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1.
Pest Manag Sci ; 80(5): 2383-2392, 2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-37899495

RESUMO

BACKGROUND: Current European Union and United Kingdom legislation prohibits the use of neonicotinoid insecticidal seed treatments in oilseed rape (OSR, Brassica napus). This ban, and the reduction in efficacy of pyrethroid insecticide sprays due to resistance, has exacerbated pest pressure from the cabbage stem flea beetle (Psylliodes chrysocephala) in winter OSR. We quantified the direct impact of P. chrysocephala injury on the productivity of OSR. Leaf area was removed from young plants to simulate differing intensities of adult feeding injury alone or in combination with varying larval infestation levels. RESULTS: OSR can compensate for up to 90% leaf area loss at early growth stages, with no meaningful effect on yield. Significant impacts were observed with high infestations of more than five larvae per plant; plants were shorter, produced fewer flowers and pods, with fewer seeds per pod which had lower oil content and higher glucosinolate content. Such effects were not recorded when five larvae or fewer were present. CONCLUSION: These data confirm the yield-limiting potential of the larval stages of P. chrysocephala but suggest that the current action thresholds which trigger insecticide application for both adult and larval stages (25% leaf area loss and five larvae/plant, respectively) are potentially too low as they are below the physiological injury level where plants can fully compensate for damage. Further research in field conditions is needed to define physiological thresholds more accurately as disparity may result in insecticide applications that are unnecessary to protect yield and may in turn exacerbate the development and spread of insecticide resistance in P. chrysocephala. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.


Assuntos
Brassica napus , Besouros , Inseticidas , Piretrinas , Animais , Inseticidas/farmacologia , Piretrinas/farmacologia , Resistência a Inseticidas , Larva
2.
Pestic Biochem Physiol ; 191: 105339, 2023 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-36963921

RESUMO

There are many insect pests worldwide that damage agricultural crop and reduce yield either by direct feeding or by the transmission of plant diseases. To date, control of pest insects has been achieved largely by applying synthetic insecticides. However, insecticide use can be seriously impacted by legislation that limits their use or by the evolution of resistance in the target pest. Thus, there is a move towards less use of insecticides and increased adoption of integrated pest management strategies using a wide range of non-chemical and chemical control methods. For good pest control there is a need to understand the mode of action and selectivity of insecticides, the life cycles of the pests and their biology and behaviours, all of which can benefit from good quality genome data. Here we present the complete assembled (chromosome level) genomes (incl. mtDNA) of 19 insect pests, Agriotes lineatus (click beetle/wireworm), Aphis gossypii (melon/cotton aphid), Bemisia tabaci (cotton whitefly), Brassicogethes aeneus (pollen beetle), Ceutorhynchus obstrictus (seedpod weevil), Chilo suppressalis (striped rice stem borer), Chrysodeixis includens (soybean looper), Diabrotica balteata (cucumber beetle), Diatraea saccharalis (sugar cane borer), Nezara viridula (green stink bug), Nilaparvata lugens (brown plant hopper), Phaedon cochleariae (mustard beetle), Phyllotreta striolata (striped flea beetle), Psylliodes chrysocephala (cabbage stem flea beetle), Spodoptera exigua (beet army worm), Spodoptera littoralis (cotton leaf worm), Diabrotica virgifera (western corn root worm), Euschistus heros (brown stink bug) and Phyllotreta cruciferae (crucifer flea beetle). For the first 15 of these we also present the annotation of genes encoding potential xenobiotic detoxification enzymes. This public resource will aid in the elucidation and monitoring of resistance mechanisms, the development of highly selective chemistry and potential techniques to disrupt behaviour in a way that limits the effect of the pests.


Assuntos
Afídeos , Besouros , Heterópteros , Inseticidas , Mariposas , Animais , Inseticidas/farmacologia , Agricultura/métodos , Controle de Pragas , Besouros/genética , Controle de Insetos/métodos
3.
BMC Genomics ; 23(1): 198, 2022 Mar 12.
Artigo em Inglês | MEDLINE | ID: mdl-35279098

RESUMO

BACKGROUND: Sphaerophoria rueppellii, a European species of hoverfly, is a highly effective beneficial predator of hemipteran crop pests including aphids, thrips and coleopteran/lepidopteran larvae in integrated pest management (IPM) programmes. It is also a key pollinator of a wide variety of important agricultural crops. No genomic information is currently available for S. rueppellii. Without genomic information for such beneficial predator species, we are unable to perform comparative analyses of insecticide target-sites and genes encoding metabolic enzymes potentially responsible for insecticide resistance, between crop pests and their predators. These metabolic mechanisms include several gene families - cytochrome P450 monooxygenases (P450s), ATP binding cassette transporters (ABCs), glutathione-S-transferases (GSTs), UDP-glycosyltransferases (UGTs) and carboxyl/choline esterases (CCEs). METHODS AND FINDINGS: In this study, a high-quality near-chromosome level de novo genome assembly (as well as a mitochondrial genome assembly) for S. rueppellii has been generated using a hybrid approach with PacBio long-read and Illumina short-read data, followed by super scaffolding using Hi-C data. The final assembly achieved a scaffold N50 of 87Mb, a total genome size of 537.6Mb and a level of completeness of 96% using a set of 1,658 core insect genes present as full-length genes. The assembly was annotated with 14,249 protein-coding genes. Comparative analysis revealed gene expansions of CYP6Zx P450s, epsilon-class GSTs, dietary CCEs and multiple UGT families (UGT37/302/308/430/431). Conversely, ABCs, delta-class GSTs and non-CYP6Zx P450s showed limited expansion. Differences were seen in the distributions of resistance-associated gene families across subfamilies between S. rueppellii and some hemipteran crop pests. Additionally, S. rueppellii had larger numbers of detoxification genes than other pollinator species. CONCLUSION AND SIGNIFICANCE: This assembly is the first published genome for a predatory member of the Syrphidae family and will serve as a useful resource for further research into selectivity and potential tolerance of insecticides by beneficial predators. Furthermore, the expansion of some gene families often linked to insecticide resistance and selectivity may be an indicator of the capacity of this predator to detoxify IPM selective insecticides. These findings could be exploited by targeted insecticide screens and functional studies to increase effectiveness of IPM strategies, which aim to increase crop yields by sustainably and effectively controlling pests without impacting beneficial predator populations.


Assuntos
Dípteros , Inseticidas , Animais , Cromossomos , Dípteros/genética , Tamanho do Genoma , Humanos , Resistência a Inseticidas/genética , Inseticidas/farmacologia
4.
BMC Genomics ; 23(1): 45, 2022 Jan 11.
Artigo em Inglês | MEDLINE | ID: mdl-35012450

RESUMO

BACKGROUND: Orius laevigatus, a minute pirate bug, is a highly effective beneficial predator of crop pests including aphids, spider mites and thrips in integrated pest management (IPM) programmes. No genomic information is currently available for O. laevigatus, as is the case for the majority of beneficial predators which feed on crop pests. In contrast, genomic information for crop pests is far more readily available. The lack of publicly available genomes for beneficial predators to date has limited our ability to perform comparative analyses of genes encoding potential insecticide resistance mechanisms between crop pests and their predators. These mechanisms include several gene/protein families including cytochrome P450s (P450s), ATP binding cassette transporters (ABCs), glutathione S-transferases (GSTs), UDP-glucosyltransferases (UGTs) and carboxyl/cholinesterases (CCEs). METHODS AND FINDINGS: In this study, a high-quality scaffold level de novo genome assembly for O. laevigatus has been generated using a hybrid approach with PacBio long-read and Illumina short-read data. The final assembly achieved a scaffold N50 of 125,649 bp and a total genome size of 150.98 Mb. The genome assembly achieved a level of completeness of 93.6% using a set of 1658 core insect genes present as full-length genes. Genome annotation identified 15,102 protein-coding genes - 87% of which were assigned a putative function. Comparative analyses revealed gene expansions of sigma class GSTs and CYP3 P450s. Conversely the UGT gene family showed limited expansion. Differences were seen in the distributions of resistance-associated gene families at the subfamily level between O. laevigatus and some of its targeted crop pests. A target site mutation in ryanodine receptors (I4790M, PxRyR) which has strong links to diamide resistance in crop pests and had previously only been identified in lepidopteran species was found to also be present in hemipteran species, including O. laevigatus. CONCLUSION AND SIGNIFICANCE: This assembly is the first published genome for the Anthocoridae family and will serve as a useful resource for further research into target-site selectivity issues and potential resistance mechanisms in beneficial predators. Furthermore, the expansion of gene families often linked to insecticide resistance may be an indicator of the capacity of this predator to detoxify selective insecticides. These findings could be exploited by targeted pesticide screens and functional studies to increase effectiveness of IPM strategies, which aim to increase crop yields by sustainably, environmentally-friendly and effectively control pests without impacting beneficial predator populations.


Assuntos
Heterópteros , Inseticidas , Tisanópteros , Animais , Genoma , Humanos , Resistência a Inseticidas
5.
Sci Rep ; 11(1): 13495, 2021 06 29.
Artigo em Inglês | MEDLINE | ID: mdl-34188110

RESUMO

Wheat is an economically, socially, and nutritionally important crop, however, aphid infestation can often reduce wheat yield through feeding and virus transmission. Through field phenotyping, we investigated aphid resistance in ancestral wheat Triticum monococcum (L.). Aphid (Rhopalosiphum padi (L.), Sitobion avenae (F.) and Metopolophium dirhodum (Wlk.)) populations and natural enemy presence (parasitised mummified aphids, ladybird adults and larvae and lacewing eggs and larvae) on two naturally susceptible wheat varieties, Triticum aestivum (L.) var. Solstice and T. monococcum MDR037, and three potentially resistant genotypes T. monococcum MDR657, MDR045 and MDR049 were monitored across three years of field trials. Triticum monococcum MDR045 and MDR049 had smaller aphid populations, whereas MDR657 showed no resistance. Overall, natural enemy presence was positively correlated with aphid populations; however, MDR049 had similar natural enemy presence to MDR037 which is susceptible to aphid infestation. It is hypothesised that alongside reducing aphid population growth, MDR049 also confers indirect resistance by attracting natural enemies. The observed resistance to aphids in MDR045 and MDR049 has strong potential for introgression into commercial wheat varieties, which could have an important role in Integrated Pest Management strategies to reduce aphid populations and virus transmission.


Assuntos
Afídeos/crescimento & desenvolvimento , Resistência à Doença/fisiologia , Doenças das Plantas/parasitologia , Triticum , Animais , Triticum/crescimento & desenvolvimento , Triticum/parasitologia
6.
Age Ageing ; 50(2): 431-439, 2021 02 26.
Artigo em Inglês | MEDLINE | ID: mdl-32970798

RESUMO

BACKGROUND: Lower nurse staffing levels are associated with increased hospital mortality. Older patients with cognitive impairments (CI) have higher mortality rates than similar patients without CI and may be additionally vulnerable to low staffing. OBJECTIVES: To explore associations between registered nurse (RN) and nursing assistant (NA) staffing levels, mortality and readmission in older patients admitted to general medical/surgical wards. RESEARCH DESIGN: Retrospective cohort. PARTICIPANTS: All unscheduled admissions to an English hospital of people aged ≥75 with cognitive screening over 14 months. MEASURES: The exposure was defined as deviation in staffing hours from the ward daily mean, averaged across the patient stay. Outcomes were mortality in hospital/within 30 days of discharge and 30-day re-admission. Analyses were stratified by CI. RESULTS: 12,544 admissions were included. Patients with CI (33.2%) were exposed to similar levels of staffing as those without. An additional 0.5 RN hours per day was associated with 10% reduction in the odds of death overall (odds ratio 0.90 [95% CI 0.84-0.97]): 15% in patients with CI (OR 0.85 [0.74-0.98]) and 7% in patients without (OR 0.93 [0.85-1.02]). An additional 0.5 NA hours per day was associated with a 15% increase in mortality in patients with no impairment. Readmissions decreased by 6% for an additional 0.5 RN hours in patients with CI. CONCLUSIONS: Although exposure to low staffing was similar, the impact on mortality and readmission for patients with CI was greater. Increased mortality with higher NA staffing in patients without CI needs exploration.


Assuntos
Disfunção Cognitiva , Recursos Humanos de Enfermagem Hospitalar , Idoso , Disfunção Cognitiva/diagnóstico , Mortalidade Hospitalar , Hospitais , Humanos , Readmissão do Paciente , Admissão e Escalonamento de Pessoal , Estudos Retrospectivos , Recursos Humanos
7.
Crop Prot ; 138: 105316, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-33273750

RESUMO

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.

8.
Mol Ecol ; 29(14): 2661-2675, 2020 07.
Artigo em Inglês | MEDLINE | ID: mdl-32510730

RESUMO

The evolution of resistance to drugs and pesticides poses a major threat to human health and food security. Neonicotinoids are highly effective insecticides used to control agricultural pests. They target the insect nicotinic acetylcholine receptor and mutations of the receptor that confer resistance have been slow to develop, with only one field-evolved mutation being reported to date. This is an arginine-to-threonine substitution at position 81 of the nAChR_ß1 subunit in neonicotinoid-resistant aphids. To validate the role of R81T in neonicotinoid resistance and to test whether it may confer any significant fitness costs to insects, CRISPR/Cas9 was used to introduce an analogous mutation in the genome of Drosophila melanogaster. Flies carrying R81T showed an increased tolerance (resistance) to neonicotinoid insecticides, accompanied by a significant reduction in fitness. In comparison, flies carrying a deletion of the whole nAChR_α6 subunit, the target site of spinosyns, showed an increased tolerance to this class of insecticides but presented almost no fitness deficits.


Assuntos
Proteínas de Drosophila/genética , Drosophila melanogaster/genética , Aptidão Genética , Resistência a Inseticidas , Neonicotinoides , Receptores Nicotínicos/genética , Animais , Resistência a Inseticidas/genética , Inseticidas/toxicidade , Mutação , Neonicotinoides/toxicidade
9.
Insect Biochem Mol Biol ; 122: 103388, 2020 07.
Artigo em Inglês | MEDLINE | ID: mdl-32376273

RESUMO

Voltage-gated sodium channels (VGSCs) are a major target site for the action of pyrethroid insecticides and resistance to pyrethroids has been ascribed to mutations in the VGSC gene. VGSCs in insects are encoded by only one gene and their structural and functional diversity results from posttranscriptional modification, particularly, alternative splicing. Using whole cell patch clamping of neurons from pyrethroid susceptible (wild-type) and resistant strains (s-kdr) of housefly, Musca domestica, we have shown that the V50 for activation and steady state inactivation of sodium currents (INa+) is significantly depolarised in s-kdr neurons compared with wild-type and that 10 nM deltamethrin significantly hyperpolarised both of these parameters in the neurons from susceptible but not s-kdr houseflies. Similarly, tail currents were more sensitive to deltamethrin in wild-type neurons (EC15 14.5 nM) than s-kdr (EC15 133 nM). We also found that in both strains, INa+ are of two types: a strongly inactivating (to 6.8% of peak) current, and a more persistent (to 17.1% of peak) current. Analysis of tail currents showed that the persistent current in both strains (wild-type EC15 5.84 nM) was more sensitive to deltamethrin than was the inactivating type (wild-type EC15 35.1 nM). It has been shown previously, that the presence of exon l in the Drosophila melanogaster VGSC gives rise to a more persistent INa+ than does the alternative splice variant containing exon k and we used PCR with housefly head cDNA to confirm the presence of the housefly orthologues of splice variants k and l. Their effect on deltamethrin sensitivity was determined by examining INa+ in Xenopus oocytes expressing either the k or l variants of the Drosophila para VGSC. Analysis of tail currents, in the presence of various concentrations of deltamethrin, showed that the l splice variant was significantly more sensitive (EC50 42 nM) than the k splice variant (EC50 866 nM). We conclude that in addition to the presence of point mutations, target site resistance to pyrethroids may involve the differential expression of splice variants.


Assuntos
Processamento Alternativo , Drosophila melanogaster/fisiologia , Moscas Domésticas/fisiologia , Resistência a Inseticidas/genética , Mutação , Nitrilas/farmacologia , Piretrinas/farmacologia , Canais de Sódio Disparados por Voltagem/genética , Animais , Drosophila melanogaster/genética , Técnicas de Silenciamento de Genes , Moscas Domésticas/genética , Inseticidas/farmacologia , Canais de Sódio Disparados por Voltagem/metabolismo
10.
Pestic Biochem Physiol ; 166: 104562, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32448417

RESUMO

The buff-tailed bumblebee, Bombus terrestris audax is an important pollinator within both landscape ecosystems and agricultural crops. During their lifetime bumblebees are regularly challenged by various environmental stressors including insecticides. Historically the honey bee (Apis mellifera spp.) has been used as an 'indicator' species for 'standard' ecotoxicological testing, but it has been suggested that it is not always a good proxy for other eusocial or solitary bees. To investigate this, the susceptibility of B. terrestris to selected pesticides within the neonicotinoid, pyrethroid and organophosphate classes was examined using acute insecticide bioassays. Acute oral and topical LD50 values for B. terrestris against these insecticides were broadly consistent with published results for A. mellifera. For the neonicotinoids, imidacloprid was highly toxic, but thiacloprid and acetamiprid were practically non-toxic. For pyrethroids, deltamethrin was highly toxic, but tau-fluvalinate only slightly toxic. For the organophosphates, chlorpyrifos was highly toxic, but coumaphos practically non-toxic. Bioassays using insecticides with common synergists enhanced the sensitivity of B. terrestris to several insecticides, suggesting detoxification enzymes may provide a level of protection against these compounds. The sensitivity of B. terrestris to compounds within three different insecticide classes is similar to that reported for honey bees, with marked variation in sensitivity to different insecticides within the same insecticide class observed in both species. This finding highlights the need to consider each compound within an insecticide class in isolation rather than extrapolating between different insecticides in the same class or sharing the same mode of action.


Assuntos
Inseticidas , Animais , Abelhas , Combinação de Medicamentos , Ecossistema , Glicerol , Salicilatos
11.
Insect Biochem Mol Biol ; 115: 103247, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31626952

RESUMO

The diamondback moth, Plutella xylostella, is a damaging pest of cruciferous crops, and has evolved resistance to many of the insecticides used for control, including members of the diamide class. Previous work on the molecular basis of resistance to diamides has documented mutations in the target-site, the ryanodine receptor, in resistant populations of P. xylostella worldwide. In contrast the role of metabolic resistance to this insecticide class is significantly less clear. Here we show that overexpression of a flavin-dependent monooxgenase (FMO) confers resistance to the diamide chlorantraniliprole in P. xylostella. Transcriptome profiling of diamide resistant strains, with and without target-site resistance, revealed constitutive over-expression of several transcripts encoding detoxification enzymes compared to susceptible strains. Two of these, CYP6BG1, and PxFMO2 were particularly highly overexpressed (33,000 and 14,700-fold, respectively) in a resistant strain (HAW) lacking target-site resistance. After 17 generations without diamide selection the resistance of the HAW strain fell by 52-fold and the expression of PxFMO2 by > 1300-fold, however, the expression of CYP6BG1 declined by only 3-fold. Generation of transgenic Drosophila melanogaster expressing these genes demonstrated that PxFMO2, but not CYP6BG1, confers resistance in vivo. Overexpression of PxFMO2 in the HAW strain is associated with mutations, including a putative transposable element insertion, in the promoter of this gene. These enhance the expression of a reporter gene when expressed in a lepidopteran cell line suggesting they are, at least in part, responsible for the overexpression of PxFMO2 in the resistant strain. Our results provide new evidence that insect FMOs can be recruited to provide resistance to synthetic insecticides.


Assuntos
Família 6 do Citocromo P450/metabolismo , Inseticidas , Mariposas/enzimologia , Oxigenases/metabolismo , ortoaminobenzoatos , Animais , Feminino , Perfilação da Expressão Gênica , Inativação Metabólica , Resistência a Inseticidas , Masculino
12.
Insect Biochem Mol Biol ; 111: 103171, 2019 08.
Artigo em Inglês | MEDLINE | ID: mdl-31136794

RESUMO

Recent work has shown that two bumblebee (Bombus terrestris) cytochrome P450s of the CYP9Q subfamily, CYP9Q4 and CYP9Q5, are important biochemical determinants of sensitivity to neonicotinoid insecticides. Here, we report the characterisation of a third P450 gene CYP9Q6, previously mis-annotated in the genome of B. terrestris, encoding an enzyme that metabolises the N-cyanoamidine neonicotinoids thiacloprid and acetamiprid with high efficiency. The genomic location and complete ORF of CYP9Q6 was corroborated by PCR and its metabolic activity characterised in vitro by expression in an insect cell line. CYP9Q6 metabolises both thiacloprid and acetamiprid more rapidly than the previously reported CYP9Q4 and CYP9Q5. We further demonstrate a direct, in vivo correlation between the expression of the CYP9Q6 enzyme in transgenic Drosophila melanogaster and an increased tolerance to thiacloprid and acetamiprid. We conclude that CYP9Q6 is an efficient metaboliser of N-cyanoamidine neonicotinoids and likely plays a key role in the high tolerance of B. terrestris to these insecticides.


Assuntos
Abelhas/enzimologia , Sistema Enzimático do Citocromo P-450/metabolismo , Neonicotinoides/metabolismo , Tiazinas/metabolismo , Animais , Animais Geneticamente Modificados , Abelhas/genética , Abelhas/metabolismo , Linhagem Celular , Sistema Enzimático do Citocromo P-450/genética , Drosophila melanogaster/genética , Drosophila melanogaster/metabolismo , Resistência a Inseticidas/genética , Mariposas
13.
PLoS Genet ; 15(2): e1007903, 2019 02.
Artigo em Inglês | MEDLINE | ID: mdl-30716069

RESUMO

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.


Assuntos
Abelhas/efeitos dos fármacos , Abelhas/genética , Neonicotinoides/farmacologia , Animais , Evolução Biológica , Sistema Enzimático do Citocromo P-450/genética , Europa (Continente) , Genômica/métodos , Inseticidas/farmacologia , Polinização/efeitos dos fármacos , Polinização/genética , Tiazinas/farmacologia
14.
Artigo em Inglês | MEDLINE | ID: mdl-30106739

RESUMO

Many non-model organisms lack reference genomes and the sequencing and de novo assembly of an organism's transcriptome is an affordable means by which to characterize the coding component of its genome. Despite the advances that have made this possible, assembling a transcriptome without a known reference usually results in a collection of full-length and partial gene transcripts. The downstream analysis of genes represented as partial transcripts then often requires further experimental work in the laboratory in order to obtain full- length sequences. We have explored whether partial transcripts, encoding genes of interest present in de novo assembled transcriptomes of a model and non-model insect species, could be further extended by iterative mapping against the raw transcriptome sequencing reads. Partial sequences encoding cytochrome P450s and carboxyl/cholinesterase were used in this analysis because they are large multigene families and exhibit significant variation in expression. We present an effective method to improve the continuity of partial transcripts in silico that, in the absence of a reference genome, maybe a quick and cost-effective alternative to their extension by laboratory experimentation. Our approach resulted in the successful extension of incompletely assembled transcripts, often to full length. We experimentally validated these results \textit{in silico} and using real-time PCR and sequencing.

15.
Curr Biol ; 28(14): R772-R773, 2018 07 23.
Artigo em Inglês | MEDLINE | ID: mdl-30040932

RESUMO

Neonicotinoids are a class of pesticides widely used in agriculture. In this Quick Guide, Bass and Field explain their mechanism of action and discuss the evidence supporting a potential role in pollinator declines.


Assuntos
Controle de Insetos , Insetos/efeitos dos fármacos , Resistência a Inseticidas , Inseticidas/farmacologia , Neonicotinoides/farmacologia , Agricultura , Animais , Abelhas/efeitos dos fármacos , Evolução Biológica , Inseticidas/efeitos adversos , Neonicotinoides/efeitos adversos , Polinização , Dinâmica Populacional
16.
Curr Biol ; 28(7): 1137-1143.e5, 2018 04 02.
Artigo em Inglês | MEDLINE | ID: mdl-29576476

RESUMO

The impact of neonicotinoid insecticides on the health of bee pollinators is a topic of intensive research and considerable current debate [1]. As insecticides, certain neonicotinoids, i.e., N-nitroguanidine compounds such as imidacloprid and thiamethoxam, are as intrinsically toxic to bees as to the insect pests they target. However, this is not the case for all neonicotinoids, with honeybees orders of magnitude less sensitive to N-cyanoamidine compounds such as thiacloprid [2]. Although previous work has suggested that this is due to rapid metabolism of these compounds [2-5], the specific gene(s) or enzyme(s) involved remain unknown. Here, we show that the sensitivity of the two most economically important bee species to neonicotinoids is determined by cytochrome P450s of the CYP9Q subfamily. Radioligand binding and inhibitor assays showed that variation in honeybee sensitivity to N-nitroguanidine and N-cyanoamidine neonicotinoids does not reside in differences in their affinity for the receptor but rather in divergent metabolism by P450s. Functional expression of the entire CYP3 clade of P450s from honeybees identified a single P450, CYP9Q3, that metabolizes thiacloprid with high efficiency but has little activity against imidacloprid. We demonstrate that bumble bees also exhibit profound differences in their sensitivity to different neonicotinoids, and we identify CYP9Q4 as a functional ortholog of honeybee CYP9Q3 and a key metabolic determinant of neonicotinoid sensitivity in this species. Our results demonstrate that bee pollinators are equipped with biochemical defense systems that define their sensitivity to insecticides and this knowledge can be leveraged to safeguard bee health.


Assuntos
Abelhas/fisiologia , Sistema Enzimático do Citocromo P-450/efeitos dos fármacos , Inseticidas/toxicidade , Neonicotinoides/toxicidade , Animais , Abelhas/efeitos dos fármacos , Abelhas/metabolismo
17.
Pest Manag Sci ; 73(9): 1873-1882, 2017 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-28195678

RESUMO

BACKGROUND: Metabolic resistance is an important consideration in the whitefly Bemisia tabaci, where an esterase-based mechanism has been attributed to pyrethroid resistance and over-expression of the cytochrome P450, CYP6CM1, has been correlated to resistance to imidacloprid and other neonicotinoids. RESULTS: In vitro interactions between putative synergists and CYP6CM1, B and Q-type esterases were investigated, and structure-activity relationship analyses allowed the identification of chemical structures capable of acting as inhibitors of esterase and oxidase activities. Specifically, methylenedioxyphenyl (MDP) moieties with a polyether chain were preferable for optimum inhibition of B-type esterase, whilst corresponding dihydrobenzofuran structures were potent for the Q-esterase variation. Potent inhibition of CYP6CM1 resulted from structures which contained an alkynyl chain with a terminal methyl group. CONCLUSIONS: Synergist candidates could be considered for field control of B. tabaci, especially to abrogate neonicotinoid resistance. © 2017 Society of Chemical Industry.


Assuntos
Hemípteros/enzimologia , Resistência a Inseticidas , Desintoxicação Metabólica Fase I , Animais , Inibidores Enzimáticos/farmacologia , Esterases/antagonistas & inibidores , Esterases/metabolismo , Hemípteros/efeitos dos fármacos , Hemípteros/metabolismo , Ligação Proteica
18.
Eur Biophys J ; 46(7): 675-679, 2017 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-28070661

RESUMO

The pyrethroid insecticides are a very successful group of compounds that have been used extensively for the control of arthropod pests of agricultural crops and vectors of animal and human disease. Unfortunately, this has led to the development of resistance to the compounds in many species. The mode of action of pyrethroids is known to be via interactions with the voltage-gated sodium channel. Understanding how binding to the channel is affected by amino acid substitutions that give rise to resistance has helped to elucidate the mode of action of the compounds and the molecular basis of their selectivity for insects vs mammals and between insects and other arthropods. Modelling of the channel/pyrethroid interactions, coupled with the ability to express mutant channels in oocytes and study function, has led to knowledge of both how the channels function and potentially how to design novel insecticides with greater species selectivity.


Assuntos
Inseticidas/farmacologia , Piretrinas/farmacologia , Agonistas do Canal de Sódio Disparado por Voltagem/metabolismo , Animais , Proteínas de Insetos/química , Proteínas de Insetos/metabolismo , Agonistas do Canal de Sódio Disparado por Voltagem/química
19.
Pest Manag Sci ; 73(2): 371-379, 2017 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-27174317

RESUMO

BACKGROUND: Piperonyl butoxide (PBO) is a well-known insecticide synergist capable of interacting with phase 1 metabolic enzymes, specifically esterases and cytochrome P450s. In this study, structure-activity relationship analyses were used to characterise the interaction of around 30 analogues of PBO with the esterase FE4 and the P450 CYP6CY3 from insecticide-resistant Myzus persicae (Sulzer), in order to predict the synthesis of more potent inhibitors. RESULTS: Enzyme inhibition studies were performed against esterase and oxidase activities and, together with in silico modelling, key activity determinants of the analogues were identified and optimised. Novel analogues were then designed and synthesised, some of which showed greater inhibition against both enzymatic systems: specifically, dihydrobenzofuran moieties containing an alkynyl side chain and a butyl side chain against FE4, and benzodioxole derivatives with a propyl/butyl side chain and an alkynyl ether moiety for CYP6CY3. CONCLUSIONS: In vitro assays identified potential candidate synergists with high inhibitory potency. The in vivo confirmation of such results will allow consideration for a possible use in agriculture. © 2016 Society of Chemical Industry.


Assuntos
Afídeos/enzimologia , Sinergistas de Praguicidas , Butóxido de Piperonila/análogos & derivados , Animais , Simulação por Computador , Sistema Enzimático do Citocromo P-450/metabolismo , Esterases/antagonistas & inibidores , Resistência a Inseticidas , Sinergistas de Praguicidas/química , Butóxido de Piperonila/química , Relação Estrutura-Atividade
20.
Neurotoxicology ; 60: 224-233, 2017 May.
Artigo em Inglês | MEDLINE | ID: mdl-27246647

RESUMO

Diamide insecticides, such as flubendiamide and chlorantraniliprole, are a new class of insecticide with a novel mode of action, selectively activating the insect ryanodine receptor (RyR). They are particularly active against lepidopteran pests of cruciferous vegetable crops, including the diamondback moth, Plutella xylostella. However, within a relatively short period following their commercialisation, a comparatively large number of control failures have been reported in the field. In this review we summarise the current body of knowledge regarding the molecular mechanisms of diamide resistance in P. xylostella. Resistant phenotypes collected from different countries can often be linked to specific target-site mutation(s) in the ryanodine receptors' transmembrane domain. Metabolic mechanisms of resistance have also been proposed. Rapid resistance development is probably a consequence of over-reliance on this one class of chemistry for diamondback moth control.


Assuntos
Diamida/farmacologia , Proteínas de Insetos/genética , Resistência a Inseticidas , Inseticidas/farmacologia , Mariposas/efeitos dos fármacos , Canal de Liberação de Cálcio do Receptor de Rianodina/genética , Aminoácidos/genética , Animais , Proteínas de Insetos/metabolismo , Mariposas/genética , Polimorfismo Genético , RNA Mensageiro/metabolismo , Canal de Liberação de Cálcio do Receptor de Rianodina/metabolismo
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