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  • About
  • The Global ETD Search service is a free service for researchers to find electronic theses and dissertations. This service is provided by the Networked Digital Library of Theses and Dissertations.
    Our metadata is collected from universities around the world. If you manage a university/consortium/country archive and want to be added, details can be found on the NDLTD website.
1

Efeito do manejo de solo nos parâmetros agronômicos e energéticos de híbridos de milho transgênico e não transgênico /

Riquetti, Neilor Bugoni, 1985. January 2011 (has links)
Resumo: Os diferentes tipos de preparo de solo para a implantação de culturas devem oferecer condições favoráveis para a semeadura, emergência e desenvolvimento das plantas, porém, apresentam diferenças em relação ao consumo de combustível das máquinas e na compactação do solo, além de poderem interferir nas características agronômicas e na produtividade das culturas instaladas. Diferentes sistemas de produção têm diferentes demandas por energia dependendo do tempo de uso de máquinas e implementos, consumo de combustível, herbicidas e inseticidas. As culturas modificadas geneticamente, seja para resistência a pragas ou herbicidas, geralmente, requerem menor quantidade de energia, devido à redução no número de aplicações de agrotóxicos. O presente trabalho teve por objetivo avaliar os efeitos de diferentes preparos de solo quanto a produtividade de grãos e algumas características das plantas da cultura do milho transgênico e de seu isogênico não transgênico, além da demanda energética exigida por diferentes sistemas de produção. O estudo foi conduzido na safra agrícola 2009/2010 na Faculdade de Ciências Agronômicas - UNESP, localizada no município de Botucatu, SP, em área cultivada com plantio direto há 11 anos. O delineamento utilizado foi o fatorial 3x2, constituído por três métodos de preparo de solo e dois híbridos de milho, transgênico e não transgênico, totalizando seis tratamentos cada um com quatro repetições. Os dados obtidos foram submetidos à análise de variância a 5% de probabilidade e as médias estatisticamente diferentes foram comparadas pelo teste de Tukey a 5% de probabilidade. Os resultados obtidos mostram que, quanto ao tipo de preparo de solo, o sistema de plantio direto e o cultivo mínimo foram os que resultaram em maior produtividade... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The different types of soil tillage for crop implementation should offer favorable conditions for sowing, emergence and development of plants, however, differ in relation to fuel consumption of machinery and soil compaction, and may also interfere with the agronomic characteristics and crop productivity installed. Different production systems have different demands for energy depending on time of use of machinery and implements, fuel, herbicides and insecticides. The genetically modified crops, either for resistance to pests or herbicides generally require less energy due to the reduction in the number of agrochemicals applications. This study aimed to evaluate the effects of different soil tillage systems on grain yield and some characteristics of crop plants of transgenic maize and its isogenic non-transgenic, as well energy demands required by different production systems. The study was performed in the 2009/2010 agricultural season in the College of Agronomic Sciences - UNESP, located in Botucatu, SP, in area cultivated with no-tillage for 11 years. The design was 3x2 factorial, consisting of three methods of tillage and two corn hybrids, transgenic and non-transgenic, with six treatments each with four replications. The obtained data were submitted to analysis of variance at 5% probability and statistically different averages were compared by Tukey test at 5% probability. The obtained results show that, the type of tillage, the no-tillage and the minimum tillage were those that resulted in higher productivity differing statistically of the productivity obtained in the conventional system. Regarding hybrids, productivity showed no statistically significant difference. The systems of tillage and the different hybrids did not interfere in most other characteristics analyzed. The greater energy efficiency... (Complete abstract click electronic access below) / Orientador: Sérgio Hugo Benez / Coorientador: Paulo Roberto Arbex Silva / Banca: Silvio José Bicudo / Banca: Marco Antonio Gandolfo / Mestre
2

Over-Expression of the Cucumber Expansin Gene (Cs-EXPA1) in Transgenic Maize Seed for Cellulose Deconstruction

Yoon, Sangwoong, Devaiah, Shivakumar P., Choi, Seo eun, Bray, Jeff, Love, Robert, Lane, Jeffrey, Drees, Carol, Howard, John H., Hood, Elizabeth E. 01 April 2016 (has links)
Plant cell wall degradation into fermentable sugars by cellulases is one of the greatest barriers to biofuel production. Expansin protein loosens the plant cell wall by opening up the complex of cellulose microfibrils and polysaccharide matrix components thereby increasing its accessibility to cellulases. We over-expressed cucumber expansin in maize kernels to produce enough protein to assess its potential to serve as an industrial enzyme for applications particularly in biomass conversion. We used the globulin-1 embryo-preferred promoter to express the cucumber expansin gene in maize seed. Expansin protein was targeted to one of three sub-cellular locations: the cell wall, the vacuole, or the endoplasmic reticulum (ER). To assess the level of expansin accumulation in seeds of transgenic kernels, a high throughput expansin assay was developed. The highest expressing plants were chosen and enriched crude expansin extract from those plants was tested for synergistic effects with cellulase on several lignocellulosic substrates. Activity of recombinant cucumber expansin from transgenic kernels was confirmed on these pretreated substrates. The best transgenic lines (ER-targeted) can now be used for breeding to increase expansin expression for use in the biomass conversion industry. Results of these experiments show the success of expansin over-expression and accumulation in transgenic maize seed without negative impact on growth and development and confirm its synergistic effect with cellulase on deconstruction of complex cell wall substrates.
3

Comparative phenology of Lepidoptera on genetically modified BT- and non-BT maize / A. van Wyk.

Van Wyk, Annemie January 2006 (has links)
The maize stem borers, Busseola fusca (Fuller) (Lepidoptera: Noctuidae) and Chilo partellus (Swinhoe) (Lepidoptera: Pyralidae) are economically important pests of maize in South Africa. Genetically modified Bt maize (MON810) expressing Cry1Ab protein is used to control these pests on approximately 425 000 hectares in South Africa. Before this study no information was available on the diversity of Lepidoptera on maize in South Africa or the potential impact of Bt maize on non-target Lepidoptera species under field conditions. There was also no information on the susceptibility to Bt maize of another stem borer species, Sesamia calamistis (Hampson) (Lepidoptera: Noctuidae), which is not a target species of Bt maize. The aims of this study were to determine which Lepidoptera species occur and feed on maize and could be directly exposed to Bt toxin as well as to assess the levels of infestation of target stem borer species and non-target Lepidoptera species on Bt- and non-Bt maize fields. Field collections of Lepidoptera that were directly exposed to Bt toxin through feeding on Bt maize plants were done between January 2004 and May 2006. Surveys were conducted in the North-West, Free State, Gauteng and Limpopo provinces. In order to quantify infestation levels and incidence of larvae on plants, sampling was done by inspecting between 300 – 900 plants per field. Studies were also done to compare the incidence of damaged plants and larvae on plants between Bt- and adjacent non-Bt maize fields. The susceptibility of S. calamistis to several Bt maize hybrids was evaluated under laboratory and greenhouse conditions. Fifteen species of Lepidoptera were recorded on maize plants. The following six species were recorded to feed on Bt maize and were reared on Bt maize until the adult stage: Acantholeucania loreyi (Noctuidae), Agrotis segetum (Noctuidae), B. fusca (Noctuidae), Helicoverpa armigera (Noctuidae), Eublemma gayneri (Noctuidae) and Nola phaeocraspis (Nolidae). Although Bt maize was damaged by several species of leaf, stem and ear feeding Lepidoptera in this study, the incidence of damage was always significantly lower on Bt maize fields than susceptible fields. This study provided base line data on Lepidoptera that feed on Bt maize in South Africa. Non-target Lepidoptera species that are directly exposed to Bt toxin was identified. An ecological model wasused to develop a preliminary risk assessment for Bt maize through which priority species for research and monitoring was identified as well as species that are at risk of resistance development. / Thesis (M. Environmental Science (Plant Protection))--North-West University, Potchefstroom Campus, 2007.
4

Comparative phenology of Lepidoptera on genetically modified BT- and non-BT maize / A. van Wyk.

Van Wyk, Annemie January 2006 (has links)
The maize stem borers, Busseola fusca (Fuller) (Lepidoptera: Noctuidae) and Chilo partellus (Swinhoe) (Lepidoptera: Pyralidae) are economically important pests of maize in South Africa. Genetically modified Bt maize (MON810) expressing Cry1Ab protein is used to control these pests on approximately 425 000 hectares in South Africa. Before this study no information was available on the diversity of Lepidoptera on maize in South Africa or the potential impact of Bt maize on non-target Lepidoptera species under field conditions. There was also no information on the susceptibility to Bt maize of another stem borer species, Sesamia calamistis (Hampson) (Lepidoptera: Noctuidae), which is not a target species of Bt maize. The aims of this study were to determine which Lepidoptera species occur and feed on maize and could be directly exposed to Bt toxin as well as to assess the levels of infestation of target stem borer species and non-target Lepidoptera species on Bt- and non-Bt maize fields. Field collections of Lepidoptera that were directly exposed to Bt toxin through feeding on Bt maize plants were done between January 2004 and May 2006. Surveys were conducted in the North-West, Free State, Gauteng and Limpopo provinces. In order to quantify infestation levels and incidence of larvae on plants, sampling was done by inspecting between 300 – 900 plants per field. Studies were also done to compare the incidence of damaged plants and larvae on plants between Bt- and adjacent non-Bt maize fields. The susceptibility of S. calamistis to several Bt maize hybrids was evaluated under laboratory and greenhouse conditions. Fifteen species of Lepidoptera were recorded on maize plants. The following six species were recorded to feed on Bt maize and were reared on Bt maize until the adult stage: Acantholeucania loreyi (Noctuidae), Agrotis segetum (Noctuidae), B. fusca (Noctuidae), Helicoverpa armigera (Noctuidae), Eublemma gayneri (Noctuidae) and Nola phaeocraspis (Nolidae). Although Bt maize was damaged by several species of leaf, stem and ear feeding Lepidoptera in this study, the incidence of damage was always significantly lower on Bt maize fields than susceptible fields. This study provided base line data on Lepidoptera that feed on Bt maize in South Africa. Non-target Lepidoptera species that are directly exposed to Bt toxin was identified. An ecological model wasused to develop a preliminary risk assessment for Bt maize through which priority species for research and monitoring was identified as well as species that are at risk of resistance development. / Thesis (M. Environmental Science (Plant Protection))--North-West University, Potchefstroom Campus, 2007.
5

Efeito do manejo de solo nos parâmetros agronômicos e energéticos de híbridos de milho transgênico e não transgênico

Riquetti, Neilor Bugoni [UNESP] 22 July 2011 (has links) (PDF)
Made available in DSpace on 2014-06-11T19:24:40Z (GMT). No. of bitstreams: 0 Previous issue date: 2011-07-22Bitstream added on 2014-06-13T19:52:12Z : No. of bitstreams: 1 riquetti_nb_me_botfca.pdf: 374256 bytes, checksum: d2e408f5ed43d29d664171e02f12dc0b (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Os diferentes tipos de preparo de solo para a implantação de culturas devem oferecer condições favoráveis para a semeadura, emergência e desenvolvimento das plantas, porém, apresentam diferenças em relação ao consumo de combustível das máquinas e na compactação do solo, além de poderem interferir nas características agronômicas e na produtividade das culturas instaladas. Diferentes sistemas de produção têm diferentes demandas por energia dependendo do tempo de uso de máquinas e implementos, consumo de combustível, herbicidas e inseticidas. As culturas modificadas geneticamente, seja para resistência a pragas ou herbicidas, geralmente, requerem menor quantidade de energia, devido à redução no número de aplicações de agrotóxicos. O presente trabalho teve por objetivo avaliar os efeitos de diferentes preparos de solo quanto a produtividade de grãos e algumas características das plantas da cultura do milho transgênico e de seu isogênico não transgênico, além da demanda energética exigida por diferentes sistemas de produção. O estudo foi conduzido na safra agrícola 2009/2010 na Faculdade de Ciências Agronômicas – UNESP, localizada no município de Botucatu, SP, em área cultivada com plantio direto há 11 anos. O delineamento utilizado foi o fatorial 3x2, constituído por três métodos de preparo de solo e dois híbridos de milho, transgênico e não transgênico, totalizando seis tratamentos cada um com quatro repetições. Os dados obtidos foram submetidos à análise de variância a 5% de probabilidade e as médias estatisticamente diferentes foram comparadas pelo teste de Tukey a 5% de probabilidade. Os resultados obtidos mostram que, quanto ao tipo de preparo de solo, o sistema de plantio direto e o cultivo mínimo foram os que resultaram em maior produtividade... / The different types of soil tillage for crop implementation should offer favorable conditions for sowing, emergence and development of plants, however, differ in relation to fuel consumption of machinery and soil compaction, and may also interfere with the agronomic characteristics and crop productivity installed. Different production systems have different demands for energy depending on time of use of machinery and implements, fuel, herbicides and insecticides. The genetically modified crops, either for resistance to pests or herbicides generally require less energy due to the reduction in the number of agrochemicals applications. This study aimed to evaluate the effects of different soil tillage systems on grain yield and some characteristics of crop plants of transgenic maize and its isogenic non-transgenic, as well energy demands required by different production systems. The study was performed in the 2009/2010 agricultural season in the College of Agronomic Sciences - UNESP, located in Botucatu, SP, in area cultivated with no-tillage for 11 years. The design was 3x2 factorial, consisting of three methods of tillage and two corn hybrids, transgenic and non-transgenic, with six treatments each with four replications. The obtained data were submitted to analysis of variance at 5% probability and statistically different averages were compared by Tukey test at 5% probability. The obtained results show that, the type of tillage, the no-tillage and the minimum tillage were those that resulted in higher productivity differing statistically of the productivity obtained in the conventional system. Regarding hybrids, productivity showed no statistically significant difference. The systems of tillage and the different hybrids did not interfere in most other characteristics analyzed. The greater energy efficiency... (Complete abstract click electronic access below)
6

Migration patterns and survival of Busseola fusca larvae in maize plantings with different ratios of Bt and non-Bt seed / Jaco Marais

Marais, Jaco January 2014 (has links)
The high-dose/refuge strategy is used globally to manage insect resistance development in genetically modified crops with insecticidal properties (Bt crops). The “refuge in a bag” (RIB) strategy is also being considered for deployment against several pest species. Busseola fusca, the target pest of Bt maize in South Africa, evolved resistance to Cry1Ab proteins. The objective of this study was to determine whether migrating B. fusca larvae are effectively controlled using the RIB strategy. A field study with a single-gene event (Cry1Ab) and a “pyramid” event (Cry1A.105 + Cry2Ab2) was conducted in which the migration patterns of B. fusca larvae in plots with different seed mixture treatments were studied. The experiment consisted of five seed mixture ratios (5%, 10%, 15%, 20% non-Bt seed and 100 % non-Bt seed as control). Natural infestation was augmented by artificial inoculation with neonate larvae into the central non-Bt maize plant of each plot. Rate of larval survival and migration, measured in terms of increase in number of plants per plot that exhibited borer damage was recorded at weekly intervals until flowering. A laboratory study was conducted to determine larval growth and survival when simulating migration between Bt and non-Bt maize plants. A feeding experiment in which larvae were reared on different types of maize (Bt and non-Bt) was conducted and larval survival and mass recorded after a 7-day feeding period. The incidence of damaged ears, stem damage and damaged internodes per stem were recorded and relationships between these variables determined by means of correlation analyses. A review was conducted in order to identify and discuss similarities and differences between the high-dose/refuge and seed mixture strategies. This was done to determine which strategy would be the most appropriate insect resistance management (IRM) strategy against B. fusca. The rate of survival and migration of B. fusca larvae was significantly higher in the plots with maize expressing Cry1Ab and control plots, than in plots with the pyramid Bt event. Older larvae exhibited improved growth and survival in the laboratory experiment when they were transferred from non-Bt to Bt plants. Positive correlations were found between early and late season damage, although some weaker than others. Plants of the “pyramid event” suffered less late-season damage than those of the single-gene event. Since the increase in number of damaged maize plants over time is associated with migration of older and larger larvae, the observed tendencies may indicate that the assumed high-dose does not kill larvae above a certain developmental stage. The high-dose refuge strategy seems to be the better option for delaying resistance development. / MSc (Environmental Sciences), North-West University, Potchefstroom Campus, 2014
7

Genetic diversity and susceptibility to Vip3Aa20 protein in Brazilian populations of Helicoverpa armigera and Helicoverpa zea (Lepidoptera: Noctuidae) / Diversidade genética e suscetibilidade à proteína Vip3Aa20 em populações brasileiras de Helicoverpa armigera e Helicoverpa zea (Lepidoptera: Noctuidae)

Leite, Natália Alves 12 April 2016 (has links)
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.
8

Migration patterns and survival of Busseola fusca larvae in maize plantings with different ratios of Bt and non-Bt seed / Jaco Marais

Marais, Jaco January 2014 (has links)
The high-dose/refuge strategy is used globally to manage insect resistance development in genetically modified crops with insecticidal properties (Bt crops). The “refuge in a bag” (RIB) strategy is also being considered for deployment against several pest species. Busseola fusca, the target pest of Bt maize in South Africa, evolved resistance to Cry1Ab proteins. The objective of this study was to determine whether migrating B. fusca larvae are effectively controlled using the RIB strategy. A field study with a single-gene event (Cry1Ab) and a “pyramid” event (Cry1A.105 + Cry2Ab2) was conducted in which the migration patterns of B. fusca larvae in plots with different seed mixture treatments were studied. The experiment consisted of five seed mixture ratios (5%, 10%, 15%, 20% non-Bt seed and 100 % non-Bt seed as control). Natural infestation was augmented by artificial inoculation with neonate larvae into the central non-Bt maize plant of each plot. Rate of larval survival and migration, measured in terms of increase in number of plants per plot that exhibited borer damage was recorded at weekly intervals until flowering. A laboratory study was conducted to determine larval growth and survival when simulating migration between Bt and non-Bt maize plants. A feeding experiment in which larvae were reared on different types of maize (Bt and non-Bt) was conducted and larval survival and mass recorded after a 7-day feeding period. The incidence of damaged ears, stem damage and damaged internodes per stem were recorded and relationships between these variables determined by means of correlation analyses. A review was conducted in order to identify and discuss similarities and differences between the high-dose/refuge and seed mixture strategies. This was done to determine which strategy would be the most appropriate insect resistance management (IRM) strategy against B. fusca. The rate of survival and migration of B. fusca larvae was significantly higher in the plots with maize expressing Cry1Ab and control plots, than in plots with the pyramid Bt event. Older larvae exhibited improved growth and survival in the laboratory experiment when they were transferred from non-Bt to Bt plants. Positive correlations were found between early and late season damage, although some weaker than others. Plants of the “pyramid event” suffered less late-season damage than those of the single-gene event. Since the increase in number of damaged maize plants over time is associated with migration of older and larger larvae, the observed tendencies may indicate that the assumed high-dose does not kill larvae above a certain developmental stage. The high-dose refuge strategy seems to be the better option for delaying resistance development. / MSc (Environmental Sciences), North-West University, Potchefstroom Campus, 2014
9

Genetic diversity and susceptibility to Vip3Aa20 protein in Brazilian populations of Helicoverpa armigera and Helicoverpa zea (Lepidoptera: Noctuidae) / Diversidade genética e suscetibilidade à proteína Vip3Aa20 em populações brasileiras de Helicoverpa armigera e Helicoverpa zea (Lepidoptera: Noctuidae)

Natália Alves Leite 12 April 2016 (has links)
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.

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