Tariq Mahmud Butt

Entomopathogenic fungi (EPF) are well established as biological control agents, but their emerging role as endophytes reveals a broader and more powerful function in crop protection. By colonizing plant tissues, endophytic entomopathogenic fungi (EEPF) create a dynamic tripartite interaction between plants, fungi, and herbivores, enabling systemic, plant-mediated pest suppression. This review synthesizes current knowledge on the behavioral and ecological responses of herbivorous arthropods to EEPF-colonized plants, with an emphasis on the mechanisms and implications for integrated pest management (IPM). Growing evidence indicates that EEPF consistently modify herbivore behavior and performance across diverse crops and insect taxa. Colonization frequently alters feeding, host selection, and oviposition, often deterring pests, although mediated responses may vary among fungal species, host plants, insect taxa, and environmental conditions. These responses are driven by EEPF-induced changes in plant chemistry, including shifts in volatile organic compounds (VOCs) and defensive metabolites. In parallel, EEPF impair insect fitness by delaying development, reducing survival, and lowering fecundity, thereby suppressing pest populations. These plant-mediated and behavioral changes extend to multitrophic interactions, potentially affecting associations with natural enemies and the transmission efficiency of some insect vectors of plant viruses. Despite rapid progress, critical gaps remain in resolving the mechanistic basis of these interactions and their stability under field conditions. Advancing the application of EEPF will require integrated approaches combining microbial ecology, chemical ecology, and insect behavioral biology. Harnessing these interactions offers a compelling pathway to reduce reliance on synthetic pesticides while enhancing the resilience and sustainability of agricultural systems.

The invasive tomato leafminer, Tuta absoluta, poses a severe global threat to solanaceous crops, necessitating sustainable biocontrol solutions. Through systematic bioprospecting across several Moroccan soils, we constructed a novel library of indigenous fungal isolates using complementary Tenebrio molitor baiting and selective media methods. High-throughput phenotyping identified 49 highly pathogenic isolates, which were characterized for conidial production, thermotolerance, and virulence against T. absoluta. We discovered two lead isolates, Beauveria bassiana UCA-350 and Metarhizium robertsii UCA-329, that demonstrated superior virulence, reducing median survival time and achieving lower LC₅₀ values than most commercial reference strains. Notably, virulence was positively correlated with in vitro conidial yield, revealing a key trait linkage for strain selection. Furthermore, genus-level divergence in thermotolerance was observed, with Beauveria isolates exhibiting significantly higher heat resilience. Our integrated multi-trait screening pipeline not only delivers two potent, regionally sourced biocontrol candidates but also establishes a phenotypic selection framework that prioritizes both efficacy and production scalability, advancing the rational development of next-generation mycoinsecticides.

The cabbage stem flea beetle (CSFB, Psylliodes chrysocephala L) is a major pest of oilseed rape (OSR, Brassica napus L), especially in temperate regions where it is challenging the sustainable production of the crop. Conventional management has relied almost exclusively on synthetic insecticides, yet increasing regulatory restrictions, pest resistance and environmental concerns require the urgent development of alternative strategies. Semiochemicals, chemical cues that mediate inter‐ and intraspecific behavioural interactions, offer considerable potential for integration into push–pull pest management programmes, where combinations of attractant and deterrent or repellent stimuli are used synergistically to reduce crop pest infestation. This review synthesises knowledge of olfaction in CSFB and their behavioural responses to plant‐derived and other bioactive compounds, as well as to synthetic analogues that attract or repel the pest. Glucosinolates, which are non‐volatile, and their volatile derivatives, isothiocyanates, have been shown to act as phagostimulants and attractants, respectively. Furthermore, companion planting with attractive host plant ‘trap crops’ has potential to suppress and divert pest pressure from the main OSR crop. By contrast, companion planting with non‐host plant species, as well as the application of plant solvent extracts and plant‐derived hormones, has been demonstrated to repel, deter and/or induce antifeedant effects in CSFB, respectively. Although these approaches show great promise, significant knowledge gaps remain regarding the identity of the chemicals involved, dose‐dependent behavioural responses, and the interplay between visual, olfactory and tactile cues in CSFB host selection, feeding and oviposition behaviours. Addressing these gaps could inform the design and development of targeted semiochemical‐based interventions that are effective, environmentally benign and compatible with integrated pest management. © 2026 The Author(s). Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Western flower thrips (Frankliniella occidentalis) are major pests of horticultural crops worldwide, creating a need for sensitive monitoring tools to support integrated pest management. Semiochemical lures are widely used to enhance sticky trap capture, but their effectiveness depends on both attractant composition and dispenser design. In this study, the plant-derived volatiles p-anisaldehyde (PANI) and S(−)-verbenone were evaluated as individual and blended attractants, together with the development of practical dispenser systems, across field trials conducted between 2018 and 2021 in commercial strawberry production systems. Initial short-term trials in 2018 showed that both compounds increased trap capture relative to controls, with the PANI–verbenone blend providing the greatest enhancement across repeated 48 h assessments. Follow-on trials in 2019 supported these findings and introduced ethylene vinyl acetate (EVA) tubing as a controlled-release dispenser, improving lure practicality and durability without loss of efficacy. Expanded multi-site trials in 2021, conducted across four farms over four weeks, showed that although early capture dynamics were similar among treatments, differences emerged over time. By day 28, blended attractants, particularly when delivered via polymer-based dispensers, consistently exceeded controls and performed comparably to, or sometimes better than, the commercial standard Lurem-TR. These findings show that combining plant-derived volatile blends with optimised controlled-release dispensers can improve monitoring sensitivity for F. occidentalis under commercial growing conditions.

Introduction: Fungi have been a rich source of pharmaceuticals such as antibiotics, immunosuppressants, and cholesterol-lowering drugs; however, their therapeutic potential remains largely untapped due to difficulties in culturing and elucidating the genetic basis of beneficial traits. Fungi contain ‘cryptic’ genes that are expressed under certain, and often obscure, growth conditions and can produce complex compounds that are difficult to synthesize economically. Developments in genome sequencing and DNA-synthesis technologies offer new opportunities to produce such compounds using biotechnological techniques, however, accurately identifying useful and novel genes, a prerequisite for such approaches, remains challenging. Methods: We present a novel ‘two-by-two’ comparative genomics pipeline for comprehensive gene analysis of selected fungal groups, enabling more confident identification of unique genes across the analyzed species. The approach compares gene sets from two strains of the same species with those from two strains of different species or families within a fungal order. Self-clustering orthologs that are unique to strains from the same species provide higher confidence in identifying species-specific proteins and help reduce noise from low-quality genome assemblies and gene prediction errors. Results: We validated our method in a well-studied fungal group, correctly identifying known species-specific genes and gene clusters, and discovering a novel functional myco-serpin and a fungal Tc toxin complex in Metarhizium . Discussion: Elucidating the genes underlying beneficial traits in fungi presents significant challenges, largely due to the unique and relatively complex aspects of their lifestyles. The two-by-two approach shows broad potential for fungal genome mining and bioprospecting, demonstrated by our discovery of the first fungal Tc toxin complex and a functional myco-serpin in Metarhizium (Hypocreales), and is applicable to other fungal orders such as Eurotiales and Xylariales. Furthermore, the two-by-two approach can be adapted to other organisms with genome architectures similar to fungi.

Root-knot nematodes (RKNs) of the genus Meloidogyne are major plant pests causing severe crop losses. Microbial volatile organic compounds (VOCs) have emerged as promising biopesticides. In this study, we evaluated the effects of two fungal VOCs, 1-octen-3-ol and 3-octanone, on nematode survival in five Meloidogyne species (M. incognita, M. javanica, M. hapla, M. arenaria, and M. luci) in plate assays. Results showed near-complete (95–100%) J2 mortality at 500–1000 ppm within 24 h. At lower concentrations, mobility declined, and species-dependent differences were observed: 1-octen-3-ol was more effective against M. arenaria. Meanwhile, 3-octanone showed stronger effects only on M. hapla and moderate effects on M. incognita and M. javanica. Further experiments using solely M. javanica showed that egg differentiation was significantly inhibited at 7, 14, and 21 days, with up to an 80% reduction at 1000 ppm, and the effects persisted at 125 ppm. Egg hatching from egg masses was reduced by up to 95% in a concentration-dependent manner, irrespective of compound type. Soil-like microcosm assays resulted in substantial reductions in recovered juveniles, with over 90% reduction at 125 ppm after 24 h, suggesting sustained effects under the tested conditions. In more complex plant–soil greenhouse conditions, effects were reduced, although decreasing trends in nematode infection were observed. Overall, these results indicate that fungal VOCs exhibit strong effects on different nematode life stages under controlled conditions, highlighting 1-octen-3-ol and 3-octanone as promising candidates for further evaluation in nematode management strategies.

Entomopathogenic fungi (EPF), including Beauveria spp., Metarhizium spp., Lecanicillium spp., and Isaria spp., help manage pest populations through direct infestation and insecticidal toxin production. Beyond pest control, they enhance plant growth and overall health, which makes them essential components of sustainable agriculture. EPF orchestrates systemic acquired resistance (SAR) and induced systemic resistance (ISR) via salicylic acid (SA) and jasmonic acid (JA) signalling pathways, respectively. Through these mechanisms, EPF promotes plant resilience against both biotic (pathogens and pests) and abiotic stresses (drought, nutrients, heat, and salinity), while improving nutrient uptake, hormone production, and soil quality. In addition, the secondary metabolites produced by EPF strengthen plant defense and stress tolerance. Their ability to persist as endophytes ensures long‐term crop protection and growth stimulation. Incorporating EPF into Integrated pest management as a biocontrol agent reduces the reliance on chemical pesticides, enhances plant resistance, and supports environmental sustainability. This review examines the multifaceted contributions of EPF as a bioinoculant that promotes crop health and tolerance to biotic and abiotic stresses, and discusses the prospects for large‐scale commercial applications.

Root-knot nematodes (RKNs) of the genus Meloidogyne are major plant pests causing severe crop losses. Microbial volatile organic compounds (VOCs) have emerged as promising biopesticides. In this work, we evaluated the nematicidal activity of two fungal VOCs, 1-octen-3-ol and 3-octanone, against five Meloidogyne species (M. incognita, M. javanica, M. hapla, M. arenaria, and M. luci). Plate assays showed near-complete (95–100%) juvenile mortality at 500–1000 ppm within 24 h. At lower concentrations, mobility declined and species-specific effects appeared: 1-octen-3-ol was more effective against M. arenaria, while 3-octanone strongly affected M. hapla and moderately M. incognita and M. javanica. Egg differentiation was significantly inhibited across time points, with up to 80% reduction at 1000 ppm and effects persisting at 125 ppm. Egg hatching from egg masses was reduced up to 95% in a concentration-dependent manner, irrespective of compound type. Soil-like assays confirmed enhanced efficacy at low doses, with >90% mortality at 125 ppm after 24 h, suggesting prolonged activity. These results show that fungal VOCs are potent nematicides at low doses and short exposures, highlighting 1-octen-3-ol and 3-octanone as eco-friendly candidates for sustainable nematode control.