Exploratory comparative transcriptomic analysis reveals potential gene targets associated with Cry1A.105 and Cry2Ab2 resistance in fall armyworm (Spodoptera frugiperda).

Genetically modified (GM) crops, expressing Bacillus thuringiensis (Bt) insecticidal toxins, have substantially transformed agriculture. Despite rapid adoption, their environmental and economic benefits face scrutiny due to unsustainable agricultural practices and the emergence of resistant pests like Spodoptera frugiperda, known as the fall armyworm (FAW). FAW's adaptation to Bt technology in corn and cotton compromises the long-term efficacy of Bt crops. To advance the understanding of the genetic foundations of resistance mechanisms, we conducted an exploratory comparative transcriptomic analysis of two divergent FAW populations. One population exhibited practical resistance to the Bt insecticidal proteins Cry1A.105 and Cry2Ab2, expressed in the genetically engineered MON-89Ø34 - 3 maize, while the other population remained susceptible to these proteins. Differential expression analysis supported that Cry1A.105 and Cry2Ab2 significantly affect the FAW physiology. A total of 247 and 254 differentially expressed genes were identified in the Cry-resistant and susceptible populations, respectively. By integrating our findings with established literature and databases, we underscored 53 gene targets potentially involved in FAW's resistance to Cry1A.105 and Cry2Ab2. In particular, we considered and discussed the potential roles of the differentially expressed genes encoding ABC transporters, G protein-coupled receptors, the P450 enzymatic system, and other Bt-related detoxification genes. Based on these findings, we emphasize the importance of exploratory transcriptomic analyses to uncover potential gene targets involved with Bt insecticidal proteins resistance, and to support the advantages of GM crops in the face of emerging challenges.

Fall armyworm (Lepidoptera: Noctuidae): practical resistance of 2 Brazilian populations to Cry1A.105†+†Cry2Ab and Cry1F Bt maize.

In the Americas, transgenic crops producing insecticidal proteins from Bacillus thuringiensis Berliner (Bt, Bacillales: Bacillaceae) have been used widely to manage fall armyworm (FAW, Spodoptera frugiperda [J.E. Smith]). As resistance to Cry1 single-gene Bt maize (Zea mays L.) rapidly evolved in some FAW populations, pyramided Bt maize hybrids producing Cry1, Cry2, or Vip3Aa proteins were introduced in the 2010s. We examined field-evolved resistance to single- and dual-protein Bt maize hybrids in 2 locations in southeastern Brazil, where plant damage by FAW larvae far exceeded the economic threshold in 2017. We collected late-instar larvae in Cry1A.105 + Cry2Ab and Cry1F maize fields and established 2 FAW populations in the laboratory. The F1 offspring reared on the foliage of Bt and non-Bt maize plants (Cry1A.105 + Cry2Ab and Cry1F) showed neonate-to-adult survival rates as high as 70% for both populations. There was no significant difference in the life-table parameters of armyworms reared on non-Bt and Bt maize foliage, indicating complete resistance to Cry1A.105 + Cry2Ab maize. Larval survival rates of reciprocal crosses of a susceptible laboratory strain and the field-collected populations indicated nonrecessive resistance to Cry1F and a recessive resistance to Cry1A.105 + Cry2Ab maize. When relaxing the selection pressure, the armyworm fitness varied on Cry1A.105 + Cry2Ab and non-Bt maize; the resistance was somewhat stable across 12 generations, without strong fitness costs, although one of the lines died confounded by a depleted-quality, artificial rearing diet. To our knowledge, this is the first report documenting the practical resistance of FAW to a pyramided Bt crop. We discuss the implications for resistance management.

Comparative analysis of Spodoptera frugiperda (J. E. Smith) (Lepidoptera, Noctuidae) corn and rice strains microbiota revealed minor changes across life cycle and strain endosymbiont association.

Background: Spodoptera frugiperda (FAW) is a pest that poses a significant threat to corn production worldwide, causing millions of dollars in losses. The species has evolved into two strains (corn and rice) that differ in their genetics, reproductive isolation, and resistance to insecticides and Bacillus thuringiensis endotoxins. The microbiota plays an important role in insects' physiology, nutrient acquisition, and response to chemical and biological controls. Several studies have been carried out on FAW microbiota from larvae guts using laboratory or field samples and a couple of studies have analyzed the corn strain microbiota across its life cycle. This investigation reveals the first comparison between corn strain (CS) and rice strain (RS) of FAW during different developmental insect stages and, more importantly, endosymbiont detection in both strains, highlighting the importance of studying both FAW populations and samples from different stages. Methods: The composition of microbiota during the life cycle of the FAW corn and rice strains was analyzed through high-throughput sequencing of the bacterial 16S rRNA gene using the MiSeq system. Additionally, culture-dependent techniques were used to isolate gut bacteria and the Transcribed Internal Spacer-ITS, 16S rRNA, and gyrB genes were examined to enhance bacterial identification. Results: Richness, diversity, and bacterial composition changed significantly across the life cycle of FAW. Most diversity was observed in eggs and males. Differences in gut microbiota diversity between CS and RS were minor. However, Leuconostoc, A2, Klebsiella, Lachnoclostridium, Spiroplasma, and Mucispirilum were mainly associated with RS and Colidextribacter, Pelomonas, Weissella, and Arsenophonus to CS, suggesting that FAW strains differ in several genera according to the host plant. Firmicutes and Proteobacteria were the dominant phyla during FAW metamorphosis. Illeobacterium, Ralstonia, and Burkholderia exhibited similar abundancies in both strains. Enterococcus was identified as a conserved taxon across the entire FAW life cycle. Microbiota core communities mainly consisted of Enterococcus and Illeobacterium. A positive correlation was found between Spiroplasma with RS (sampled from eggs, larvae, pupae, and adults) and Arsenophonus (sampled from eggs, larvae, and adults) with CS. Enterococcus mundtii was predominant in all developmental stages. Previous studies have suggested its importance in FAW response to B. thuringensis. Our results are relevant for the characterization of FAW corn and rice strains microbiota to develop new strategies for their control. Detection of Arsenophonus in CS and Spiroplasma in RS are promising for the improvement of this pest management, as these bacteria induce male killing and larvae fitness reduction in other Lepidoptera species.

Characterization of the inheritance of field-evolved resistance to diamides in the fall armyworm (Spodoptera frugiperda) (Lepidoptera: Noctuidae) population from Puerto Rico.

The fall armyworm (Spodoptera frugiperda) is one of the most destructive pests of corn. New infestations have been reported in the East Hemisphere, reaching India, China, Malaysia, and Australia, causing severe destruction to corn and other crops. In Puerto Rico, practical resistance to different mode of action compounds has been reported in cornfields. In this study, we characterized the inheritance of resistance to chlorantraniliprole and flubendiamide and identified the possible cross-resistance to cyantraniliprole and cyclaniliprole. The Puerto Rican (PR) strain showed high levels of resistance to flubendiamide (RR50 = 2,762-fold) and chlorantraniliprole (RR50 = 96-fold). The inheritance of resistance showed an autosomal inheritance for chlorantraniliprole and an X-linked inheritance for flubendiamide. The trend of the dominance of resistance demonstrated an incompletely recessive trait for H1 (♂ SUS × â™€ PR) × and an incompletely dominant trait for H2 (♀ SUS × â™‚ PR) × for flubendiamide and chlorantraniliprole. The PR strain showed no significant presence of detoxification enzymes (using synergists: PBO, DEF, DEM, and VER) to chlorantraniliprole; however, for flubendiamide the SR = 2.7 (DEM), SR = 3.2 (DEF) and SR = 7.6 (VER) indicated the role of esterases, glutathione S- transferases and ABC transporters in the metabolism of flubendiamide. The PR strain showed high and low cross-resistance to cyantraniliprole (74-fold) and cyclaniliprole (11-fold), respectively. Incomplete recessiveness might lead to the survival of heterozygous individuals when the decay of diamide residue occurs in plant tissues. These results highlight the importance of adopting diverse pest management strategies, including insecticide rotating to manage FAW populations in Puerto Rico and other continents.

Transcriptomics and interactomics during the primary infection of an SfNPV baculovirus on Spodoptera frugiperda larvae.

The fall armyworm (FAW), Spodoptera frugiperda, has been the most devastating pest of corn as well as of other crops in America, and more recently in Africa and Asia. The development of resistance to chemical insecticides led the search for environmentally friendly biological alternatives such as baculoviruses. This study focuses on the primary infection of the baculovirus SfNPV-Ar in the FAW's midgut epithelium, by analyzing the differential expression of transcripts in excised midguts at 6, 12, and 24 h post-infection (hpi), and predicted their interactions. Interaction of viral factors with the infected midgut tissue could alters various cellular processes, such as the apoptotic system due to the up-regulation observed of FABP at 6 hpi and of HSP90 at 24 hpi, along with the down-regulated PRX at 6 hpi and FABP transcripts between 12 and 24 hpi. Changes in transcript regulation could affect the cellular architecture of infected cells due to up-regulation of ARP 2/3 at 6 and 12 hpi, followed by down-regulation at 24 hpi. In relation to protein folding proteins, HSP90 was up-regulated at 24 hpi and PDI was down-regulated between 6 and 12 hpi. With respect to metabolism and cellular transport, AcilBP and ATPS0 were up regulated at 6 hpi and 12 hpi, respectively. In reference to transcription and translation up-regulation of RPL11 at 6 hpi and of FPN32 and RPL19 at 24 hpi was detected, as well as the down-regulation of RPL19 at 6 hpi, of PDI and RPL7 at 12 hpi, and of FABP at 24 hpi. In conclusion, gene regulation induced by viral infection could be related to the cytoskeleton and cellular metabolism as well as to oxidative stress, apoptosis, protein folding, translation, and ribosomal structure. The results presented in this work are an approach to understanding how the virus takes control of the general metabolism of the insect host during the primary infection period.

Using genetic comparisons of populations from Arizona, Mexico, and Texas to investigate fall armyworm migration in the American southwest.

Fall armyworm (FAW) is a global agricultural pest, causing substantial economic losses in corn and many other crops. Complicating efforts to control this pest is its capacity for long distance flights, which has been described in greatest detail for the central and eastern sections of the United States. FAW infestations are also routinely found in agricultural areas in southern Arizona, which lie beyond the western limits of the mapped migratory pathways. Climate suitability analysis found that the affected Arizona locations cannot support permanent FAW populations, indicating that these FAW most likely arise from annual migrations. A better understanding of this migration would provide insights into how large moth populations can move across desert habitats as well as the degree of gene flow occurring between FAW populations across the North American continent. In this study the Arizona populations were genetically characterized and compared to a selection of permanent and migratory FAW from multiple sites in the United States and Mexico. The results are consistent with migratory contributions from permanent populations in the states of Texas (United States) and Sinaloa (Mexico), while also providing evidence of significant barriers to gene flow between populations within Mexico. An unexpected finding was that two genetically distinct FAW subpopulations known as "host strains" have a differential distribution in the southwest that may indicate significant differences in their migration behavior in this region. These findings indicate that the combination of mitochondrial and Z-linked markers have advantages in comparing FAW populations that can complement and extend the findings from other methods.

Biological basis for adoption of an isoline of Telenomus remus (Hymenoptera: Scelionidae) for an augmentative biological-control program for Spodoptera frugiperda (Lepidoptera: Noctuidae).

The widely distributed, polyphagous fall armyworm, Spodoptera frugiperda (J.E. Smith, 1797), is one of the most important crop pests worldwide. The egg-parasitoid wasp, Telenomus remus Nixon, 1937, is frequently described as a possible control agent for S. frugiperda. We selected an isoline of T. remus and evaluated its parasitism potential (for 24 h) in S. frugiperda eggs, in laboratory conditions, and also its ability to fly at different temperatures and relative humidity levels, aiming to provide basic information about this isoline. The selected isoline maintained good flight capacity without affecting its parasitism efficiency or developing inefficient haplotypes for biological-control programs, compared across generations to a regularline laboratory-reared for more than 60 generations. The flight capacity of the isoline was best at 25-30 °C and relative humidity 70-90%.

Comparative Genomics of Pesticide-Degrading Enterococcus Symbionts of Spodoptera frugiperda (Lepidoptera: Noctuidae) Leads to the Identification of Two New Species and the Reappraisal of Insect-Associated Enterococcus Species.

Enterococcus species have been described as core members of the microbial community of Spodoptera frugiperda (Lepidoptera: Noctuidae) and have been previously reported as insecticide degrading agents. This study aimed to investigate the molecular composition of these microbial symbionts of S. frugiperda to better understand their association with the host and their potential for insecticide metabolization. Through phenotypic assays and comparative genomic analyses of several pesticide-degrading Enterococcus isolated from the gut of S. frugiperda larvae, we identified two new species: Enterococcus entomosocium n. sp. and Enterococcus spodopteracolus n. sp. Their identities as new species were confirmed by whole genome alignment, utilizing cut-offs of 95-96% for the average nucleotide identity (ANI) and 70% for the digital DNA: DNA hybridization (dDDH) values. The systematic positioning of these new species within the genus Enterococcus was resolved using genome-based analysis, revealing Enterococcus casseliflavus as a sister group of E. entomosocium n. sp., and Enterococcus mundtii as a sister group of E. spodopteracolus n. sp. Comparative genomic analyses of several isolates of E. entomosocium n. sp. and E. spodopteracolus n. sp. provided a better assessment of the interactions established in the symbiotic association with S. frugiperda and led to the discovery of misidentified new species of Enterococcus associated with insects. Our analyses indicated that the potential of E. entomosocium n. sp. and E. spodopteracolus n. sp. to metabolize different pesticides arises from molecular mechanisms that result in rapid evolution of new phenotypes in response to environmental stressors, in this case, the pesticides their host insect is exposed to.

Population genomics of fall armyworm by genotyping-by-sequencing: Implications for pest management.

The fall armyworm (FAW), Spodoptera frugiperda, is a significant pest of many crops in the world and it is native to the Americas, where the species has shown the ability to rapidly evolve resistance to insecticides and transgenic plants. Despite the importance of this species, there is a gap in the knowledge regarding the genetic structure of FAW in South America. Here, we examined the genetic diversity of FAW populations across a wide agricultural area of Brazil and Argentina using a Genotyping-by-Sequencing (GBS) approach. We also characterized samples by their host strain based on mitochondrial and Z-linked genetic markers. The GBS methodology enabled us to discover 3309 SNPs, including neutral and outlier markers. Data showed significant genetic structure between Brazil and Argentina populations, and also among the Argentinian ecoregions. Populations inside Brazil showed little genetic differentiation indicating high gene flow among locations and confirming that structure is related to the presence of corn and rice strains. Outlier analysis indicated 456 loci putatively under selection, including genes possibly related to resistance evolution. This study provides clarification of the population genetic structure of FAW in South America and highlights the importance of genomic research to understand the risks of spread of resistance genes.

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.

Bacillus thuringiensis-Based Bioproduct: Characterization and Performance Against Spodoptera frugiperda Strains in Maize Under Different Environmental Temperatures.

Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae) is an important pest in several regions being the use of Bacillus thuringiensis-based bioproducts an alternative for its control. Firstly, 3 L of an aqueous bioproduct suspension was produced and characterized. Its 50% lethal concentration against molecularly identified corn and rice S. frugiperda strains using an artificial diet were 77.01% (95% CL, 68.16-90.47) and 2.22% (95% CL, 0.01-6.68), respectively. The next objective of this work was to evaluate the performance of this bioproduct in maize against S. frugiperda strains under different simulated agrological regions mimicking their corresponding periodic day/night temperatures. Thus, the impact of environmental temperature on the bioproduct efficacy (E) was studied. It was observed that a warmer scenario (35 °C day/30 °C night) could favor the tolerance of corn S. frugiperda strain to the bioproduct (E = 56.36 ± 0.61%) maintaining a high efficacy (92.44 ± 6.55%) when it was tested against rice S. frugiperda strain. Conversely, under temperate conditions, efficacy values ranged from 84 to 95% for both S. frugiperda strains. On the other hand, based on a foliar feeding damage analysis, our bioproduct displayed a significant foliar protection in maize plants infested with either corn or rice S. frugiperda strains.

Spodoptera frugiperda: Ecology, Evolution, and Management Options of an Invasive Species.

The fall armyworm (FAW), Spodoptera frugiperda (Lepidoptera, Noctuidae), is a well-known agricultural pest in its native range, North and South America, and has become a major invasive pest around the globe in the past decade. In this review, we provide an overview to update what is known about S. frugiperda in its native geographic ranges. This is followed by discussion of studies from the invaded areas to gain insights into S. frugiperda's ecology, specifically its reproductive biology, host plant use, status of insecticide resistance alleles, and biocontrol methods in native and invasive regions. We show that reference to host strains is uninformative in the invasive populations because multidirectional introduction events likely underpinned its recent rapid spread. Given that recent genomic analyses show that FAW is much more diverse than was previously assumed, and natural selection forces likely differ geographically, region-specific approaches will be needed to control this global pest.

Non-targeted metabolomics reveals differences in the gut metabolic profile of the fall armyworm strains when feeding different food sources.

Spodoptera frugiperda (fall armyworm - FAW) is an important polyphagous agricultural pest feeding on nearly 350 host plants. FAW is undergoing incipient speciation with two well-characterized host-adapted strains, the "corn" (CS) and "rice" (RS) strains, which are morphologically identical but carry several genes under positive selection for host adaptation. We used non-targeted metabolomics based on gas chromatography/mass spectrometry to identify differences in metabolite profiles of the larval gut of CS and RS feeding on different host plants. Larvae were fed on artificial diet, maize, rice, or cotton leaves from eclosion to the sixth instar, when they had their midgut dissected for analysis. This study revealed that the midgut metabolome of FAW varied due to larval diet and differed between the FAW host-adapted strains. Additionally, we identified several candidate metabolites that may be involved in the adaptation of CS and RS to their host plants. Our findings provide clues toward the gut metabolic activities of the FAW strains.

Characterization and genomic analyses of a novel alphabaculovirus isolated from the black armyworm, Spodoptera cosmioides (Lepidoptera: Noctuidae).

The black armyworm Spodoptera cosmioides is a pest of increasing importance in Cry1Ac-Bt toxin crops and non-Bt crops of soybean and cotton in Brazil. Here we characterized a baculovirus isolated from extracts of S. cosmioides that died with symptoms of nuclear polyhedrosis. The putative novel virus exhibited polyhedral occlusion bodies (OBs) with virions containing multiple rod-shaped nucleocapsids, characteristic of alphabaculoviruses. The virus isolate was named Spodoptera cosmioides nucleopolyhedrovirus isolate CNPSo-72 (SpcoNPV-CNPSo-72). SpcoNPV-CNPSo-72 was lethal to third-instar S. cosmioides caterpillars but not to S. frugiperda under the tested viral concentrations. Moreover, SpcoNPV-CNPSo-72 contained a circular 147,763 bp long genome and a G + C content of 44.8% with 151 annotated ORFs (10 unique for baculovirus) and five homologous regions (hrs). The 38 currently defined baculovirus core genes were found in the SpcoNPV-CNPSo-72 genome. After phylogenetic analysis, the novel virus was found to be closely related to other members of Alphabaculovirus, especially to the Spodoptera-infecting viruses, which included Spodoptera eridania nucleopolyhedrovirus isolate 251, Spodoptera litura nucleopolyhedrovirus isolate II, Spodoptera exigua multiple nucleopolyhedrovirus isolate US-1, Spodoptera eridania nucleopolyhedrovirus isolate CNPSo-165, and Spodoptera frugiperda multiple nucleopolyhedrovirus isolate 19. Surprisingly, the new baculoviral genome was found to code for a putative arginine-associated tRNA gene with a predicted intronic sequence of 105 nt. The gene was found inside the bjdp CDS. Overall, baculoviruses are pathogens that lethally infect insect larvae and their study allows a better understanding of large DNA virus evolution, which provides important insights for the development and improvement of biological control agents.

The role of genetically engineered soybean and Amaranthus weeds on biological and reproductive parameters of Spodoptera cosmioides (Lepidoptera: Noctuidae).

BACKGROUND: In soybean fields containing insecticide- and herbicide-resistant genetically engineered varieties, some weed species have increasingly become difficult to manage and may favor the population growth of secondary pests like Spodoptera cosmioides (Walker) (Lepidoptera: Noctuidae). To test this hypothesis, we measured life-history traits, population growth parameters and adult nutrient content of S. cosmioides reared on foliage from four Amaranthus species, from Cry1Ac Bt and non-Bt soybean varieties, and on meridic artificial diet. RESULTS: Larvae reared on A. palmeri and A. spinosus had a shorter development time (5-7 days) than larvae raised on the soybean varieties and A. hybridus. Armyworm survival probability was zero on A. viridis and highest (80% and 71%) on soybeans and A. palmeri. The latter and the artificial diet produced the heaviest larvae and pupae, in contrast to the non-Bt soybean variety. Body nutrient content diverged mostly for adults reared on artificial diet compared with those raised on the soybean varieties. The intrinsic rate of population increase (overall fitness) was 27.88% higher for the armyworms on A. palmeri, Cry1Ac Bt soybean and artificial diet compared with those on non-Bt soybean, A. spinosus and A. hybridus. CONCLUSIONS: Cry1Ac soybean fields infested by some Amaranthus weeds, especially A. palmeri, are conducive to the population growth of S. cosmioides. Integrated pest management programs may be needed to properly manage S. cosmioides in soybean fields, with surveillance for population peaks and judicious control measures when needed. © 2022 Society of Chemical Industry.

Recombinant Expression of ABCC2 Variants Confirms the Importance of Mutations in Extracellular Loop 4 for Cry1F Resistance in Fall Armyworm.

Fall armyworm (FAW), Spodoptera frugiperda, is a highly destructive and invasive global noctuid pest. Its control is based on insecticide applications and Bacillus thuringiensis (Bt) insecticidal Cry toxins expressed in transgenic crops, such as Cry1F in Bt corn. Continuous selection pressure has resulted in populations that are resistant to Bt corn, particularly in Brazil. FAW resistance to Cry1F was recently shown to be conferred by mutations of ATP-binding cassette transporter C2 (ABCC2), but several mutations, particularly indels in extracellular loop 4 (ECL4), are not yet functionally validated. We addressed this knowledge gap by baculovirus-free insect cell expression of ABCC2 variants (and ABCC3) by electroporation technology and tested their response to Cry1F, Cry1A.105 and Cry1Ab. We employed a SYTOXTM orange cell viability test measuring ABCC2-mediated Bt toxin pore formation. In total, we tested seven different FAW ABCC2 variants mutated in ECL4, two mutants modified in nucleotide binding domain (NBD) 2, including a deletion mutant lacking NBD2, and S. frugiperda ABCC3. All tested ECL4 mutations conferred high resistance to Cry1F, but much less to Cry1A.105 and Cry1Ab, whereas mutations in NBD2 hardly affected Bt toxin activity. Our study confirms the importance of indels in ECL4 for Cry1F resistance in S. frugiperda ABCC2.

Assessing fitness costs of the resistance of Spodoptera frugiperda (Lepidoptera: Noctuidae) to pyramided Cry1 and Cry2 insecticidal proteins on different host plants.

Fall armyworm (FAW), Spodoptera frugiperda (Smith), is one of the major pests targeted by transgenic crops expressing insecticidal proteins from Bacillus thuringiensis (Bt) Berliner. However, FAW presents a high capacity to develop resistance to Bt protein-expressing crop lines, as reported in Brazil, Argentina, Puerto Rico and the southeastern U.S. Here, FAW genotypes resistant to pyramided maize events expressing Cry1F/Cry1A.105/Cry2Ab2 (P-R genotype) and Cry1A.105/Cry2Ab2 (Y-R genotype) from Brazil were used to investigate the interactions between non-Bt hosts (non-Bt maize, non-Bt cotton, millet and sorghum) and fitness costs. We also tested a FAW genotype susceptible to Bt maize and F1 hybrids of the resistant and susceptible genotypes (heterozygotes). Recessive fitness costs (i.e., costs affecting the resistant insects) were observed for pupal and neonate to adult survival of the P-R genotype on non-Bt cotton; larval developmental time of the P-R genotype on millet and sorghum; larval and neonate-to-adult developmental time of the Y-R genotype on non-Bt cotton and sorghum; the fecundity of the Y-R genotype on non-Bt cotton; and mean generation time of both resistant genotypes. However, on non-Bt cotton and non-Bt maize, the P-R genotype had a higher fitness (i.e., fitness benefits), displaying greater fecundity and rates of population increases than the Sus genotype. Non-recessive fitness costs (i.e., costs affecting heterozygotes) were found for fecundity and population increases on millet and sorghum. These findings suggest that, regardless of the disadvantages of the resistant genotypes in some hosts, the resistance of FAW to Cry1 and Cry2 Bt proteins is not linked with substantial fitness costs, and may persist in field conditions once present.

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.

Pathogenic Assessment of SfMNPV-Based Biopesticide on Spodoptera frugiperda (Lepidoptera: Noctuidae) Developing on Transgenic Soybean Expressing Cry1Ac Insecticidal Protein.

Pathogenic assessment of a baculovirus-based biopesticide containing Spodoptera frugiperda multiple nucleopolyhedrovirus (SfMNPV: Baculoviridae: Alphabaculovirus) infecting fall armyworm, Spodoptera frugiperda (J. E. Smith, 1797) (Lepidoptera: Noctuidae) is reported. In the bioassays, neonates were infected with different doses of SfMNPV applied on Cry1Ac Bt soybean and non-Bt soybean. Our findings indicated that S. frugiperda neonates did not survive at 10 d post infection or develop into adults on Bt and non-Bt soybean sprayed with the field recommended dose of SfMNPV. In contrast, a proportion of the infected neonates developed into adults when infected with lower doses of SfMNPV (50%, 25%, and 10% of field dose) in both Bt and non-Bt soybean. However, S. frugiperda neonates surviving infection at the lowest virus doses on both soybean varieties showed longer neonate-to-pupa and neonate-to-adult periods, lower larval and pupal weights, reduced fecundity, and increased population suppression. Nevertheless, more pronounced pathogenicity of SfMNPV infecting neonates of S. frugiperda were verified on larvae that developed on Bt soybean. These findings revealed that, beyond mortality, the biopesticide containing SfMNPV also causes significant sublethal pathogenic effects on neonates of S. frugiperda developing on Bt and non-Bt soybean and suggested an additive effect among SfMNPV and Cry1Ac insecticidal protein expressed in Bt soybean.

Field-evolved resistance to chlorpyrifos by Spodoptera frugiperda (Lepidoptera: Noctuidae): Inheritance mode, cross-resistance patterns, and synergism.

BACKGROUND: Fall armyworm (FAW), Spodoptera frugiperda (Smith), is an economically important pest worldwide. In this study, we selected a genotype of FAW resistant to chlorpyrifos from a field-collected population, characterized the genetic basis of resistance, and evaluated cross-resistance and mechanisms of resistance using synergists. RESULTS: The LD50 values of chlorpyrifos for the resistant (Clorp-R) and susceptible (Sus) FAW genotypes were 24.26 and 0.023 µg per larva, respectively, representing a resistance ratio > 1050-fold. The LD50 values of chlorpyrifos against heterozygotes were 3.34 and 4.00 µg per larva, suggesting that resistance is autosomally inherited. The chlorpyrifos resistance in FAW was influenced by few genes, with the minimum numbers of segregations being 1.74 and 1.88. On chlorpyrifos-sprayed plants and leaves, Clorp-R and heterozygote genotypes showed >95% and >52% survival, respectively, whereas the Sus genotype had no survival, indicating that the resistance is incompletely dominant at the field rate of chlorpyrifos. The Clorp-R genotype presented some cross-resistance to acephate, but low cross-resistance to thiodicarb, methomyl, chlorfenapyr, flubendiamide, methoxyfenozide, spinetoram, and teflubenzuron. The synergists piperonyl butoxide, diethyl maleate, and S,S,S-tributyl phosphorotrithiotate did not have relevant effects on the Clorp-R genotype, suggesting a minor role for metabolic resistance. CONCLUSIONS: The inheritance of chlorpyrifos resistance in FAW was characterized as autosomal, incompletely dominant, and polygenic, with metabolic resistance playing a small role in the detoxification of chlorpyrifos. Low cross-resistance between chlorpyrifos and other mode of action (MoA) insecticides occurs in FAW, highlighting the importance of considering the rotation of MoA as a strategy to delay resistance. © 2021 Society of Chemical Industry.