BACKGROUND:Genetically engineered crops that produce insecticidal proteins from Bacillus thuringiensis (Bt) have many benefits and are used globally to manage key insect pests, including Helicoverpa zea (Lepidoptera: Noctuidae), a major pest of crops in the Americas. However, pests of at least 11 species, including H. zea, have evolved resistance to Bt crops, diminishing their effectiveness and benefits. For H. zea in the United States, practical resistance to Bt corn and cotton producing crystalline (Cry) Bt proteins is widespread and early warning of resistance to the vegetative insecticidal protein Vip3Aa has been reported. Thus, a better understanding of the genetic basis of resistance to Vip3Aa is needed to monitor, manage and counter resistance. In some strains of lepidopteran pests, resistance to Vip3Aa is associated with disruptive mutations in the chitin synthase 2 (CHS2) gene but this association had not been investigated previously in H. zea. RESULTS:Here, we show that mutations introduced by CRISPR/Cas9 editing of the CHS2 gene can cause resistance to Vip3A in H. zea. Disruptive mutations in CHS2 facilitated the creation of strain CHS2-KO that had 29 000-fold resistance to Vip3Aa relative to its unedited parental susceptible strain. Resistance to Vip3Aa in CHS2-KO was autosomal, recessive, and did not cause cross-resistance to Cry1Ac or Cry2Ab. CONCLUSION:Results of this study indicate that CHS2 plays an important role in Vip3Aa intoxication in H. zea. It will be important to determine if mutations in CHS2 contribute to field-evolved resistance to Vip3Aa in H. zea and other pests. © 2025 Society of Chemical Industry. This article has been contributed to by U.S. Government employees and their work is in the public domain in the USA.

Crops genetically engineered to produce insecticidal proteins from the bacterium Bacillus thuringiensis (Bt) have revolutionized pest management, but their benefits have been reduced by evolution of practical resistance in at least 31 cases. To delay evolution of resistance, farmers have shifted from crops producing one Bt protein to crops called pyramids that produce two or more Bt proteins or other traits targeting each pest. Here, we focus on resistance to transgenic corn pyramids in the western corn rootworm (Diabrotica virgifera virgifera) and northern corn rootworm (Diabrotica barberi), which cost farmers in the United States $2 billion yearly in yield losses. We analyzed 998 relevant data values for 2005 to 2023 from 12 published field studies. The results support the hypothesis that rootworm resistance to Bt proteins Cry3Bb and Gpp34/Tpp35Ab reduces the efficacy of pyramids that produce one or both of these proteins and an RNA interference trait (DvSnf7) targeting rootworms, despite no apparent strong cross-resistance between the Bt proteins and DvSnf7. The reduced efficacy of pyramids producing Bt proteins and DvSnf7 entails increased root injury and emergence of adult beetles. Efficacy of DvSnf7 was substantially lower for reducing root injury than emergence. Because root injury decreases yield, the increased root injury has immediate practical consequences. More sustainable control may be achieved by deploying pyramids with traits that are each highly effective against rootworms, increasing the abundance of host plants that do not target rootworms, and combining transgenic corn with crop rotation and other control tactics in integrated pest management programs.

Crops genetically engineered to produce insecticidal proteins from the bacterium Bacillus thuringiensis (Bt) have been used extensively to control some major crop pests, but their benefits decrease when pests evolve resistance. Better understanding of the genetic basis of resistance is needed to effectively monitor, manage, and counter pest resistance to Bt crops. Resistance to Bt proteins in at least 11 species of Lepidoptera, including many important crop pests, is associated with naturally occurring mutations that disrupt one or more of three larval midgut proteins: cadherin and ATP-binding cassette proteins ABCC2 and ABCC3. Here, we determined how CRISPR/Cas9-mediated mutations disrupting cadherin, ABCC2, and ABCC3 singly and in pairs affect resistance to Bt proteins Cry1Ab and Cry1Fa in the Asian corn borer (Ostrinia furnacalis), which is the most damaging pest of corn in Asia and is closely related to the European corn borer (Ostrinia nubilalis), a major pest in Europe and North America. The results from bioassays of six knockout strains and their parent susceptible strain support a model in which Cry1Ab can kill larvae via one path requiring ABCC2 or another path requiring cadherin and ABCC3, whereas Cry1Fa uses only the first path. The model's predictions are generally supported by results from genetic linkage analyses and responses to Cry1Ab and Cry1Fa of Sf9 cells and Xenopus oocytes modified to produce cadherin, ABCC2, and ABCC3 singly or in pairs. The functional redundancy identified here for Cry1Ab could sustain its efficacy against O. furnacalis and may exemplify a widespread natural strategy for delaying resistance.

The vegetative insecticidal protein Vip3Aa from Bacillus thuringiensis (Bt) has been produced by transgenic crops to counter pest resistance to the widely used crystalline (Cry) insecticidal proteins from Bt. To proactively manage pest resistance, there is an urgent need to better understand the genetic basis of resistance to Vip3Aa, which has been largely unknown. We discovered that retrotransposon-mediated alternative splicing of a midgut-specific chitin synthase gene was associated with 5,560-fold resistance to Vip3Aa in a laboratory-selected strain of the fall armyworm, a globally important crop pest. The same mutation in this gene was also detected in a field population. Knockout of this gene via CRISPR/Cas9 caused high levels of resistance to Vip3Aa in fall armyworm and 2 other lepidopteran pests. The insights provided by these results could help to advance monitoring and management of pest resistance to Vip3Aa.

>Insect pest damage to crops is a threat to global food security(Tilman et al.,2011).Climate change,the evolution of insecticide resistance,and the phasing out of insecticides due to environmental and safety concerns exacerbate this problem.Farmers urgently need safe and effective crop protection tools to sustainably generate yields that meet ever-increasing global demand.

Biopesticides based on RNA interference (RNAi) took a major step forward with the first registration of a sprayable RNAi product, which targets the world's most damaging potato pest. Proactive resistance management is needed to delay the evolution of resistance by pests and sustain the efficacy of RNAi biopesticides.

Transgenic crops producing Bacillus thuringiensis (Bt) toxins are commonly used for controlling insect pests. Nearby refuges of non-Bt host plants play a central role in delaying the evolution of resistance to Bt toxins by pests. Pervasive fitness costs associated with resistance, which entail lower fitness of resistant than susceptible individuals in refuges, can increase the ability of refuges to delay resistance. Moreover, these costs are affected by environmental factors such as host plant suitability, implying that manipulating refuge plant suitability could improve the success of the refuge strategy. Based on results from a previous study of Trichoplusia ni resistant to Bt sprays, it was proposed that low-suitability host plants could magnify costs. To test this hypothesis, we investigated the association between host plant suitability and fitness costs for 80 observations from 30 cases reported in 18 studies of 8 pest species from 5 countries. Consistent with the hypothesis, the association between plant suitability and fitness cost was negative. With plant suitability scaled to range from 0 (low) to 1 (high), the expected cost was 20.7% with a suitability of 1 and the fitness cost increased 2.5% for each 0.1 decrease in suitability. The most common type of resistance to Bt toxins involves mutations affecting a few types of midgut proteins to which Bt toxins bind to kill insects. A better understanding of how such mutations interact with host plant suitability to generate fitness costs could be useful for enhancing the refuge strategy and sustaining the efficacy of Bt crops.

Transgenic crops producing crystalline (Cry) proteins from the bacteriumBacillus thuringiensis(Bt) have been used extensively to control some major crop pests. However, many populations of the noctuid mothHelicoverpa zea, one of the most important crop pests in the United States, have evolved practical resistance to several Cry proteins including Cry1Ac. Although mutations in single genes that confer resistance to Cry proteins have been identified in lab-selected and gene-edited strains ofH. zeaand other lepidopteran pests, the genetic basis of field-evolved resistance to Cry proteins inH. zeahas remained elusive. We used a genomic approach to analyze the genetic basis of field-evolved resistance to Cry1Ac in 937H. zeaderived from 17 sites in seven states of the southern United States. We found evidence for extensive gene flow among all populations studied. Field-evolved resistance was not associated with mutations in 20 single candidate genes previously implicated in resistance or susceptibility to Cry proteins inH. zeaor other lepidopterans. Instead, resistance in field samples was associated with increased copy number of a cluster of nine trypsin genes. However, trypsin gene amplification occurred in a susceptible sample and not in all resistant samples, implying that this amplification does not always confer resistance and mutations in other genes also contribute to field-evolved resistance to Cry1Ac inH. zea. The mismatch between lab-generated and field-evolved resistance inH. zeais unlike other cases of Bt resistance and reflects challenges for managing this pest.

Transgenic crops that produce insecticidal proteins from Bacillus thuringiensis (Bt) have provided control of some key pests since 1996. However, the evolution of resistance by pests reduces the benefits of Bt crops. Resistance to Bt crops that is not recessively inherited is especially challenging to manage. Here we analyzed nonrecessive resistance to Bt toxin Cry1Ac in eight field populations of Helicoverpa armigera sampled in 2018 from northern China, where this global pest has been exposed to Cry1Ac in Bt cotton since 1997. Bioassays revealed 7.5% of field-derived larvae were resistant to Cry1Ac of which 87% had at least one allele conferring nonrecessive resistance. To analyze this nonrecessive resistance, we developed and applied a variant of a genomic mapping approach called quantitative trait locus (QTL)-seq. This analysis identified a region on chromosome 10 associated with nonrecessive resistance to Cry1Ac in all 21 backcross families derived from field-collected moths. Individual sequencing revealed that all 21 field-collected resistant grandparents of the backcross families had a previously identified dominant point mutation in the tetraspanin gene HaTSPAN1 that occurs in the region of chromosome 10 identified by QTL-seq. QTL-seq also revealed a region on chromosome 26 associated with nonrecessive resistance in at most 14% of the backcross families. Overall, the results imply the point mutation in HaTSPAN1 is the primary genetic basis of nonrecessive resistance to Cry1Ac in field populations of H. armigera from northern China. Moreover, because nonrecessive resistance is predominant, tracking the frequency of this point mutation could facilitate resistance monitoring in the region.

The polyphagous pest Helicoverpa zea (Lepidoptera: Noctuidae) has evolved practical resistance to transgenic corn and cotton producing Cry1 and Cry2 crystal proteins from Bacillus thuringiensis (Bt) in several regions of the United States. However, the Bt vegetative insecticidal protein Vip3Aa produced by Bt corn and cotton remains effective against this pest. To advance knowledge of resistance to Vip3Aa, we selected a strain of H. zea for resistance to Vip3Aa in the laboratory. After 28 generations of continuous selection, the resistance ratio was 267 for the selected strain (GA-R3) relative to a strain not selected with Vip3Aa (GA). Resistance was autosomal and almost completely recessive at a concentration killing all individuals from GA. Declines in resistance in heterogeneous strains containing a mixture of susceptible and resistant individuals reared in the absence of Vip3Aa indicate a fitness cost was associated with resistance. Previously reported cases of laboratory-selected resistance to Vip3Aa in lepidopteran pests often show partially or completely recessive resistance at high concentrations and fitness costs. Abundant refuges of non-Bt host plants can maximize the benefits of such costs for sustaining the efficacy of Vip3Aa against target pests.

Metacognition, the ability to reflect upon, and evaluate our own beliefs, can help us avoid making decisions based on unreliable evidence. Here, we provide empirical tests of the importance of human metacognition during the COVID-19 pandemic. ...Metacognition, our ability to reflect on our own beliefs, manifests itself in the confidence we have in these beliefs, and helps us guide our behavior in complex and uncertain environments. Here, we provide empirical tests of the importance of ...

Transgenic crops have revolutionized insect pest control, but evolution of resistance by pests threatens their continued success. The primary strategy for combating pest resistance to crops producing insecticidal proteins from Bacillus thuringiensis (Bt) uses refuges of non-Bt host plants to allow survival of susceptible insects. The prevailing paradigm is that refuges delay resistance that is rare and recessively inherited. However, we discovered refuges countered resistance to Bt cotton that was neither rare nor recessive. In a 15-year field study of the cotton bollworm, the frequency of a mutation conferring dominant resistance to Bt cotton surged 100-fold from 2006 to 2016 yet did not rise from 2016 to 2020. Computer simulations indicate the increased refuge percentage from 2016 to 2020 is sufficient to explain the observed halt in the evolution of resistance. The results also demonstrate the efficacy of a Bt crop can be sustained by non-Bt refuges of other crops.

Transgenic crops producing insecticidal proteins from the bacterium Bacillus thuringiensis (Bt) control some important insect pests. However, evolution of resistance by pests reduces the efficacy of Bt crops. Here we review resistance to Bt cotton in the pink bollworm, Pectinophora gossypiella, one of the world's most damaging pests of cotton. Field outcomes with Bt cotton and pink bollworm during the past quarter century differ markedly among the world's top three cotton-producing countries: practical resistance in India, sustained susceptibility in China, and eradication of this invasive lepidopteran pest from the United States achieved with Bt cotton and other tactics. We compared the molecular genetic basis of pink bollworm resistance between lab-selected strains from the U.S. and China and field-selected populations from India for two Bt proteins (Cry1Ac and Cry2Ab) produced in widely adopted Bt cotton. Both lab- and field-selected resistance are associated with mutations affecting the cadherin protein PgCad1 for Cry1Ac and the ATP-binding cassette transporter protein PgABCA2 for Cry2Ab. The results imply lab selection is useful for identifying genes important in field-evolved resistance to Bt crops, but not necessarily the specific mutations in those genes. The results also suggest that differences in management practices, rather than genetic constraints, caused the strikingly different outcomes among countries.

Crops genetically engineered to produce insecticidal proteins from the bacterium Bacillus thuringiensis (Bt) have improved pest management and reduced reliance on insecticide sprays. However, evolution of practical resistance by some pests has reduced the efficacy of Bt crops. We analyzed global resistance monitoring data for 24 pest species based on the first 25 yr of cultivation of Bt crops including corn, cotton, soybean, and sugarcane. Each of the 73 cases examined represents the response of one pest species in one country to one Bt toxin produced by one or more Bt crops. The cases of practical resistance rose from 3 in 2005 to 26 in 2020. Practical resistance has been documented in some populations of 11 pest species (nine lepidopterans and two coleopterans), collectively affecting nine widely used crystalline (Cry) Bt toxins in seven countries. Conversely, 30 cases reflect no decrease in susceptibility to Bt crops in populations of 16 pest species in 10 countries. The remaining 17 cases provide early warnings of resistance, which entail genetically based decreases in susceptibility without evidence of reduced field efficacy. The early warnings involve four Cry toxins and the Bt vegetative insecticidal protein Vip3Aa. Factors expected to favor sustained susceptibility include abundant refuges of non-Bt host plants, recessive inheritance of resistance, low resistance allele frequency, fitness costs, incomplete resistance, and redundant killing by multi-toxin Bt crops. Also, sufficiently abundant refuges can overcome some unfavorable conditions for other factors. These insights may help to increase the sustainability of current and future transgenic insecticidal crops.

Insect pests are increasingly evolving practical resistance to insecticidal transgenic crops that produce Bacillus thuringiensis (Bt) proteins. Here, we analyzed data from the literature to evaluate the association between practical resistance to Bt crops and two pest traits: fitness costs and incomplete resistance. Fitness costs are negative effects of resistance alleles on fitness in the absence of Bt toxins. Incomplete resistance entails a lower fitness of resistant individuals on a Bt crop relative to a comparable non-Bt crop. In 66 studies evaluating strains of nine pest species from six countries, costs in resistant strains were lower in cases with practical resistance (14%) than without practical resistance (30%). Costs in F1 progeny from crosses between resistant and susceptible strains did not differ between cases with and without practical resistance. In 24 studies examining seven pest species from four countries, survival on the Bt crop relative to its non-Bt crop counterpart was higher in cases with practical resistance (0.76) than without practical resistance (0.43). Together with previous findings showing that the nonrecessive inheritance of resistance is associated with practical resistance, these results identify a syndrome associated with practical resistance to Bt crops. Further research on this resistance syndrome could help sustain the efficacy of Bt crops.

Crops genetically engineered to produce insect-killing proteins from Bacillus thuringiensis (Bt) have revolutionized management of some major pests, but their efficacy is reduced when pests evolve resistance. Practical resistance, which is field-evolved resistance that reduces the efficacy of Bt crops and has practical implications for pest management, has been reported in 26 cases in seven countries involving 11 pest species. This special collection includes six original papers that present a global perspective on field-evolved resistance to Bt crops. One is a synthetic review providing a comprehensive global summary of the status of the resistance or susceptibility to Bt crops of 24 pest species in 12 countries. Another evaluates the inheritance and fitness costs of resistance of Diabrotica virgifera virgifera to Gpp34/Tpp35Ab (formerly called Cry34/35Ab). Two papers describe and demonstrate advances in techniques for monitoring field-evolved resistance. One uses a modified F2 screen for resistance to Cry1Ac and Cry2Ab in Helicoverpa zea in the United States. The other uses genomics to analyze nonrecessive resistance to Cry1Ac in Helicoverpa armigera in China. Two papers provide multi-year monitoring data for resistance to Bt corn in Spain and Canada, respectively. The monitoring data from Spain evaluate responses to Cry1Ab of the corn borers Sesamia nonagrioides and Ostrinia nubilalis, whereas the data from Canada track responses of O. nubilalis to Cry1Ab, Cry1Fa, Cry1A.105, and Cry2Ab. We hope the new methods, results, and conclusions reported here will spur additional research and help to enhance the sustainability of current and future transgenic insecticidal crops.

Transgenic crops producing insecticidal proteins from Bacillus thuringiensis (Bt) have revolutionized control of some major pests. However, more than 25 cases of field-evolved practical resistance have reduced the efficacy of transgenic crops producing crystalline (Cry) Bt proteins, spurring adoption of alternatives including crops producing the Bt vegetative insecticidal protein Vip3Aa. Although practical resistance to Vip3Aa has not been reported yet, better understanding of the genetic basis of resistance to Vip3Aa is urgently needed to proactively monitor, delay, and counter pest resistance. This is especially important for fall armyworm ( Spodoptera frugiperda ), which has evolved practical resistance to Cry proteins and is one of the world’s most damaging pests. Here, we report the identification of an association between downregulation of the transcription factor gene SfMyb and resistance to Vip3Aa in S. frugiperda . Results from a genome-wide association study, fine-scale mapping, and RNA-Seq identified this gene as a compelling candidate for contributing to the 206-fold resistance to Vip3Aa in a laboratory-selected strain. Experimental reduction of SfMyb expression in a susceptible strain using RNA interference (RNAi) or CRISPR/Cas9 gene editing decreased susceptibility to Vip3Aa, confirming that reduced expression of this gene can cause resistance to Vip3Aa. Relative to the wild-type promoter for SfMyb , the promoter in the resistant strain has deletions and lower activity. Data from yeast one-hybrid assays, genomics, RNA-Seq, RNAi, and proteomics identified genes that are strong candidates for mediating the effects of SfMyb on Vip3Aa resistance. The results reported here may facilitate progress in understanding and managing pest resistance to Vip3Aa.

The pink bollworm ( Pectinophora gossypiella ) is one of the world’s most destructive pests of cotton. This invasive lepidopteran occurs in nearly all cotton-growing countries. Its presence in the Ord Valley of North West Australia poses a potential threat to the expanding cotton industry there. To assess this threat and better understand population structure of pink bollworm, we analysed genomic data from individuals collected in the field from North West Australia, India, and Pakistan, as well as from four laboratory colonies that originated in the United States. We identified single nucleotide polymorphisms (SNPs) using a reduced-representation, genotyping-by-sequencing technique (DArTseq). The final filtered dataset included 6355 SNPs and 88 individual genomes that clustered into five groups: Australia, India-Pakistan, and three groups from the United States. We also analysed sequences from Genbank for mitochondrial DNA (mtDNA) locus cytochrome c oxidase I (COI) for pink bollworm from six countries. We found low genetic diversity within populations and high differentiation between populations from different continents. The high genetic differentiation between Australia and the other populations and colonies sampled in this study reduces concerns about gene flow to North West Australia, particularly from populations in India and Pakistan that have evolved resistance to transgenic insecticidal cotton. We attribute the observed population structure to pink bollworm’s narrow host plant range and limited dispersal between continents.