Pest Management Science

BACKGROUND: Aphids damage crops through sustained feeding and by rapid transmission of plant viruses, processes that are not effectively prevented by conventional insecticides. The sugarcane aphid (Melanaphis sacchari), an invasive pest of sorghum in the United States, can inoculate non-persistent viruses within minutes of probing. Disrupting feeding phases represents an alternative strategy in plant disease and integrated pest management. ATP-binding cassette transporters (ABC transporters) influence insect physiological performance, but their role in regulating aphid feeding behavior relevant to crop damage and virus transmission remains unknown. This study evaluated whether ABC transporter activity alters feeding phases of M. sacchari on sorghum. RESULTS: Aphids exposed to ABC transporter modulators via artificial diet and monitored using Electrical Penetration Graph (EPG) recordings showed significant reductions in salivation in E1 phase, compared with controls (P < 0.05) and an increased duration in xylem phase when treated with elacridar, an ABC transporter inhibitor. Aphids treated with dexamethasone, an ABC transporter activator, exhibited increased time spent in pathway phase compared to the controls. ATPase assays also showed significant variations in modulator-treated samples compared to the controls. CONCLUSION: These results demonstrate that transporter modulators alter feeding behaviors which might lead to reduced crop damage/virus transmission, including pathway activities, salivation, and xylem ingestion. This study opens the path to identify ABC transporters as regulators of feeding phases, and to advance the understanding of aphid-plant interactions. This new knowledge will support the development of feeding-based pest management strategies to reduce crop damage and virus transmission.

Background: Resistance to protoporphyrinogen oxidase (PPO) inhibiting herbicides is mainly driven by diverse target-site mutations, reducing the effectiveness of this site of action in row crop systems. Fendioxypyracil is a newly developed PPO inhibitor with high intrinsic grass and broadleaf activity, but its performance against resistant populations and target-site enzyme variants remains insufficiently characterized. Results: Enzyme assays using PPO2 from Amaranthus palmeri and Setaria viridis demonstrated that fendioxypyracil maintained low IC50 values across a broad range of resistance associated mutations, including dG210 deletion and G210, R128, and G399 substitutions, whereas oxadiazon, tiafenacil, and saflufenacil showed substantial loss of potency. Greenhouse dose response experiments confirmed strong fendioxypyracil efficacy, with susceptible and G399A populations controlled at <3 g ai/ha, while dG210 and R128G populations showed only moderate shifts in sensitivity but remained effectively controlled at the recommended rate. Transgenic Arabidopsis thaliana expressing resistant PPX2 alleles exhibited faster and more severe injury with fendioxypyracil compared to saflufenacil. Field trials conducted in a PPO resistant Amaranthus palmeri population demonstrated that fendioxypyracil provided consistent weed control and density reduction, matching the performance of trifludimoxazin and saflufenacil while exceeding that of fomesafen. Conclusion: Fendioxypyracil provides robust and broad-spectrum activity against PPO resistant Amaranthus populations and target mutant enzymes, maintaining efficacy across diverse mutation backgrounds. These results demonstrate its potential as an effective tool for managing PPO inhibitor resistance and sustaining weed control in row-crop production systems.

Many economically important insect pests reproduce through asexual or partially asexual life cycles, yet how reproductive mode influences insecticide resistance management remains unclear. The choice of resistance management strategy has been suggested to differ for sexual and asexual pests. For instance, current IRAC guidance suggests that pesticide mixtures are less effective in non-mating pests than in sexually reproducing populations. Here, stochastic evolutionary simulations are used to compare resistance evolution under sequences and mixtures across four reproductive modes observed in pests of economic importance: sexual reproduction, obligate parthenogenesis, cyclical parthenogenesis and haplodiploidy. Contrary to current expectations, mixtures are not disadvantaged in asexual populations and, in some cases, lead to delayed resistance evolution compared to sexually reproducing populations. These differences arise as the result of reduced genetic recombination which constrained the assembly and spread of multi-resistant genotypes. Overall, these findings suggest that mixtures remain a viable resistance management strategy for pests with asexual reproduction.

BACKGROUNDIn 2023, soybean growers in Eaton County, Michigan, reported repeated failures to control common ragweed (Ambrosia artemisiifolia L.) with (PPO)-inhibiting herbicides, sulfentrazone and fomesafen, in non-genetically modified soybeans. This study aimed to investigate resistance levels and mechanisms of resistance in two suspected resistant populations (R1 and R2). RESULTSDose-response assays revealed resistance to sulfentrazone in both populations, with LD50 values 24 and 36-fold higher than the susceptible population and reduced sensitivity to fomesafen. Nanopore sequencing identified two PPO2 target-site substitutions at codon 98: Arg- 98-Leu (R98L) and a novel Arg-98-Gln (R98Q) mutation. Both substitutions were associated with reduced herbicide binding affinity in computational modelling simulations. R98Q substitution conferred strong and selective resistance in the PPO2 enzyme inhibition assays, supporting its role as a key resistance mechanism. CONCLUSIONThis study reports the first field occurrence of the R98Q substitution in PPO2 of A. artemisiifolia populations and demonstrates its association with to PPO inhibitor resistance. These findings highlight the rapid evolution of target-site resistance and the need for continued monitoring and rapid diagnostic tools to detect emerging mutations. Further research is needed to clarify the contribution of additional resistance mechanisms.

1- Biological control of mosquitoes using aquatic predators offers a sustainable alternative to chemical insecticides, yet the specific predator and prey functional traits that govern consumption efficacy remain poorly quantified at a global scale. 2- We conducted a global meta-analysis of 755 effect sizes across 59 studies to evaluate how predator identity (fish vs. odonate naiads), body size, dietary guilds, and prey characteristics influence consumption rates. Using multilevel models and robust publication-bias corrections, we quantified predation efficiency, expressed throughout as the consumption rate (CR, larvae predator -1 h -1 ), while accounting for methodological variations across experimental designs. The primary literature itself proved geographically skewed towards Asia (chiefly India), with Africa, the Americas, and Europe markedly under-represented. 3- Grouping predators solely by broad taxonomic identity concealed the central pattern in our data. Although naiads outperformed fish when compared directly within the same studies, this taxon-level difference was driven almost entirely by non-mosquitofish species, the least efficient predator group overall. Mosquitofish (\textit{Gambusia} spp.) and dragonfly naiads were statistically indistinguishable from one another, indicating that dietary specialisation, not taxonomic identity, is the stronger predictor of predation efficacy. Predator body size strongly and positively predicted consumption rates in naiads---driven primarily by dragonflies---but showed no significant or negative relationship in fish. 4- Methodological traits heavily structured the extreme heterogeneity observed across studies; notably, exposure time acted as a severe rate-suppressor, where prolonged assays drastically underestimated per-hour consumption rates due to satiety or handling constraints. Nevertheless, a combined model incorporating all significant ecological moderators simultaneously explained a substantial share of the between-study variance, confirming that predator-prey dynamics in these systems are highly predictable from functional traits. 5- Effective biological control cannot rely on broad taxonomic assumptions but requires evidence-based trait-matching. Management programs should prioritise body size when deploying insect predators, selecting the largest individuals within species known to consume mosquito larvae and favour insectivorous fish species over generalists. Crucially, because short-term laboratory assays artificially inflate efficacy, multi-duration assessments are essential to accurately scale up biocontrol predictions from experimental arenas to complex, real-world ecosystems.

Neurotoxic chemicals, including pyrethroids, neonicotinoids, and chlorinated hydrocarbons, have been effective for mosquito control; however, their continued use has resulted in widespread insecticide resistance. This resistance has necessitated increased insecticide application, a phenomenon known as the pesticide treadmill, which poses significant risks to both environmental and human health. Consequently, there is a growing need for sustainable alternatives with minimal adverse impacts. Although plant-based repellents have shown promise, many are formulated as topical products that may be costly, inconvenient, or culturally unacceptable in some communities. Despite the documented mosquito-repellent properties of Hyptis suaveolens, its traditional use relies largely on fresh plant material, which is limited by seasonality and may not provide year-round protection. In addition, evidence on practical, sustained-release delivery systems for plant-derived mosquito repellents remains limited. Candle wax represents a readily available and culturally familiar medium that may facilitate the gradual release of volatile repellent compounds during combustion without requiring direct skin application. This study therefore evaluated the mosquito-repellent efficacy of Hyptis suaveolens extracts infused into candle wax as a simple plant-based delivery system. Leaves of H. suaveolens were collected, air-dried, ground, and extracted using ethanol and water. The crude extracts were incorporated into melted candle wax at a 5:10 concentration, then solidified and burned in a mosquito-infested room for 30 minutes. Results indicated that rooms with infused candles had significantly fewer mosquitoes than the control room. These findings support the mosquito-repellent properties of Hyptis suaveolens and demonstrate that its incorporation into candle wax is a promising plant-based delivery system for mosquito control, warranting further evaluation under field conditions.

Personal insect repellents can offer an effective means of preventing mosquito bites. Despite a wide variety of products that claim to repel mosquitoes, relatively few studies have systematically compared their performance through standardized tests. In Australia, chemical-based repellents must be registered with the Australian Pesticides and Veterinary Medicines Authority (APVMA); however, many unregistered products with unverified effectiveness remain commercially available. Here, we tested 55 insect repellents and 5 non-repellent perfumes and deodorants sold in Australia for their protection against Aedes aegypti mosquito landings (as a proxy for bites) using arm-in-cage assays across eight human subjects. We tested 43 topical repellents (containing 18 different primary active ingredients), four wristbands, three clothing stickers, three ultrasonic devices, one garment, one repellent applied to fabric, two perfumes, two deodorants and one antiperspirant. Thirteen repellents, including all wristbands, stickers and ultrasonic devices did not provide complete protection against mosquito landings for any human subject, while a further sixteen provided complete protection for 1 hr or less on average. Repellents containing DEET, oil of lemon eucalyptus, picaridin and IR3535 as the primary active ingredient were the only products that provided more than 2 hours of complete protection, but with significant variation among products with the same active ingredient. Only 7/30 essential oil-based topical repellents and perfumes provided complete protection for greater than 1 hr. Nearly half (25/52) of the repellents subject to registration with the APVMA were unregistered despite being advertised or labelled as mosquito repellents, and these tended to be less effective than registered products. Only 5/28 repellents met or exceeded their claimed protection time. This study highlights the prevalence of unregistered and ineffective mosquito repellents on the Australian market while providing evidence for products and active ingredients that do protect against mosquito bites under laboratory conditions.

The rapid spread of herbicide-resistant Amaranthus palmeri S. Watson across the Mediterranean region poses a severe threat to crop production. This study characterizes the resistance profile of an A. palmeri population from Gonen, Israel, which survived multiple post-emergence herbicide applications in maize. Greenhouse dose-response and screening trials were conducted across four modes of action (MOAs). The Gonen population exhibited three-way multiple resistance to acetolactate synthase (ALS), photosystem II (PSII) and 4-hydroxyphenylpyruvate dioxygenase (HPPD) inhibitors, marking the first confirmed case outside the Americas. High-level resistance was demonstrated for the ALS inhibitor foramsulfuron and the PSII inhibitor metribuzin, maintaining 40-70% and 80-100% survival, respectively, at double the labelled field rate. Sequencing of the ALS gene revealed a Trp574 to Leu substitution in resistant plants. The population showed differential PSII sensitivity, as atrazine provided complete control despite high metribuzin survival. Sequencing revealed no known psbA mutations, and malathion pre-treatment caused only a slight reduction in metribuzin survival. Partial resistance was observed for the HPPD inhibitor tembotrione (30-50% survival), where malathion pre-treatment reduced survival by 20-30%. In contrast, pre-mergence applications of S-metolachlor (very long chain fatty acid inhibitor), isoxaflutole (HPPD inhibitor), and a fomesafen + terbutryn mixture [protoporphyrinogen oxidase (PPO) + PSII inhibitors] significantly reduced emergence. These findings confirm multiple-resistant A. palmeri in the region and highlight the urgent need for integrated weed management incorporating effective pre-emergence herbicides rather than relying solely on post-emergence chemical control in summer cereal rotations.

Fungal and oomycete plant pathogens are major drivers of yield losses worldwide and are spurring the search for sustainable alternatives to synthetic pesticides. Algae, in general, and microalgae, in particular, represent a promising source of bioactive compounds for crop protection. Despite this, their efficacy across different host-pathogen systems remains poorly characterised. This study evaluated the biocontrol potential of the aqueous extract of Chlorella sorokoniana against three economically important phytopathogens using complementary in vitro, ex vivo, and in planta assays. The strongest activity was observed against Magnaporthe oryzae, with the extract reducing fungal growth by approximately 70% in vitro, inhibiting appressorium formation by 55%, suppressing lesion development on detached rice leaves by more than 75%, and reducing rice blast severity by 64.5% as a preventive foliar treatment. In contrast, the extract showed little or no direct in vitro antifungal activity against Pythium ultimum and Rhizoctonia solani, yet it significantly reduced disease severity in planta by 13-37% and 48-55%, respectively. The contrasting responses among pathosystems suggest that C. sorokoniana aqueous extracts act through different mechanisms depending on the pathogen and the crop, combining direct antifungal activity against M. oryzae with plant-associated protective effects against soil-borne pathogens. These findings highlight the importance of evaluating candidate biocontrol products using complementary in vitro and in planta approaches, as laboratory antimicrobial assays alone may substantially underestimate their agricultural potential. The broad-spectrum protection achieved with an unrefined aqueous extract further supports C. sorokoniana as a promising source of sustainable crop protection products and provides a strong foundation for future mechanistic studies, formulation development, and field validation. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=115 SRC="FIGDIR/small/742703v1_ufig1.gif" ALT="Figure 1"> View larger version (45K): org.highwire.dtl.DTLVardef@394716org.highwire.dtl.DTLVardef@6a208corg.highwire.dtl.DTLVardef@17f0724org.highwire.dtl.DTLVardef@adda96_HPS_FORMAT_FIGEXP M_FIG C_FIG

IntroductionChanges in vector control strategies alter the selection pressures exerted on natural populations of Anopheles gambiae s.l. and may influence the dynamics of insecticide resistance mechanisms. However, the consequences of discontinuing indoor residual spraying (IRS) campaigns on the evolution of Ace-1-mediated resistance remain poorly documented under natural conditions. This study aimed to assess the spatiotemporal evolution of resistance to pirimiphos-methyl following the cessation of IRS and to investigate evidence consistent with the existence of a biological cost associated with Ace-1-mediated resistance. MethodsNatural populations of An. gambiae s.l. were collected between 2017 and 2023 in three districts in Burkina Faso that had undergone IRS campaigns (Kampti, Solenzo, and Kongoussi). Susceptibility tests with pirimiphos-methyl (0.25%) were conducted out following WHO protocols, and a subsample of exposed mosquitoes was genotyped to detect the Ace-1 G119S mutation. Spatiotemporal trends in mortality, allele frequencies and genotypes were analyzed according to the pre-IRS, IRS, and post-IRS periods. The association between the Ace-1 genotype and survival following exposure to pirimiphos-methyl was assessed using logistic regression, while the concordance between phenotypic and molecular indicators of resistance was examined using Spearmans correlation. ResultsThe susceptibility of An. gambiae s.l. populations to pirimiphos-methyl was gradually restored after the discontinuation of IRS at all sites. At the same time, the frequencies of the Ace-1 119S resistance allele declined sharply, particularly in Kampti and Solenzo, while they remained low in Kongoussi throughout the study period. Mosquitoes carrying resistant genotypes had a significantly higher probability of survival after exposure to pirimiphos-methyl than susceptible homozygotes, with resistant homozygotes (RR) exhibiting the greatest survival advantage (OR = 27.62; 95% CI: 6.91-110.45; p < 0.001). A significant negative correlation was observed between the frequency of the Ace-1 119S allele and phenotypic mortality ({rho} = -0.48; p = 0.033), indicating a concordance between the two indicators of resistance. The progressive decline in allele frequencies, the decreasing prevalence of resistant genotypes, and the concomitant restoration of susceptibility are field observations consistent with the existence of biological costs associated with Ace-1-mediated resistance. ConclusionThis study provides field evidence consistent with the existence of biological costs associated with Ace-1-mediated resistance in natural populations of An. gambiae s.l. These results underscore the value of an adaptive resistance management strategy based on alternating selection pressures and could guide future vector control strategies, particularly if indoor residual spraying campaigns or other interventions relying on organophosphates were reintroduced.

In Germany, insecticide use is recommended during mosquito-borne disease outbreaks, yet nationwide data on insecticide resistance in key vector species are scarce. This is particularly relevant for Culex pipiens s.l., a vector of West Nile virus. Here, we provide the first broad-scale assessment of molecular insecticide resistance (kdr and ace-1 mutations) in Cx. pipiens f. pipiens across Germany. We associated insecticide resistances with environmental conditions, potential links with mitochondrial variation of the cytochrome c oxidase I (COI) gene as well as phenotypic resistance (CDC bottle assay with permethrin) of the Frankfurt population with elevated kdr frequencies. Kdr resistance alleles were detected in German Cx. pipiens f. pipiens reaching frequencies of up to 50% in the metropolitan areas of Berlin and Frankfurt am Main. Annual precipitation was negatively associated with kdr allele frequency. Interestingly, ace-1 resistance mutations were only present in the metropolitan area of Frankfurt am Main and in Muggensturm (Rhine valley, southwestern Germany). COI haplotype analyses revealed low genetic variability within German Cx. pipiens f. pipiens populations with no association to the insecticide resistance genotype. Phenotypic assays of the Frankfurt population showed early signs of permethrin resistance, but unmatched genotypic and phenotypic resistance state. This study provides the first broad-scale assessment of molecular insecticide resistance in Cx. pipiens f. pipiens across Germany, establishing a baseline for resistance surveillance and vector control. Further research is needed to identify factors shaping phenotypic resistance, as molecular markers alone may not reliably predict resistance. Since sampling was restricted to urban green spaces, broader sampling across environmental gradients is needed to identify additional environmental effectors.

Maize (Zea mays) is a staple food for millions in sub-Saharan Africa (SSA), contributing to both caloric intake and essential micronutrients. Recent breeding efforts have focused on enhancing its nutritional value by increasing provitamin A (PVA) carotenoid content to address vitamin A deficiency (VAD), a common problem among children under 5 years, pregnant and lactating mothers across SSA. However, high rainfall, and both warm and humid conditions in various SSA regions result in devastating foliar diseases such as maize streak virus (MSV), northern corn leaf blight (NCLB), southern corn leaf blight (SCLB), southern corn rust, grey leaf spot (GLS), and Curvularia leaf spot (CLS). Breeding for resistance can aid in mitigating yield losses caused by those diseases. Rapid, efficient screening methods can allow for examining large germplasm collections. The current study evaluated 21 maize inbred lines with contrasting PVA content, along with two commercial inbred controls, for resistance to Exserohilum turcicum, Bipolaris maydis, and Curvularia lunata, causal agents of NCLB, SCLB, and CLS, respectively, using a detached leaf assay (DLA) and under natural field infestation. Significant variation was detected among the inbreds, seven were classified as resistant to certain diseases, ten as moderately resistant, and six as susceptible to all foliar diseases. Overall, high PVA inbred lines had less susceptibility to NCLB, SCLB, and CLS. The results suggest that PVA-enriched maize inbred lines possess improved resistance to multiple foliar diseases. The field assessments of disease severity validated the effectiveness of the DLA in distinguishing resistant from susceptible inbred lines, and resistance to other diseases in field conditions was detected in parallel. The results indicate that high PVA maize has the potential to simultaneously address VAD, mycotoxin contamination in SSA with improved resistance to multiple foliar diseases.

Chemotaxis assays are widely used to quantify sensory-driven behavioral responses in Caenorhabditis elegans. Sodium azide is commonly used to immobilize animals at assay endpoints, although concerns have been raised that diffusion beyond the endpoint region could affect behavior before a choice is completed. We compared sodium azide-mediated immobilization with post-assay cooling using the repellent 1-octanol and attractant diacetyl. Wild-type animals showed robust avoidance of 30% 1-octanol and attraction to 0.5% diacetyl with either method, across multiple assay durations, and with or without worm food Escherichia coli OP50. These results support post-assay cooling as an alternative endpoint-scoring method.

Accurately determining the mating status of the agricultural fruit fly pest Ceratitis capitata, commonly known as Medfly, is essential for timely and effective eradication efforts. To overcome the limitations of subjective DAPI-based staining assessments of females captured in Jackson dry traps and Multilure liquid traps, we developed a multi-tier molecular diagnostic method that unequivocally detects mating status using DNA probes targeting the male-specific Y114 locus on the Y-chromosome of the species. Our protocol integrates morphological evaluation with increasingly sensitive molecular assays through the following steps: 1) A preliminary quality assessment of the specimens physical condition, DNA preservation, and mating status using conventional PCR followed by agarose electrophoresis (cPCR); 2) Quantification and real-time detection of sperm presence via quantitative PCR (qPCR); and 3) Detection of trace sperm amounts through droplet digital PCR (ddPCR). This PCR-based framework is designed for samples collected in the field, enabling accurate analysis of specimens exposed to adverse environmental conditions and varying levels of preservation after 2- and 3-weeks weathering times in traps. It allows quantitative determination of mating status even when sperm concentrations are extremely low, such as during transient copulation, and achieves detection limits down to approximately 14 spermatozoa in a mated female. By accounting for variable specimen quality and the performance characteristics of each molecular platform, this tiered approach ensures highly sensitive and unequivocal detection of mated females. The methodology can be used to assist eradication efforts across the C. capitata geographic range through the timely detection of mated females, halting their expansion and establishment into novel regions reducing control and eradication costs.

Calonectria ilicicola, an emerging and economically important soilborne pathogen in the U.S. Midwest, causes red crown rot (RCR) disease of soybean. It is essential to have an early and accurate detection method for effective management of RCR, since disease symptoms are nearly identical to those of other soybean diseases. In this study, we developed and validated a high resolution melting (HRM) assay targeting the translation elongation factor 1&alpha (TEF-1&alpha) gene for the specific detection of C. ilicicola. The assay was evaluated using a specificity panel of 78 DNA samples, including 67 fungal and oomycete DNA samples and 11 soybean host-DNA samples, as well as DNA from 56 field-collected symptomatic and asymptomatic soybean tissues. The HRM assay generated distinct melting profiles and, where applicable, absence or delayed amplification that reliably differentiated C. ilicicola from non target fungi and oomycetes. Additionally, the marker distinguished three haplotypes among C. ilicicola isolates, consistent with nucleotide polymorphisms in the TEF-1&alpha target region. The assay demonstrated strong quantitative performance, exhibiting high linearity (R^2= 0.9992) and acceptable amplification efficiency (94.05%) across a broad DNA concentration range. In field samples, HRM based detection aligned with disease status, accurately identifying C. ilicicola in symptomatic plants and no detection in asymptomatic tissues. Overall, this HRM assay provides a rapid, sensitive, and reliable diagnostic tool for routine molecular detection of C. ilicicola.

Stink bugs (Hemiptera Pentatomidae) are major agricultural pests worldwide, including both native and invasive species. In France, for instance, Halyomorpha halys has rapidly expanded its range, affecting key crops such as hazelnuts, apples, and vegetables. Meanwhile, several native, polyphagous species - Nezara viridula, Palomena prasina and, to a lesser extent, Graphosoma italicum - are re-emerging and impacting diverse crops because of changes in agricultural practices and climatic conditions. Egg parasitoids represent promising biological control agents for these pests, be it through conservation, augmentation or introduction biological control. However, their actual potential is not precisely documented due to a lack of ambitious surveys and possible taxonomic issues. This study aims to address this gap by compiling results obtained from various of egg parasitism surveys of Pentatomidae in France and implementing a large-scale DNA Barcoding approach (2000 individuals) for the emerged parasitoids. Our results indicate a high diversity of egg parasitoids, and detected some taxonomic issues and molecular inconsistencies that warrant further study. Among the three families of egg parasitoids recovered (Encyrtidae, Eupelmide, and Scelionidae), Scelionidae was the most abundant and diverse with nine species of Trissolcus (including the two adventive species, Tr. japonicus and Tr. mitsukurii) and four Telenomus species (including two unidentified to species). Scelionidae dominated the parasitoid assemblages associated with most of the Pentatomidae host species, except for H. halys for which the eupelmid Anastatus bifasciatus was the most common species.

The black bean aphid, Aphis fabae is a crop pest and vector of insect-transmitted pathogens, comprising closely related sub-species with overlapping host ranges. In other Aphis species, over-expression of specific detoxification genes has been linked to insecticide tolerance. We present two chromosome-scale assemblies for a clonal A. fabae line, representing two phased haplotypes, generated using HiFi and Hi-C sequencing technologies. A comprehensive genome annotation, built with PacBio Iso-Seq data, was used to investigate genes underlying insecticide tolerance. Both genomes are comprised of four chromosomal blocks (haplotype 1: 427 Mb; haplotype 2: 396 Mb) with high BUSCO completeness (98.7%). Comparative genomics revealed an expansion of UDP-glycosyltransferases, whose expression is linked to insecticide detoxification in other Aphis species. These high-quality references provide a foundation for studying A. fabae sub-species and a genomic resource for investigating insecticide tolerance across the Aphis genus. Author summaryHere we have provided a comprehensive assembly and annotation for further study into the Black bean aphid, Aphis fabae, using up to date long-range sequencing technologies. The final assemblies for both haplotypes are chromosome length and consist of 4 main chromosome blocks, consistent with the literature. The A. fabae genome was found to contain an increase in copy number of UDP-glycosyltransferases, which have previously been linked to insecticide resistance. The work here will be a resource to those studying insecticide tolerance in crop pests, as well as the differences between A. fabae sub-species.

The generalist Helicoverpa armigera and the specialist Helicoverpa assulta are closely related noctuid pests and are among the few insect herbivores capable of feeding on and damaging hot pepper fruits, which contain the defensive compound capsaicin. Cytochrome P450 monooxygenases (P450s) contribute to the metabolism of plant defensive compounds and can facilitate insect adaptation to chemically defended host plants. Here, we identified CYP6AN1 in H. assulta (HassCYP6AN1) and comparatively characterized the CYP6AN1 orthologs from H. armigera (HarmCYP6AN1) and H. assulta. RACE identified one full-length HarmCYP6AN1 transcript and three full-length HassCYP6AN1 transcript isoforms. The HassCYP6AN1 isoforms contained distinct 5' UTRs generated by alternative transcription initiation and splicing but shared an identical coding sequence. Sequence comparisons and phylogenetic analysis supported their assignment as orthologs. Constitutive CYP6AN1 expression was higher in the H. assulta midgut, whereas dietary capsaicin significantly induced CYP6AN1 expression in the H. armigera midgut. Recombinant CYP6AN1 proteins were co-expressed with NADPH-cytochrome P450 reductase in Escherichia coli, and their substrate-metabolizing activities were evaluated using HPLC-based depletion assays. Both orthologs metabolized capsaicin, but HarmCYP6AN1 exhibited an approximately 2.3-fold higher depletion activity than HassCYP6AN1 under the conditions tested. HarmCYP6AN1 also showed P450-content-dependent xanthotoxin depletion, whereas no detectable xanthotoxin metabolism was observed for HassCYP6AN1. These findings establish CYP6AN1 as a component of the capsaicin-metabolizing repertoire of both species and reveal substantial divergence between the orthologs in transcript organization, expression regulation, catalytic activity, and detectable substrate range.

Despite their fundamental role in sustaining terrestrial biodiversity and ecosystem functioning, soils are increasingly contaminated by pesticides, often occurring as complex mixtures. Understanding chemical uptake, distribution, and elimination in soil organisms is essential for reliable risk assessment. We investigated the toxicokinetics of a binary mixture of imidacloprid and epoxiconazole in the earthworm Aporrectodea caliginosa. Single-substance experiments were conducted to estimate uptake and elimination rates, and allowed the implementation of single substance toxicokinetic models. A mixture experiment tested five concentration ratios to assess potential interactions. Epoxiconazole, an azole fungicide, exhibited rapid uptake and elimination (k_u = 1.9 gsoil*gworm-1*d-1; k_e = 1.7 d-1), resulting in low bioaccumulation (BAF = 0.9), whereas imidacloprid, a neonicotinoid insecticide, accumulated slowly with low elimination (k_u = 1.6 gsoil*gworm-1* d-1; k_e = 0.12 d-1), producing high bioaccumulation (BAF = 20). The explicit inclusion of the 24-hour Petri dish depuration phase proved to be critical, especially for epoxiconazole, highlighting the influence of gut-clearance procedures on apparent kinetics. In accordance with only a small synergistic effect in the mixture, no toxicokinetic interactions were detected in the mixture; internal concentrations of each compound were consistent across ratios. Concentration-dependent patterns were observed, with indications of a saturation process at high exposure levels for imidacloprid, and an increased BAF at high concentrations for epoxiconazole. This finding questions the use of single, concentration-independent bioaccumulation factors for hazard assessment and highlights the value of integrated TKTD approaches.

To reduce reliance on pesticides in potato production, alternative partial control methods are needed, yet their efficacy and reliability are seldom quantified over multiple years. Canopy architecture is a potential lever against late blight, caused by Phytophthora infestans, a pathogen whose infection and development are highly driven by humidity and temperature. However, its practical value under field conditions in pure crop stands remains poorly documented. We compared two commercial potato cultivars with contrasting canopy architectures but similar leaf tissue susceptibility to P. infestans, in replicated field trials conducted over six years (2011 2016) under irrigation, and over three of these years also without irrigation. Disease and canopy development (height, closure, leaf area, stem number) were monitored throughout the growing seasons and analysed using non linear growth models. Monalisa, an erect and ramose cultivar, consistently slowed epidemic progress relative to Bintje, a semi-erect and leafy cultivar. This effect was significant in every non irrigated trial but in only half of the irrigated ones. No single canopy trait explained this suppression across all years and conditions. However, canopy closure at the time of inoculation was the most consistent correlate, supporting the hypothesis of a microclimate mediated effect. Disease reduction seldom translated into a yield benefit, except under irrigation, where Monalisa occasionally out yielded Bintje. Our results confirm that canopy architecture can complement other levers within integrated late blight management. Nevertheless, its significant year to year variability and dependence on environmental conditions must be accounted for if it is to be deployed as a reliable control method.