CRISPR/Cas9-mediated generation of a mutant library of cotton CDPK gene family for identifying insect-resistant genes.

Calcium-dependent protein kinases (CDPKs) are pivotal signaling transduction enzymes in plants, especially responsive to diverse stress, including herbivory. In this study, through comprehensive analysis of CDPK gene family in upland cotton, we showed that GhCPKs are widely expressed in multiple tissues of cotton and positively respond to various biotic and abiotic stress. We developed a strategy for screening insect-resistant genes based on the CRISPR/Cas9 mutant library of GhCPKs. The library contains 82 members of the GhCPKs using 246 sgRNAs to generate 518 independent T0 plants. The coverage rate of target genes reached to 86.18%, the genome editing rate reached to 89.49%, and the editing heritability reached 82%. Through field insect bioassay, 14 GhCPK mutants resistant or susceptible to insect were identified. The most obvious insect-resistant mutant, cpk33/74 (simultaneously knocking out the homologous genes GhCPK33 and GhCPK74), was selected for further study. Oral secretions (OS) from Spodoptera litura induced a rapid influx of Ca2+ in cpk33/74 leaves, resulting in a significant increase in jasmonic acid (JA) content. S-adenosylmethionine synthase (SAMS) is an important protein involved in plant stress response, protein interaction experiments provided evidence of interactions between GhCPK33 and GhCPK74 with GhSAMS1 and GhSAM2, respectively. Additionally, silencing GhSAMS1 and GhSAM2 in cotton using VIGS resulted in decreased defense against S. litura. This study provides an effective strategy for constructing a mutant library of gene families in polyploid plant species and valuable insights into the role of CDPKs in the interaction between plants and herbivorous insects.

PdPLR1 effectively enhances resistance of Populus deltoides 'shalinyang' to Anoplophora glabripennis by positive regulation of lignan synthesis.

Anoplophora glabripennis (ALB) is one of the most devastating wood boring insects of poplars. Populus deltoides 'Shalinyang (PdS), a new poplar variety, shows strong resistance to ALB infestation. However, the molecular mechanism of insect resistance in PdS is unclear. Here, we found that lignan content was much higher in PdS phloem after ALB infestation than in healthy trees, and that adding lignan to artificial diet significantly reduced: larval weight; digestive enzyme activity (cellulase [CL], polygalacturonase [PG]); detoxification enzyme activity (carboxylesterase [CarE], glutathione S-transferase [GSH-ST]); and defense enzyme activity (Catalase [CAT]). We further identified the lignan biosynthesis-related PdPLR1 gene (Pinoresinol-lariciresinol reductase, PLR) based on transcriptome analysis, and it was significantly up-regulated in the PdS phloem attacked by ALB. Overexpression of PdPLR1 in Arabidopsis increased th lignan content. In contrast, silencing PdPLR1 in PdS significantly decreased expression levels of PdPLR1 and lignan content by 82.45% and 56.85%. However, silencing PdPLR1 increased the number of adults ovipositions and eggs hatching. The activity of CL, PG, CarE, GSH-ST and CAT and the biomass of larvae fed on phloem of PdS with silenced PdPLR1 were significantly higher than in the control. Taken together, up regulation of PdPLR1 enhanced PdS resistance to ALB by regulating lignan synthesis. Our findings provide in-depth insights into the molecular mechanisms of PdS-ALB interactions, which lay the foundation for understanding of defense in poplars to pest infection.

Is insecticide resistance a factor contributing to the increasing problems with Dalbulus maidis (Hemiptera: Cicadellidae) in Brazil?

BACKGROUND: The corn leafhopper, Dalbulus maidis, is an important pest in Brazil. While chemical control has traditionally been the cornerstone for managing this pest, field control failures have been reported for some insecticides. To understand if these failures are due to resistance, we evaluated the susceptibility of 11 field-collected populations of D. maidis to major insecticides during the 2021-2022 crop seasons in Brazil using concentration-mortality bioassays. Additionally, we employed diagnostic concentration bioassays and foliar sprays at label-recommended rates in 8-10 populations collected during the 2022-2023 crop seasons. RESULTS: High susceptibility to methomyl, carbosulfan and acephate was observed on concentration-mortality bioassays across all populations tested with resistance ratio (RR) based on LC50 <10-fold, except for one population from Bahia State that exhibited reduced susceptibility to methomyl (RR = 17.5). On the other hand, all populations exhibited reduced susceptibility to bifenthrin, acetamiprid, and imidacloprid, with RR ranging from 90 to 2000-fold. This reduced susceptibility to neonicotinoid and pyrethroid insecticides was further confirmed at diagnostic concentrations based on LC99 of the susceptible strain, with survival rates >20% and in foliar sprays with mortality rates <80%. Most populations exposed to acephate and carbosulfan exhibited low survival rates at diagnostic concentrations (<5%) and high mortality rates in foliar sprays (>80%). CONCLUSIONS: The reduced susceptibility to pyrethroid and neonicotinoid insecticides likely explain the field failures in controlling D. maidis populations in Brazil. This study represents the first large-scale susceptibility monitoring of D. maidis to insecticides, and the results will contribute to decision-making regarding the management of this pest. © 2024 Society of Chemical Industry.

Epigenetic remodeling in insect immune memory.

The innate immune system of insects can respond more swiftly and efficiently to pathogens based on previous experience of encountering antigens. The understanding of molecular mechanisms governing immune priming, a form of immune memory in insects, including its transgenerational inheritance, remains elusive. It is still unclear if the enhanced expression of immune genes observed in primed insects can persist and be regulated through changes in chromatin structure via epigenetic modifications of DNA or histones, mirroring observations in mammals. Increasing experimental evidence suggests that epigenetic changes at the level of DNA/RNA methylation and histone acetylation can modulate the activation of insects' immune responses to pathogen exposure. Moreover, transgenerational inheritance of certain epigenetic modifications in model insect hosts can influence the transmission of pre-programmed immune responses to the offspring, leading to the development of evolved resistance. Epigenetic research in model insect hosts is on the brink of significant progress in the mechanistic understanding of chromatin remodeling within innate immunity, particularly the direct relationships between immunological priming and epigenetic alterations. In this review, we discuss the latest discoveries concerning the involvement of DNA methylation and histone acetylation in shaping the development, maintenance, and inheritance of immune memory in insects, culminating in the evolution of resistance against pathogens.

Maize yields have stagnated in sub-Sahara Africa: a possible transgenic solution to weed, pathogen and insect constraints.

Despite major breeding efforts by various national and international agencies, yields for the ~40 million hectares of maize, the major food crop in sub-Saharan Africa, have stagnated at <2 tons/ha/year for the past decade, one-third the global average. Breeders have succeeded in breeding increased yield with a modicum of tolerance to some single-weed or pathogen stresses. There has been minimal adoption of these varieties because introgressing polygenic yield and tolerance traits into locally adapted material is very challenging. Multiple traits to deal with pests (weeds, pathogens, and insects) are needed for farmer acceptance, because African fields typically encounter multiple pest constraints. Also, maize has no inherent resistance to some of these pest constraints, rendering them intractable to traditional breeding. The proposed solution is to simultaneously engineer multiple traits into one genetic locus. The dominantly inherited multi-pest resistance trait single locus can be bred simply into locally adapted, elite high-yielding material, and would be valuable for farmers, vastly increasing maize yields, and allowing for more than regional maize sufficiency. © 2024 The Author(s). Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Physiological responses and transcriptome analysis of Hemerocallis citrina Baroni exposed to Thrips palmi feeding stress.

Thrips are serious pests of Hemerocallis citrina Baroni (daylily), affecting crop yield and quality. To defend against pests, daylily has evolved a set of sophisticated defense mechanisms. In the present study, induction of systemic resistance in Hemerocallis citrina 'Datong Huanghua' by Thrips palmi feeding was investigated at both biochemical and molecular levels. The soluble sugar content of daylily leaves was significantly lower than that in control check (CK) at all time points of feeding by T. palmi, whereas the amino acid and free fatty acid contents started to be significantly lower than those in CK after 7 days. Secondary metabolites such as tannins, flavonoids, and total phenols, which are harmful to the growth and reproduction of T. palmi, were increased significantly. The activities of defense enzymes such as peroxidase (POD), phenylalanine ammonia lyase (PAL), and polyphenol oxidase (PPO) were significantly increased, and the degree of damage to plants was reduced. The significant increase in protease inhibitor (PI) activity may lead to disrupted digestion and slower growth in T. palmi. Using RNA sequencing, 1,894 differentially expressed genes (DEGs) were identified between control and treatment groups at five timepoints. DEGs were mainly enriched in secondary metabolite synthesis, jasmonic acid (JA), salicylic acid (SA), and other defense hormone signal transduction pathways, defense enzyme synthesis, MAPK signaling, cell wall thickening, carbohydrate metabolism, photosynthesis, and other insect resistance pathways. Subsequently, 698 DEGs were predicted to be transcription factors, including bHLH and WRKY members related to biotic stress. WGCNA identified 18 hub genes in four key modules (Purple, Midnight blue, Blue, and Red) including MYB-like DNA-binding domain (TRINITY_DN2391_c0_g1, TRINITY_DN3285_c0_g1), zinc-finger of the FCS-type, C2-C2 (TRINITY_DN21050_c0_g2), and NPR1 (TRINITY_DN13045_c0_g1, TRINITY_DN855_c0_g2). The results indicate that biosynthesis of secondary metabolites, phenylalanine metabolism, PIs, and defense hormones pathways are involved in the induced resistance to T. palmi in daylily.

Proactive resistance management for sustaining the efficacy of RNA interference for pest control.

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.

Genetically engineered eucalyptus expressing pesticidal proteins from Bacillus thuringiensis for insect resistance: a risk assessment evaluation perspective.

Eucalyptus covers approximately 7.5 million hectares in Brazil and serves as the primary woody species cultivated for commercial purposes. However, native insects and invasive pests pose a significant threat to eucalyptus trees, resulting in substantial economic losses and reduced forest productivity. One of the primary lepidopteran pests affecting eucalyptus is Thyrinteina arnobia (Stoll, 1782) (Lepidoptera: Geometridae), commonly referred to as the brown looper caterpillar. To address this issue, FuturaGene, the biotech division of Suzano S.A., has developed an insect-resistant (IR) eucalyptus variety, which expresses Cry pesticidal proteins (Cry1Ab, Cry1Bb, and Cry2Aa), derived from Bacillus thuringiensis (Bt). Following extensive safety assessments, including field trials across various biomes in Brazil, the Brazilian National Technical Commission of Biosafety (CTNBio) recently approved the commercialization of IR eucalyptus. The biosafety assessments involved the analysis of molecular genomics, digestibility, thermostability, non-target organism exposure, degradability in the field, and effects on soil microbial communities and arthropod communities. In addition, in silico studies were conducted to evaluate allergenicity and toxicity. Results from both laboratory and field studies indicated that Bt eucalyptus is as safe as the conventional eucalyptus clone for humans, animals, and the environment, ensuring the secure use of this insect-resistant trait in wood production.

Larval density-dependent mortality of western corn rootworm (Coleoptera: Chrysomelidae) in Bt and non-Bt maize and implications on dose calculations†.

BACKGROUND: Transgenic crops producing insecticidal toxins from the bacterium Bacillus thuringiensis (Bt) have been used to manage insect pests for nearly 30 years. Dose of a Bt crop is key to assessing the risk of resistance evolution because it affects the heritability of resistance traits. Western corn rootworm (Diabrotica virgifera virgifera, LeConte), a major pest of maize, has evolved resistance to all commercially available Bt traits targeting it, and threatens resistance to future transgenic traits. Past research shows the dose of Bt maize targeting western corn rootworm can be confounded by larval density-dependent mortality. We conducted a 2-year field study at two locations to quantify larval density-dependent mortality in Bt and non-Bt maize. We used these results to calculate dose for our method and compared it to three previously published methods. Additionally, adult emergence and root injury were analyzed for predicting initial egg density. RESULTS: Increased pest density caused greater proportions of larvae to die in Bt maize than in non-Bt maize. All methods for calculating dose produced values less than high-dose, and stochastic variation had the greatest impact on dose at high and low pest densities. Our method for calculating dose did not produce values positively correlated with pest density while the three other methods did. CONCLUSION: To achieve the most accurate calculation of dose for transgenic maize targeting western corn rootworm, density-dependent mortality should be taken into account for both transgenic and non-transgenic maize and assessed at moderate pest densities. © 2024 Society of Chemical Industry.

Critical Facets of European Corn Borer Adult Movement Ecology Relevant to Mitigating Field Resistance to Bt-Corn.

The European corn borer (Ostrinia nubilalis, Hübner) has been managed successfully in North America since 1996 with transgenic Bt-corn. However, field-evolved resistance to all four available insecticidal Bt proteins has been detected in four provinces of Canada since 2018. Evidence suggests resistance may be spreading and evolving independently in scattered hotspots. Evolution and spread of resistance are functions of gene flow, and therefore dispersal, so design of effective resistance management and mitigation plans must take insect movement into account. Recent advances in characterizing European corn borer movement ecology have revealed a number of surprises, chief among them that a large percentage of adults disperse from the natal field via true migratory behavior, most before mating. This undermines a number of common key assumptions about adult behavior, patterns of movement, and gene flow, and stresses the need to reassess how ecological data are interpreted and how movement in models should be parameterized. While many questions remain concerning adult European corn borer movement ecology, the information currently available is coherent enough to construct a generalized framework useful for estimating the spatial scale required to implement possible Bt-resistance prevention, remediation, and mitigation strategies, and to assess their realistic chances of success.

Comprehensive analysis of MAPK gene family in upland cotton (Gossypium hirsutum) and functional characterization of GhMPK31 in regulating defense response to insect infestation.

KEY MESSAGE: The transcriptomic, phenotypic and metabolomic analysis of transgenic plants overexpressing GhMPK31 in upland cotton revealed the regulation of H2O2 burst and the synthesis of defensive metabolites by GhMPK31. Mitogen-activated protein kinases (MAPKs) are a crucial class of protein kinases, which play an essential role in various biological processes in plants. Upland cotton (G. hirsutum) is the most widely cultivated cotton species with high economic value. To gain a better understanding of the role of the MAPK gene family, we conducted a comprehensive analysis of the MAPK gene family in cotton. In this study, a total of 55 GhMPK genes were identified from the whole genome of G. hirsutum. Through an investigation of the expression patterns under diverse stress conditions, we discovered that the majority of GhMPK family members demonstrated robust responses to abiotic stress, pathogen stress and pest stress. Furthermore, the overexpression of GhMPK31 in cotton leaves led to a hypersensitive response (HR)-like cell death phenotype and impaired the defense capability of cotton against herbivorous insects. Transcriptome and metabolomics data analysis showed that overexpression of GhMPK31 enhanced the expression of H2O2-related genes and reduced the accumulation of defensive related metabolites. The direct evidence of GhMPK31 interacting with GhRBOHB (H2O2-generating protein) were found by Y2H, BiFC, and LCI. Therefore, we propose that the increase of H2O2 content caused by overexpression of GhMPK31 resulted in HR-like cell death in cotton leaves while reducing the accumulation of defensive metabolites, ultimately leading to a decrease in the defense ability of cotton against herbivorous insects. This study provides valuable insights into the function of MAPK genes in plant resistance to herbivorous insects.

Rice varietal resistance to the vector Sogatella furcifera hinders transmission of Southern rice black-streaked dwarf virus.

BACKGROUND: The Southern rice black-streaked dwarf virus (SRBSDV) transmitted by Sogatella furcifera constitutes a threat to sustainable rice production. However, most rice varieties are highly vulnerable to SRBSDV, whereas the occurrence of the viral disease varies significantly under field conditions. This study aimed to evaluate the potential of rice varietal resistance to S. furcifera in reducing SRBSDV transmission. RESULTS: Among the five rice varieties, Zhongzheyou8 and Deyou108 exhibited high resistance to S. furcifera, Baixiangnuo33 was susceptible, and TN1 and Diantun502 were highly susceptible. The S. furcifera generally showed non-preference for and low feeding on the Zhongzheyou8 and Deyou108 plants, which may explain the resistance of these varieties to S. furcifera. Transmission of SRBSDV by S. furcifera was significantly impaired on the resistant varieties, both inoculation and acquisition rates were much lower on Zhongzheyou8 than on TN1. The short durations of S. furcifera salivation and phloem-related activities and the low S. furcifera feeding amount may explain the reduced SRBSDV inoculation and acquisition rates associated with Zhongzheyou8. Spearman's rank correlation revealed a significant negative correlation between S. furcifera resistance and SRBSDV transmission among the tested varieties. CONCLUSION: The results indicate that rice varietal resistance to the vector S. furcifera hinders SRBSDV transmission, which is largely associated with the host plant selection and feeding behaviors of the vector. The current findings shed light on the management of the SRBSDV viral disease through incorporation of S. furcifera resistant rice varieties in the management protocol. © 2024 Society of Chemical Industry.

Insecticide mixtures-uses, benefits and considerations.

Insecticides remain an important tool for the control of many insect pests. There has long been an interest in insecticide mixtures (in-can and tank-mix) as a means to provide the needed efficacy and/or spectrum to control many insect public health, crop pests or crop pest complexes. This aspect has become more important since insecticides developed in the last 30 years tend to be narrower in spectrum with many primarily focused on either sap-feeding or chewing insect pests. Insecticide mixtures are also seen as an important approach to insect resistance management (IRM) with certain requirements for optimal implementation. Additionally, insecticide mixtures can also address certain agronomic, commercial and intellectual property needs and opportunities. This perspective will review some of the drivers and considerations for insecticide mixtures and their potential uses. © 2024 Society of Chemical Industry.

Singlet oxygen signalling and its potential roles in plant biotic interactions.

Singlet oxygen (SO) is among the most potent reactive oxygen species, and readily oxidizes proteins, lipids and DNA. It can be generated at the plant surface by phototoxins in the epidermis, acting as a direct defense against pathogens and herbivores (including humans). SO can also accumulate within mitochondria, peroxisomes, cytosol and the nucleus through multiple enzymatic and nonenzymatic processes. However, the majority of research on intracellular SO generation in plants has focused on transfer of light energy to triplet oxygen by photopigments from the chloroplast. SO accumulates in response to diverse stresses that perturb chloroplast metabolism, and while its high reactivity limits diffusion distances, it participates in retrograde signalling through the EXECUTER1 sensor, generation of carotenoid metabolites and possibly other unknown pathways. SO thereby reprogrammes nuclear gene expression and modulates hormone signalling and programmed cell death. While SO signalling has long been known to regulate plant responses to high-light stress, recent literature also suggests a role in plant interactions with insects, bacteria and fungi. The goals of this review are to provide a brief overview of SO, summarize evidence for its involvement in biotic stress responses and discuss future directions for the study of SO in defense signalling.

Effector-triggered and self-regulated plant resistance to insects.

Despite many years of research, the molecular mechanisms underlying the activation and regulation of host plant resistance (HPR) to insects remain elusive. Recently, Guo et al. reported that a nucleotide-binding leucine-rich repeat NLR protein activates HPR through direct recognition of an insect effector and that autophagy-mediated degradation of this effector negatively regulates HPR.

Larvae of Colorado potato beetle (Leptinotarsa decemlineata Say) resistant to double-stranded RNA (dsRNA) remain susceptible to small-molecule pesticides.

BACKGROUND: Implementation of resistance management tools is crucial for the continued efficacy of insect control technologies. An important aspect of insect resistance management (IRM) is the combined or sequential use of different modes-of-action to reduce selection pressure and delay evolution of resistance. This is especially important for insect pests with established ability to develop resistance to insecticides, such as the Colorado potato beetle (Leptinotarsa decemlineata, CPB). A new class of insecticides, based on double-stranded RNA (dsRNA) activating the gene silencing RNA-interference (RNAi) pathway, are currently under review for regulatory approval and commercial use in the USA against CPB. However, there is no information available on the potential for cross-resistance between RNAi insecticides and other classes of insecticides used against CPB. Herein, we aim to fill this knowledge gap by capitalizing on the availability of a CPB strain highly resistant to dsRNAs and test its susceptibility to diverse small-molecule insecticide classes compared to reference dsRNA-susceptible CPB strains. RESULTS: Differences in activity were observed among the four insecticides tested, with abamectin demonstrating highest activity against all three strains of CPB. However, no differences were observed among the dsRNA-resistant and susceptible CPB strains for any of the tested compounds. Overall, these results demonstrate lack of cross-resistance to commonly used chemical insecticides in the dsRNA-resistant strain of CPB. CONCLUSION: These data support the use of these different insecticide classes along with RNAi-based insecticides as part of an effective insect resistance management framework aimed at delaying resistance in CPB. © 2023 Society of Chemical Industry.

Interaction of Whitefly Effector G4 with Tomato Proteins Impacts Whitefly Performance.

The phloem-feeding insect Bemisia tabaci is an important pest, responsible for the transmission of several crop-threatening virus species. While feeding, the insect secretes a cocktail of effectors to modulate plant defense responses. Here, we present a set of proteins identified in an artificial diet on which B. tabaci was salivating. We subsequently studied whether these candidate effectors can play a role in plant immune suppression. Effector G4 was the most robust suppressor of an induced- reactive oxygen species (ROS) response in Nicotiana benthamiana. In addition, G4 was able to suppress ROS production in Solanum lycopersicum (tomato) and Capsicum annuum (pepper). G4 localized predominantly in the endoplasmic reticulum in N. benthamiana leaves and colocalized with two identified target proteins in tomato: REF-like stress related protein 1 (RSP1) and meloidogyne-induced giant cell protein DB141 (MIPDB141). Silencing of MIPDB141 in tomato reduced whitefly fecundity up to 40%, demonstrating that the protein is involved in susceptibility to B. tabaci. Together, our data demonstrate that effector G4 impairs tomato immunity to whiteflies by interfering with ROS production and via an interaction with tomato susceptibility protein MIPDB141. [Formula: see text] Copyright © 2024 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

Effect of herbivore stress on transgene behaviour in maize crosses with different genetic backgrounds: cry1Ab transgene transcription, insecticidal protein expression and bioactivity against insect pests.

Background: Decades after their first commercial release, many theoretical assumptions are still taken for granted in the deployment of genetically modified (GM) crops. Theoretically, in the case of maize, active transcription of the cry1Ab transgene would result in dose-dependent production of the insecticidal Cry1Ab protein, which would in turn induce dose-dependent mortality on lepidopteran pests. We produced data to realistically approach this question by using a model that includes two genetic background contexts from two geographical provenances in Brazil and South Africa, and two lepidopteran pests (Helicoverpa armigera and Spodoptera littoralis). However, in this study, the effect of insect herbivory was superimposed to investigate possible stress-induced effects in transgene expression at three levels: mRNA, protein and bioactivity. Results: Overall, we found that herbivore damage by H. armigera was reflected only at the translational level, with a higher level of Cry1Ab protein measured in the Brazilian crosses under herbivore stress. On the other hand, compared to non-stress growing conditions, the herbivore damage by S. littoralis was not directly reflected in mRNA, protein or bioactivity in the South African crosses. Conclusions: The differences between South African and Brazilian genetic backgrounds, and between the stressor effect of the two herbivores used, highlight the complexity of transgene expression at the agroecological level. Supplementary Information: The online version contains supplementary material available at 10.1186/s12302-023-00815-3.

Transcriptome Dynamics of Brassica juncea Leaves in Response to Omnivorous Beet Armyworm (Spodoptera exigua, Hübner).

Cruciferous plants manufacture glucosinolates (GSLs) as special and important defense compounds against insects. However, how insect feeding induces glucosinolates in Brassica to mediate insect resistance, and how plants regulate the strength of anti-insect defense response during insect feeding, remains unclear. Here, mustard (Brassica juncea), a widely cultivated Brassica plant, and beet armyworm (Spodoptera exigua), an economically important polyphagous pest of many crops, were used to analyze the changes in GSLs and transcriptome of Brassica during insect feeding, thereby revealing the plant-insect interaction in Brassica plants. The results showed that the content of GSLs began to significantly increase after 48 h of herbivory by S. exigua, with sinigrin as the main component. Transcriptome analysis showed that a total of 8940 DEGs were identified in mustard challenged with beet armyworm larvae. The functional enrichment results revealed that the pathways related to the biosynthesis of glucosinolate and jasmonic acid were significantly enriched by upregulated DEGs, suggesting that mustard might provide a defense against herbivory by inducing JA biosynthesis and then promoting GSL accumulation. Surprisingly, genes regulating JA catabolism and inactivation were also activated, and both JA signaling repressors (JAZs and JAMs) and activators (MYCs and NACs) were upregulated during herbivory. Taken together, our results indicate that the accumulation of GSLs regulated by JA signaling, and the regulation of active and inactive JA compound conversion, as well as the activation of JA signaling repressors and activators, collectively control the anti-insect defense response and avoid over-stunted growth in mustard during insect feeding.

Assessment of the 2021 post-market environmental monitoring report on the cultivation of genetically modified maize MON 810 in the EU.

Following a request from the European Commission, the European Food Safety Authority (EFSA) assessed the 2021 post-market environmental monitoring (PMEM) report on the cultivation of Cry1Ab-expressing maize event MON 810. Evidence provided in the PMEM report shows that farmers growing maize MON 810 in Spain complied partially with refuge requirements, while full compliance was achieved in Portugal. Cry1Ab susceptibility tests performed on European and Mediterranean corn borer populations collected from north-eastern Spain in 2021 indicated no symptoms of resistance evolution to maize MON 810. However, unexpected damage to maize MON 810 plants was observed in a field trial in the province of Girona (north-eastern Spain), which may point to the presence of resistance alleles in this region. Information retrieved through farmer questionnaires and the scientific literature reveals no unanticipated adverse effects on human and animal health or the environment arising from the cultivation of maize MON 810. Overall, EFSA concludes that the evidence reported in the 2021 PMEM report does not invalidate its previous conclusions on the safety of maize MON 810. The possible presence of Cry1Ab resistance alleles at frequencies leading to damage to maize MON 810 plants in Girona requires twofold actions: (1) increase monitoring efforts in this area; and (2) implement remedial measures to limit the suspected evolution and spread of resistance. As in previous years, EFSA identified shortcomings on resistance monitoring that need revision. In particular, full refuge compliance must be achieved in Spain. Moreover, the sensitivity of the monitoring plan must be increased, which can be achieved by replacing the current susceptibility assays by periodic F2 screens. EFSA also recommends the consent holder to revise the farmer questionnaires to account for the emergence of teosinte as a noxious agricultural weed in maize MON 810-growing areas in Spain.