/p\u003e\u003cp\u003eThe yield advantage of T3 (inorganic N) have emphasised the centrality of nitrogen in supporting kernel development, even under pest stress. However, T2's comparable performance indicated that biochar-borne microbial inoculants can substitute partially for synthetic N while sustaining productivity. This synergy is consistent with the findings of Kohira et al. (\u003cspan citationid=\"CR85\" class=\"CitationRef\"\u003e2025\u003c/span\u003e) and Beilei et al. (2023), who demonstrated that biochar improves nutrient and moisture retention critical to grain filling.\u003c/p\u003e\u003cp\u003eThe modest yield reductions in sprayed T2 and T3 treatments also suggested a potential trade-off between pest control and plant health, due to phytotoxicity or chemical interference with microbial colonization (Yu et al., \u003cspan citationid=\"CR89\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). Hence these findings emphasized the importance of balancing chemical inputs with biological amendments for optimized yield.(Rajiv et el.,2024)\u003c/p\u003e\u003cp\u003eMoreover, the gains observed in lower-yielding treatments following spraying (T6 and T7) confirm that insecticide application remains effective where biological support is lacking. However, their limited ecological benefits and dependence on external inputs highlighted the superior agronomic efficiency of integrated approaches in T2.Treatment 2 (moderate N\u0026thinsp;+\u0026thinsp;biochar\u0026thinsp;+\u0026thinsp;Mycorrhiza) emerged as the most efficient and sustainable strategy, delivering yields comparable to full chemical regimes with fewer ecological trade-offs. While biopesticide and biochar-based treatments (T1, T2, T7) were effective in the reduction of insect pest pressure of Spodoptera frugiperda, their impact on disease suppression was less pronounced. Disease severity scores across most treatments and conditions remained relatively low, but a consistent pattern emerged\u0026mdash;sprayed plots tended to exhibit slightly elevated disease severity, particularly for Stripe.(Rajiv et al., 2024)\u003c/p\u003e\u003cp\u003eThis disease increase may be attributed to foliar spray effects such as physical cuticle damage or increased leaf moisture retention, which promoted opportunistic fungal colonization (Abdelaaziz et al., 2025). The pronounced rise in Stripe under foliar sprays (+\u0026thinsp;0.7 units on average) signals a potential trade-off in integrated pest management (IPM)\u0026mdash;while insecticides effectively suppress pests, they may inadvertently create favorable microclimates for disease development.(Golan et al., \u003cspan citationid=\"CR99\" class=\"CitationRef\"\u003e2023\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eMoreover, biochar\u0026ndash;microbial amendments (T1, T2, T7) did not consistently suppress foliar diseases when compared to the untreated control (T5). This could suggest that under moderate pest pressure, the antagonistic activity of endophytes against foliar pathogens may be insufficient unless augmented by fungicides (Waqas et al., \u003cspan citationid=\"CR100\" class=\"CitationRef\"\u003e2017\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eThe analysis reveals that chemical insecticide application, while effective against pests, may unintentionally elevate foliar disease risk, particularly under humid conditions. This finding supports a refined IPM approach\u0026mdash;integrating microbial Mycorrhiza with low-phytotoxicity or botanically derived insecticides, and pairing them with targeted fungicide applications when disease pressure is high(Cucu et al., \u003cspan citationid=\"CR101\" class=\"CitationRef\"\u003e2025\u003c/span\u003e) Such a strategy could optimize both pest and disease management while minimizing environmental impacts in maize cultivation systems(Henri et al., \u003cspan citationid=\"CR102\" class=\"CitationRef\"\u003e2025\u003c/span\u003e)\u003c/p\u003e\u003cp\u003eSpray Interaction Effects\u003c/p\u003e\u003cp\u003eThe overall, sprayed plots exhibited elevated trait values, but the differential between sprayed and unsprayed conditions was smallest in T2 and T1. Thus suggesting that microbial inoculants and biochar build resilience even in the absence of synthetic chemical inputs. The stable performance of non-sprayed T2 plots hence supported the findings by Zheng et al. (\u003cspan citationid=\"CR105\" class=\"CitationRef\"\u003e2025\u003c/span\u003e), who noted that biochar\u0026ndash;microbial systems sustained physiological function under biotic stress. Thus, reaffirming the synergistic effect of microbial Mycorrhiza and biochar on water and nutrient uptake (Li et al.,, 2023). The biochar matrix may have contributed to improved soil aeration and microbial colonization, which then resulted in superior physiological support(Khan, et al., \u003cspan citationid=\"CR107\" class=\"CitationRef\"\u003e2024\u003c/span\u003e)\u003c/p\u003e\u003cp\u003eThe PCA effectively differentiated treatment impacts based on composite trait performance:\u003c/p\u003e\u003cp\u003ePC1 Interpretation \u0026ndash; Growth and Yield Efficiency shows that the treatments with integrated inputs\u0026mdash;particularly T2 (50 kg N/ha\u0026thinsp;+\u0026thinsp;Mycorrhiza\u0026thinsp;+\u0026thinsp;Biochar)\u0026mdash;demonstrated the most balanced trait enhancement, scoring high on PC1. This supports the previous findings that moderate inorganic fertilization supplemented with microbial consortia and biochar enhances nutrient use efficiency, early vigour, and yield accumulation (Faloye et al., \u003cspan citationid=\"CR108\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). In contrast, lower PC1 values in T5 and T7 shows the limitations of untreated or Trichoderma-only strategies in the absence of nutrient support.(Sanjay et al.,2018)\u003c/p\u003e\u003cp\u003ePC2 Interpretation \u0026ndash; Physiological Integrity and Disease Mitigation:shows the High PC2 scores in T1 and T2 which highlighted their role in sustaining physiological traits\u0026mdash;such as chlorophyll content, moisture retention, and disease resistance\u0026mdash;independent of yield-centric gains.(Lopes and Reynolds \u003cspan citationid=\"CR110\" class=\"CitationRef\"\u003e2012\u003c/span\u003e) This also reinforces the benefits of microbe-driven resilience, as corroborated by Muaz et al. (2024) and Chaudhary et al. (\u003cspan citationid=\"CR112\" class=\"CitationRef\"\u003e2022\u003c/span\u003e), who reported that endophyte-rich systems mitigate stress effects and promote systemic resistance.(Muaz et al., Hassan 2025)\u003c/p\u003e\u003cp\u003eThe Trade-offs with Synthetic Control:show from the observation thatT6 (Ampligo), while enhancing yield metrics (PC1), consistently underperformed on PC2. This indicates potential disruption of beneficial plant\u0026ndash;microbe interactions or physiological processes by chemical inputs\u0026mdash;an ecological trade-off also noted by Burlakoti et al. (\u003cspan citationid=\"CR113\" class=\"CitationRef\"\u003e2024\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eConclusion and recommendation\u003c/p\u003e\u003cp\u003eTherefore future trials should focus on refining spray timing and exploring microbial consortia capable of dual pest and disease suppression to fully realize the potential of integrated pest management (IPM) systems in maize cultivation.\u003c/p\u003e\u003cp\u003eThe PCA evidently showed the multidimensional efficacy of integrated treatments like T2, which maximized both productivity (PC1) and physiological health (PC2). Unlike purely chemical control, biochar-based microbial strategies offered holistic improvements without compromising plant integrity. These results advocates for the inclusion of such biological amendments in sustainable Integrated Pest Management (IPM) frameworks for maize cultivation in Sub-Saharan Africa.\u003c/p\u003e"},{"header":"Declarations","content":"\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003eEdet Iwebaffa Amos and Akinbode Oluwafolake Adenike wrote the main manuscript text and prepared figures 1-12. All authors reviewed the manuscript.\"\u003c/p\u003e\u003ch2\u003eData Availability\u003c/h2\u003e\u003cp\u003ePlease where do I deposit the research data\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eSwati P, Rasane P, Kaur J, Kaur S, Ercisli S, Assouguem A, Ullah R, Alqahtani A, Singh J (2024) The nutritional, phytochemical composition, and utilisation of different parts of maize: A comparative analysis. 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Appl Microbiol 4(3):1000\u0026ndash;1015. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3390/applmicrobiol4030068\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\n\u003cli\u003eHassan, Etesami The dual nature of plant growth-promoting bacteria: Benefits, risks, and pathways to sustainable deployment,Current Research in Microbial Sciences Volume 9,2025,100421,ISSN 2666\u0026ndash;5174,\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.crmicr.2025.100421\u0026nbsp;\u003c/span\u003e\u003c/span\u003e(\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://www.sciencedirect.com/science/article/pii/S2666517425000835\u003c/span\u003e\u003c/span\u003e)\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Spodoptera frugiperda, maize, biochar, microbial inoculants","lastPublishedDoi":"10.21203/rs.3.rs-7022858/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7022858/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eFall armyworm (\u003cem\u003eSpodoptera frugiperda\u003c/em\u003e) poses a significant threat to maize production in sub-Saharan Africa, particularly in Nigeria, where infestation levels continue to disrupt food security. This study investigates the efficacy of entomopathogenic microbial inoculants, delivered through biochar-based formulations, as a sustainable strategy for managing \u003cem\u003eS. frugiperda\u003c/em\u003e in maize cultivation. Field and screenhouse trials were conducted to evaluate the effects of various treatment combinations involving \u003cem\u003eBacillus thuringiensis, Trichoderma\u003c/em\u003e spp., and synthetic insecticide(Ampligo) under sprayed and non-sprayed conditions. Agronomic traits disease incidence, and yield parameterswere assessed. Results revealed that the combination of NPK (50 kg/ha), Mycorrhiza, and biochar (T2) significantly improved plant growth, reduced armyworm damage, and enhanced yield performance, closely rivaling chemical control method. Principal Component Analysis (PCA) confirmed that T2 contributed to superior vegetative vigour (PC1) and physiological stability (PC2). These findings supported the integration of microbial inoculants and biochar as part of an environmentally friendly and scalable Integrated Pest Management (IPM) approach.\u003c/p\u003e","manuscriptTitle":"Entomopathogenic microbial potential in the management of Fall Armyworm, Spodoptera frugiperda (J.E. Smith) in Maize Production","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-07-07 14:20:58","doi":"10.21203/rs.3.rs-7022858/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"4f7cefa0-020a-4788-9e5e-46da61576193","owner":[],"postedDate":"July 7th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2025-09-07T15:23:28+00:00","versionOfRecord":[],"versionCreatedAt":"2025-07-07 14:20:58","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-7022858","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7022858","identity":"rs-7022858","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}