RESUMEN
The sustainable control of many highly damaging insect crop pests and disease vectors is threatened by the evolution of insecticide resistance. As a consequence, strategies have been developed that aim to prevent or delay resistance development by rotating or mixing insecticides with different modes of action (MoA). However, these approaches can be compromised by the emergence of mechanisms that confer cross-resistance to insecticides with different MoA. Despite the applied importance of cross-resistance, its evolutionary underpinnings remain poorly understood. Here we reveal how a single gene evolved the capacity to detoxify two structurally unrelated insecticides with different MoA. Using transgenic approaches we demonstrate that a specific variant of the cytochrome P450 CYP6ER1, previously shown to confer resistance to the neonicotinoid imidacloprid in the brown planthopper, N. lugens, also confers cross-resistance to the phenylpyrazole ethiprole. CYP6ER1 is duplicated in resistant strains, and we show that while the acquisition of mutations in two encoded substrate recognition sites (SRS) of one of the parologs led to resistance to imidacloprid, a different set of mutations, outside of known SRS, are primarily responsible for resistance to ethiprole. Epistatic interactions between these mutations and their genetic background suggest that the evolution of dual resistance from the same gene copy involved functional trade-offs in respect to CYP6ER1 catalytic activity for ethiprole versus imidacloprid. Surprisingly, the mutations leading to ethiprole and imidacloprid resistance do not confer the ability to detoxify the insecticide fipronil, another phenylpyrazole with close structural similarity to ethiprole. Taken together, these findings reveal how gene duplication and divergence can lead to the evolution of multiple novel functions from a single gene. From an applied perspective they also demonstrate how cross-resistance to structurally unrelated insecticides can evolve, and illustrate the difficulty in predicting cross-resistance profiles mediated by metabolic mechanisms.
Asunto(s)
Hemípteros , Insecticidas , Animales , Sistema Enzimático del Citocromo P-450/genética , Sistema Enzimático del Citocromo P-450/metabolismo , Duplicación de Gen , Resistencia a los Insecticidas/genética , Insecticidas/metabolismo , Insecticidas/farmacologíaRESUMEN
The alkaloid, nicotine, produced by tobacco and other Solanaceae as an anti-herbivore defence chemical is one of the most toxic natural insecticides in nature. However, some insects, such as the whitefly species, Trialeurodes vaporariorum and Bemisia tabaci show strong tolerance to this allelochemical and can utilise tobacco as a host. Here, we used biological, molecular and functional approaches to investigate the role of cytochrome P450 enzymes in nicotine tolerance in T. vaporariorum and B. tabaci. Insecticide bioassays revealed that feeding on tobacco resulted in strong induced tolerance to nicotine in both species. Transcriptome profiling of both species reared on tobacco and bean hosts revealed profound differences in the transcriptional response these host plants. Interrogation of the expression of P450 genes in the host-adapted lines revealed that P450 genes belonging to the CYP6DP subfamily are strongly upregulated in lines reared on tobacco. Functional characterisation of these P450s revealed that CYP6DP1 and CYP6DP2 of T. vaporariorum and CYP6DP3 of B. tabaci confer resistance to nicotine in vivo. These three genes, in addition to the B. tabaci P450 CYP6DP5, were also found to confer resistance to the neonicotinoid imidacloprid. Our data provide new insight into the molecular basis of nicotine resistance in insects and illustrates how divergence in the evolution of P450 genes in this subfamily in whiteflies may have impacted the extent to which different species can tolerate a potent natural insecticide.
Asunto(s)
Hemípteros , Insecticidas , Animales , Nicotina/farmacología , Nicotina/metabolismo , Insecticidas/farmacología , Insecticidas/metabolismo , Resistencia a los Insecticidas/genética , Neonicotinoides/farmacología , Neonicotinoides/metabolismo , Sistema Enzimático del Citocromo P-450/genética , Sistema Enzimático del Citocromo P-450/metabolismo , Nicotiana/genética , Hemípteros/metabolismo , Nitrocompuestos/farmacología , Nitrocompuestos/metabolismoRESUMEN
The impact of pesticides on the health of bee pollinators is determined in part by the capacity of bee detoxification systems to convert these compounds to less toxic forms. For example, recent work has shown that cytochrome P450s of the CYP9Q subfamily are critically important in defining the sensitivity of honey bees and bumblebees to pesticides, including neonicotinoid insecticides. However, it is currently unclear if solitary bees have functional equivalents of these enzymes with potentially serious implications in relation to their capacity to metabolise certain insecticides. To address this question, we sequenced the genome of the red mason bee, Osmia bicornis, the most abundant and economically important solitary bee species in Central Europe. We show that O. bicornis lacks the CYP9Q subfamily of P450s but, despite this, exhibits low acute toxicity to the N-cyanoamidine neonicotinoid thiacloprid. Functional studies revealed that variation in the sensitivity of O. bicornis to N-cyanoamidine and N-nitroguanidine neonicotinoids does not reside in differences in their affinity for the nicotinic acetylcholine receptor or speed of cuticular penetration. Rather, a P450 within the CYP9BU subfamily, with recent shared ancestry to the Apidae CYP9Q subfamily, metabolises thiacloprid in vitro and confers tolerance in vivo. Our data reveal conserved detoxification pathways in model solitary and eusocial bees despite key differences in the evolution of specific pesticide-metabolising enzymes in the two species groups. The discovery that P450 enzymes of solitary bees can act as metabolic defence systems against certain pesticides can be leveraged to avoid negative pesticide impacts on these important pollinators.
Asunto(s)
Abejas/efectos de los fármacos , Abejas/genética , Neonicotinoides/farmacología , Animales , Evolución Biológica , Sistema Enzimático del Citocromo P-450/genética , Europa (Continente) , Genómica/métodos , Insecticidas/farmacología , Polinización/efectos de los fármacos , Polinización/genética , Tiazinas/farmacologíaRESUMEN
The cytochrome P450 family (P450s) of arthropods includes diverse enzymes involved in endogenous essential physiological functions and in the oxidative metabolism of xenobiotics, insecticides and plant allelochemicals. P450s can also establish insecticide selectivity in bees and pollinators. Several arthropod P450s, distributed in different phylogenetic groups, have been associated with xenobiotic metabolism, and some of them have been functionally characterized, using different in vitro and in vivo systems. The purpose of this review is to summarize scientific publications on arthropod P450s from major insect and mite agricultural pests, pollinators and Papilio sp, which have been functionally characterized and shown to metabolize xenobiotics and/or their role (direct or indirect) in pesticide toxicity or resistance has been functionally validated. The phylogenetic relationships among these P450s, the functional systems employed for their characterization and their xenobiotic catalytic properties are presented, in a systematic approach, including critical aspects and limitations. The potential of the primary P450-based metabolic pathway of target and non-target organisms for the development of highly selective insecticides and resistance-breaking formulations may help to improve the efficiency and sustainability of pest control.
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Artrópodos , Insecticidas , Animales , Artrópodos/metabolismo , Abejas , Sistema Enzimático del Citocromo P-450/genética , Sistema Enzimático del Citocromo P-450/metabolismo , Inactivación Metabólica , Insecticidas/toxicidad , Filogenia , Xenobióticos/toxicidadRESUMEN
The cabbage stem flea beetle, Psylliodes chrysocephala L. is a major pest of winter oilseed rape in several European countries. Traditionally, neonicotinoid and pyrethroid insecticides have been widely used for control of P. chrysocephala, but in recent years, following the withdrawal of neonicotinoid insecticide seed treatments, control failures have occurred due to an over reliance on pyrethroids. In line with previous surveys, UK populations of P. chrysocephala were found to exhibit high levels of resistance to the pyrethroid lambda-cyhalothrin. This resistance was suppressed by pre-treatment with the cytochrome P450 inhibitor PBO under laboratory conditions, suggesting that the resistance has a strong metabolic component. The L1014F (kdr) mutation in the voltage-gated sodium channel, which confers relatively low levels (10-20 fold) of resistance to pyrethroids, was also found to be widespread across the UK regions sampled, whereas the L925I (s-kdr) mutation was much less common. The current survey also suggests that higher levels of pyrethroid resistance have spread to the North and West of England, and that resistance levels continue to remain high in the South East.
RESUMEN
BACKGROUND: The glasshouse whitefly, Trialeurodes vaporariorum, is a damaging crop pest and an invasive generalist capable of feeding on a broad range of host plants. As such this species has evolved mechanisms to circumvent the wide spectrum of anti-herbivore allelochemicals produced by its host range. T. vaporariorum has also demonstrated a remarkable ability to evolve resistance to many of the synthetic insecticides used for control. RESULTS: To gain insight into the molecular mechanisms that underpin the polyphagy of T. vaporariorum and its resistance to natural and synthetic xenobiotics, we sequenced and assembled a reference genome for this species. Curation of genes putatively involved in the detoxification of natural and synthetic xenobiotics revealed a marked reduction in specific gene families between this species and another generalist whitefly, Bemisia tabaci. Transcriptome profiling of T. vaporariorum upon transfer to a range of different host plants revealed profound differences in the transcriptional response to more or less challenging hosts. Large scale changes in gene expression (> 20% of genes) were observed during adaptation to challenging hosts with a range of genes involved in gene regulation, signalling, and detoxification differentially expressed. Remarkably, these changes in gene expression were associated with significant shifts in the tolerance of host-adapted T. vaporariorum lines to natural and synthetic insecticides. CONCLUSIONS: Our findings provide further insights into the ability of polyphagous insects to extensively reprogram gene expression during host adaptation and illustrate the potential implications of this on their sensitivity to synthetic insecticides.
Asunto(s)
Regulación de la Expresión Génica de las Plantas , Hemípteros/genética , Resistencia a los Insecticidas/genética , Adaptación Fisiológica/genética , Animales , Proteasas de Cisteína/genética , Proteasas de Cisteína/metabolismo , Sistema Enzimático del Citocromo P-450/genética , Sistema Enzimático del Citocromo P-450/metabolismo , Genes de Insecto , Genoma de los Insectos , Hemípteros/enzimología , Hemípteros/metabolismo , Interacciones Huésped-Patógeno/genética , Proteínas de Insectos/genética , Proteínas de Insectos/metabolismo , Insecticidas , Plantas , RNA-Seq , Transducción de Señal/genética , Transcriptoma , Xenobióticos/metabolismoRESUMEN
We discovered the A301S mutation in the RDL GABA-gated chloride channel of fiprole resistant rice brown planthopper, Nilaparvata lugens populations by DNA sequencing and SNP calling via RNASeq. Ethiprole selection of two field N. lugens populations resulted in strong resistance to both ethiprole and fipronil and resulted in fixation of the A301S mutation, as well as the emergence of another mutation, Q359E in one of the selected strains. To analyse the roles of these mutations in resistance to phenylpyrazoles, three Rdl constructs: wild type, A301S and A301S+Q359E were expressed in Xenopus laevis oocytes and assessed for their sensitivity to ethiprole and fipronil using two-electrode voltage-clamp electrophysiology. Neither of the mutant Rdl subtypes significantly reduced the antagonistic action of fipronil, however there was a significant reduction in response to ethiprole in the two mutated subtypes compared with the wild type. Bioassays with a Drosophila melanogaster strain carrying the A301S mutation showed strong resistance to ethiprole but not fipronil compared to a strain without this mutation, thus further supporting a causal role for the A301S mutation in resistance to ethiprole. Homology modelling of the N. lugens RDL channel did not suggest implications of Q359E for fiprole binding in contrast to A301S located in transmembrane domain M2 forming the channel pore. Synergist bioassays provided no evidence of a role for cytochrome P450s in N. lugens resistance to fipronil and the molecular basis of resistance to this compound remains unknown. In summary this study provides strong evidence that target-site resistance underlies widespread ethiprole resistance in N. lugens populations.
Asunto(s)
Hemípteros/efectos de los fármacos , Hemípteros/genética , Proteínas de Insectos/genética , Resistencia a los Insecticidas , Insecticidas/farmacología , Mutación Missense , Receptores de GABA-A/genética , Animales , Secuencia de Bases , Hemípteros/metabolismo , Proteínas de Insectos/metabolismo , Pirazoles/farmacología , Piretrinas/farmacología , Receptores de GABA-A/metabolismoRESUMEN
Many genes increase coding capacity by alternate exon usage. The gene encoding the insect nicotinic acetylcholine receptor (nAChR) α6 subunit, target of the bio-insecticide spinosad, is one example of this and expands protein diversity via alternative splicing of mutually exclusive exons. Here, we show that spinosad resistance in the tomato leaf miner, Tuta absoluta is associated with aberrant regulation of splicing of Taα6 resulting in a novel form of insecticide resistance mediated by exon skipping. Sequencing of the α6 subunit cDNA from spinosad selected and unselected strains of T. absoluta revealed all Taα6 transcripts of the selected strain were devoid of exon 3, with comparison of genomic DNA and mRNA revealing this is a result of exon skipping. Exon skipping cosegregated with spinosad resistance in survival bioassays, and functional characterization of this alteration using modified human nAChR α7, a model of insect α6, demonstrated that exon 3 is essential for receptor function and hence spinosad sensitivity. DNA and RNA sequencing analyses suggested that exon skipping did not result from genetic alterations in intronic or exonic cis-regulatory elements, but rather was associated with a single epigenetic modification downstream of exon 3a, and quantitative changes in the expression of trans-acting proteins that have known roles in the regulation of alternative splicing. Our results demonstrate that the intrinsic capacity of the α6 gene to generate transcript diversity via alternative splicing can be readily exploited during the evolution of resistance and identifies exon skipping as a molecular alteration conferring insecticide resistance.
Asunto(s)
Empalme Alternativo , Exones , Insectos/genética , Resistencia a los Insecticidas/genética , Receptor Nicotínico de Acetilcolina alfa 7/genética , Animales , Combinación de Medicamentos , Humanos , MacrólidosRESUMEN
Host plant shifts of herbivorous insects may be a first step toward sympatric speciation and can create new pests of agriculturally important crops; however, the molecular mechanisms that mediate this process are poorly understood. Certain races of the polyphagous aphid Myzus persicae have recently adapted to feed on tobacco (Myzus persicae nicotianae) and show a reduced sensitivity to the plant alkaloid nicotine and cross-resistance to neonicotinoids a class of synthetic insecticides widely used for control. Here we show constitutive overexpression of a cytochrome P450 (CYP6CY3) allows tobacco-adapted races of M. persicae to efficiently detoxify nicotine and has preadapted them to resist neonicotinoid insecticides. CYP6CY3, is highly overexpressed in M. persicae nicotianae clones from three continents compared with M. persicae s.s. and expression level is significantly correlated with tolerance to nicotine. CYP6CY3 is highly efficient (compared with the primary human nicotine-metabolizing P450) at metabolizing nicotine and neonicotinoids to less toxic metabolites in vitro and generation of transgenic Drosophila expressing CYP6CY3 demonstrate that it confers resistance to both compounds in vivo. Overexpression of CYP6CY3 results from the expansion of a dinucleotide microsatellite in the promoter region and a recent gene amplification, with some aphid clones carrying up to 100 copies. We conclude that the mutations leading to overexpression of CYP6CY3 were a prerequisite for the host shift of M. persicae to tobacco and that gene amplification and microsatellite polymorphism are evolutionary drivers in insect host adaptation.
Asunto(s)
Adaptación Biológica/genética , Áfidos/enzimología , Repeticiones de Dinucleótido/genética , Amplificación de Genes/genética , Nicotiana/parasitología , Polimorfismo Genético/genética , Animales , Áfidos/efectos de los fármacos , Hidrocarburo de Aril Hidroxilasas/metabolismo , Secuencia de Bases , Cromatografía Liquida , Interacciones Huésped-Parásitos , Datos de Secuencia Molecular , Mutación/genética , Nicotina/toxicidad , Reacción en Cadena en Tiempo Real de la Polimerasa , Análisis de Secuencia de ADN , Espectrometría de Masas en TándemRESUMEN
Cabbage stem flea beetle, Psylliodes chrysocephala L. (Coleoptera: Chrysomelidae) is a major pest of winter oilseed rape in several European countries particularly attacking young emerging plants in autumn. Over the last several decades, pyrethroid insecticides have been foliarly applied to control flea beetle outbreaks. Recent control failures in northern Germany suggested pyrethroid resistance development in cabbage stem flea beetles, which were confirmed by resistance monitoring bioassays using lambda-cyhalothrin in an adult vial test. The purpose of this study was to investigate the presence of polymorphisms in the para-type voltage-gated sodium channel gene of P. chrysocephala known to be involved in knock-down resistance (kdr). By using a degenerate primer approach we PCR amplified part of the para-type sodium channel gene and identified in resistant flea beetles a single nucleotide polymorphism resulting in an L1014F (kdr) mutation within domain IIS6 of the channel protein, known as one of the chief pyrethroid target-site resistance mechanisms in several other pest insects. Twenty populations including four archived museum samples collected between 1945 and 1958 were analyzed using a newly developed pyrosequencing diagnostic assay. The assay revealed a kdr allele frequency of 90-100% in those flea beetle populations expressing high-level cross-resistance in discriminating dose bioassays against different pyrethroids such as lambda-cyhalothrin, tau-fluvalinate, etofenprox and bifenthrin. The presence of target-site resistance to pyrethroids in cabbage stem flea beetle is extremely worrying considering the lack of effective alternative modes of action to control this pest in Germany and other European countries, and is likely to result in major control problems once it expands to other geographies. The striking fact that cabbage stem flea beetle is next to pollen beetle, Meligethes aeneus the second coleopteran pest in European winter oilseed rape resisting pyrethroid treatments by expressing a target-site mutation, underpins the importance of diversity in available chemistry for resistance management tactics based on mode of action rotation in order to guarantee sustainable winter oilseed rape cultivation in Europe.
Asunto(s)
Brassica napus/parasitología , Escarabajos/efectos de los fármacos , Escarabajos/genética , Proteínas de Insectos/genética , Resistencia a los Insecticidas , Insecticidas/farmacología , Enfermedades de las Plantas/parasitología , Piretrinas/farmacología , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Escarabajos/metabolismo , Alemania , Proteínas de Insectos/química , Proteínas de Insectos/metabolismo , Datos de Secuencia Molecular , Mutación , Alineación de Secuencia , Canales de Sodio/química , Canales de Sodio/genética , Canales de Sodio/metabolismoRESUMEN
BACKGROUND: Plutella xylostella (L.) is a destructive pest of cruciferous crops worldwide that has evolved resistance to many insecticides. Here we examined the mode of inheritance, cross-resistance profile, and potential mechanisms of emamectin benzoate resistance in a field-derived strain of P. xylostella from Japan. RESULTS: A field-collected population of P. xylostella, was found to exhibit strong (> 150-fold) resistance to emamectin benzoate in insecticide bioassays when compared with a laboratory susceptible strain. Genetic analysis showed that resistance is inherited as an autosomal, recessive trait, and is conferred by a single or a few closely linked loci. The emamectin benzoate resistant strain also exhibited resistance to abamectin, lepimectin, chlorantraniliprole, lufenuron, spinetoram, indoxacarb, fipronil, dieldrin, endosulfan and lambda-cyhalothrin, demonstrating a remarkable multi-resistance profile. Insecticide bioassays employing inhibitors of detoxification enzymes revealed that piperonyl butoxide (PBO) increased the toxicity of emamectin benzoate in the resistant strain by ten-fold indicating the potential involvement of cytochrome P450 monooxygenases in avermectin resistance. Furthermore, cloning and sequencing of the primary receptor of avermectins, the GluCl channel, revealed the absence of target-site mutations in the resistant strain. CONCLUSIONS: Our data on the mode of inheritance and mechanisms of resistance to emamectin benzoate in a P. xylostella strain from Japan provide a foundation for the development of regional resistance management strategies. However, the high levels of phenotypic resistance in this strain to a diverse range of other insecticide classes available for control illustrate the challenges associated with the sustainable control of this important pest. © 2023 Society of Chemical Industry.
Asunto(s)
Insecticidas , Ivermectina/análogos & derivados , Mariposas Nocturnas , Animales , Insecticidas/farmacología , Resistencia a los Insecticidas/genéticaRESUMEN
The tobacco whitefly, Bemisia tabaci, is a polyphagous crop pest which causes high levels of economic damage across the globe. Insecticides are often required for the effective control of this species, among which the neonicotinoid class have been particularly widely used. Deciphering the mechanisms responsible for resistance to these chemicals is therefore critical to maintain control of B. tabaci and limit the damage it causes. An important mechanism of resistance to neonicotinoids in B. tabaci is the overexpression of the cytochrome P450 gene CYP6CM1 which leads to the enhanced detoxification of several neonicotinoids. In this study we show that qualitative changes in this P450 dramatically alter its metabolic capacity to detoxify neonicotinoids. CYP6CM1 was significantly over-expressed in two strains of B. tabaci which displayed differing levels of resistance to the neonicotinoids imidacloprid and thiamethoxam. Sequencing of the CYP6CM1 coding sequence from these strains revealed four different alleles encoding isoforms carrying several amino acid changes. Expression of these alleles in vitro and in vivo provided compelling evidence that a mutation (A387G), present in two of the CYP6CM1 alleles, results in enhanced resistance to several neonicotinoids. These data demonstrate the importance of both qualitative and quantitative changes in genes encoding detoxification enzymes in the evolution of insecticide resistance and have applied implications for resistance monitoring programs.
Asunto(s)
Hemípteros , Insecticidas , Animales , Mutación Puntual , Neonicotinoides/farmacología , Neonicotinoides/metabolismo , Insecticidas/farmacología , Insecticidas/metabolismo , Nitrocompuestos/farmacología , Nitrocompuestos/metabolismo , Resistencia a los Insecticidas/genética , Sistema Enzimático del Citocromo P-450/genética , Sistema Enzimático del Citocromo P-450/metabolismo , Hemípteros/genética , Hemípteros/metabolismoRESUMEN
The Neotropical brown stink bug, Euschistus heros, is a major pest of soybean in South America. The importance of E. heros as a pest has grown significantly in recent times due to increases in its abundance and range, and the evolution of insecticide resistance. Recent work has begun to examine the genetic diversity, population structure, and genetic mechanisms of insecticide resistance in E. heros. However, to date, investigation of these topics has been hampered by a lack of genomic resources for this species. Here we address this need by assembling a high-quality draft genome for E. heros. We used a combination of short and long read sequencing to assemble an E. heros genome of 1.4 Gb comprising 906 contigs with a contig N50 of 3.5 MB. We leveraged this new genomic resource, in combination with genotyping by sequencing, to explore genetic diversity in populations of this species in Brazil and identify genetic loci in the genome which are under selection. Our genome-wide analyses, confirm that there are two populations of E. heros co-occurring in different geographical regions in Brazil, and that, in certain regions of the country these populations are hybridizing. We identify several regions of the genome as under selection, including markers associated with putative insecticide resistance genes. Taken together, the new genomic resources generated in this study will accelerate research into fundamental aspects of stinkbug biology and applied aspects relating to the sustainable control of a highly damaging crop pest.
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Heterópteros , Insecticidas , Animales , Estudio de Asociación del Genoma Completo , Heterópteros/genética , Brasil , DemografíaRESUMEN
The tomato leafminer, Tuta absoluta, is an invasive crop pest that has evolved resistance to many of the insecticides used for its control. To facilitate the investigation of the underpinning mechanisms of resistance in this species we generated a contiguous genome assembly using long-read sequencing data. We leveraged this genomic resource to investigate the genetic basis of resistance to the diamide insecticide chlorantraniliprole in Spanish strains of T. absoluta that exhibit high levels of resistance to this insecticide. Transcriptomic analyses revealed that, in these strains, resistance is not associated with previously reported target-site mutations in the diamide target-site, the ryanodine receptor, but rather is associated with the marked overexpression (20- to >100-fold) of a gene encoding a UDP-glycosyltransferase (UGT). Functional expression of this UGT, UGT34A23, via ectopic expression in Drosophila melanogaster demonstrated that it confers strong and significant resistance in vivo. The genomic resources generated in this study provide a powerful resource for further research on T. absoluta. Our findings on the mechanisms underpinning resistance to chlorantraniliprole will inform the development of sustainable management strategies for this important pest.
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Insecticidas , Lepidópteros , Mariposas Nocturnas , Solanum lycopersicum , Animales , Insecticidas/farmacología , Diamida , Resistencia a los Insecticidas/genética , Drosophila melanogaster , Uridina DifosfatoRESUMEN
The development of novel and safe insecticides remains an important need for a growing world population to protect crops and animal and human health. New chemotypes modulating the insect nicotinic acetylcholine receptors have been recently brought to the agricultural market, yet with limited understanding of their molecular interactions at their target receptor. Herein, we disclose the first crystal structures of these insecticides, namely, sulfoxaflor, flupyradifurone, triflumezopyrim, flupyrimin, and the experimental compound, dicloromezotiaz, in a double-mutated acetylcholine-binding protein which mimics the insect-ion-channel orthosteric site. Enabled by these findings, we discovered novel pharmacophores with a related mode of action, and we describe herein their design, synthesis, and biological evaluation.
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Diseño de Fármacos , Proteínas de Insectos/metabolismo , Insecticidas/síntesis química , Receptores Nicotínicos/metabolismo , 4-Butirolactona/análogos & derivados , 4-Butirolactona/química , 4-Butirolactona/metabolismo , Animales , Sitios de Unión , Escarabajos/efectos de los fármacos , Escarabajos/metabolismo , Cristalografía por Rayos X , Humanos , Control de Insectos/métodos , Proteínas de Insectos/química , Proteínas de Insectos/genética , Insecticidas/metabolismo , Insecticidas/farmacología , Conformación Molecular , Simulación de Dinámica Molecular , Mutagénesis Sitio-Dirigida , Piridinas/química , Piridinas/metabolismo , Pirimidinonas/química , Pirimidinonas/metabolismo , Receptores Nicotínicos/química , Receptores Nicotínicos/genética , Compuestos de Azufre/química , Compuestos de Azufre/metabolismoRESUMEN
Insect cytochrome P450-monooxygenases (P450s) are an enzyme superfamily involved in the oxidative transformation of endogenous and exogenous substrates, including insecticides. They were also shown to determine insecticide selectivity in beneficial arthropods such as bee pollinators, and to detoxify plant secondary metabolites. The recent explosion in numbers of P450s due to increased invertebrate genomes sequenced, allowed researchers to study their functional relevance for xenobiotic metabolism by recombinant expression using different expression systems. Troubleshooting strategies, including different systems and protein modifications typically adapted from mammalian P450s, have been applied to improve the functional expression, with partial success. The aim of this mini review is to critically summarize different strategies recently developed and used to produce recombinant insect P450s for xenobiotic metabolism studies.
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Sistema Enzimático del Citocromo P-450/metabolismo , Inactivación Metabólica , Insectos/enzimología , Animales , Sistema Enzimático del Citocromo P-450/genética , Insectos/genética , Xenobióticos/metabolismoRESUMEN
The green peach aphid, Myzus persicae, is a globally distributed highly damaging crop pest. This species has demonstrated an exceptional ability to evolve resistance to both synthetic insecticides used for control, and natural insecticides produced by certain plants as a chemical defense against insect attack. Here we review work characterizing the evolution of resistance in M. persicae to the natural insecticide nicotine and the structurally related class of synthetic neonicotinoid insecticides. We outline how research on this topic has provided insights into long-standing questions of both evolutionary and applied importance. These include questions pertaining to the origins of novel traits, the number and nature of mutational events or 'adaptive steps' underlying the evolution of new phenotypes, and whether host plant adaptations can be co-opted to confer resistance to synthetic insecticides. Finally, research on the molecular mechanisms underlying insecticide resistance in M. persicae has generated several outstanding questions on the genetic architecture of resistance to both natural and synthetic xenobiotics, and we conclude by identifying key knowledge gaps for future research. © 2021 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.
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Áfidos , Insecticidas , Animales , Áfidos/genética , Resistencia a los Insecticidas/genética , Insecticidas/farmacología , Neonicotinoides , NicotinaRESUMEN
The aphid Myzus persicae is a destructive agricultural pest that displays an exceptional ability to develop resistance to both natural and synthetic insecticides. To investigate the evolution of resistance in this species we generated a chromosome-scale genome assembly and living panel of >110 fully sequenced globally sampled clonal lines. Our analyses reveal a remarkable diversity of resistance mutations segregating in global populations of M. persicae. We show that the emergence and spread of these mechanisms is influenced by host-plant associations, uncovering the widespread co-option of a host-plant adaptation that also offers resistance against synthetic insecticides. We identify both the repeated evolution of independent resistance mutations at the same locus, and multiple instances of the evolution of novel resistance mechanisms against key insecticides. Our findings provide fundamental insights into the genomic responses of global insect populations to strong selective forces, and hold practical relevance for the control of pests and parasites.
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Áfidos/genética , Evolución Molecular , Variación Genética , Genoma de los Insectos/genética , Resistencia a los Insecticidas/genética , Insecticidas/farmacología , Animales , Áfidos/clasificación , Áfidos/fisiología , Secuencia de Bases , Genómica/métodos , Geografía , Interacciones Huésped-Parásitos/efectos de los fármacos , Mutación , Filogenia , Plantas/parasitología , Polimorfismo de Nucleótido Simple , Homología de Secuencia de Ácido NucleicoRESUMEN
Host shifts can lead to ecological speciation and the emergence of new pests and pathogens. However, the mutational events that facilitate the exploitation of novel hosts are poorly understood. Here, we characterize an adaptive walk underpinning the host shift of the aphid Myzus persicae to tobacco, including evolution of mechanisms that overcame tobacco chemical defenses. A series of mutational events added as many as 1.5 million nucleotides to the genome of the tobacco-adapted subspecies, M. p. nicotianae, and yielded profound increases in expression of an enzyme that efficiently detoxifies nicotine, both in aphid gut tissue and in the bacteriocytes housing the obligate aphid symbiont Buchnera aphidicola. This dual evolutionary solution overcame the challenge of preserving fitness of a mutualistic symbiosis during adaptation to a toxic novel host. Our results reveal the intricate processes by which genetic novelty can arise and drive the evolution of key innovations required for ecological adaptation.
RESUMEN
BACKGROUND: The tomato leafminer, Tuta absoluta, is an economically important pest of tomatoes in Europe, Africa, Asia and South America. In the UK this species is controlled using an integrated pest management (IPM) programme which incorporates the insecticides spinosad and chlorantraniliprole. In response to UK grower concerns of loss of efficacy of these compounds at certain sites, insecticide bioassays were performed on five populations collected from four commercial glasshouses and potential mechanisms of resistance investigated. RESULTS: We observed high levels of resistance to spinosad in four of the strains, and in two of these tolerance to chlorantraniliprole. Selection of one of these strains with chlorantraniliprole rapidly resulted in a line exhibiting potent resistance to this compound. Sequencing of messenger RNA encoding the nicotinic acetylcholine receptor (nAChR) α6 subunit, target of spinosad, revealed Taα6 transcripts in the spinosad-resistant strains that lack exon 4 and encode a highly truncated protein, or contain a triplet deletion in the predicted first transmembrane domain resulting in the loss of a highly conserved amino acid. Sequencing of the ryanodine receptor gene, encoding the target of diamide insecticides, of the chlorantraniliprole-selected line revealed an amino acid substitution (G4903V) that has been previously linked to diamide resistance in populations of T. absoluta in the Mediterranean and South America. CONCLUSION: Taken together our results reveal emerging resistance in UK populations of T. absoluta to two of the most important insecticides used as part of IPM, with significant implications for the control of this species in the UK. © 2019 Society of Chemical Industry.