Genome-Wide Exploration of Long Non-Coding RNAs of Helicoverpa armigera in Response to Pyrethroid Insecticide Resistance.

Genome-wide long non-coding RNAs (lncRNAs) in low, moderate, and high pyrethroid insecticide-resistant and -susceptible strains of Helicoverpa armigera were identified in this study. Using 45 illumina-based RNA-sequencing datasets, 8394 lncRNAs were identified. In addition, a sublethal dose of deltamethrin was administered to a Korean-resistant strain (Kor-T). The average length of lncRNAs was approximately 531 bp, and the expression ratio of lncRNAs was 28% of the total RNA. The identified lncRNAs were divided into six categories-intronic, intergenic, sense, antisense, cis-RNA, and trans-RNA-based on their location and mechanism of action. Intergenic and intronic lncRNA transcripts were the most abundant (38% and 33%, respectively). Further, 828 detoxification-related lncRNAs were selected using the Gene Ontology analysis. The cytochrome P450-related lncRNA expression levels were significantly higher in susceptible strains than in resistant strains. In contrast, cuticle protein-related lncRNA expression levels were significantly higher in all resistant strains than in susceptible strains. Our findings suggest that certain lncRNAs contribute to the downregulation of insecticide resistance-related P450 genes in susceptible strains, whereas other lncRNAs may be involved in the overexpression of cuticle protein genes, potentially affecting the pyrethroid resistance mechanism.

Transcriptomic investigation of the molecular mechanisms underlying resistance to the neonicotinoid thiamethoxam and the pyrethroid lambda-cyhalothrin in Euschistus heros (Hemiptera: Pentatomidae).

BACKGROUND: Laboratory-selected resistant strains of Euschistus heros to thiamethoxam (NEO) and lambda-cyhalothrin (PYR) were recently reported in Brazil. However, the mechanisms conferring resistance to these insecticides in E. heros remain unresolved. We utilized comparative transcriptome profiling and single nucleotide polymorphism (SNP) calling of susceptible and resistant strains of E. heros to investigate the molecular mechanism(s) underlying resistance. RESULTS: The E. heros transcriptome was assembled, generating 91 673 transcripts with a mean length of 720 bp and N50 of 1795 bp. Comparative gene expression analysis between the susceptible (SUS) and NEO strains identified 215 significantly differentially expressed (DE) transcripts. DE transcripts associated with the xenobiotic metabolism were all up-regulated in the NEO strain. The comparative analysis of the SUS and PYR strains identified 204 DE transcripts, including an esterase (esterase FE4), a glutathione-S-transferase, an ABC transporter (ABCC1) and aquaporins that were up-regulated in the PYR strain. We identified 9588 and 15 043 nonsynonymous SNPs in the PYR and NEO strains. One of the SNPs (D70N) detected in the NEO strain occurs in a subunit (α5) of the nAChRs, the target site of neonicotinoid insecticides. Nevertheless, this residue position in α5 is not conserved among insects. CONCLUSIONS: Neonicotinoid and pyrethroid resistance in laboratory-selected E. heros is associated with a potential metabolic resistance mechanism by the overexpression of proteins commonly involved in the three phases of xenobiotic metabolism. Together these findings provide insight into the potential basis of resistance in E. heros and will inform the development and implementation of resistance management strategies against this important pest. © 2023 Society of Chemical Industry.

Susceptibility monitoring and comparative gene expression of susceptible and resistant strains of Spodoptera frugiperda to lambda-cyhalothrin and chlorpyrifos.

BACKGROUND: Spodoptera frugiperda (J. E. Smith) is a widespread agricultural pest with several records of resistance to different insecticides and Bt proteins, including the neurotoxic insecticides chlorpyrifos (organophosphate) and lambda-cyhalothrin (pyrethroid). Here, we (i) characterized and monitored the susceptibility of field populations of S. frugiperda to chlorpyrifos (194 populations) and lambda-cyhalothrin (197 populations) collected from major maize-growing regions of Brazil from 2003 to 2016, and (ii) compared gene expression levels of laboratory-selected, chlorpyrifos- and lambda-cyhalothrin-resistant strains to a susceptible reference strain (Sf-ss) of S. frugiperda. RESULTS: The susceptibility monitoring detected average survival ranging from 29.3% to 36.0% for chlorpyrifos, and 23.1% to 68.0% for lambda-cyhalothrin. The resistance ratio of the chlorpyrifos-resistant strain (Clo-rr) was 25.4-fold and of the lambda-cyhalothrin-resistant strain (Lam-rr) was 21.5-fold. We identified 1098 differentially expressed genes (DEGs) between Clo-rr and Sf-ss, and 303 DEGs between Lam-rr and Sf-ss. Functional analyses of the DEGs revealed the up-regulation of several detoxification enzymes, mainly cytochrome P450 belonging to CYP3 and CYP6 clans. Genes associated with regulatory processes, such as the forkhead box class O (FoxO) transcription factor were also up-regulated. Variant analysis of target-site mutations for both pesticides identified the A201S and F290V mutations in acetylcholinesterase-1, both occurring in heterozigosis in the Clo-rr S. frugiperda strain. CONCLUSION: Our data show that the overexpression of the enzymatic detoxification machinery is the main difference to explain the resistance of Clo-rr and Lam-rr strains of S. frugiperda to chlorpyrifos and lambda-cyhalothrin, although a target-site mutation also contributes to the Clo-rr resistance to chlorpyrifos. © 2023 Society of Chemical Industry.

Trends towards Lower Susceptibility of Spodoptera frugiperda (Lepidoptera: Noctuidae) to Teflubenzuron in Brazil: An Evidence for Field-Evolved Resistance.

Susceptibility monitoring to insecticides is a key component to implementing insecticide resistance management (IRM) programs. In this research, the susceptibility to teflubenzuron in Spodoptera frugiperda (J.E Smith) was monitored in more than 200 field-collected populations from major corn-growing regions of Brazil, from 2004 to 2020. Initially, we defined a diagnostic concentration of 10 µg mL-1 of teflubenzuron using a diet-overlay bioassay for monitoring the susceptibility. A variation in the susceptibility to teflubenzuron in S. frugiperda was detected among populations from different locations. We also detected a significant reduction in the susceptibility to teflubenzuron throughout time in all the populations of S. frugiperda evaluated, with larval survival at diagnostic concentration varying from values of <5% in 2004 to up 80% in 2020. Thus, this research provides evidence of field-evolved resistance of S. frugiperda to teflubenzuron and reinforces that IRM practices are urgently needed to be implemented in Brazil.

The genome sequence of the Neotropical brown stink bug, Euschistus heros provides insights into population structure, demographic history and signatures of adaptation.

The Neotropical brown stink bug, Euschistus heros, is a major pest of soybean in South America. The importance of E. heros as a pest has grown significantly in recent times due to increases in its abundance and range, and the evolution of insecticide resistance. Recent work has begun to examine the genetic diversity, population structure, and genetic mechanisms of insecticide resistance in E. heros. However, to date, investigation of these topics has been hampered by a lack of genomic resources for this species. Here we address this need by assembling a high-quality draft genome for E. heros. We used a combination of short and long read sequencing to assemble an E. heros genome of 1.4 Gb comprising 906 contigs with a contig N50 of 3.5 MB. We leveraged this new genomic resource, in combination with genotyping by sequencing, to explore genetic diversity in populations of this species in Brazil and identify genetic loci in the genome which are under selection. Our genome-wide analyses, confirm that there are two populations of E. heros co-occurring in different geographical regions in Brazil, and that, in certain regions of the country these populations are hybridizing. We identify several regions of the genome as under selection, including markers associated with putative insecticide resistance genes. Taken together, the new genomic resources generated in this study will accelerate research into fundamental aspects of stinkbug biology and applied aspects relating to the sustainable control of a highly damaging crop pest.

The Cost of Resistance to Diamide Insecticide Varies With the Host Plant in Spodoptera frugiperda (Lepidoptera: Noctuidae).

Fitness costs associated with insect resistance to insecticides can be exploited to implement resistance management programs. However, most of these studies are restricted to evaluating biological traits on artificial diets. Here, we investigated the fitness cost associated with chlorantraniliprole in Spodoptera frugiperda (J.E. Smith) feeding on corn, soybean, and cotton plants. We used a near-isogenic strain of S. frugiperda resistant to chlorantraniliprole (Iso-RR), a susceptible strain (SS), and heterozygotes strains (H1 and H2) to evaluate several biological and population growth parameters. Larval survival of the Iso-RR strain was on average 90% on corn, 65% on soybean, and 57% on cotton plants. Development time of the larval stage also differed among host plants, Iso-RR strain took on average 14, 17, and 26 days to reach the pupal stage on corn, soybean, and cotton plants respectively. Net reproductive rate, intrinsic rate of population increase, and finite rate of population increase were higher for Iso-RR strain feeding on corn plants than other host plants. The relative fitness, based on the intrinsic rate of population increase, of S. frugiperda resistant strain on corn, soybean, and cotton plants were 1.04, 0.85, and 0.88, respectively. Therefore, no fitness cost was observed for S. frugiperda feeding on corn plants, but a significant fitness cost was observed when this pest fed on soybean and cotton plants. We showed that the food source influences the fitness cost of S. frugiperda resistant to diamide. Such information may help to implement resistance management strategies based on each crop.

Baseline Susceptibility and Cross-Resistance of HearNPV in Helicoverpa armigera (Lepidoptera: Noctuidae) in Brazil.

The marked adoption of bioinsecticides in Brazilian agriculture in recent years is, at least partially, explained by the increasingly higher levels of insect pest resistance to synthetic insecticides. In particular, several baculovirus-based products have been registered in the last 5 years, including Helicoverpa armigera nucleopolyhedrovirus (HearNPV: Baculoviridae: Alphabaculovirus (Armigen®)). Understanding the susceptibility of Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) to HearNPV is an important step toward development of robust Integrated Pest Management (IPM) and Insect Resistance Management programs (IRM) aimed at managing this serious insect pest. In this study, droplet feeding bioassays were used to characterize the baseline susceptibility to HearNPV (Armigen®) in H. armigera populations collected from major soybean and cotton-growing regions in Brazil. We defined and validated a diagnostic concentration for susceptibility monitoring of H. armigera populations to HearNPV. Additionally, cross-resistance between HearNPV and the insecticides flubendiamide and indoxacarb was evaluated by testing HearNPV in a susceptible strain and in resistant strains of H. armigera to these insecticides. A low interpopulation variation of H. armigera to HearNPV was detected. The LC50 values ranged from 1.5 × 105 to 1.1 × 106 occlusion bodies (OBs) per mL (7.3-fold variation). The mortality rate at the identified diagnostic concentration of 6.3 × 108 OBs/mL, based on the calculated LC99, ranged from 98.6 to 100% in populations of H. armigera collected from 2018 to 2020. No cross-resistance was detected between HearNPV and flubendiamide or indoxacarb. These results suggest that HearNPV (Armigen®) can be an effective tool in IPM and IRM programs to control H. armigera in Brazil.

Modeling the Resistance Evolution to Insecticides Driven by Lepidopteran Species Competition in Cotton, Soybean, and Corn Crops.

Intra- and interspecific competition is considered a fundamental phenomenon in ecology. It acts as one of the most powerful selective forces that drives ecological diversity, the spatiotemporal distribution of organisms, fitness, and evolutionary aspects. Spodoptera frugiperda and Helicoverpa armigera are devastating pests and can co-occur in systems consisting of multiple agricultural crops and compete for food resources. Insecticide resistance in populations of these species has been a major threat to the sustainability of agroecosystems. No study to date has shown the effect of intra- and interspecific competition as a selective pressure agent on the evolution of insecticide resistance in lepidopteran pests in an experimental and theoretical way. Our study developed a parameterized computational model with experimental results for S. frugiperda and H. armigera competition. We simulated the behavior of heterozygous individuals with a competition capacity 100% equal to homozygous individuals resistant (100 RR) or susceptible to insecticides (00 RR), and intermediate between them (50 RR). Competition involving strains of these insect species can accelerate the evolution of their resistance to insecticides in agricultural crops. We found that competitive processes can result in a high probability of competitive exclusion for individuals with the susceptibility allele of these lepidopteran species. The results of this study are of paramount importance for understanding the impact of ecological factor competition on the evolution of insecticide resistance in lepidopteran pests, which until now has been neglected in these types of evolutionary dynamics studies.

Large-Scale Monitoring of the Frequency of Ryanodine Receptor Target-Site Mutations Conferring Diamide Resistance in Brazilian Field Populations of Fall Armyworm, Spodoptera frugiperda (Lepidoptera: Noctuidae).

Fall armyworm (FAW), Spodoptera frugiperda, is an important lepidopteran pest in the Americas, and recently invaded the Eastern Hemisphere. In Brazil, FAW is considered the most destructive pest of corn and cotton. FAW has evolved resistance to many insecticides and Bacillus thuringiensis (Bt) proteins. Here, a large-scale monitoring was performed between 2019 and 2021 to assess diamide insecticide susceptibility in more than 65 FAW populations sampled in corn and cotton. We did not detect a significant shift in FAW susceptibility to flubendiamide, but a few populations were less affected by a discriminating rate. F2 screen results of 31 selected FAW populations across regions confirmed that the frequency of diamide resistance alleles remained rather stable. Two laboratory-selected strains exhibited high resistance ratios against flubendiamide, and cross-resistance to anthranilic diamides. Reciprocal crosses indicated that resistance is autosomal and (incompletely) recessive in both strains. F1 backcrosses suggested monogenic resistance, supported by the identification of an I4734M/K target-site mutation in the ryanodine receptor (RyR). Subsequent genotyping of field-collected samples employing a TaqMan-based allelic discrimination assay, revealed a low frequency of RyR I4790M/K mutations significantly correlated with phenotypic diamide resistance. Our findings will help to sustainably employ diamides in FAW resistance management strategies across crops.

Regional pest suppression associated with adoption of Cry1Ac soybean benefits pest management in tropical agriculture.

BACKGROUND: Bacillus thuringiensis (Bt) crops have been adopted worldwide, providing high-level protection from insect pests. Furthermore, Bt crops preserve natural enemies, promote higher yield, and economically benefit farmers. Although regional pest suppression by widespread Bt crop adoption has been observed in temperate regions, this possibility remains uncertain in tropical areas due to the high diversity of alternative hosts and mild winters. RESULTS: Evidence of regional reduction in insecticide use across areas was observed in Brazil where Cry1Ac soybean has been grown since 2013, with up to 50% reduction in the number of insecticide sprays for managing lepidopteran pests on non-Bt soybean observed at specific locations from 2012 to 2019. Pest monitoring data from four mesoregions across 5 years of commercial plantings of Cry1Ac soybean from December 2014 to July 2019 showed reduced numbers of Chrysodeixis includens moths captured in pheromone traps across years at all locations. The number of Helicoverpa spp. moths captured also was reduced at three locations. CONCLUSION: We provide evidence for regional suppression of lepidopteran pests and reduced insecticide use with the widespread adoption of Cry1Ac soybean in Brazil, bringing economic, social and environmental benefits. © 2022 Society of Chemical Industry.

Interspecific Variation in Susceptibility to Insecticides by Lepidopteran Pests of Soybean, Cotton, and Maize Crops From Brazil.

The interspecific variation in susceptibility to insecticides by lepidopteran species of soybean [Glycine max L. (Merr.)], cotton (Gossypium hirsutum L.), and maize (Zea mays L.) crops from Brazil were evaluated. Populations of Anticarsia gemmatalis (Hübner) (Lepidoptera: Erebidae), Chrysodeixis includens (Walker), Helicoverpa armigera (Hübner), Spodoptera frugiperda (Smith), Spodoptera eridania (Stoll), Spodoptera cosmioides (Walker), and Spodoptera albula (Walker) (Lepidoptera: Noctuidae) were collected from 2019 to 2021. Early L3 larvae (F2 generation) were exposed to the formulated insecticides methoxyfenozide, indoxacarb, spinetoram, flubendiamide, and chlorfenapyr in diet-overlay bioassays. The median lethal concentrations (LC50) were used to calculate tolerance ratios (TR) of each species in relation to the most susceptible species to each insecticide. The lowest LC50 values were verified for A. gemmatalis to all insecticides tested. Chrysodeixis includens and most of the Spodoptera species were moderately tolerant to methoxyfenozide (TR < 8.0-fold) and indoxacarb (TR < 39.4-fold), whereas H. armigera was the most tolerant species to methoxyfenozide (TR = 21.5-fold), and indoxacarb (TR = 106.4-fold). Spodoptera cosmioides, S. eridania, and S. albula showed highest tolerance to spinetoram (TR > 1270-fold), S. eridania, S. frugiperda, and S. albula to flubendiamide (TR from 38- to 547-fold), and S. albula to indoxacarb (TR = 138.6-fold). A small variation in susceptibility to chlorfenapyr (TR < 4.4-fold) was found among the lepidopteran evaluated. Our findings indicate a large variation in susceptibility to indoxacarb, spinetoram, and flubendiamide and a relatively low variation in susceptibility to methoxyfenozide and chlorfenapyr by lepidopteran species of soybean, cotton, and maize from Brazil.

Fitness costs associated with chlorantraniliprole resistance in Spodoptera frugiperda (Lepidoptera: Noctuidae) strains with different genetic backgrounds.

BACKGROUND: Spodoptera frugiperda (J.E. Smith) is a difficult pest to manage mainly because of its resistance to insecticides and Bt proteins. We evaluated fitness costs of S. frugiperda resistant strains to diamide insecticides with different genetic backgrounds aiming to highlight the importance of using isogenic strains. We established a near-isogenic strain of S. frugiperda resistant to diamides (Iso-RR), using a chlorantraniliprole resistant strain (RR) selected from a field-collected population and a susceptible reference strain (SS). Fitness costs were assayed using strains with close-related genetic backgrounds (Iso-RR and SS) and strains with distant-related genetic backgrounds (RR and SS). RESULTS: No fitness cost associated with chlorantraniliprole resistance in S. frugiperda was observed using the Iso-RR strain, based on life history traits. The only parameter that differs between Iso-RR and SS strains was the mean length of a generation (T), whereas the Iso-RR strain presented T = 35.8 and SS strain showed T = 34.6. On the other hand, a significant fitness cost was detected using the RR strain. All population growth parameters differ between RR and SS strains. Based on the intrinsic rate of population increase (rm ) parameter, the relative fitness estimated was 1.02 for the Iso-RR strain and 0.64 for the RR strain. CONCLUSION: The genetic background of the resistant strains alters fitness cost outcomes. The RR strain showed fitness costs associated with resistance, but the Iso-RR did not. Our work supports the decision-making process of resistance management programs and adds to the growing body of research that enlightens the importance of strain genetics in fitness cost experiments.

Lime sulfur to control citrus flat mite and its interactions with the entomopathogenic fungus Lecanicillium muscarium

Lime sulfur is one of the few products indicated to control Brevipalpus yothersi in Brazilian organic citrus orchards. Other strategies, such as the use of entomopathogenic fungi should be evaluated, and Lecanicillium muscarium is one of the basic choices for pest management. Knowledge of the interactions between lime sulfur and this entomopathogen is critical for developing control strategies. With this goal, it was conducted the toxicological characterization of lime sulfur to B. yothersi and the compatibility evaluation with L. muscarium. Finally, the effects of L. muscarium and lime sulfur mixtures on B. yothersi control were evaluated. Product evaluation for B. yothersi was done through direct and residual contact bioassay, and different concentrations of lime sulfur mixed in potato dextrose agar culture medium were used to evaluate compatibility with L. muscarium. Lime sulfur was effective against adults of B. yothersi and caused eggs unviability of up to 71.0%, at a dose of 80 L per 2,000 L of H2O. The lethal concentration (LC50 and LC99) of lime sulfur estimated for mite adults were 246.62 and 858.5 µg of sulfur per mL of H2O (ppm a.i.). Lime sulfur concentrations of 180 to 560 ppm a.i. showed promise for use in combination with L. muscarium. However, concentrations of 1,000 and 5,600 ppm significantly reduced colony size and the number of spores/colony. The mixture of 100 and 180 ppm a.i. of lime sulfur with L. muscarium (108 conidia·mL­1) was not able to reduce the lethal time of entomopathogen on B. yothersi.

Resistance status of lepidopteran soybean pests following large-scale use of MON 87701 × MON 89788 soybean in Brazil.

Widespread adoption of MON 87701 × MON 89788 soybean, expressing Cry1Ac Bt protein and glyphosate tolerance, has been observed in Brazil. A proactive program was implemented to phenotypically and genotypically monitor Cry1Ac resistance in Chrysodeixis includens (Walker). Recent cases of unexpected injury in MON 87701 × MON 89788 soybean were investigated and a large-scale sampling of larvae on commercial soybean fields was performed to assess the efficacy of this technology and the distribution of lepidopteran pests in Brazil. No significant shift in C. includens susceptibility to Cry1Ac was observed eight years after commercial introduction of this technology in Brazil. F2 screen results confirmed that the frequency of Cry1Ac resistance alleles remains low and stable in C. includens. Unexpected injury caused by Rachiplusia nu (Guenée) and Crocidosema aporema (Walsingham) in MON 87701 × MON 89788 soybean was detected during the 2020/21 season, and studies confirmed a genetically based alteration in their susceptibility to Cry1Ac. MON 87701 × MON 89788 soybean remains effective against Anticarsia gemmatalis (Hübner), C. includens, Chloridea virescents (Fabricius) and Helicoverpa armigera (Hübner) in Brazil. However, there is evidence of field-evolved resistance to MON 87701 × MON 89788 soybean by the secondary soybean pests R. nu and C. aporema.

Monitoring Resistance of Euschistus heros (Fabricius) (Hemiptera: Pentatomidae) to Insecticides by Using Encapsulated Artificial Diet Bioassay.

The novel ingestion bioassay method was developed for detecting and monitoring resistance of Euschistus heros by encapsulating an artificial liquid diet using Parafilm®. This methodology was compared with the tarsal contact (vial test) and topical application methods for thiamethoxam, imidacloprid, and lambda-cyhalothrin. The best bioassay method for the neonicotinoid insecticides thiamethoxam and imidacloprid was ingestion. For pyrethroid insecticide lambda-cyhalothrin, the best result was obtained by topical application. Using the best bioassay method for each insecticide, the susceptibility to these insecticides was monitored in 30 populations of E. heros collected from soybean crops in Brazil from 2018 to 2020. High variations in susceptibility to thiamethoxam (resistance ratios, 1.6-22 times), imidacloprid (resistance ratios, 1.6-22 times), and lambda-cyhalothrin (resistance ratios, 5-40 times) were detected among the evaluated E. heros populations. In order to monitor the susceptibility of E. heros to insecticides, diagnostic concentrations were defined based on the LC99 of the susceptible reference population: 5.65 µL of a.i./mL for thiamethoxam, 12.45 µL of a.i./mL for imidacloprid, and 0.20 µg of a.i./insect for lambda-cyhalothrin. Subsequently, we select an E. heros strain resistant to neonicotinoid insecticides and another to lambda-cyhalothrin. The resistance ratios obtained after seven selection cycles were 66, 41 and 44 times for thiamethoxam, imidacloprid and lambda-cyhalothrin, respectively.

Large-scale assessment of lepidopteran soybean pests and efficacy of Cry1Ac soybean in Brazil.

The soybean technology MON 87701 × MON 89788, expressing Cry1Ac and conferring tolerance to glyphosate, has been widely adopted in Brazil since 2013. However, pest shifts or resistance evolution could reduce the benefits of this technology. To assess Cry1Ac soybean performance and understand the composition of lepidopteran pest species attacking soybeans, we implemented large-scale sampling of larvae on commercial soybean fields during the 2019 and 2020 crop seasons to compare with data collected prior to the introduction of Cry1Ac soybeans. Chrysodeixis includens was the main lepidopteran pest in non-Bt fields. More than 98% of larvae found in Cry1Ac soybean were Spodoptera spp., although the numbers of Spodoptera were similar between Cry1Ac soybean and non-Bt fields. Cry1Ac soybean provided a high level of protection against Anticarsia gemmatalis, C. includens, Chloridea virescens and Helicoverpa spp. Significant reductions in insecticide sprays for lepidopteran control in soybean were observed from 2012 to 2019. Our study showed that C. includens and A. gemmatalis continue to be primary lepidopteran pests of soybean in Brazil and that Cry1Ac soybean continues to effectively manage the target lepidopteran pests. However, there was an increase in the relative abundance of non-target Spodoptera spp. larvae in both non-Bt and Cry1Ac soybeans.

Inheritance patterns, cross-resistance and synergism in Spodoptera frugiperda (Lepidoptera: Noctuidae) resistant to emamectin benzoate.

BACKGROUND: Fall armyworm, Spodoptera frugiperda (J. E. Smith), is a relevant global pest due to severe damage caused on agricultural crops and its capacity to evolve resistance to insecticides. Here, we selected a strain of S. frugiperda resistant to emamectin benzoate under laboratory conditions to understand the inheritance patterns, cross-resistance and synergism involved in the resistance. RESULTS: The emamectin benzoate-resistant (Ben-R) strain was isolated by using F2 screen in a field population collected in Lucas do Rio Verde, Mato Grasso state, Brazil. After ten generations of selection pressure with emamectin benzoate, the estimated LC50 of the Ben-R strain was 678.38 µg a.i. mL-1 whereas that of the susceptible (Sus) strain was 0.29 µg a.i.mL-1 , resulting in a resistance ratio (RR) of ~ 2340-fold. The LC50 values of the offspring from reciprocal crosses of Sus and Ben-R strains were 93.37 and 105.32 µg a.i. mL-1 , suggesting that resistance is an autosomal incompletely dominant trait. The high survival of heterozygous and Ben-R strains (>92%) on non-Bt maize sprayed with the field rate of emamectin benzoate confirmed that resistance is functionally dominant. The minimum number of segregations influencing resistance was 3.55, suggesting a polygenic effect. Low cross-resistance was detected between emamectin benzoate and the insecticides methomyl, chlorpyrifos, lambda-cyhalothrin, spinetoram, indoxacarb and chlorantraniliprole (RR <5.75-fold). There was no effect of synergists piperonyl butoxide, diethyl maleate and S, S, S-tributyl phosphorotrithiotate on the Ben-R strain, suggesting a minor role of metabolic resistance. CONCLUSIONS: Our results showed a high risk of resistance evolution of S. frugiperda to emamectin benzoate, based on incompletely dominant inheritance. Rotation of insecticides with different modes of action can be one of the resistance management strategies to be implemented to delay the evolution of resistance of S. frugiperda to emamectin benzoate in Brazil.

Shifts in Ecological Dominance between Two Lepidopteran Species in Refuge Areas of Bt Cotton.

Competition behavior involving agricultural pest species has long been viewed as a powerful selective force that drives ecological and phenotypic diversity. In this context, a Game Theory-based approach may be useful to describe the decision-making dilemma of a competitor with impacts to guarantee its superiority in terms of ecological dominance or sharing of the food resource with its competitor. In an attempt to elucidate the consequences of competitive dynamics for the ecological dominance of these species in refuge areas of Bt cotton, we conducted a study that was divided into two parts. The first study consisted of an evaluation of interactions involving Spodoptera frugiperda (JE Smith, 1797) and Helicoverpa armigera (Hübner, 1808) on non-Bt cotton plants in a field trial. In the second study, we explored the data matrix collected in the field to parameterize a model of Cellular Automata (CA) with update rules inspired by Game Theory. Computer simulations were analyzed in hypothetical scenarios involving the application (or not) of insecticides in the refuge areas in combination with the resistance factor of one or both pest species to the insecticides used in the refuge areas. H. armigera had superior competitive performance in relation to S. frugiperda only at high densities. According to the density-mediated shift in dominance of the species, the resistance of S. frugiperda to insecticides is seen as a risk factor for the production of susceptible individuals of H. armigera on a large scale in the refuge areas. Additionally, S. frugiperda insecticide resistance may potentially impact the resistance evolution of the H. armigera population to Bt cotton. Thus, ecological dominance could diverge by the presence of a resistance allele to insecticides with interspecific competition perhaps subordinate to evolutionary processes.

Resistance to spirodiclofen in Brevipalpus yothersi (Acari: Tenuipalpidae) from Brazilian citrus groves: detection, monitoring, and population performance.

BACKGROUND: Brevipalpus yothersi mite is the main vector of Citrus leprosis virus (CiLV), the causal agent of citrus leprosis disease. The acaricide spirodiclofen has been widely used to control this mite. However, failures in control using spirodiclofen have been frequently reported by citrus growers. In this study, we estimated the diagnostic concentration to monitor the resistance to spirodiclofen of B. yothersi populations collected in nine citrus groves in Brazil. We then selected the B. yothersi population that showed lowest mortality with the estimated diagnostic concentration of spirodiclofen to characterize the frequency of resistant individuals, as well as demographic and life table parameters. RESULTS: Variability was higher between populations in terms of susceptibility to spirodiclofen. The frequency of resistant eggs between populations ranged from 0.7% to 85.8%. The resistance ratio of B. yothersi to spirodiclofen was low to moderate. Survival rates of the immature stage, total adult longevity, oviposition days, and female fecundity were lower in the resistant strain. Furthermore, net reproduction rate, intrinsic rate of increase, finite rate of increase, and mean length of a generation were also lower in the R strain. CONCLUSION: Although variations in resistance to spirodiclofen were observed between populations, the resistance ratio was low to moderate. Such data can be useful in the development of resistance management strategies for B. yothersi in Brazilian citrus groves. © 2021 Society of Chemical Industry.

Baseline Response, Monitoring, and Cross-Resistance of Spodoptera frugiperda (Lepidoptera: Noctuidae) to Sodium Channel Blocker Insecticides in Brazil.

Spodoptera frugiperda (J.E. Smith) is one of the key cross-crop pests in Brazilian agroecosystems. Field-evolved resistance of S. frugiperda to some conventional insecticides and Bt proteins has already been reported. Thus, the use of insecticides with new mode of action such as sodium channel blockers (indoxacarb and metaflumizone) could be an important tool in insecticide resistance management (IRM) programs. To implement a proactive IRM, we conducted baseline response and monitoring to indoxacarb and metaflumizone in 87 field populations of S. frugiperda collected from major maize-growing regions of Brazil from 2017 to 2020, estimated the frequency of resistance alleles to indoxacarb, and evaluated cross-resistance of indoxacarb and metaflumizone to some selected insecticides and Bt proteins. Low variation in susceptibility to indoxacarb (4.6-fold) and metaflumizone (2.6-fold) was detected in populations of S. frugiperda in 2017. The frequency of the resistance allele to indoxacarb was 0.0452 (0.0382-0.0527 CI 95%), by using F2 screen method. The mean survival at diagnostic concentration, based on CL99, varied from 0.2 to 12.2% for indoxacarb and from 0.0 to 12.7% for metaflumizone, confirming high susceptibility of S. frugiperda to these insecticides in Brazil. No cross-resistance was detected between sodium channel blocker insecticides and other insecticides (organophosphate, pyrethroid, benzoylurea, spinosyn, and diamide) and Bt proteins. These findings showed that sodium channel blocker insecticides are important candidates to be exploited in IRM strategies of S. frugiperda in Brazil.