Helicoverpa armigera (Hübner) was officially reported in Brazil in 2013. This species is closely related to Helicoverpa zea (Boddie) and has caused significant crop damage in Brazil. The use of genetically modified crops expressing insecticidal protein from Bacillus thuringiensis (Berliner) has been one of the control tactics for managing these pests. Genetically modified maize expressing Vip3Aa20 was approved to commercial use in Brazil in 2009. Understanding the genetic diversity and the susceptibility to B. thuringiensis proteins in H. armigera and H. zea populations in Brazil are crucial for establishing Insect Resistance Management (IRM) programs in Brazil. Therefore, the objectives of this study were: (a) to infer demographic parameters and genetic structure of H. armigera and H. zea Brazil; (b) to assess the intra and interspecific gene flow and genetic diversity of H. armigera and H. zea; and (c) to evaluate the susceptibility to Vip3Aa20 protein in H. armigera and H. zea populations of Brazil. A phylogeographic analysis of field H. armigera and H. zea populations was performed using a partial sequence data from the cytochrome c oxidase I (COI) gene. H. armigera individuals were most prevalent on dicotyledonous hosts and H. zea individuals were most prevalent on maize crops. Both species showed signs of demographic expansion and no genetic structure. High genetic diversity and wide distribution were observed for H. armigera. A joint analysis indicated the presence of Chinese, Indian, and European lineages within the Brazilian populations of H. armigera. In the cross-species amplification study, seven microsatellite loci were amplified; and showed a potential hybrid offspring in natural conditions. Interespecific analyses using the same microsatellite loci with Brazilian H. armigera and H. zea in compare to the USA H. zea were also conducted. When analyses were performed within each species, 10 microsatellites were used for H. armigera, and eight for H. zea. We detected high intraspecific gene flow in populations of H. armigera and H. zea from Brazil and H. zea from the USA. Genetic diversity was similar for both species. However, H. armigera was more similar to H. zea from Brazil than H. zea from the USA and some putative hybrid individuals were found in Brazilian populations.Tthere was low gene flow between Brazilian and USA H. zea. The baseline susceptibility to Vip3Aa20 resulted in low interpopulation variation for H. zea (3-fold) and for H. armigera (5-fold), based on LC50. H. armigera was more tolerant to Vip3Aa20 than H. zea (≈ 40 to 75-fold, based on CL50). The diagnostic concentration for susceptibility monitoring, based on CL99, was fairly high (6,400 ng Vip3Aa20/cm2) for H. zea and not validated for H. armigera due to the high amount of protein needed for bioassays. Implementing IRM strategies to Vip3Aa20 in H. armigera and H. zea will be of a great challenge in Brazil, mainly due to the low susceptibility to Vip3Aa20 and high genetic diversity and gene flow in both species, besides a potential of hybrid individuals between H. armigera and H. zea under field conditions. / Helicoverpa armigera (Hübner) foi oficialmente reportada no Brasil em 2013. Esta espécie é estreitamente relacionada a Helicoverpa zea (Boddie) e tem causado danos significativos nas culturas no Brasil. O uso de plantas geneticamente modificadas, que expressam proteínas inseticidas de Bacillus thuringiensis (Berliner), tem sido uma das táticas de controle para o manejo dessas pragas. O milho geneticamente modificado que expressa Vip3Aa20 foi aprovado para comercialização no Brasil em 2009. O entendimento da diversidade genética e da suscetibilidade às proteínas de B. thuringiensis em populações de H. armigera e H. zea no Brasil são cruciais para o estabelecimento de programas de Manejo da Resistência de Insetos (MRI). Assim, os objetivos desse estudo foram: (a) inferir parâmetros demográficos e estrutura genética de H. armigera e H. zea no Brasil; (b) avaliar o fluxo gênico intra e interespecífico e a diversidade genética em H. armigera e H. zea; e (c) aferir a suscetibilidade de populações brasileiras de H. armigera e H. zea a proteína Vip3Aa20. Uma análise filogeográfica de populações de campo de H. armigera e H. zea foi realizada com o uso de sequências do gene citocromo c oxidase I (COI). Indivíduos de H. armigera foram mais prevalentes em dicotiledôneas e H. zea na cultura do milho. Ambas as espécies mostraram sinais de expansão demográfica e ausência de estrutura genética. Alta diversidade genética e ampla distribuição foram observadas em H. armigera. Análises conjuntas indicaram a presença de linhagens da China, Índia e Europa em populações brasileiras de H. armigera. A partir de um estudo de amplificação cruzada de microssatélites, sete locos amplificaram em ambas as espécies e evidenciaram a possibilidade de hibridização no campo. Estes mesmos locos foram usados para análises interespecíficas de H. armigera e H. zea do Brasil em comparação a H. zea dos EUA. Nas análises para cada espécie, 10 microssatélites foram usados para H. armigera e oito para H. zea. Alto fluxo gênico intraespecífico foi detectado em populações de H. armigera e H. zea. A diversidade genética foi similar em ambas as espécies. H. armigera foi mais similar a H. zea do Brasil que dos EUA e possíveis híbridos foram encontrados nas populações brasileiras. Houve um baixo fluxo gênico entre populações brasileiras e americanas de H. zea. A linha-básica de suscetibilidade a Vip3Aa20 resultou numa variação interpopulacional baixa em H. zea (3 vezes) e em H. armigera (5 vezes), baseada na CL50. H. armigera foi mais tolerante a Vip3Aa20 que H. zea (≈ 40 to 75 vezes, baseado na CL50). A concentração diagnóstica, baseada na CL99, foi bastante alta (6.400 ng Vip3Aa20/cm2) para H. zea e não validada para H. armigera devido à alta quantidade de proteína necessária para os bioensaios. A implementação de estratégias de MRI a Vip3Aa20 em H. armigera e H. zea serão um grande desafio no Brasil, principalmente devido à baixa suscetibilidade a Vip3Aa20 e alta diversidade genética e fluxo gênico em ambas as espécies, além da possibilidade de indivíduos híbridos entre H. armigera e H. zea nas condições de campo.

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.

O cultivo sucessivo de soja em grandes áreas agrícolas contribuíram significativamente para o aumento da incidência de pragas nessa cultura. Os percevejos sugadores são responsáveis por significativas perdas de rendimento, qualidade e potencial germinativo, além de causar a retenção foliar. Sendo assim, os objetivos deste trabalho foram i) avaliar a adaptabilidade e estabilidade em progênies de soja com excelente desempenho agronômico e resistência ao complexo de percevejos em três ambientes; e ii) explorar a viabilidade do uso de diferentes métodos de melhoramento para no desenvolvimento de progênies resistentes ao complexo de percevejos. Para a compreensão da interação genótipo por ambiente, 28 progênies e cinco testemunhas foram avaliados em três ambientes, e os resultados foram submetidos à análise de GGE Biplot. Os caracteres avaliados foram: número de dias para a maturidade, período de granação, produtividade de grãos, peso de cem sementes, retenção foliar e peso sementes boas. A análise gráfica biplot demonstrou que existe interação do tipo complexa entre as progênies avaliadas, e as progênies 7, 9, 8 e 23 foram as que tiveram a maior estabilidade nos três ambientes. Para a análise de viabilidade dos métodos genealógico e MSD (descendência de múltiplas sementes), foram avaliadas duas populações de soja, respectivamente à cada método. Ambas populações são oriundas do cruzamento entre as cultivares IAC-100 (fonte de resistência ao complexo de percevejos) e CD- 215 (suscetível). Os caracteres utilizados para essa análise foram a produtividade de grãos e peso de sementes boas. Foram realizadas análises descritiva de cada método e uma análise de modelos mistos para a verificação da diferença entre eles. A maior média e variabilidade genética foi encontrada na população oriunda do método MSD. Todavia, o método genealógico apresentou 10 progênies maiores que a média da melhor testemunha. Os resultados foram influenciados pela presença da interação genótipo por ambiente, e a diferença entre os métodos foi detectada pelo teste Wald-F. Ambos os métodos foram eficientes para a geração de progênies com alto potencial agronômico e com resistência ao complexo de percevejos. Com isso, é necessário submeter as melhores progênies à ensaios de competição para um futuro lançamento de novas cultivares de soja que reúnam alta produtividade e resistência ao complexo de percevejos. / The continuous cultivation of a given crop in large areas contributes significantly to the increased incidence of pests. One such pest are the sucking insects that can cause significant losses. Stink bugs will feed on developing soybean seeds by piercing and immature pods, causing seed damage and reductions in yield, quality and germination potential, in addition to causing leaf retention. Thus, the objectives of this study were i) to evaluate the adaptability and stability in soybean progenies with excellent agronomic performance and resistance to stink bugs in three environments; and ii) to study the viability to use different breeding methods for development of resistant progeny to stink bugs. To evaluate the genotype-byenvironment interaction, 28 progenies and five experimental checks were evaluated in three environments, and the results submitted to analysis of GGE Biplot. The following traits were evaluated in the field: number of days to maturity, grain filling period, grain yield, weight of one hundred seeds, leaf retention, and weight of healthy seeds. The graphical biplot analysis showed that there is complex interaction between progenies, and the progenies 7, 9, 8 and 23 were those that had the highest stability in all three environments. To analyze the feasibility of pedigree and multiple seeds descent (MSD) methods were evaluated two soybean populations respectively for each method. Both populations were from the cross between IAC-100 (resistance source to stink bug complex) and CD-215 (susceptible). The traits used for this analysis were grain yield and weight of healthy seeds. Descriptive analyses of each method and a mixed model analysis were made to verify the difference between them. The highest average and genetic variability was found in the MSD method derived from the population. However, the pedigree method showed 10 progenies higher than the average of the best experimental check. The results were influenced by the presence of genotype-by-environment and the difference between the methods was detected by the Wald-F test. Both methods were efficient to generated progenies with high agronomic potential and resistance to stink bugs. Thus, it is necessary to submit the best progeny to competition tests for a future release of new soybean cultivars with high productivity and resistance.

Les plantes possèdent de multiples systèmes de défense contre les insectes phytophages. Dans un contexte agronomique, manipuler ces défenses au moyen de la sélection pourrait contribuer à réduire les dommages causés par les insectes ravageurs en augmentant la résistance naturelle des plantes. Cette stratégie se heurte cependant à des contraintes très fortes lorsqu'il s'agit de l'appliquer aux insectes. Après avoir détaillé ces contraintes, nous proposons une démarche alternative aux méthodes classiques. Celle-ci vise à identifier des traits-clés de la plante qui modulent son interaction avec le ravageur. Si de tels traits sont identifiés et validés expérimentalement, ils permettront ensuite de conduire la sélection sans nécessiter d'insecte. Nous avons testé cette démarche dans un système composé du colza (Brassica napus) et du méligèthe Meligethes aeneus, un ravageur majeur de cette culture. Le méligèthe est un coléoptère pollinivore dont les adultes sont généralistes mais ne pondent que sur certaines brassicacées. Les dégâts agronomiques sont causés par ces adultes qui, avant que la floraison ne démarre, détruisent les boutons floraux pour atteindre le pollen qu'ils contiennent. Quatre étapes cruciales de l'interaction ont été étudiées : l'attraction à distance, l'alimentation des adultes, la production et la ponte des œufs, et le développement larvaire. Six génotypes de colza ont été comparés dans une série d'expérimentations au laboratoire. La mise en relation des résultats de préférence/performance de l'insecte avec des profilages métaboliques larges de tissus floraux a permis d'identifier des traits-clés candidats. Les conclusions principales de ce travail sont (i) que la composition biochimique du périanthe est déterminante dans la stimulation de l'alimentation des adultes, et que cette stimulation pourrait être largement sous l'influence d'un petit nombre de composés dont le saccharose ; (ii) que cette stimulation détermine de façon majeure, par un effet domino, la production d'œufs en contraignant l'ovogenèse ; (iii) que la qualité nutritionnelle du pollen impacte à la fois les larves et les adultes, et que cette qualité pourrait être déterminée en bonne partie par la concentration en amidon et en certains glucosinolates (des métabolites secondaires typiques de quelques familles végétales dont les brassicacées). La combinaison des différents résultats obtenus permet également de proposer des hypothèses plus générales, parmi lesquelles le fait que le contexte agronomique dans lequel l'interaction a lieu ait largement influencé, voire perturbé, l'interaction qui liait le méligèthe et les brassicacées sauvages avant que les cultures de colza ne se généralisent. Cette thèse a montré qu'une nouvelle voie était peut-être envisageable pour contribuer à protéger les cultures de façon durable contre les insectes ravageurs, en particulier pour les systèmes agronomiques où les dégâts sont causés à un stade temporairement sensible de la culture. / Plants display multiple defense systems against phytophagous insects. Manipulating these defenses by means of selection could contribute to decrease damages caused by insect pests, by increasing natural resistance of crops. This strategy faces great constraints when applied to insects. We first detail these constraints and then propose an alternative approach to classical methods. It consists in identifying key plant traits that determine the intensity of the interaction between the plant and the pest. If such traits are identified, selection could be conducted on the basis if these sole traits, without needing any insect. We tested this approach in a system composed of oilseed rape (Brassica napus) and the pollen beetle (Meligethes aeneus), a major pest of this culture. This coleopteran generalist pollen feeder lays eggs only on certain brassicaceous plants. Agronomical damage arise before plant flowering and are caused by adults, which destroy flower buds to get the pollen they contain. Four crucial steps of the interaction were studied: attraction, adult feeding, egg production and oviposition, and larval development. Six oilseed rape genotypes were compared in a series of experiments conducted in the laboratory. By linking insect preference/performance to large metabolic profiling of bud tissues, we identified candidate key traits. Main conclusions of this work are (i) that biochemical composition of the perianth, especially a few compounds among which sucrose, is determinant for feeding intensity; (ii) that feeding stimulation has an important impact on egg production by constraining oogenesis; (iii) that pollen nutritional quality, probably mostly determined by starch and some glucosinolates, interacts with both pollen beetle larvae and adults. Combination of several results also allows drawing more general hypotheses about the oilseed rape - pollen beetle interaction. One of these is that the agronomical context in which the interaction takes place may have largely influenced, or even disturbed, the interaction that linked this insect and wild brassicaceous plants before oilseed rape cultivation. This thesis showed that a new way might be possible to protect cultures against insect pests. It could be both efficient and sustainable, especially in systems where agronomical damage is caused at a temporary vulnerable plant stage.

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.

Please read the abstract in the section 07chapter7. / Dissertation (MSc (Genetics))--University of Pretoria, 2002. / Genetics / unrestricted

The green peach aphid (GPA; Myzus persicae Sülzer) is a damaging pest that has a broad host range that includes plants in the Brassicaceae, Solanaceae, Cucurbitaceae, and Rosaceae families. It also vectors several important viral diseases. However, how plants perceive GPA to limit infestation is poorly understood. This study demonstrates an important role for the HOMOLOG OF RPW8 4 (HR4), which encodes a protein that contains the RESISTANCE TO POWDERY MILDEW 8 (RPW8) motif that is found in some intracellular receptor proteins that are involved in defense against pathogens. In the accession Moscow of Arabidopsis thaliana, a Brassicaceae family plant, located at the RPW8 locus on chromosome 3 are RPW8.1 and RPW8.2, which confer resistance against a broad spectrum of powdery mildew pathogens. In comparison, in the powdery mildew-susceptible accession Columbia (Col-0), RPW8.1 and RPW8.2 are replaced by the homologous HR4Col-0. HR4Col-0, but not its homologs, was found to be involved in limiting GPA infestation. The presence of HR4Col-0 adversely impacted GPA fecundity, feeding from the phloem, and host selection. Transcriptomic analysis revealed that HR4Col-0 influences biological processes such as response to chitin, amino acid metabolism, and lipid metabolism. The resistance-enhancement ability of HR4Col-0 towards GPA infestation could be transferred to the Arabidopsis accession Wassilewskija and the oil-seed crop Camelina sativa, both of which naturally lack HR4. These findings position HR4Col-0 as a promising candidate for enhancing resistance to the GPA in plants other than Arabidopsis. This research significantly advances our understanding of plant immune mechanisms against an important agricultural pest.

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.

Hunger ist nach wie vor ein weit verbreitetes Problem. Nach der jüngsten Schätzung der FAO hungern weltweit noch immer über 800 Mio. Menschen. Prognosen gehen davon aus, dass die Agrarproduktion bis 2050 verdoppelt werden muss, um der steigenden Nachfrage nach Agrarprodukten durch Bevölkerungswachstum, Bioenergieproduktion und sich verändernde Ernährungsgewohnheiten zu begegnen. Unter vielen Möglichkeiten, das Welternährungsproblem zu beschreiben, wird in dieser Dissertation das Konzept der Ernährungssicherheit nach der Definition der FAO herangezogen. Ernährungssicherheit umfasst als wichtigste Aspekte die Verfügbarkeit von Nahrungsmitteln und den ökonomischen Zugang zu Nahrungsmitteln. Nahrungsmittelverfügbarkeit ist in erster Linie von einer global ausreichenden Produktion abhängig, der Zugang zu Nahrung von der Kaufkraft der Haushalte. Die Kaufkraft wird sowohl durch das Einkommen wie auch die Agrarpreise beeinflusst. Die Nahrungsmittelverfügbarkeit kann vor allem durch Ertragssteigerungen erhöht werden. Sollen die Ertragssteigerungen ressourcenschonend erfolgen, muss die Produktivität der Agrarproduktion bezogen auf alle Produktionsfaktoren inkl. Umweltgütern steigen. Nur durch technische und organisatorische Innovation kann dies realisiert werden. Ertragssteigerungen in Entwicklungsländern erhöhen gleichzeitig die Einkommen von potenziell von Hunger Betroffenen. Eine Begrenzung der Nachfrage nach Agrargütern kann global in erster Linie in den Bereichen Bioenergie und Konsum tierischer Produkte erfolgen. Simulationen können helfen, den Einfluss einzelner Faktoren der globalen Nachfrage und des Angebots nach Agrarrohstoffen und -produkten auf die Welternährungslage genauer abzuschätzen. Ein Großteil der Simulationen in der Literatur kommt mithilfe von partiellen oder allgemeinen Gleichgewichtsmodellen zu Aussagen über gehandelte Mengen und Preise oder Wohlfahrtseffekte. In dieser Arbeit wird ein einfaches partielles Gleichgewichtsmodell für den Weltmarkt für Biotreibstoffe, Fleisch und Getreide entwickelt und mit der Methode der FAO zur Schätzung der Zahl der Hungernden gekoppelt. Dies ermöglicht die Abschätzung des Einflusses verschiedener Szenarien auf die Zahl der Hungernden. Für das Referenzjahr 2011 werden Szenarien alternativer Nachfrage nach Fleisch und Biotreibstoffen simuliert, ebenso wie ein Szenario alternativer landwirtschaftlicher Produktivität in der Europäischen Union. Die Schätzungen ergeben einen signifikanten Einfluss der simulierten Nachfrage- und Angebotsverschiebungen. Im Vergleich mit dem Status Quo führt eine Reduktion des Fleischkonsums in den OECD-Ländern um 50% zu einer Reduktion der Zahl der Hungernden um 5%. Eine ii Flächenproduktivität im Getreideanbau in der EU auf dem Niveau von 1980 steigert die Zahl der Hungernden um mehr als 5%, ein Anstieg der Biotreibstoffquote in der OECD auf 10% erhöht die Zahl der Hungernden gar um 10%. Die Durchsetzung geeigneter politischer Maßnahmen zur Verbesserung der Welternährungslage kann durch eine breite öffentliche Unterstützung beschleunigt oder erst ermöglicht werden. Über die öffentliche Meinung zu Fragen der Welternährung in Deutschland existieren bisher keine expliziten Studien. In den Themenbereichen landwirtschaftliche Produktion und Nahrungsmittelkonsum stellt das Thema Welternährung nur einen Aspekt dar, der neben Fragen nach z.B. der Lebensmittelsicherheit und Gesundheit sowie den Auswirkungen von Agrarproduktion auf Landschaft und Umwelt steht. Hier werden Ergebnisse einer Online-Befragung zum Thema Welternährung präsentiert, die im März 2012 mit einer repräsentativen Stichprobe 1.200 deutscher Internetnutzer durchgeführt wurde. Die Befragung beinhaltete im Wesentlichen die Bewertung der Wirkung verschiedener potenzieller Ursachen der weltweiten Unterernährung, sowie Maßnahmen und Entwicklungen zu ihrer Lösung. Die Ergebnisse zeigen, dass das Thema Welthunger in erster Linie mit Merkmalen akuter (Hunger-) Krisen in Verbindung gebracht wird. Die vielversprechendsten Lösungsmaßnahmen sind in den Augen der Befragten der Ausbau des Fairen Handels, Absatzmärkte für Kleinbauern sowie ein Lebensmittelspekulationsverbot. Der intensiven Landwirtschaft in der Welt wie auch in Europa sowie insbesondere der Grünen Gentechnik wird kein Potenzial zur Lösung des Problems zugesprochen. Die Antwortmuster lassen auf die Präsenz eines Halo-Effekts mit einer Übertragung unterstellter negativer Umweltwirkungen auf das Thema Welternährung schließen. Die Grüne Gentechnik ist eine Agrartechnologie, die potenziell zur Angebotsausweitung von Agrarprodukten genauso beitragen kann wie zu einer ressourcenschonenderen Landwirtschaft. Bisherige Anwendungen, hauptsächlich Herbizidtoleranz sowie Insektenresistenz, haben u.a. zu einer Steigerung der effektiven Erträge, einer Einsparung von Insektiziden und einer Steigerung der Profite vor allem im Baumwollanbau beigetragen. Trotz zahlreicher Belege besteht über die Auswirkungen von gentechnisch veränderten (GV) Nutzpflanzen eine anhaltende Kontroverse. Ein Großteil der europäischen Bevölkerung lehnt den Anbau ebenso ab wie einen Verzehr von Produkten aus GV-Pflanzen. Unterstellte sozio-ökonomische Auswirkungen oder Auswirkungen auf die Welternährung sind nur zwei Determinanten von vielen, die zu den Einstellungen zur Grünen Gentechnik beitragen. Zur Konsolidierung des Wissens im Bereich agronomischer sowie sozio-ökonomischer Auswirkungen von GV-Pflanzen wurde eine Meta-Analyse durchgeführt. In wissenschaftlichen Datenbanken wurden iii 147 Primärstudien identifiziert, die für die Meta-Analyse verwendet wurden. Die Analyse beschränkt sich auf gentechnisch veränderte Sojabohnen, Mais und Baumwolle. Als mittlere Effekte des Anbaus von GV-Pflanzen ergibt die Meta-Analyse eine Steigerung des Ertrages um 22%, eine Reduktion der Menge der angewendeten Pflanzenschutzmittel um 37% und eine Steigerung der Profite der Bauern um 68%. Die Höhe der Effekte hängt stark von verschiedenen Faktoren ab. Die Effekte sind für alle drei genannten Variablen in Entwicklungsländern betragsmäßig größer als in Industrieländern. Auch sind die Effekte des Anbaus von GV-Pflanzen bei Erträgen und beim Pflanzenschutzmitteleinsatz betragsmäßig höher für insektenresistente Pflanzen als bei solchen mit Herbizidtoleranz. Aus den in der Arbeit gewonnenen Erkenntnissen kann geschlussfolgert werden, dass sowohl das Nahrungsmittelangebot gesteigert als auch die Nachfrage begrenzt werden sollte. Dies verlangt eine standortangepasste Ausnutzung vorhandener und die Entwicklung neuer Technologien. Auch die Gentechnik kann einen Beitrag leisten. Der Kleinbauernsektor muss vordringlich gefördert werden. Zur Begrenzung der Nachfrage können neben der Politik, welche vor allem die Biotreibstoffnachfrage beeinflussen kann, auch Konsumenten durch Verzicht auf tierische Produkte beitragen. Dies wie auch die Durchführung effektiver politischer Maßnahmen sind auf eine unverzerrte Sichtweise der Öffentlichkeit auf das Thema Welternährung angewiesen. Dazu muss das Thema in Wissenschaftskommunikation und Bildung stärker thematisiert werden.

The Russian wheat aphid (RWA) (Diuraphis noxia Kurdjumov) is one of the important insect pests of wheat (Triticum aestivum L.), barley (Hordeum vulgare L.) and other grasses. To date, there are four RWA biotypes identified in South Africa. The virulent biotypes emerged, partly due to climate change and new genetic variations within populations of RWA; hence there is a need to improve host-plant resistance, as an effective control measure. Bread wheat (Triticum aestivum L.) accession Cereal Introduction (CItr) 2401 is known to be resistant to all RWA biotypes worldwide. The goal of this study was to use a backcrossed near-isogenic line (NIL) BC5F5 mapping population, developed from a cross between CItr 2401 and susceptible Kavkaz, to identify and validate single nucleotide polymorphism (SNP) markers linked to the resistance phenotype in CItr 2401. This was achieved by (i) conducting a preliminary study that evaluated the suitability of simple sequence repeat (SSR) markers previously reported in literature for discriminating stacked RWA resistance genes and, (ii) employing SNP markers for the first time in a RWA resistance study as a future alternative to the widely used SSR markers. None of the tested SSR markers showed potential use in marker-assisted selection (MAS). The mapping population was phenotypically evaluated for RWA resistance using the four South African biotypes, viz. RWASA1, RWASA2, RWASA3 and RWASA4. Analysis of variance (ANOVA) showed significant (P<0.001) differences of genotypes after confirming the normality of residuals and homogeneity of variance. The Illumina iSelect 9,000 wheat SNP platform was used to genotype the two crossing parents and a selection of 24 NIL genotypes from the mapping population. Eight SNP markers found to be linked to the phenotype were converted to breeder-friendly and high-throughput Kompetitive allele-specific polymerase chain reaction (KASP) markers. The designed KASP markers were validated on the two crossing parents, the 24 NIL sent for SNP genotyping, on the mapping population and on the preliminary study genotypes for their effectiveness. The KASP assays developed in this study will be useful for stacking the RWA resistance from CItr 2401 with other Dn genes effective against the RWA. / Life and Consumer Sciences / M. Sc. (Life Sciences)