Um dos inseticidas mais utilizados para o controle do psilídeo Diaphorina citri (Hemiptera: Liviidae) em pomares de citros no Brasil tem sido o neonicotinoide imidacloprid. Para subsidiar um programa de Manejo da Resistência de D. citri a imidacloprid foram realizados estudos de monitoramento da suscetibilidade a inseticidas em populações de D. citri provenientes das principais regiões citrícolas do Estado de São Paulo, interações de imidacloprid com inseticidas e fungicidas, e avaliação da viabilidade de associação de imidacloprid com o parasitoide Tamarixia radiata (Hymenoptera: Eulophidae). Para a caracterização de linhas básicas de suscetibilidade de D. citri aos inseticidas imidacloprid, deltamethrin e dimethoate foram utilizados bioensaios de contato residual. Foram definidas as concentrações diagnósticas de 56 mg de imidacloprid/L água (CL95), 32 mg de deltamethrin/L água (CL90) e 56 mg de dimethoate/L água (CL95) para o monitoramento da suscetibilidade de 25 populações de D. citri entre 2010 e 1012. Não ocorreram diferenças na sobrevivência aos inseticidas testados em populações de psilídeos coletados em pomares de citros com diferentes intensidades de pulverização com inseticidas. A maior sobrevivência de insetos nas concentrações diagnósticas foi observada em 2010, com valores de sobrevivência variando de 4,7% a 24,0% para imidacloprid, de 0,9% a 11,8% para deltamethrin e de 5,2% a 13,0% para dimethoate. Não foram observados aumentos significativos na sobrevivência de D. citri nos monitoramentos realizados em 2011 e 2012. As interações de imidacloprid com deltamethrin ou dimethoate foram testadas com as CL25 dos respectivos inseticidas em bioensaio de contato residual. As interações foram aditivas sobre a mortalidade de adultos em condições de campo e de casa de vegetação. A mistura de imidacloprid com buprofezin e pyriproxyfen sobre ninfas de 3º instar também foi aditiva. A atividade biológica das concentrações campo de imidacloprid (40?L/mL) e buprofezin (375?g/mL) e da mistura dos mesmos mostrou uma degradação da atividade similar para o controle de D. citri em condições de campo e casa de vegetação. A avaliação da persistência da mistura de imidacloprid e pyriproxyfen (avaliado nas concentrações de 6,25?g/mL e 100?g/mL) indicou que pyriproxyfen teve degradação mais rápida que imidacloprid. A mistura de imidacloprid na CL25 para D. citri com a concentração de campo dos fungicidas Benzimidazol (500?g/mL de tiofanato-metilico), estrubilurina (37,5?g /mL de piraclostrobina) e triazol (50?g/mL de difenenoconazol) não afetaram o desempenho de imidacloprid. Houve significativa redução na taxa instantânea de crescimento (ri) de D. citri quando expostas a diferentes idades de resíduos de imidacloprid pulverizado na concentração de 40?g/mL, mesmo sobre resíduos com 56 dias de idade, levando à extinção da população ou em processo de extinção. A exposição a CL5 (0,501?g/mL), CL10 (0,804?g/mL), CL25 (1,995?g/mL) e CL50 (5,213?g/mL) de imidacloprid para D. citri reduziu o crescimento populacional de D. citri, mas não causou sua extinção. Contudo, essas concentrações de imidacloprid afetaram a ri do parasitoide com a CL50 causando sua extinção. / One of the most widely used insecticides for control of Diaphorina citri (Hemiptera: Liviidae) in citrus groves in Brazil has been the neonicotinoid imidacloprid. To implement an Insect Resistance Management program of D. citri to imidacloprid, studies were conducted to monitor the susceptibility to insecticides in D. citri populations collected from main citrus production regions of São Paulo State, to evaluate the interactions of imidacloprid with insecticides and fungicides, and to assess the feasibility of imidacloprid association with the parasitoid Tamarixia radiata (Hymenoptera: Eulophidae). Residual contact bioassays were used to characterize the baseline susceptibility of D. citri to the insecticides imidacloprid, deltamethrin and dimethoate. Diagnostic concentrations of 56 mg of imidacloprid/L water (LC95), 32 mg of deltamethrin/L water (LC90) and 56 mg of dimethoate/L water (LC95) were defined for monitoring the susceptibility in 25 populations of D. citri from 2010 to 2012. There were no differences in survival to the insecticides tested in D. citri populations collected in citrus groves with different regimes of insecticide use. The highest survival of insects at diagnostic concentrations was observed in 2010, with survival values ranging from 4.7% to 24.0% for imidacloprid, from 0.9% to 11.8% for deltamethrin and 5.2% to 13.0% dimethoate. There were no significant increases in survival of D. citri on monitoring conducted in 2011 and 2012. The interactions of imidacloprid with deltamethrin or dimethoate were tested with the LC25 of each insecticide with residual contact bioassays. The interactions of these insecticides were additive on D. citri adult mortality under field or greenhouse conditions. The interactions of imidacloprid with buprofezin and pyriproxyfen were additive on 3rd instar nymphs. The biological activity at field rates of imidacloprid (40?l/mL) and buprofezin (375?g/mL) as well as the mixture of these insecticides showed a similar degradation in the activity to control D. citri under field and greenhouse conditions. The evaluation of the persistence of the mixture of pyriproxyfen and imidacloprid (evaluated at concentrations of 6.25 ?g/mL and 100?g/mL) indicated that pyriproxyfen degradation was faster than imidacloprid. The mixture of LC25 of imidacloprid to D. citri with fungicides field rates of benzimidazole (500?g/mL of thiophanate-methil), strobilurin (37.5 ?g/mL of pyraclostrobin), and triazole (50?g/mL of difenoconazole) did not affect the performance of imidacloprid. A significant reduction in the instantaneous rate of increase (ri) of D. citri was observed when exposed to different residue ages of imidacloprid sprayed at concentration of 40?g/mL, even when exposed to residues of 56-day old, by leading to their extinction or in extinction process. The exposure to CL5 (0.501 ?g/mL), CL10 (0.804 ?g/mL), LC25 (1.995 ?g/mL) and LC50 (5.213 ?g/mL) of imidacloprid to D. citri did not cause their extinction. However, these concentrations of imidacloprid affected the ri of the parasitoid causing their extinction at LC50.

Transgenic plants containing genes from Bacillus thuringiensis have been used as an alternative to chemical insecticides for insect pest control. The vegetative insecticidal proteins (Vip) secreted during the vegetative growth phase of bacteria are considered a second generation of insecticidal proteins since they do not share any structural or sequence homology with previously used crystal proteins (Cry) as well as having a wide insecticidal spectrum. One of the target pests for this protein is the fall armyworm (FAW) (Spodoptera frugiperda), the most important corn pest in South America. Previously it has been controlled by insecticides and maize expressing Cry proteins, but has rapidly evolved resistance to many control practices and remains a top concern for sustainable biotechnology control efforts. Thus, resistance characterization involving mode of action and genetics of resistance can help with Insect Resistance Management strategies, and improve the durability of control. In this dissertation, using two selected FAW population resistant to Vip3Aa20 Bt protein (Vip-R1and Vip-R2) we generated comparative proteomic and transcriptomic data among resistant and susceptible colonies. In the chapter 2, we bring FAW biology/ecology and Brazilian agriculture landscape data to support the high adaptive potential of this pest to genetically modified maize expressing Bt Cry proteins in Brazil. Proteomics studies in the chapter 3 revealed that neither Vip-R1 nor Vip-R2 showed difference between resistant and susceptible colonies either for Vip3Aa20 activation through proteolysis assay nor protein binding to the receptor. Transcriptomic sequencing and RNA-seq analysis in the chapter 4 showed strong evidence of ABC transporter genes associated with resistance as well as genes related to G-protein signaling pathway as downregulated. These results will be discussed in context of providing best management practices for managing FAW resistance to Vip, and extending the durability of Vip technology. / Plantas Transgênicas expressando genes de Bacillus thuringiensis (Bt) tem sido usadas como alternativa ao controle químico para controle de insetos praga. A proteina Vip (Vegetative Insecticide Protein) cuja secreção é realizada durante fase de crescimento da bacteria é considerada como segunda geração de proteinas inseticidas em função desta não apresentar similaridade de sequencias com todas as outras proteinas cristal (Cry), apresentando ainda maior espectro de controle de pragas. Uma das pragas alvo desta proteina é a lagarta-do-cartucho do milho (Spodoptera frugiperda), considerada a mais importante na cultura do milho na América do Sul. Larvas desta espécie foram sempre controladas com inseticidas e mais recentemente, milho expressando proteínas Cry. No entanto, esta praga tem desenvolvido resistência para várias ferramentas de controle, trazendo preocupação para a sustentabilidade das taticas de controle geradas através da biotecnologia. Dessa forma, estudos de caracterização da resistencia envolvendo modo de ação e characteristicas genéticas envolvidas com resistência pode contribuir para melhorar estratégias de Manejo de Resistencia de Insetos (IRM) e aumentar a durabilidade destas tecnologias para o controle. Nesta dissertação, foi gerado dados proteômicos e de transcriptoma comparando uma população de S. frugiperda resistente a Vip3Aa20 com a susceptivel. No capítulo 2, abordamos as características de bio-ecologia da praga associado ao sistema de cultivo suportando o alto potencial adaptativo desta espécie para hibridos de milho expressando proteinas Bt no Brazil. No capitulo 3, estudos de proteômica mostrou que Vip-R1 e Vip-R2 quando comparado com SUS, não demostraram diferenças para ativação da proteina nem ausencia de ligação da proteína com receptor de membrana no intestino do inseto. Dados de transcriptoma descritos no capitulo 5 mostrou forte evidências de que a baixa expressão de genes relacionados ao sistema transportador ABC pode estar associado com resistência bem como genes da via de sinalização das proteínas G. Estes resultados serão discutidos em um contexto para suportar boas praticas de manejo de resistência para lagarta-do-cartucho e assim estender a durabilidade da tecnologia Viptera® no campo.

<p dir="ltr">Palmer amaranth (Amaranthus palmeri S. Watts) and waterhemp [Amaranthus tuberculatus (Moq.) J. D. Sauer] are two of the most troublesome weeds in U.S. soybean production and the auxin herbicides dicamba and 2,4-D, are currently used extensively for postemergence control of these species. In some cases, complete control of weeds at the time of auxin application is not achieved due to adverse environmental conditions, plant factors or misapplications. In these instances, a subsequent postemergence herbicide may be required to control any plant that survived the initial auxin herbicide application. This research was conducted to determine the efficacy and optimal successive time interval between applications of viable postemergence herbicides in soybean on Palmer amaranth and waterhemp plants surviving a previous application of 2,4-D or dicamba. Results from this research indicated that respraying a failed auxin herbicide application with a subsequent auxin herbicide, especially dicamba, was less effective than respraying with glufosinate or fomesafen to control waterhemp in addition to being a less desirable approach for resistance management. Additionally, respray herbicide applications should target 7 to 14 d after the initial failed herbicide application on waterhemp for optimal overall efficacy. When dicamba was the initial herbicide sprayed on Palmer amaranth, 94% or greater control was achieved with glufosinate in 2019 on a 7- and 14-d respray interval, which was greater than the efficacy observed with either dicamba or 2,4-D (< 82%). In 2020, these differences in herbicide efficacy were not evident within these time intervals. These general trends in treatment differences were also manifested in the data for plant height, biomass and viable apical meristems. When 2,4-D was the initial herbicide sprayed on Palmer amaranth, fomesafen and glufosinate applications on a 7-d respray interval and glufosinate on a 14-d respray interval resulted in greater control than 2,4-D in 2019. All other trends in herbicide treatments and time intervals were consistent with dicamba applied as the initial herbicide. Another part of his research was conducted with the objective of quantifying herbicide spray solution deposition and herbicide efficacy on waterhemp growing in different densities plant densities: low, high-thinned and high densities. In both field and greenhouse experiments, spray deposition (µl cm-2) on waterhemp leaves was up to 53% less on plants grown in high density compared to the other density treatments. Even though no differences in herbicide deposition between low and the high-thinned densities were observed, there were differences in herbicide efficacy. When applied to plants growing in low density, applications of glufosinate, fomesafen and topramezone reduced weed biomass to a greater extent than plants growing in the high-thinned density. Following herbicide damage to the apical meristem, plants growing in high-density produced new branches from axillary buds that were previously dormant, whereas plants growing in the low density already had axillary branches initiated from these buds and did not provide an opportunity for new shoot growth. In general, no differences in herbicide efficacy were observed across weed densities following dicamba applications. Source-to-sink translocation of dicamba to previously active meristems, or axillary buds that broke dormancy after the herbicide application, may have compensated for less spray solution interception on plants growing in high density. Lower levels of postemergence herbicide efficacy in high density weed populations are not only influenced by spray deposition differences, but also by changes in plant growth and apical dominance. This research provides further evidence that justifies the need for weed managers to reduce weed densities as much as possible, via non-chemical or soil residual herbicides (preemergence), as much as possible to optimize the efficacy of foliar herbicide applications.</p>

Roughly 79-100% of maize in the United States (US) is treated with a neonicotinoid seed treatment (NST), and transgenic (GMO) maize, Zea mays L. (Poaceae), that produces insecticidal toxins by way of genes derived from Bacillus thuringiensis (Bt), occupies more than 75% of maize acreage. Among a variety of secondary pests targeted by NSTs, the primary soil-dwelling pest targeted by Bt maize is the western corn rootworm (WCR), Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysomelidae). Transgenic Bt technology has dramatically reduced insecticide use for WCR, and insect resistance poses the greatest threat to its utility. To delay resistance to Bt traits, in 2010 the US Environmental Protection Agency (EPA) approved a "refuge-in-a-bag" (RIB) insect resistance management (IRM) strategy, where 5% of seeds do not express Bt toxins (i.e., "refuge" maize). The RIB strategy is intended to preserve Bt trait effectiveness if mating between 'resistant' insects from Bt plants and 'susceptible' insects from refuge plants occurs at a high enough frequency. Investigations into the effectiveness of RIB for WCR have shown that beetles emerged from Bt plants tend to vastly outnumber beetles emerged from refuge plants, which contributes to low rates of mixed mating. Large proportions of Bt beetles is one of several factors that contributes to resistance development, and resistance to all currently-available WCR-Bt traits has been documented. I conducted field experiments in two regions (Indiana and Virginia) comparing refuge beetle proportions in NST-treated (NST+) and NST-untreated (NST-) 5% RIB maize, to determine whether NSTs may be limiting refuge beetle emergence. To assess advantages of combining use of Bt and NSTs, I compared stand, root injury rating, and yield between NST+, NST-, Bt and non-Bt maize in both states. I also measured neonicotinoid residues in soil, water, and stream sediment within and surrounding fields of maize, to study the off-site movement and soil residence time of these compounds. I found that 5% seed blends did not produce large populations of refuge beetles in any site-year, and that NSTs showed inconsistent effects on refuge beetle populations. Treatment comparisons showed inconsistent benefits of NSTs when combined with Bt traits. I detected neonicotinoid residues in soil matrices throughout the growing season (range: 0 – 417.42 ppb), including prior to planting, suggesting year-round presence of these compounds. My results suggest that, while the effects of NSTs on Bt IRM may be inconsistent, the benefits of universally applying NSTs to Bt maize for soil pests may not be worth the ecological costs of doing so in all cases. / Doctor of Philosophy / About 75% of maize planted in the United States is encoded with genetic traits allowing them to manufacture insecticidal proteins which are toxic to key pests. These insecticidal proteins, known as "Bt toxins," are highly specific to targeted insects, and are virtually non-toxic all other animals and non-target insects. One target insect is the western corn rootworm (WCR), which feeds on and damages maize roots. In the past, WCR was controlled by applying millions of kilograms of chemical insecticides every year, at ever-increasing rates, to compensate for reduced effectiveness due to pest resistance. "Bt" plants, were released for commercial use in the late 1990s; they reduced the need for growers to apply chemical insecticides for managing key pests. The Environmental Protection Agency established regulations aiming to maintain the effectiveness of Bt technology, which they declared have "provided substantial human health, environmental, and economic benefit." This Insect Resistance Management (IRM) strategy, also known as the "refuge" strategy, is meant to preserve Bt-susceptible insects so they can pass on their Bt-susceptible genes to successive generations. The refuge strategy works by incorporating a certain percentage of non-Bt plants per every field of Bt plants; if enough insects survive on these "refuge" plants to outnumber "resistant" insects, population-wide Bt-susceptibility may be maintained. While this strategy has been effective for other key maize pests, it is failing for WCR, evidenced by WCR populations that have developed Bt-resistance. We know current refuge configurations aren't producing enough refuge-WCR to mix sufficiently with resistant WCR, likely due to insufficient refuge sizes. I wanted to know whether the refuge strategy could be improved, if increasing refuge proportions is not an option. Considering that Bt toxins are only effective against a narrow range of insects, seed manufacturers always coat seeds with an insecticide to protect seedlings against other insects. These coatings, or neonicotinoid seed treatments (NSTs), are included on nearly all seed, including those reserved for Bt refuges. I conducted two years of field trials to investigate whether removing NSTs from refuge seeds would improve WCR-IRM by providing an insecticide-free "refuge." My results suggest that removing NSTs may increase refuge beetle proportions, however my results also show that refuge plant proportions are simply too small to support large enough refuge-WCR populations to delay resistance, regardless of whether NSTs are present or not. While NSTs may provide extra comfort to growers at little additional cost, questions regarding their necessity at current use patterns have been raised. Several studies have shown inconsistent benefits, and others have shown longer-than-expected persistence in the environment, movement into streams and groundwater, and even alterations to insect and non-insect animal communities associated with their use. I conducted research comparing their relative effectiveness against WCR and non-WCR insect pests in fields of Bt maize. Additionally, I scouted for their residues in soil collected in field margins, forests buffering streams, and in water collected from agricultural ditches and waterways neighboring fields. I found that, while NSTs produced higher plant populations, they rarely resulted in greater yields. I detected neonicotinoid residues in soil matrices throughout the growing season, including prior to planting, suggesting year-round presence of these compounds. My results suggest that, while the effects of NSTs on Bt IRM may be inconsistent, the benefits of universally applying NSTs to Bt maize for soil pests may not be worth the ecological costs of doing so in all cases.

<p>Bed bugs (<i>Cimex lectularius</i> L.) are globally important human ectoparasites. Their management necessitates the use of multiple control techniques. Plant-derived essential oils are extracts from aromatic plants that represent one of the alternative control measures for bed bug control, in addition to mechanical options and synthetic pesticides. However, there is limited information available on the efficacy and toxicology of plant essential oils against bed bugs. This project was designed with the aim to provide in-depth information on efficacy, toxicology and mode-of-action of essential oils and their insecticidal constituents in bed bugs. Initially, I evaluated topical and fumigant toxicity of fifteen essential oil components against adult male bed bugs of the Harlan strain (an insecticide susceptible strain). Neurological effects of the six most toxicologically active compounds were also determined. In both topical and fumigant bioassays, carvacrol and thymol were the most active compounds. Spontaneous electrical activity measurements of the bed bug nervous system demonstrated neuroinhibitory effects of carvacrol, thymol and eugenol, whereas linalool and bifenthrin (a pyrethroid class insecticide) produced excitatory effects. Further, I evaluated the efficacy and neurological impacts of a mixture of three neuroinhibitory compounds; carvacrol, eugenol and thymol in 1:1:1 ratio against adult male bed bugs of the Harlan strain. This mixture of monoterpenoids as well as the mixture of synthetic insecticides exhibited a synergistic affect in topical bioassays. In electrophysiology experiments, the monoterpenoid mixture led to higher neuroinhibitory effects, whereas a mixture of synthetic insecticides caused higher neuroexcitatory effects in comparison to single compounds. </p> <p>In the next objective of my dissertation, I compared the efficacy of five plant essential oils (thyme, oregano, clove, geranium and coriander), their major components (thymol, carvacrol, eugenol, geraniol and linalool) and EcoRaider^® (commercial product) between pyrethroid susceptible (Harlan) and field collected (Knoxville) bed bug populations. Initially, I found that the Knoxville strain was 72,893 and 291,626 fold resistant to topically applied deltamethrin (a pyrethroid class insecticide) compared to the susceptible Harlan strain at the LD₂₅ and LD₅₀ lethal dose levels, respectively. Synergist bioassays and detoxification enzyme assays showed that the Knoxville strain possesses significantly higher activity of cytochrome P450 and esterase enzymes. Further, Sanger sequencing revealed the presence of the L925I mutation in the voltage gated sodium channel gene. The Knoxville strain, however, did not show any resistance to plant essential oils, their major components or EcoRaider^® in topical bioassays (resistance ratios of ~ 1). In the final objective, I evaluated the efficacy of binary mixtures of above-mentioned essential oils or their major components or EcoRaider^® with deltamethrin in susceptible and resistant bed bugs. In topical application bioassays, binary mixtures of essential oils or their major components or EcoRaider^® and deltamethrin at the LD₂₅ dose caused a synergistic increase in toxicity in resistant bed bugs. Further, I studied the inhibitory effects of major essential oil components on detoxification enzyme activities (cytochrome P450s, esterases and glutathione transferases). Detoxification enzyme assays conducted using protein extracts from bed bugs pre-treated with essential oil constituents showed that these compounds significantly inhibited cytochrome P450 activity in the resistant strain, but esterase and glutathione transferase activity were unaffected. No inhibition of detoxification enzyme activities was observed in the Harlan strain bed bugs pre-treated with essential oil constituents.</p> <p>In conclusion, my dissertation research has created the foundation for utilization of natural products for bed bug management by (i) describing the efficacy of plant essential oils and their components against bed bugs, (ii) discovering synergistic interactions between essential oil components at the nervous system level, (iii) determining susceptibility of deltamethrin-resistant bed bugs to plant essential oils and their constituents and (iv) identifying synergistic effects of essential oils or their components on toxicity of pyrethroid insecticides and underlying mechanisms of this synergistic interaction. </p> <br>

Commercialisées depuis 1996, les plantes génétiquement modifiées produisant une toxine insecticide (toxine Cry) dérivée de Bacillus thuringiensis et appelées plantes Bt ciblent certains Lépidoptères ou Coléoptères ravageurs. Au fil des ans, les surfaces cultivées en plantes Bt sont de plus en plus importantes et contrôlent de larges populations d'insectes. Pour limiter le risque de développement de populations résistantes, une stratégie agricole appelée 'Haute Dose / Zone Refuge' est actuellement recommandée aux Etats-Unis par l'Environmental Protection Agency. Cette stratégie préventive nécessite la plantation d'une 'zone refuge' composée de plantes non-Bt utilisables par le ravageur ciblé et plantée à proximité de la 'zone Bt' qui synthétise une haute dose de toxine Cry. Mon projet de recherche s’inscrit dans le cadre de l’évaluation de l'efficacité de cette stratégie et s’articule en deux phases : une phase expérimentale et une phase théorique. La première se concentre sur la caractérisation en laboratoire de l'impact des toxines Cry sur la biologie d'un ravageur. Cette phase constitue un support au volet théorique : la mise au point d’un modèle mathématique évaluant l'efficacité de la stratégie HD/R. L'originalité de ce projet repose entre autre sur l'interactivité entre ces deux volets. Volet expérimental. Impact des toxines Cry sur la biologie de Plodia interpunctella. Nous évaluons séparément l'impact d'une gamme de concentrations de deux toxines Cry (CryXX et CryYY) sur une série de paramètres comportementaux et biologiques d'un insecte commun des denrées stockées: Plodia interpunctella (Hübner) (Lepidoptera : Pyralidae). Ces paramètres sont sélectionnés car leur variation pourrait avoir un impact sur l'efficacité de la stratégie HD/R dans le contrôle de la résistance. Il est donc pertinent de les quantifier pour intégrer dans le modèle des ordres de grandeur réalistes et générer des résultats qui ne sont pas uniquement basés sur des spéculations théoriques. Volet théorique A. Efficacité de la stratégie HD/R pour des plantes Bt synthétisant une ou deux toxines simultanément. La stratégie 'HD/R' a été développée pour prévenir la résistance envers les plantes Bt synthétisant une seule toxine. Or, depuis 2003, de nouvelles variétés de coton Bt synthétisant simultanément deux toxines Cry sont commercialisées (BollgardII® et WidestrikeTM). Nous évaluons, grâce au modèle que nous avons développé, l'efficacité de cette stratégie lors d'une utilisation exclusive de plantes Bt synthétisant une ou deux toxines. Volet théorique B. Impact du ralentissement du développement des insectes sur les plantes Bt sur l'efficacité de la stratégie HD/R. Le volet expérimental met en évidence un allongement de la durée du développement des larves se nourrissant sur une diète contaminée en toxine Cry. Ce ralentissement induit une séparation temporelle entre l'émergence des adultes de la zone Bt et de la zone refuge et perturbe une hypothèse principale de la stratégie HD/R: le croisement aléatoire entre adultes, indépendamment du génotype et de la zone d'origine. Dans ce troisième chapitre, nous étudions l'impact de la perturbation du croisement aléatoire sur l'efficacité de la stratégie HD/R. Nous testons également deux options pour optimiser la stratégie en cas d'asynchronie: l'utilisation de plantes Bt synthétisant une faible concentration en toxine (atténuant le décalage entre l'émergence des adultes) ou l'augmentation de la taille de la zone refuge (favorisant la survie des individus porteurs d'allèle de sensibilité et donc optimisant la dilution de la résistance à la génération suivante). Ce travail s'intègre dans une problématique actuelle et utilise des outils de biologie théorique (théories de la dynamique et de la génétique des populations) ainsi que le développement d'un modèle mathématique. Il apporte des éléments de réponse et de réflexion sur l'optimisation de la gestion de la résistance des insectes mais c'est aussi une illustration de la complémentarité entre la biologie expérimentale et théorique. / On the market since 1996, genetically modified plants synthesizing an insecticidal toxin (Cry toxin) stemmed from Bacillus thuringiensis, called Bt plants, target several insect pests (Lepidoptera or Coleoptera). Bt crops cover increasingly larger areas and control important pest populations The Insect Resistance Management Strategy (IRM) strategy currently recommended in the U.S.A. to limit the development of resistant populations is the High Dose / Refuge zone (HD/R) strategy. This pre-emptive strategy requires a refuge zone composed by non-Bt plants, usable by the target insect and in close proximity of the Bt zone synthesizing a high toxin concentration. My research project contributes to the effectiveness assessment of this HD/R strategy. It is structured on two main parts: an experimental, and a theoretical section. The first part characterizes the impact of Cry toxins on the biology of an insect pest. It is the basis of the theoretical part: the implementation of a mathematical model, which evaluates the effectiveness of the HD/R strategy. The originality of this project is based on the interactivity of these two components. Experimental section. Impact of the Cry toxins on the biology of Plodia interpunctella. We assess the impact of a range of concentrations of two Cry toxins (CryXX et CryYY) on several behavioural and biological parameters of a common pest of stored products: Plodia interpunctella (Hübner) (Lepidoptera : Pyralidae). These parameters are selected because their variation could influence the effectiveness of a HD/R strategy. So, it is important to quantify these parameters so that realistic values can be integrated in our model. The results of the model are thus not based on theoretical assumptions alone. Theoretical section A. Effectiveness of a HD/R strategy with Bt plants synthesizing one or two toxins. Initially, the HD/R strategy has been developed to limit the resistance towards Bt plants synthesizing one toxin. However, since 2003, new Bt cotton varieties synthesize two toxins simultaneously (BollgardII® et WidestrikeTM). We assess, with our model, the effectiveness of this strategy for Bt plants synthesizing one or two toxins. Theoretical section B. Impact of the slowing down of the insect development reared on Bt plants on the effectiveness of the HD/R strategy. The experimental part demonstrates that larvae reared on a Bt diet have a protracted development duration. The consequence of this is a temporal separation between adult emergence in the two zones (Bt zone and refuge zone). This could affect the main assumption of the HD/R strategy, i. e. random mating independently of the genotype and of the native zone. In this third chapter, we study the impact of random mating disruption on the effectiveness of a HD/R strategy. We test two options to optimise the strategy in case of asynchrony: the use of Bt plants synthesizing a lower toxin concentration (limiting emergence asynchrony) or increasing the refuge zone size (favouring the survival of insect carrying one or two susceptible allele and thus optimising the dilution of resistance at the next generation). This work is applied to a current issue. It uses some of the tools of theoretical biology (theories of population dynamics and population genetics) and develops a mathematical model. It provides some responses and some elements of thought about insect resistance management. It is also an illustration of the complementarity between experimental and theoretical biology.

Commercialisées depuis 1996, les plantes génétiquement modifiées produisant une toxine insecticide (toxine Cry) dérivée de Bacillus thuringiensis et appelées plantes Bt ciblent certains Lépidoptères ou Coléoptères ravageurs. Au fil des ans, les surfaces cultivées en plantes Bt sont de plus en plus importantes et contrôlent de larges populations d'insectes. Pour limiter le risque de développement de populations résistantes, une stratégie agricole appelée 'Haute Dose / Zone Refuge' est actuellement recommandée aux Etats-Unis par l'Environmental Protection Agency. Cette stratégie préventive nécessite la plantation d'une 'zone refuge' composée de plantes non-Bt utilisables par le ravageur ciblé et plantée à proximité de la 'zone Bt' qui synthétise une haute dose de toxine Cry. <p><p>Mon projet de recherche s’inscrit dans le cadre de l’évaluation de l'efficacité de cette stratégie et s’articule en deux phases :une phase expérimentale et une phase théorique. La première se concentre sur la caractérisation en laboratoire de l'impact des toxines Cry sur la biologie d'un ravageur. Cette phase constitue un support au volet théorique :la mise au point d’un modèle mathématique évaluant l'efficacité de la stratégie HD/R. L'originalité de ce projet repose entre autre sur l'interactivité entre ces deux volets.<p><p>Volet expérimental. Impact des toxines Cry sur la biologie de Plodia interpunctella. Nous évaluons séparément l'impact d'une gamme de concentrations de deux toxines Cry (CryXX et CryYY) sur une série de paramètres comportementaux et biologiques d'un insecte commun des denrées stockées: Plodia interpunctella (Hübner) (Lepidoptera :Pyralidae). Ces paramètres sont sélectionnés car leur variation pourrait avoir un impact sur l'efficacité de la stratégie HD/R dans le contrôle de la résistance. Il est donc pertinent de les quantifier pour intégrer dans le modèle des ordres de grandeur réalistes et générer des résultats qui ne sont pas uniquement basés sur des spéculations théoriques.<p><p>Volet théorique A. Efficacité de la stratégie HD/R pour des plantes Bt synthétisant une ou deux toxines simultanément. La stratégie 'HD/R' a été développée pour prévenir la résistance envers les plantes Bt synthétisant une seule toxine. Or, depuis 2003, de nouvelles variétés de coton Bt synthétisant simultanément deux toxines Cry sont commercialisées (BollgardII® et WidestrikeTM). Nous évaluons, grâce au modèle que nous avons développé, l'efficacité de cette stratégie lors d'une utilisation exclusive de plantes Bt synthétisant une ou deux toxines.<p><p>Volet théorique B. Impact du ralentissement du développement des insectes sur les plantes Bt sur l'efficacité de la stratégie HD/R. Le volet expérimental met en évidence un allongement de la durée du développement des larves se nourrissant sur une diète contaminée en toxine Cry. Ce ralentissement induit une séparation temporelle entre l'émergence des adultes de la zone Bt et de la zone refuge et perturbe une hypothèse principale de la stratégie HD/R: le croisement aléatoire entre adultes, indépendamment du génotype et de la zone d'origine. Dans ce troisième chapitre, nous étudions l'impact de la perturbation du croisement aléatoire sur l'efficacité de la stratégie HD/R. Nous testons également deux options pour optimiser la stratégie en cas d'asynchronie: l'utilisation de plantes Bt synthétisant une faible concentration en toxine (atténuant le décalage entre l'émergence des adultes) ou l'augmentation de la taille de la zone refuge (favorisant la survie des individus porteurs d'allèle de sensibilité et donc optimisant la dilution de la résistance à la génération suivante). <p><p>Ce travail s'intègre dans une problématique actuelle et utilise des outils de biologie théorique (théories de la dynamique et de la génétique des populations) ainsi que le développement d'un modèle mathématique. Il apporte des éléments de réponse et de réflexion sur l'optimisation de la gestion de la résistance des insectes mais c'est aussi une illustration de la complémentarité entre la biologie expérimentale et théorique.<p><p><p>/<p><p>On the market since 1996, genetically modified plants synthesizing an insecticidal toxin (Cry toxin) stemmed from Bacillus thuringiensis, called Bt plants, target several insect pests (Lepidoptera or Coleoptera). Bt crops cover increasingly larger areas and control important pest populations The Insect Resistance Management Strategy (IRM) strategy currently recommended in the U.S.A. to limit the development of resistant populations is the High Dose / Refuge zone (HD/R) strategy. This pre-emptive strategy requires a refuge zone composed by non-Bt plants, usable by the target insect and in close proximity of the Bt zone synthesizing a high toxin concentration.<p><p>My research project contributes to the effectiveness assessment of this HD/R strategy. It is structured on two main parts: an experimental, and a theoretical section. The first part characterizes the impact of Cry toxins on the biology of an insect pest. It is the basis of the theoretical part: the implementation of a mathematical model, which evaluates the effectiveness of the HD/R strategy.<p>The originality of this project is based on the interactivity of these two components.<p><p>Experimental section. Impact of the Cry toxins on the biology of Plodia interpunctella. We assess the impact of a range of concentrations of two Cry toxins (CryXX et CryYY) on several behavioural and biological parameters of a common pest of stored products: Plodia interpunctella (Hübner) (Lepidoptera :Pyralidae). These parameters are selected because their variation could influence the effectiveness of a HD/R strategy. So, it is important to quantify these parameters so that realistic values can be integrated in our model. The results of the model are thus not based on theoretical assumptions alone.<p> <p>Theoretical section A. Effectiveness of a HD/R strategy with Bt plants synthesizing one or two toxins. Initially, the HD/R strategy has been developed to limit the resistance towards Bt plants synthesizing one toxin. However, since 2003, new Bt cotton varieties synthesize two toxins simultaneously (BollgardII® et WidestrikeTM). We assess, with our model, the effectiveness of this strategy for Bt plants synthesizing one or two toxins.<p><p>Theoretical section B. Impact of the slowing down of the insect development reared on Bt plants on the effectiveness of the HD/R strategy. The experimental part demonstrates that larvae reared on a Bt diet have a protracted development duration. The consequence of this is a temporal separation between adult emergence in the two zones (Bt zone and refuge zone). This could affect the main assumption of the HD/R strategy, i. e. random mating independently of the genotype and of the native zone. In this third chapter, we study the impact of random mating disruption on the effectiveness of a HD/R strategy. We test two options to optimise the strategy in case of asynchrony: the use of Bt plants synthesizing a lower toxin concentration (limiting emergence asynchrony) or increasing the refuge zone size (favouring the survival of insect carrying one or two susceptible allele and thus optimising the dilution of resistance at the next generation). <p><p>This work is applied to a current issue. It uses some of the tools of theoretical biology (theories of population dynamics and population genetics) and develops a mathematical model. It provides some responses and some elements of thought about insect resistance management. It is also an illustration of the complementarity between experimental and theoretical biology.<p> / Doctorat en Sciences / info:eu-repo/semantics/nonPublished