Insect Resistance Management Programs Research Articles

Oviposition patterns of primary lepidopteran defoliators in soybean and the impact on structured refuge recommendations.

Transgenic Bt technology in soybean, with plants expressing Cry1Ac, has been adopted as an insect pest management tool. It was first adopted in large areas of South America and Asia in 2013. The risk of resistance in target pests to this technology demands insect resistance management (IRM) programs. In Brazil, a structured refuge (area of non-Bt soybean) planted adjacent to the Bt soybean crop has been an important IRM recommendation, particularly for the primary lepidopteran defoliators Anticarsia gemmatalis (Lepidoptera: Erebidae) and Chrysodeixis includens (Lepidoptera: Noctuidae). The overall goal of this study was to validate IRM recommendations to Bt soybean. The objectives were to document the impact of soybean phenology, cultivar choice and non-Bt soybean defoliation on moth oviposition. In addition, a mark-release-recapture study estimated the dispersal capacity of these species. Five field experiments per species were performed for 3 years. Our results revealed an increase in A. gemmatalis and C. includens oviposition, respectively, on Bt plants as a consequence of the difference in plant growth stage at the time of oviposition. Defoliation of non-Bt plants significantly increased the oviposition preference of both moth species for Bt plants. The mark-release-recapture experiment indicated an average dispersal distance of ~300 m from the release point for A. gemmatalis, with maximum recapture at 1000 m. Overall, our findings emphasize the importance of planting synchronization of Bt soybean and the structured refuge. In addition, when operational aspects in large soybean areas challenge this recommendation, the priority should be for planting the refuge area first. This approach will minimize the impact of selective oviposition of A. gemmatalis and C. includens. © 2024 Society of Chemical Industry.

Continental-scale migration patterns and origin of Helicoverpa zea (Lepidoptera: Noctuidae) based on a biogeochemical marker.

Insect migrations have ecological and economic impacts, particularly in agriculture. However, there is limited knowledge about the migratory movements of pests at the continental scale, which is an important factor influencing the spread of resistance genes. Understanding the migratory patterns of economic pests, like Helicoverpa zea (Boddie), is essential for improving Integrated Pest Management (IPM) and Insect Resistance Management (IRM) strategies. In this study, we used stable hydrogen isotopic ratios in wing tissue as a biogeochemical marker to examine migratory patterns and estimate the native origins of H. zea individuals collected across a wide latitudinal gradient in North America. Samples collected at higher latitudes (Ontario, Canada and Minnesota, USA) exhibited a greater proportion (60%-96%) of nonlocal individuals, with an increased probability of origin from the southeastern United States. Populations from mid-latitudes (Florida, North Carolina, and South Carolina) showed a blend of local and nonlocal (40%-60%) individuals. Finally, 15% of the southernmost population individuals (Puerto Rico) were classified as migratory, with some having a probability of origin at higher latitudes (>30°). Overall, our results provide evidence of a northward spring/summer migration of H. zea in North America and underscore the significance of the southeastern United States as a hub for genetic flow. In addition, based on stable hydrogen isotopic ratios, there is strong evidence of reverse (southward) migration of H. zea from the continental United States to Puerto Rico. Our study highlights the implications for IPM and IRM programs and the need for management strategies that account for both northward and southward migration patterns.

Toxicity and residual effects of spinetoram on adults and larvae of Anastrepha fraterculus and Ceratitis capitata (Diptera: Tephritidae)

The use of organophosphate insecticides is one of the main strategies for the chemical control of Anastrepha fraterculus and Ceratitis capitata (Diptera: Tephritidae) in Brazil. Although organophosphate insecticides are effective in population suppression, their constant use (up to 5 applications/harvest) may favor pest resistance. Nevertheless, some organophosphate insecticides are highly toxic to natural enemies and thus their use has been banned. In this study, we investigated acute and residual toxicity of different doses of insecticide spinetoram for the control of larvae and adults of A. fraterculus and C. capitata under laboratory and field conditions. Spinetoram (Delegate 250WG™) showed high toxicity to adults and larvae of A. fraterculus and C. capitata (mortality>80%), based on topical application and ingestion bioassays. At the highest dose evaluated, 5.0 g a.i. (active ingredient) per 100 L of water, spinetoram was statistically similar to organophosphate (Lorbans 480BR™), but superior to neonicotinoid (Eleitto™), standard products for fruit fly control in Brazil. In addition, at doses 3.75 and 5 g a.i.100 L-1, spinetoram showed significant toxicity (mortality>70%) via tarsal contact for both species. Furthermore, toxic effects at depth were observed on larvae of 1st, 2nd, and 3rd instars of fly species in apple and peach fruits, reducing adult emergence by up to 90%. In the field, the highest dose evaluated (spinetoram 5 g a.i.100 L-1) reduced infestation of A. fraterculus by 90%. The results show that spinetoram could be used in the management of A. fraterculus and C. capitata, also as an alternative control for insect resistance management programs.

Within-field spatial patterns of Helicoverpa zea (Lepidoptera: Noctuidae) and spatial associations with stink bugs and their injury in field corn.

The corn earworm, Helicoverpa zea (Boddie) (Lepidoptera: Noctuidae), is a cosmopolitan pest in the field crop landscape in the southeastern United States. Field corn (Zea mays L.) is the most important midseason host for H. zea where intensive selection pressure occurs for resistance to insecticidal toxins from Bacillus thuringiensis (Bt). Because spatial patterns of H. zea in field corn have not been extensively studied, field corn was sampled for H. zea larvae and injury in 2021 and 2022. Patterns of spatial aggregation were identified in a number of fields in both larval populations and injury. Aggregation of H. zea larvae was less common at R5 than at R2. Associations between the spatial patterns of H. zea and the variability in crop phenology were identified in some fields, with positive associations between plant height and H. zea larvae, indicating that ovipositing H. zea moths avoid areas with reduced plant height and delayed reproductive maturity. Additionally, negative spatial associations between stink bug ear injury and H. zea larvae and their injury were found in a small number of cases, indicating some spatial interactions between the two pest complexes and their injury. Results from these studies provide valuable insight into the spatial patterns of H. zea in field corn. An understanding of the local dispersal and population dynamics of H. zea can be used to help further improve integrated pest management and insect resistance management programs for this major polyphagous pest.

Impact of Seed Blend and Structured Maize Refuge on Helicoverpa zea (Lepidoptera: Noctuidae) Potential Phenological Resistance Development Parameters in Pupae and Adults.

Helicoverpa zea, an economic pest in the southeastern U.S., has evolved practical resistance to Bt Cry toxins in maize and cotton. Insect resistance management (IRM) programs have historically required planting of structured non-Bt maize, but because of its low adoption, the use of seed blends has been considered. To generate knowledge on target pest biology and ecology to help improve IRM strategies, nine field trials were conducted in 2019 and 2020 in Florida, Georgia, North Carolina, and South Carolina to evaluate the impact of Bt (Cry1Ab + Cry1F or Cry1Ab + Cry1F + Vip3A) and non-Bt maize plants in blended and structured refuge treatments on H. zea pupal survival, weight, soil pupation depth, adult flight parameters, and adult time to eclosion. From a very large sample size and geography, we found a significant difference in pupal mortality and weight among treatments in seed blends with Vip3A, implying that cross-pollination occurred between Bt and non-Bt maize ears. There was no treatment effect for pupation depth, adult flight distance, and eclosion time. Results of this study demonstrate the potential impact of different refuge strategies on phenological development and survival of an important pest species of regulatory concern. This article is protected by copyright. All rights reserved.

Association of growth-regulating insecticides and limonoid-based formulations: physicochemical compatibility and toxicity against Spodoptera frugiperda (Lepidoptera: Noctuidae).

The objective of this study was to evaluate the physicochemical compatibility of mixtures of synthetic and botanical limonoid-based insecticides, as well as the toxicity of these associations, in the management of Spodoptera frugiperda (J.E. Smith) under laboratory and field conditions. For this, the associations of 4 commercial botanical insecticides based on neem registered in Brazil (Azamax, Agroneem, Azact CE, and Fitoneem) were tested with synthetic insecticides from the group of growth regulators (IGRs [triflumuron, lufenuron, methoxyfenozide and tebufenozide]). When mixed, all combinations caused a significant reduction in the pH of the mixture and a significant increase in electrical conductivity. However, all tested combinations showed similar stability behavior to the negative control (distilled water), which demonstrated their physicochemical compatibility. Furthermore, in laboratory and field bioassays, mixtures of IRGs with limonoid-based formulations provided satisfactory effects in the management of S. frugiperda. However, binary mixtures of insecticide Intrepid 240 SC with Azamax or Azact CE (at LC25 previously estimated) showed the highest toxicities on S. frugiperda larvae in laboratory bioassays and damage reduction caused by S. frugiperda in a 2-yr field experiments. Therefore, mixtures of IGRs with limonoid-based botanical insecticides are promising alternatives for the management of S. frugiperda and important component of integrated pest management and insect resistance management programs.

Helicoverpa genus on the edge of the continental U.S.: Flight phenology, analysis of hybrid presence, and insecticide performance in high-input field crops in Puerto Rico.

The genus Helicoverpa includes several agricultural pests globally. Helicoverpa armigera was reported in several countries in South America in 2013, and in Puerto Rico, in 2014. This territory is considered an agricultural hub, with a high-input system of seed production in the southern region of the island, and also at the edge of the continental U.S. Possible natural dispersion of populations of H. armigera from the Caribbean or other Central American regions poses a continuing risk to the U.S. This study was performed during the post-detection scenario of H. armigera in Puerto Rico, from 2018 to 2021. A year-round pheromone trapping program of adult males indicated an increase in the population from October to March and differences in the occurrence of Helicoverpa spp. between the municipalities Juan Diaz and Salinas. The proportion of H. armigera/H. zea and detection of congeneric hybrids between these species were assessed based on genital morphology and DNA analysis. Interestingly, neither H. armigera nor expected hybrids were detected in the present study. The susceptibility of H. zea populations to the insecticides Spinetoram, Emamectin benzoate, Chlorantraniliprole, and Esfenvalerate was assessed, and an overall significant effect of insecticide susceptibility was detected. Chlorantraniliprole and Emamectin benzoate had the highest efficacy. These results contribute to the Integrated Pest Management and Insect resistance management programs to Helicoverpa spp. in Puerto Rico. In addition, provide validated information to be considered in mitigation plans, in the scenario of an invasion of H. armigera in the continental U.S.

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

Simple SummaryHelicoverpa armigera nucleopolyhedrovirus (HearNPV: Baculoviridae: Alphabaculovirus (Armigen®)) is a registered insecticide for the management of cotton bollworm, Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) in Brazil. We conducted studies of baseline susceptibility of Brazilian populations of H. armigera to HearNPV (Armigen®, AgBiTech, Fort Worth, TX, USA) and cross-resistance between HearNPV and insecticides as valuable knowledge in support of integrated pest management and insect resistance management programs.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.

Best Management Practices to Delay the Evolution of Bt Resistance in Lepidopteran Pests Without High Susceptibility to Bt Toxins in North America.

Canadian and United States (US) insect resistance management (IRM) programs for lepidopteran pests in Bacillus thuriengiensis (Bt)-expressing crops are optimally designed for Ostrinia nubilalis Hübner in corn (Zea mays L.) and Chloridea virescens Fabricius in cotton (Gossypium hirsutum L.). Both Bt corn and cotton express a high dose for these pests; however, there are many other target pests for which Bt crops do not express high doses (commonly referred to as nonhigh dose pests). Two important lepidopteran nonhigh dose (low susceptibility) pests are Helicoverpa zea Boddie (Lepidoptera: Noctuidae) and Striacosta albicosta Smith (Lepidoptera: Noctuidae). We highlight both pests as cautionary examples of exposure to nonhigh dose levels of Bt toxins when the IRM plan was not followed. Moreover, IRM practices to delay Bt resistance that are designed for these two ecologically challenging and important pests should apply to species that are more susceptible to Bt toxins. The purpose of this article is to propose five best management practices to delay the evolution of Bt resistance in lepidopteran pests with low susceptibility to Bt toxins in Canada and the US: 1) better understand resistance potential before commercialization, 2) strengthen IRM based on regional pest pressure by restricting Bt usage where it is of little benefit, 3) require and incentivize planting of structured corn refuge everywhere for single toxin cultivars and in the southern US for pyramids, 4) integrate field and laboratory resistance monitoring programs, and 5) effectively use unexpected injury thresholds.

Fitness Cost of Chlorpyrifos Resistance in Spodoptera frugiperda (Lepidoptera: Noctuidae) on Different Host Plants.

Spodoptera frugiperda (J. E. Smith, 1797) is a polyphagous pest of global relevance due to the damage it inflicts on agricultural crops. In South American countries, this species is one of the principal pests of maize and cotton. Currently, S. frugiperda is also emerging as an important pest of soybeans and winter cereals in Brazil. Chemical control is one of the main control tactics against S. frugiperda, even though resistance against numerous modes of action insecticides has been reported. To support insect resistance management programs, we evaluated the fitness costs of resistance of S. frugiperda to the acetylcholinesterase inhibitor chlorpyrifos. Fitness costs were quantified by comparing biological parameters of chlorpyrifos-resistant and -susceptible S. frugiperda and their F1 hybrids (heterozygotes) on non-Bt cotton, non-Bt maize, non-Bt soybean, and oats. The results revealed that the chlorpyrifos-resistant genotype showed lower pupa-to-adult and egg-to-adult survivorship and reduced larval weights on oats; longer neonate-to-pupa and egg-to-adult developmental periods, and lower pupal weights and fecundity on maize; lower pupal weights on soybean; and reduced fecundity on cotton compared with the chlorpyrifos-susceptible genotype. Fitness costs also affected fertility life table parameters of the resistant genotype, increasing the mean length of a generation on cotton and maize and reducing the potential for population growth on all hosts. These findings suggest fitness costs at the individual and population levels of chlorpyrifos resistance in S. frugiperda, indicating that removal of the selective agent from the environment would result in reduced resistance and opportunities for the restoration of susceptibility.

Transgene behavior in Zea mays L. crosses across different genetic backgrounds: Segregation patterns, cry1Ab transgene expression, insecticidal protein concentration and bioactivity against insect pests.

Brazil and South Africa, countries with economies in transition, are characterized by a dual agrarian structure co-occurring, sometimes, alongside in the same region. Large-scale commercial farming produces crops for export to global markets on the one hand, and small-scale farming, on the other hand, provides for subsistence and local markets. In both systems, maize (Zea mays) is a key crop for these two countries. For the commercial system, maize is a commodity crop while for the small-scale system it is the prime staple crop. In commercial systems, farmers predominantly grow genetically modified (GM) hybrid maize. In small-scale systems, farmers rely on open pollinated varieties (OPVs) and/or landraces and are largely dependent on seed saving systems. The aim of this study was to understand the relationship between transgene expression rates, the resulting concentrations of the transgene product (Bt protein) and its bioactivity in insect pests following transgene flow from GM hybrid maize into non-genetically modified, non-GM near-isogenic maize hybrid (ISO) and OPVs. We modeled segregation patterns and measured cry1Ab transgene expression (mRNA quantification), Cry1Ab protein concentration and pest survival. Two groups of F1, F2 crosses and backcrosses with GM, ISO and OPV maize varieties from Brazil and South Africa were used. Bioassays with the larvae of two lepidopteran maize pest species, Helicoverpa armigera and Spodoptera littoralis, were carried out. Overall, the cry1Ab transgene outcrossed effectively into the genetic backgrounds tested. The cry1Ab transgene was stably expressed in both ISO and OPV genetic backgrounds. Transgene introgression led to consistent, though highly variable, concentrations of Cry1Ab toxins that were similar to those observed in GM parental maize. Most crosses, but not all, suggested the expected Mendelian segregation pattern. Transgene expression rates were significantly higher than expected from purely Mendelian segregation in the South African crosses. In South African materials, ISO and OPV crosses produced significantly lower Cry1Ab concentrations compared to the GM parental maize. The Cry1Ab toxins from crosses were bioactive and induced mortality rates of ≥92.19% in H. armigera and ≥40.63% in S. littoralis after a period of only 4 days. However, no correlations were observed between the quantitation of mRNA for cry1Ab and the corresponding Cry1Ab protein concentrations, nor between the Cry1Ab concentrations and insect mortality rates across different genetic backgrounds. We therefore suggest that while transcription of the cry1Ab transgene reliably determines the presence of Cry1Ab protein, mRNA levels do not reflect, by themselves, the end Cry1Ab protein concentrations found in the plant. Because predictably high Cry1Ab concentrations are a key prerequisite for effective insect resistance management (IRM) programs, this observation raises questions about the effectiveness of such programs in scenarios with complex crop genetic backgrounds. On the other hand, confirmed bioactivity in all crosses should be expected to impact small farmer’s selection behavior, unknowingly favoring the insecticidal trait. This may lead to a fixation of the trait in the wider population, and may influence the genetic diversity of varieties maintained by small-scale farmers.

A Framework for Effective Bt Maize IRM Programs: Incorporation of Lessons Learned From Busseola fusca Resistance Development.

Bt maize is genetically engineered to express insecticidal proteins from the bacterium Bacillus thuringiensis. Bt maize is used extensively by South African farmers to reduce yield losses caused by lepidopteran larvae. Starting in the 2004/2005 season, severe Busseola fusca-associated damage to Cry1Ab-expressing Bt maize was noted by South African farmers. The unsatisfactory pest control was eventually attributed to the development of insect resistance to the Cry1Ab protein in the Bt maize hybrids. An assessment of the historical events surrounding the development of resistance by B. fusca showed that there was room for improvement both in the insect resistance management (IRM) strategy selected and the implementation of the strategy. With the recent arrival of fall armyworm (Spodoptera frugiperda) in Africa, it is important to have IRM programs that are appropriate for all of the pests that constitute the maize lepidopteran pest complex. After the identification of shortcomings in the IRM programs implemented in South Africa, a framework is proposed for effective Bt maize IRM programs. The IRM framework integrates pre-marketing research, post-marketing monitoring, and two-level remedial action plans (RAPs). The core of the framework is a regulator-approved IRM strategy that is based on comprehensive pre-marketing research and serves to guide stakeholders during the post-marketing phase. The framework will assist technology developers and regulators, especially those with nascent regulatory systems, to select and implement IRM strategies that facilitate sustainable pest management.

Cross-crop resistance of Spodoptera frugiperda selected on Bt maize to genetically-modified soybean expressing Cry1Ac and Cry1F proteins in Brazil

Spodoptera frugiperda is one of the main pests of maize and cotton in Brazil and has increased its occurrence on soybean. Field-evolved resistance of this species to Cry1 Bacillus thuringiensis (Bt) proteins expressed in maize has been characterized in Brazil, Argentina, Puerto Rico and southeastern U.S. Here, we conducted studies to evaluate the survival and development of S. frugiperda strains that are susceptible, selected for resistance to Bt-maize single (Cry1F) or pyramided (Cry1F/Cry1A.105/Cry2Ab2) events and F1 hybrids of the selected and susceptible strains (heterozygotes) on DAS-444Ø6-6 × DAS-81419-2 soybean with tolerance to 2,4-d, glyphosate and ammonium glufosinate herbicides (event DAS-444Ø6-6) and insect-resistant due to expression of Cry1Ac and Cry1F Bt proteins (event DAS-81419-2). Susceptible insects of S. frugiperda did not survive on Cry1Ac/Cry1F-soybean. However, homozygous-resistant and heterozygous insects were able to survive and emerge as fertile adults when fed on Cry1Ac/Cry1F-soybean, suggesting that the resistance is partially recessive. Life history studies revealed that homozygous-resistant insects had similar development, reproductive performance, net reproductive rate, intrinsic and finite rates of population increase on Cry1Ac/Cry1F-soybean and non-Bt soybean. In contrast, heterozygotes had their fertility life table parameters significantly reduced on Cry1Ac/Cry1F-soybean. Therefore, the selection of S. frugiperda for resistance to single and pyramided Bt maize can result in cross-crop resistance to DAS-444Ø6-6 × DAS-81419-2 soybean. The importance of these results to integrated pest management (IPM) and insect resistance management (IRM) programs is discussed.

Resistance to insecticides in the field and baseline susceptibility to cyclaniliprole of whitefly Bemisia tabaci (Gennadius) in China

Resistance to multiple insecticides has become the biggest challenge to controlling Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae). In attempts to remedy the problem, four insecticides with novel modes of action (cyclaniliprole, cyantraniliprole, sulfoxaflor, and flupyradifurone) have recently been introduced to control B. tabaci. In the present study, we collected 18 B. tabaci populations in China from 2018 to 2019 to determine their resistance levels to six insecticides including the above four insecticides and two representative traditional insecticides (bifenthrin and imidacloprid). The results showed cyclaniliprole with LC50 values ranging from 3.4 to 21.2 mg/L was the most toxic insecticide to B. tabaci field populations and their average LC50 of 13.5 mg/L was used for the susceptible baseline to cyclaniliprole. The other three newly introduced insecticides (cyantraniliprole, sulfoxaflor, and flupyradifurone) also exhibited high toxicity to the field populations, however, the two traditional insecticides tested were of low toxicity with LC50 values ranging from 16.9 to 296.6 mg/L and from 33.1 to 397.0 mg/L for bifenthrin and imidacloprid, respectively. These results demonstrate that the newly introduced insecticides can currently be considered as effective alternative insecticides in insect resistance management programs for B. tabaci in China.

Linha Básica de Suscetibilidade de Chrysodeixis includens (Walker, [1858]) (Lepidoptera: Noctuidae) a Benzoato de Emamectina

Emamectin Benzoate was registered recently in Brazil to control Chrysodeixis includens (Walker, [1858]) (Lepidoptera: Noctuidae) in soybean. We studied the baseline susceptibility of Mato Grosso populations of C. includes to support Insect Resistance Management programs by standard feeding assays on insecticide-surface of artificial diet. Variation in susceptibility between populations was low. The LC50 values from 11 populations varied between 0.015 a 0.065 µg i.a. cm-2. Based on our results, we propose the discriminant concentration of 2.867 µg i.a. cm-2 of emamectin benzoate to monitor insecticide resistance in C. includes.

Control of western corn rootworm via RNAi traits in maize: lethal and sublethal effects of Sec23 dsRNA.

RNA interference (RNAi) triggered by maize plants expressing RNA hairpins against specific western corn rootworm (WCR) transcripts have proven to be effective at controlling this pest. To provide robust crop protection, mRNA transcripts targeted by double-stranded RNA must be sensitive to knockdown and encode essential proteins. Using WCR adult feeding assays, we identified Sec23 as a highly lethal RNAi target. Sec23 encodes a coatomer protein, a component of the coat protein (COPII) complex that mediates ER-Golgi transport. The lethality detected in WCR adults was also observed in early instar larvae, the life stage causing most of the crop damage, suggesting that WCR adults can serve as an alternative to larvae for dsRNA screening. Surprisingly, over 85% transcript inhibition resulted in less than 40% protein knockdown, suggesting that complete protein knockdown is not necessary for Sec23 RNAi-mediated mortality. The efficacy of Sec23 dsRNA for rootworm control was confirmed in planta; T0 maize events carrying rootworm Sec23 hairpin transgenes showed high levels of root protection in greenhouse assays. A reduction in larval survival and weight were observed in the offspring of WCR females exposed to Sec23 dsRNA LC25 in diet bioassays. We describe Sec23 as RNAi target for in planta rootworm control. High mortality in exposed adult and larvae and moderate sublethal effects in the offspring of females exposed to Sec23 dsRNA LC25 , suggest the potential for field application of this RNAi trait and the need to factor in responses to sublethal exposure into insect resistance management programs. © 2019 Society of Chemical Industry.

Low risk of resistance evolution of Spodoptera frugiperda to chlorfenapyr in Brazil

Insecticide resistance is serious problem for Spodoptera frugiperda (Lepidoptera: Noctuidae). Due to intense selection pressure, the evolution of S. frugiperda resistance to major insecticides groups and Bt proteins has already been documented. Therefore, studies to preserve the lifetime of insecticides with unique modes of action such as chlorfenapyr are important in insect resistance management programs. For implementing a proactive resistance management program of S. frugiperda to chlorfenapyr, studies were conducted with field populations collected from major corn-growing regions in Brazil from 2016 to 2018. We aimed to (i) characterize and monitor the susceptibility of field populations, (ii) estimate the frequency of resistance alleles and (iii) evaluate cross-resistance patterns between insecticides and Bt-resistant strains of S. frugiperda to chlorfenapyr. Baseline susceptibility of field population showed a LC50 ranged from 18.3 to 25.1 µg ml−1. The survival rate at the diagnostic concentration ranged from 0 to 8.4%. Using the F2 screen method, the overall estimated resistance allele frequency was 0.0003. No cross-resistance was observed between chlorfenapyr and other insecticides and Bt proteins. Results demonstrated a high susceptibility, low frequency of resistance alleles and lack of cross-resistance. Thus, we concluded there is a low risk of resistance evolution of S. frugiperda to chlorfenapyr in Brazil using the current pest management strategies.

Geographical distribution of Vip3Aa20 resistance allele frequencies in Spodoptera frugiperda (Lepidoptera: Noctuidae) populations in Brazil.

The use of Bt plants has been the main strategy for controlling the fall armyworm Spodoptera frugiperda (J. E. Smith) in Brazil. However, many resistance cases were already registered. The resistance of S. frugiperda to the Vip3Aa20 protein was recently characterized under laboratory conditions but it is still efficient under field conditions. Here, resistance monitoring studies were conducted using phenotypic (purified protein and Bt maize leaves) and genotypic (F1 and F2 screen) methods to support insect resistance management (IRM) programs and preserve Vip3Aa20 technology on maize. Phenotypic monitoring with purified protein showed two populations significantly different from the susceptible strain on the second crop season in 2016. This number increased for the first and second crop seasons in 2017 in several regions. The genotypic monitoring estimated a mean frequency of the resistance allele of 0.0027 for the F1 screen and 0.0033 for the F2 screen. Three new resistant strains to Vip3Aa20 were selected from F2 screen assays. Complementation tests on these new resistant strains were positive with the previous resistant strain. Here we showed that the resistance allele of S. frugiperda to Vip3Aa20 protein is widely distributed in maize-producing regions in Brazil and its frequency increases throughout crop seasons. © 2019 Society of Chemical Industry.

Population genetic structure and demographic history of Spodoptera frugiperda (Lepidoptera: Noctuidae): implications for insect resistance management programs.

Spodoptera frugiperda is a destructive pest that often imposes difficult management due to its high polyphagy and rapid insecticide resistance evolution. Knowledge of species diversification, population structure, and host preference can aid efforts to manage pest populations. Here, we investigated the patterns of hybridization, genetic structure, and gene flow in S. frugiperda populations, discussing how we can apply this knowledge to insect resistance management programs in South America. The corn-strain CS-h2 of S. frugiperda was the most frequent haplotype in all sampled populations. Spodoptera frugiperda populations are experiencing demographic expansion, and the ecoregions partially explain the genetic structure and not strains. We did not find a correlation between gene flow and susceptibility levels to flubendiamide and lufenuron insecticides, but populations with high LC50 sent a great number of migrants to all other locations, maintaining resistance alleles in the geographic range. High levels of population admixture, including between corn- and rice-strains, were found in sampled populations. We showed that S. frugiperda immigrants will not necessarily cause an increase in LC50 upon arrival in a new location but will assure the constant presence of resistance alleles in the area. Increases in LC50 largely depend on the local pesticide management adopted in the areas. Our results indicate that pesticide resistance management must be adopted on a local or small regional scale. © 2019 Society of Chemical Industry.

Susceptibility of Brazilian Populations of Chrysodeixis includens (Lepidoptera: Noctuidae) to Selected Insecticides.

Soybean looper (SBL), Chrysodeixis includens (Walker) (Lepidoptera: Noctuidae), is an important pest of soybean and cotton in Brazil. The use of insecticides is one of the main control tactics against this pest. To support Integrated Pest Management (IPM) and Insect Resistance Management (IRM) programs, we characterized the susceptibility of Brazilian populations of SBL to insecticides. Field populations were collected from soybean fields during the 2016-2017 and 2017-2018 crop seasons. In the laboratory, late L2/early L3 larvae were exposed to insecticides in diet-overlay or topical bioassays. Field populations of SBL showed high susceptibility to spinetoram (LC50 = 0.074-0.25 μg a.i. per cm2), indoxacarb (LC50 = 0.46-0.94 μg a.i. per cm2), thiodicarb (LC50 = 9.14-36.61 μg a.i. per cm2), chlorantraniliprole (LC50 = 0.15-0.57 μg a.i. per cm2), flubendiamide (LC50 = 0.45-2.01 μg a.i. per cm2), and chlorfenapyr (LC50 = 0.15-0.25 μg a.i. per cm2); the resistance ratios were less than 16-fold. In contrast, SBL populations have reduced susceptibility to lambda-cyhalothrin (LC50 = 3.71-9.54 μg a.i./cm2), methoxyfenozide (LC50 = 0.67-4.23 μg a.i. per cm2), novaluron (LC50 = 27.52-77.63 μg a.i. per cm2), and teflubenzuron (LC50 = 13.41-73.02 μg a.i. per cm2). The resistance ratios relative to a Lab population (susceptible of reference) was up to 38-, 63-, 1,553-, and 5,215-fold, respectively. These high resistance ratios can be associated with resistance evolution. Implications of these findings to IPM and IRM programs are discussed.