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1.
Pestic Biochem Physiol ; 200: 105837, 2024 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-38582599

RESUMO

Susceptibility to insecticides is one of the limiting factors preventing wider adoption of natural enemies to control insect pest populations. Identification and selective breeding of insecticide tolerant strains of commercially used biological control agents (BCAs) is one of the approaches to overcome this constraint. Although a number of beneficial insects have been selected for increased tolerance to insecticides the molecular mechanisms underpinning these shifts in tolerance are not well characterised. Here we investigated the molecular mechanisms of enhanced tolerance of a lab selected strain of Orius laevigatus (Fieber) to the commonly used biopesticide spinosad. Transcriptomic analysis showed that spinosad tolerance is not a result of overexpressed detoxification genes. Molecular analysis of the target site for spinosyns, the nicotinic acetylcholine receptor (nAChR), revealed increased expression of truncated transcripts of the nAChR α6 subunit in the spinosad selected strain, a mechanism of resistance which was described previously in insect pest species. Collectively, our results demonstrate the mechanisms by which some beneficial biological control agents can evolve insecticide tolerance and will inform the development and deployment of insecticide-tolerant natural enemies in integrated pest management strategies.


Assuntos
Inseticidas , Receptores Nicotínicos , Tisanópteros , Animais , Tisanópteros/metabolismo , Inseticidas/toxicidade , Resistência a Inseticidas/genética , Agentes de Controle Biológico/farmacologia , Receptores Nicotínicos/genética , Receptores Nicotínicos/metabolismo , Insetos/genética , Macrolídeos/farmacologia , Combinação de Medicamentos
2.
Insect Biochem Mol Biol ; 159: 103983, 2023 08.
Artigo em Inglês | MEDLINE | ID: mdl-37380137

RESUMO

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.


Assuntos
Inseticidas , Lepidópteros , Mariposas , Solanum lycopersicum , Animais , Inseticidas/farmacologia , Diamida , Resistência a Inseticidas/genética , Drosophila melanogaster , Difosfato de Uridina
3.
Insects ; 13(11)2022 Oct 30.
Artigo em Inglês | MEDLINE | ID: mdl-36354820

RESUMO

A mutation showing a distinct orange color in the nymph stages was found in Orius laevigatus (Fieber) (Hemiptera: Anthocoridae), a key biological control agent in protected crops, used to control small pests, especially thrips. A laboratory strain carrying this body color mutation ambar was established. Genetic analysis determined that the mutation (ambar) was controlled by a single autosomal recessive allele. Some biological and ecological characteristics of this orange strain were compared to a normal population. Longevity, fecundity and fertility were similar in both populations, but immature survival, development rate, body size, starvation tolerance and predation capacity were inferior in the orange strain. The utility of the orange mutant as a visible marker for biological and ecological studies of this important biological control agent is discussed.

4.
Insects ; 13(9)2022 Sep 12.
Artigo em Inglês | MEDLINE | ID: mdl-36135528

RESUMO

The reproductive potential of biological control agents (BCAs) is crucial for efficient mass-rearing and field performance, and it all begins with mating. Fecundity can be strongly influenced by intrinsic conditions, such as female age and, often neglected, male age and mating status. However, little is known about the impact of parental status at mating on female reproductive outcomes in BCAs. Orius laevigatus (Fieber) (Hemiptera: Anthocoridae) is widely used to control thrips in protected crops. We evaluated how many consecutive females a male could successfully mate and the effect on a female's reproductive output. In addition, we studied the effects of male and female age on mating. In the multiple mating experiment, the males showed a high capacity to fertilize females successively, not reducing fecundity until the sixth mated female. In the delayed mating experiment, copulation duration and fecundity increased with male age but decreased with female age. In contrast, fertility followed an opposite pattern, increasing with female age but decreasing with male age. However, fecundity gains outweighed fertility declines in both sexes. Therefore, reproductive capacity is increased when mating newly emerged females with males a few days old. The implications of our results for mass rearing and field performance are discussed.

5.
Insects ; 13(3)2022 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-35323549

RESUMO

Orius laevigatus is a key tool for the success of augmentative biological control programs in protected crops. This biological control agent is mass-reared feeding on eggs of Ephestia kuehniella. However, this factitious prey is expensive, accounting for a significant percentage of the rearing costs. Therefore, there is a need to optimize the amount of Ephestia eggs needed per individual leading to a cost reduction, which in turn will favor biological control adoption. This study investigated the effect of the amount of Ephestia eggs provided on the developmental and reproductive fitness of O. laevigatus. At least a daily supply of 1 and 3 Ephestia eggs was needed for optimal development of the first two nymphal instars, respectively, although for maximum survival, 1 egg was enough for both instars. For subsequent development until adulthood, a minimum of 8 eggs per day were needed to fully support growth, but only 3 eggs for optimal survival. Similarly, male body size was also maximized by feeding 8 eggs, but for maximum female body size 10 eggs per day were required. Oviposition rate of females increased with the daily number of Ephestia eggs provided, until a plateau was reached at 8 eggs/day. Benefits and savings for industrial production of O. laevigatus are discussed.

6.
Front Plant Sci ; 12: 647347, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34497617

RESUMO

Studies on the selection of floral traits usually consider pollinators and sometimes herbivores. However, humans also exert selection on floral traits of ornamental plants. We compared the preferences of bumblebees (Bombus terrestris), thrips (Frankliniella occidentalis), and humans for flowers of snapdragon. From a cross of two species, Antirrhinum majus and Antirrhinum linkianum, we selected four Recombinant Inbred Lines (RILs). We characterised scent emission from whole flowers and stamens, pollen content and viability, trichome density, floral shape, size and colour of floral parts. We tested the preferences of bumblebees, thrips, and humans for whole flowers, floral scent bouquets, stamen scent, and individual scent compounds. Humans and bumblebees showed preferences for parental species, whereas thrips preferred RILs. Colour and floral scent, in combination with other floral traits, seem relevant phenotypes for all organisms. Remarkably, visual traits override scent cues for bumblebees, although, scent is an important trait when bumblebees cannot see the flowers, and methyl benzoate was identified as a key attractant for them. The evolutionary trajectory of flowers is the result of multiple floral traits interacting with different organisms with different habits and modes of interaction.

7.
Sci Rep ; 11(1): 18651, 2021 09 20.
Artigo em Inglês | MEDLINE | ID: mdl-34545138

RESUMO

The African citrus psyllid Trioza erytreae is one of the major threats to citrus industry as the vector of the incurable disease known as huanglongbing (HLB) or citrus greening. The psyllid invaded the northwest of the Iberian Peninsula 6 years ago. The invasion alarmed citrus growers in the Mediterranean basin, the largest citrus producing area in Europe, which is still free of HLB. Before our study, no research had been carried out on the genetic diversity of T. erytreae populations that have invaded the Iberian Peninsula and the archipelagos of the Macaronesia (Madeira and the Canary Islands). In this study, combining microsatellites markers and mtDNA barcoding analysis, we characterize the genetic diversity, structure and maternal relationship of these new invasive populations of T. erytreae and those from Africa. Our results suggest that the outbreaks of T. erytreae in the Iberian Peninsula may have derived from the Canary Islands. The populations of T. erytreae that invaded Macaronesia and the Iberian Peninsula are likely to have originated from southern Africa. We anticipate our results to be a starting point for tracking the spread of this invasive pest outside of Africa and to be important for optimizing contingency and eradication plans in newly invaded and free areas.


Assuntos
DNA Mitocondrial/genética , Hemípteros/genética , Repetições de Microssatélites/genética , Animais , Citrus/química , Código de Barras de DNA Taxonômico/métodos , Europa (Continente) , Insetos Vetores , Espécies Introduzidas/tendências , Mitocôndrias/genética , Filogenia , Doenças das Plantas
8.
Commun Biol ; 4(1): 847, 2021 07 07.
Artigo em Inglês | MEDLINE | ID: mdl-34234279

RESUMO

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.


Assuntos
Afídeos/genética , Evolução Molecular , Variação Genética , Genoma de Inseto/genética , Resistência a Inseticidas/genética , Inseticidas/farmacologia , Animais , Afídeos/classificação , Afídeos/fisiologia , Sequência de Bases , Genômica/métodos , Geografia , Interações Hospedeiro-Parasita/efeitos dos fármacos , Mutação , Filogenia , Plantas/parasitologia , Polimorfismo de Nucleotídeo Único , Homologia de Sequência do Ácido Nucleico
9.
Pest Manag Sci ; 77(5): 2539-2546, 2021 May.
Artigo em Inglês | MEDLINE | ID: mdl-33481316

RESUMO

BACKGROUND: Insecticide resistance in the natural enemies of pest species is a very desirable trait. It allows better integration of biological control with the synthetic and natural compounds applied to manage certain pests, especially secondary pest outbreaks. Insecticide resistance in predatory insects has been documented for lady beetles and lacewings, but intriguingly no cases of field-evolved resistance have been noted in any heteropteran predator. In this work, we first explored the variation in susceptibility to pyrethroids in wild and commercial populations of Orius laevigatus (Fieber). Second, we exploited this genetic variation to artificially select a strain of O. laevigatus resistant to pyrethroids. RESULTS: We found significant variation [median lethal dose (LC50 ) 1.6-77.0 mg L-1 ] in susceptibility to pyrethroids in wild populations of the heteropteran predator O. laevigatus, with a baseline LC50 value of 14.6 mg L-1 . We successfully selected a strain of O. laevigatus highly resistant to pyrethroids (LC50 = 1059.9 mg L-1 ). In addition, such resistance was expressed in every instar, particularly in the last nymphal stages, increasing the resilience of the whole population present in the crop facing pyrethroid application. CONCLUSION: The level of resistance achieved may be sufficient to allow survival of adults and nymphs of O. laevigatus exposed to the maximum field rate of several pyrethroids and natural pyrethrins, widely used to control a number of pests in organic and integrated pest management crops. Therefore, this strain resistant to pyrethrins and pyrethroids would improve the resilience of biocontrol protocols, which is an urgent requirement for wider adoption of biological control. © 2021 Society of Chemical Industry.


Assuntos
Heterópteros , Inseticidas , Piretrinas , Animais , Resistência a Inseticidas/genética , Inseticidas/farmacologia , Controle Biológico de Vetores , Comportamento Predatório , Piretrinas/farmacologia
10.
Pestic Biochem Physiol ; 166: 104583, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32448413

RESUMO

Insecticides of the tetronic/tetramic acid family (cyclic ketoenols) are widely used to control sucking pests such as whiteflies, aphids and mites. They act as inhibitors of acetyl-CoA carboxylase (ACC), a key enzyme for lipid biosynthesis across taxa. While it is well documented that plant ACCs targeted by herbicides have developed resistance associated with mutations at the carboxyltransferase (CT) domain, resistance to ketoenols in invertebrate pests has been previously associated either with metabolic resistance or with non-validated candidate mutations in different ACC domains. A recent study revealed high levels of spiromesifen and spirotetramat resistance in Spanish field populations of the whitefly Bemisia tabaci that was not thought to be associated with metabolic resistance. We confirm the presence of high resistance levels (up to >640-fold) against ketoenol insecticides in both Spanish and Australian B. tabaci strains of the MED and MEAM1 species, respectively. RNAseq analysis revealed the presence of an ACC variant bearing a mutation that results in an amino acid substitution, A2083V, in a highly conserved region of the CT domain. F1 progeny resulting from reciprocal crosses between susceptible and resistant lines are almost fully resistant, suggesting an autosomal dominant mode of inheritance. In order to functionally investigate the contribution of this mutation and other candidate mutations previously reported in resistance phenotypes, we used CRISPR/Cas9 to generate genome modified Drosophila lines. Toxicity bioassays using multiple transgenic fly lines confirmed that A2083V causes high levels of resistance to commercial ketoenols. We therefore developed a pyrosequencing-based diagnostic assay to map the spread of the resistance alleles in field-collected samples from Spain. Our screening confirmed the presence of target-site resistance in numerous field-populations collected in Sevilla, Murcia and Almeria. This emphasizes the importance of implementing appropriate resistance management strategies to prevent or slow the spread of resistance through global whitefly populations.


Assuntos
Acetil-CoA Carboxilase , Resistência a Inseticidas , Animais , Austrália , Mutação , Espanha
11.
Pestic Biochem Physiol ; 166: 104582, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32448428

RESUMO

A key to effective insect pest management and insecticide resistance management is to provide growers with a range of new tools as potential alternatives to existing compounds or approaches. Sulfoxaflor (Isoclast™ active) is a new sulfoximine insecticide which is active on a broad range of sap-feeding insects, including species that have reduced susceptibility to currently used insecticides, such as imidacloprid from the neonicotinoid class. Sulfoxaflor (SFX) and imidacloprid (IMI) were tested in laboratory bioassays to compare the susceptibility of field populations of green peach aphid, Myzus persicae (Sulzer), exhibiting varying degrees of resistance involving an alteration (R81T) to the insect nicotinic acetylcholine receptor. The LC50 values for M. persicae exposed to SFX ranged from 0.09 to 1.31 (mg litre-1), whereas when the same populations were exposed to IMI the LC50 values ranged from 0.6 to 76.2 (mg litre-1). M. persicae were significantly more sensitive to SFX as compared to IMI for nine of the 13 populations tested. For M. persicae populations confirmed to be homozygous susceptible (ss) or heterozygous rs) for the R81T point mutation, there was no significant differences in the observed LC50 values for either SFX or IMI relative to the susceptible reference population (15LP1). However, in all M persicae populations that were homozygous (rr) for the R81T point mutation, susceptibility was significantly less to IMI as compared to the reference population with resistance ratios ranging from 22.1 to 63.5-fold. In contrast, only one homozygous resistant population (15MP9) exhibited a statistically significant change in susceptibility (RR = 10-fold) to SFX as compared to the reference population, which was far less than the 56-fold observed for imidacloprid in that same population. Thus, this study indicates there is no specific correlation between the laboratory efficacy of SFX and IMI in field collected populations in Spain displaying varying degrees of resistance to IMI. Furthermore, the presence of target site resistance in M. persicae to IMI, in the form of the R81T mutation, does not a priori translate to a reduction in sensitivity to sulfoxaflor. Consequently, SFX can be an effective tool for use in insect pest management programs for green peach aphid. These data also serve as a baseline reference for green peach aphid sensitivity to SFX prior to commercial uses in Spain.


Assuntos
Afídeos , Prunus persica , Receptores Nicotínicos , Animais , Mutação , Neonicotinoides , Nitrocompostos , Piridinas , Espanha , Compostos de Enxofre
12.
Pest Manag Sci ; 76(11): 3517-3526, 2020 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-32281233

RESUMO

Biological control is an efficient pest control method but there are still limitations that are hindering its wider adoption. Genetic improvement of biological control agents (BCAs) can help to overcome these constraints, but the choice of key attributes for better performance that need to be selected is still an open question. Several characteristics have been suggested but the harsh reality is that selective breeding of BCAs has received a lot of attention but resulted in very little progress. Identifying the appropriate traits to be prioritized may be the first step to reverse this situation. In our opinion, the best way is to look at the factors limiting the performance of key BCAs, especially generalist predators (pesticide compatibility, prey-density dependence, non-suitable crops, and extreme environmental conditions), and according to these challenges, to choose the attributes that would allow BCAs to overcome those limitations. The benefits of selection for higher resistance to toxins, whether artificially applied (pesticides) or plant produced (plant defenses); increased fitness when feeding on non-prey food (supplemented or plant-derived); and better adaptation to extreme temperature and humidity are discussed. In conclusion, genetic improvement of BCAs can bring about new opportunities to biocontrol industry and users to enhance biocontrol resilience. © 2020 Society of Chemical Industry.


Assuntos
Artrópodes , Animais , Artrópodes/genética , Agentes de Controle Biológico , Produtos Agrícolas/genética , Controle Biológico de Vetores
13.
Pest Manag Sci ; 75(8): 2079-2085, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-30785238

RESUMO

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.


Assuntos
Evolução Molecular , Resistência a Inseticidas/genética , Inseticidas/farmacologia , Mariposas/efeitos dos fármacos , Fenótipo , Sequência de Aminoácidos , Substituição de Aminoácidos , Animais , Sequência de Bases , Combinação de Medicamentos , Inglaterra , Proteínas de Insetos/química , Proteínas de Insetos/genética , Proteínas de Insetos/metabolismo , Larva/efeitos dos fármacos , Larva/genética , Larva/crescimento & desenvolvimento , Macrolídeos/farmacologia , Mariposas/genética , Mariposas/crescimento & desenvolvimento , Alinhamento de Sequência , ortoaminobenzoatos/farmacologia
14.
Pest Manag Sci ; 75(1): 45-52, 2019 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-30009510

RESUMO

BACKGROUND: Spiromesifen and spirotetramat are novel insecticides belonging to the chemical class of tetronic and tetramic acid derivatives. Both compounds have proven very effective against field populations of Bemisia tabaci around the world. However, several growers have recently reported control failures in Spain. Therefore, we studied the resistance level to these insecticides in field populations reporting control failures. In addition, we further selected a spiromesifen-resistant strain to study the mechanisms involved and the cross-resistance pattern. RESULTS: All the new field populations collected were significantly more resistant to spiromesifen than the susceptible population, confirming the presence of resistance. Several populations showing high levels of resistance to spiromesifen (>10 000-fold), exhibited cross-resistance to spirotetramat, but resistance ratios were much lower (130-fold). The spiromesifen laboratory-selected strain was very resistant to spiromesifen (LC50 > 30 000 mg L-1 ) and spirotetramat (LC50 = 368.1 mg L-1 ), but lacks any cross-resistance to other insecticides, thus providing options for resistance management. None of the synergists tested significantly restored the susceptibility of B. tabaci to either spiromesifen or spirotetramat. CONCLUSION: This is the first report of resistance to spiromesifen and spirotetramat in B. tabaci, and such high levels of resistance have not been reported before in any field collected pest. Our results suggest that enhanced detoxification does not critically contribute to resistance to ketoenols in B. tabaci. The obvious lack of a metabolic resistance mechanism either suggests a target-site resistance mechanism or a metabolic mechanism insensitive to the synergists tested. © 2018 Society of Chemical Industry.


Assuntos
Compostos Aza/farmacologia , Hemípteros/efeitos dos fármacos , Resistência a Inseticidas , Inseticidas/farmacologia , Compostos de Espiro/farmacologia , Animais , Hemípteros/crescimento & desenvolvimento , Hemípteros/fisiologia , Ninfa/efeitos dos fármacos , Ninfa/crescimento & desenvolvimento , Ninfa/fisiologia , Espanha
15.
Pest Manag Sci ; 74(7): 1552-1557, 2018 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-29377447

RESUMO

BACKGROUND: Cyantraniliprole is a novel anthranilic diamide insecticide that acts on a broad spectrum of insect pests, exclusively by activating their ryanodine receptors. Cyantraniliprole is very effective against whitefly and it presents a favorable ecotoxicological profile. In this study, the baseline susceptibility to cyantraniliprole of Trialeurodes vaporariorum populations from the Mediterranean area was established in nymphal systemic uptake bioassays. RESULTS: The bioassay data showed that the susceptibility to cyantraniliprole varied among the strains collected across the Mediterranean basin. The 50% lethal concentration (LC50 ) range of cyantraniliprole for 16 field populations was from 0.017 to 0.194 mg L-1 , a 11.4-fold natural variability between the least and most sensitive populations. These LC50 values are similar to those reported in a previous study of the use of cyantraniliprol against another species of whitefly, Bemisia tabaci [LC50  = 0.048 (0.034-0.063) mg L-1 ]. CONCLUSION: The current study confirmed the effectiveness of cyantraniliprole against T. vaporarioum strains, adding to the evidence that cyantraniliprole is a promising tool for use in integrated pest management programs. Future shifts in the susceptibility of whitefly field populations to cyantraniliprole may be documented according to the baseline susceptibility range of the populations tested in this research. © 2018 Society of Chemical Industry.


Assuntos
Hemípteros/efeitos dos fármacos , Resistência a Inseticidas , Inseticidas/farmacologia , Pirazóis/farmacologia , ortoaminobenzoatos/farmacologia , Animais , Hemípteros/crescimento & desenvolvimento , Hemípteros/fisiologia , Região do Mediterrâneo , Ninfa/efeitos dos fármacos , Ninfa/crescimento & desenvolvimento , Ninfa/fisiologia
16.
Front Microbiol ; 8: 1969, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-29067021

RESUMO

Pest control in agriculture employs diverse strategies, among which the use of predatory insects has steadily increased. The use of several species within the genus Orius in pest control is widely spread, particularly in Mediterranean Europe. Commercial mass rearing of predatory insects is costly, and research efforts have concentrated on diet manipulation and selective breeding to reduce costs and improve efficacy. The characterisation and contribution of microbial symbionts to Orius sp. fitness, behaviour, and potential impact on human health has been neglected. This paper provides the first genome sequence level description of the predominant culturable facultative bacterial symbionts associated with five Orius species (O. laevigatus, O. niger, O. pallidicornis, O. majusculus, and O. albidipennis) from several geographical locations. Two types of symbionts were broadly classified as members of the genera Serratia and Leucobacter, while a third constitutes a new genus within the Erwiniaceae. These symbionts were found to colonise all the insect specimens tested, which evidenced an ancestral symbiotic association between these bacteria and the genus Orius. Pangenome analyses of the Serratia sp. isolates offered clues linking Type VI secretion system effector-immunity proteins from the Tai4 sub-family to the symbiotic lifestyle.

17.
Insect Biochem Mol Biol ; 80: 11-20, 2017 01.
Artigo em Inglês | MEDLINE | ID: mdl-27845250

RESUMO

Insect ryanodine receptors (RyR) are the molecular target-site for the recently introduced diamide insecticides. Diamides are particularly active on Lepidoptera pests, including tomato leafminer, Tuta absoluta (Lepidoptera: Gelechiidae). High levels of diamide resistance were recently described in some European populations of T. absoluta, however, the mechanisms of resistance remained unknown. In this study the molecular basis of diamide resistance was investigated in a diamide resistant strain from Italy (IT-GELA-SD4), and additional resistant field populations collected in Greece, Spain and Brazil. The genetics of resistance was investigated by reciprocally crossing strain IT-GELA-SD4 with a susceptible strain and revealed an autosomal incompletely recessive mode of inheritance. To investigate the possible role of target-site mutations as known from diamondback moth (Plutella xylostella), we sequenced respective domains of the RyR gene of T. absoluta. Genotyping of individuals of IT-GELA-SD4 and field-collected strains showing different levels of diamide resistance revealed the presence of G4903E and I4746M RyR target-site mutations. These amino acid substitutions correspond to those recently described for diamide resistant diamondback moth, i.e. G4946E and I4790M. We also detected two novel mutations, G4903V and I4746T, in some of the resistant T. absoluta strains. Radioligand binding studies with thoracic membrane preparations of the IT-GELA-SD4 strain provided functional evidence that these mutations alter the affinity of the RyR to diamides. In combination with previous work on P. xylostella our study highlights the importance of position G4903 (G4946 in P. xylostella) of the insect RyR in defining sensitivity to diamides. The discovery of diamide resistance mutations in T. absoluta populations of diverse geographic origin has serious implications for the efficacy of diamides under applied conditions. The implementation of appropriate resistance management strategies is strongly advised to delay the further spread of resistance.


Assuntos
Proteínas de Insetos/genética , Resistência a Inseticidas/genética , Inseticidas , Mariposas/fisiologia , Mutação Puntual , Canal de Liberação de Cálcio do Receptor de Rianodina/genética , Sequência de Aminoácidos , Animais , Benzamidas/farmacologia , Clonagem Molecular , DNA Complementar/genética , DNA Complementar/metabolismo , Proteínas de Insetos/química , Proteínas de Insetos/metabolismo , Mariposas/genética , Canal de Liberação de Cálcio do Receptor de Rianodina/química , Canal de Liberação de Cálcio do Receptor de Rianodina/metabolismo , Sulfonas/farmacologia , ortoaminobenzoatos/farmacologia
18.
Mol Ecol ; 25(22): 5692-5704, 2016 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-27748560

RESUMO

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.


Assuntos
Processamento Alternativo , Éxons , Insetos/genética , Resistência a Inseticidas/genética , Receptor Nicotínico de Acetilcolina alfa7/genética , Animais , Combinação de Medicamentos , Humanos , Macrolídeos
19.
Pest Manag Sci ; 71(7): 1030-6, 2015 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-25138719

RESUMO

BACKGROUND: The whitefly Bemisia tabaci Gennadius is a severe pest in many field and greenhouse crops worldwide and has developed resistance to insecticides from most chemical classes. The ease with which this pest develops resistance makes it essential to incorporate new compounds with different modes of action and no cross-resistance with those previously used into insecticide resistance management strategies. To that end, the systemic effect of the new diamide cyantraniliprole was tested with multiresistant, selected and field populations of Q-biotype B. tabaci from the Mediterranean area. RESULTS: Bioassays with multiresistant and laboratory-selected populations indicated no cross-resistance to cyantraniliprole in the B. tabaci strains exhibiting resistance to other insecticides. The LC50 values for nymphs from 14 field populations varied between 0.011 and 0.116 mg L(-1), a 10.5-fold natural variability. The LC50 values for adults from three strains ranged from 0.060 to 0.096 mg L(-1). CONCLUSION: These baseline data will be helpful for monitoring future potential shifts in susceptibility to cyantraniliprole in Mediterranean whitefly populations within an IRM programme. Cyantraniliprole may play an important role in mitigating insecticide resistance in B. tabaci because of its high efficacy and its lack of cross-resistance to other insecticides, even in multiresistant Q-biotype populations collected from a highly problematic insecticide resistance area.


Assuntos
Hemípteros , Resistência a Inseticidas , Inseticidas , Pirazóis , ortoaminobenzoatos , Animais , Bioensaio , Região do Mediterrâneo , Ninfa
20.
Pest Manag Sci ; 71(8): 1068-74, 2015 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-25138907

RESUMO

BACKGROUND: Frankliniella occidentalis is a worldwide economically important pest. Scarcity of effective products and cross-resistance issues make resistance to existing insecticides a recurring problem that requires the development of new control tools, such as incorporating novel compounds. Lethal effects of cyantraniliprole on adults and larvae from field and insecticide-resistant populations were evaluated. In addition, the sublethal effects on biological features such as fecundity, fertility, feeding, oviposition and mating were studied. RESULTS: Results obtained for larvae produced LC50 values ranging from 33.4 to 109.2 mg L(-1) , with a low natural variability (3.3-fold) and a LC50 composite value of 52.2 mg L(-1) . The susceptibility for adults was 23-fold lower than for larvae. No evidence of cross-resistance between cyantraniliprole and established insecticides used against thrips was evident. Relevant sublethal effects of cyantraniliprole were demonstrated, including reduced fecundity, fertility, feeding, oviposition and mating success. CONCLUSION: Low variation in susceptibility across contemporary populations of F. occidentalis and a lack of cross-resistance to other insecticides indicate that cyantraniliprole is a potential candidate in rotation programmes within an insecticide resistance management strategy. The combined sublethal effect on reproduction will have an important impact on population reduction. Available data indicate that cyantraniliprole is likely to be a valuable tool for managing thrips populations.


Assuntos
Controle de Insetos , Inseticidas/farmacologia , Pirazóis/farmacologia , Tisanópteros/efeitos dos fármacos , ortoaminobenzoatos/farmacologia , Animais , Comportamento Alimentar/efeitos dos fármacos , Feminino , Larva/efeitos dos fármacos , Dose Letal Mediana , Masculino , Reprodução/efeitos dos fármacos , Tisanópteros/crescimento & desenvolvimento
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