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Insecticide resistance is a major problem in food production, environmental sustainability, and human health. The cotton bollworm Helicoverpa armigera is a globally distributed crop pest affecting over 300 crop species. H. armigera has rapidly evolved insecticide resistance, making it one of the most damaging pests worldwide. Understanding the genetic basis of insecticide resistance provides insights to develop tools, such as molecular markers, that can be used to slow or prevent the evolution of resistance. We explore the genetic architecture of H. armigera resistance to a widely used insecticide, flubendiamide, using two complementary approaches: genome-wide association studies (GWAS) in wild-caught samples and quantitative trait locus (QTL) mapping in a controlled cross of susceptible and resistant laboratory strains. Both approaches identified one locus on chromosome 2, revealing two SNPs within 976 bp that can be used to monitor field resistance to flubendiamide. This was the only region identified using linkage mapping, though GWAS revealed additional sites associated with resistance. Other loci identified by GWAS in field populations contained known insecticide detoxification genes from the ATP-binding cassette family, ABCA1, ABCA3, ABCF2 and MDR1. Our findings revealed an oligogenic genetic architecture, contrasting previous reports of monogenic resistance associated with the ryanodine receptor . This work elucidates the genetic basis of rapidly evolving insecticide resistance and will contribute to developing effective insecticide resistance management strategies.

Helicoverpa armigera is one of the primary agricultural pests in the Old World, whereas H. zea is predominant in the New World. However, H. armigera was first documented in Brazil in 2013. Therefore, the geographical distribution, range of hosts, invasion source, and dispersal routes for H. armigera are poorly understood or unknown in Brazil. In this study, we used a phylogeographic analysis of natural H. armigera and H. zea populations to (1) assess the occurrence of both species on different hosts; (2) infer the demographic parameters and genetic structure; (3) determine the potential invasion and dispersal routes for H. armigera within the Brazilian territory; and (4) infer the geographical origin of H. armigera. We analyzed partial sequence data from the cytochrome c oxidase subunit I (COI) gene. We determined that H. armigera individuals were most prevalent on dicotyledonous hosts and that H. zea were most prevalent on maize crops, based on the samples collected between May 2012 and April 2013. The populations of both species showed signs of demographic expansion, and no genetic structure. The high genetic diversity and wide distribution of H. armigera in mid-2012 are consistent with an invasion period prior to the first reports of this species in the literature and/or multiple invasion events within the Brazilian territory. It was not possible to infer the invasion and dispersal routes of H. armigera with this dataset. However, joint analyses using sequences from the Old World indicated the presence of Chinese, Indian, and European lineages within the Brazilian populations of H. armigera. These results suggest that sustainable management plans for the control of H. armigera will be challenging considering the high genetic diversity, polyphagous feeding habits, and great potential mobility of this pest on numerous hosts, which favor the adaptation of this insect to diverse environments and control strategies.

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. F 2 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 .

Competition is a driving force regulating communities often considered an intermittent phenomenon, difficult to verify and potentially driven by environmental disturbances. Insecticides are agents of environmental disturbance that can potentially change ecological relationships and competitive outcomes, but this subject has seldom been examined. As the co-existing cereal grain beetle species Sitophilus zeamais Motschulsky and Rhyzopertha dominica F. share a common realized niche, directly competing for the same resources, they were used as models in our study. Intraspecific competition experiments were performed with increasing insect densities and insecticide doses in additive and replacement series using various density combinations of both beetle species maintained on insecticide-free or -sprayed grains. Insecticide-mediated release from competitive stress was not observed in our study of intraspecific competition in grain beetles. The insecticide enhanced the effect of insect density, particularly for the maize weevil S. zeamais, further impairing population growth at high densities. Therefore, insecticide susceptibility increased with intraspecific competition favoring insecticide efficacy. However, the effect of insecticide exposure on competitive interaction extends beyond intraspecific competition, affecting interspecific competition as well. Sitophilus zeamais was the dominant species when in interspecific competition prevailing in natural conditions (without insecticide exposure), but the dominance and species prevalence shifted from S. zeamais to R. dominica under insecticide exposure. Therefore, high conspecific densities favored insecticide efficacy, but the strength of the relationship differs with the species. In addition, the insecticide mediated a shift in species dominance and competition outcome indicating that insecticides are relevant mediators of species interaction, potentially influencing community composition and raising management concerns as potential cause of secondary pest outbreaks.

Abstract Spodoptera eridania (Stoll) is a key pest of soybean crops in Brazil. However, information about the physiology, behavior, and ecology of S. eridania, including its genetic variability within the agricultural landscape, remains scarce. In this study, we conducted an exploratory analysis of the genetic diversity, population structure, and demographic patterns of S. eridania in Brazil. A cytochrome c oxidase I (COI) gene fragment of 866 bp was sequenced from 89 S. eridania individuals collected in Brazilian soybean macroregions. We identified 33 COI haplotypes with high haplotype diversity and moderate nucleotide diversity distributed throughout the country. The genetic relationships among COI haplotypes show a recent divergence with no evidence of distinct haplogroups. Spodoptera eridania collected on soybean crops showed a lack of population genetic structure associated with soybean macroregions or Brazilian states. Overall, our data suggest that S. eridania populations are undergoing demographic and spatial expansion in Brazil, with an increasing effective population size in the last 200 years. This study provides the first insights into the population diversity and demography of S. eridania in the Americas, shedding light on the dynamics and evolution of this species and further supporting integrated pest management strategies in Brazilian soybean crops.

Tropical landscapes are rapidly changing due to deforestation and agricultural expansion, entailing the loss and fragmentation of natural habitats. Understanding how these changes affect the genetic diversity and gene flow in key native pollinators is of great importance to assure their survival and provision of pollination services. In this context, we studied how landscape features influence genetic diversity and gene flow in one of the most widespread species of stingless bees in the Neotropical region, Tetragonisca angustula. We evaluated bees from 46 nests sampled across forested, agricultural and urban landscapes within the Atlantic Forest, genotyped at 745 single-nucleotide polymorphisms (SNPs). We found that forest cover negatively influenced the heterozygosity at a 500-m scale, although inbreeding and gene flow were not influenced by landscape features. Gene flow was explained mainly by geographic distance, indicating that T. angustula can disperse across heterogeneous and human-altered landscapes.

Individual traits vary among and within populations, and the co-occurrence of different endosymbiont species within a host may take place under varying endosymbiont loads in each individual host. This makes the recognition of the potential impact of such endosymbiont associations in insect species difficult, particularly in insect pest species. The maize weevil, Sitophilus zeamais Motsch. (Coleoptera: Curculionidae), a key pest species of stored cereal grains, exhibits associations with two endosymbiotic bacteria: the obligatory endosymbiont SZPE (\"Sitophilus zeamais Primary Endosymbiont\") and the facultative endosymbiont Wolbachia. The impact of the lack of SZPE in maize weevil physiology is the impairment of nutrient acquisition and energy metabolism, while Wolbachia is an important factor in reproductive incompatibility. However, the role of endosymbiont load and co-occurrence in insect behavior, grain consumption, body mass and subsequent reproductive factors has not yet been explored. Here we report on the impacts of co-occurrence and varying endosymbiont loads achieved via thermal treatment and antibiotic provision via ingested water in the maize weevil. SZPE exhibited strong effects on respiration rate, grain consumption and weevil body mass, with observed effects on weevil behavior, particularly flight activity, and potential consequences for the management of this pest species. Wolbachia directly favored weevil fertility and exhibited only mild indirect effects, usually enhancing the SZPE effect. SZPE suppression delayed weevil emergence, which reduced the insect population growth rate, and the thermal inactivation of both symbionts prevented insect reproduction. Such findings are likely important for strain divergences reported in the maize weevil and their control, aspects still deserving future attention.

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.

The egg parasitoid Telenomus remus (Hymenoptera: Scelionidae) has been investigated for classical and applied biological control of noctuid pests, especially Spodoptera (Lepidoptera: Noctuidae) species. Although T. remus was introduced into Brazil over three decades ago for classical biological control of S. frugiperda , this wasp has not been recorded as established in corn or soybean crops. We used an integrative approach to identify T. remus , combining a taxonomic key based on the male genitalia with DNA barcoding, using a cytochrome c oxidase subunit I mitochondrial gene fragment. This is the first report of natural parasitism of T. remus on S. frugiperda and S. cosmioides eggs at two locations in Brazil. We also confirmed that the T. remus lineage in Brazil derives from a strain in Venezuela (originally from Papua New Guinea and introduced into the Americas, Africa, and Asia). The occurrence of T. remus parasitizing S. frugiperda and S. cosmioides eggs in field conditions, not associated with inundative releases, suggests that the species has managed to establish itself in the field in Brazil. This opens possibilities for future biological control programs, since T. remus shows good potential for mass rearing and egg parasitism of important agricultural pests such as Spodoptera species.

Thyrinteina arnobia (Lepidoptera: Geometridae) is a native American species. Despite its historical importance as an insect pest in Eucalyptus plantations, more information is needed regarding the population diversity, demography, and climatic variables associated with its distribution in different regions of Brazil. We used a phylogeographic approach to infer the genetic diversity, genetic structure, and demographic parameters of T. arnobia . We also conducted an ecological niche modeling (ENM) to predict suitable areas for T. arnobia occurrence in Brazil and other countries worldwide. Although T. arnobia populations have low genetic diversity in Brazil, we identified mitochondrial haplogroups predominating in different Brazilian regions and high Ф ST and Ф CT values in AMOVA, suggesting a low frequency of insect movement among these regions. These results indicate that outbreaks of T. arnobia in Eucalyptus areas in different regions of Brazil are associated with local or regional populations, with no significant contribution from long-distance dispersal from different regions or biomes, suggesting that pest management strategies would be implemented on a regional scale. In Brazil, the demographic and spatial expansion signals of T. arnobia seem to be associated with the history of geographical expansion of Eucalyptus plantations, a new sustainable host for this species. ENM indicated that isothermality and annual rainfall are critical climatic factors for the occurrence of T. arnobia in tropical and subtropical areas in the Americas. ENM also suggested that T. arnobia is a potential pest in Eucalyptus areas in all Brazilian territory and in regions from Africa, Asia, and Oceania.