1 Selected issues of legal regulation of genetically modified organisms Abstract Genetically modified organisms are one of the most pressing challenges of modern agriculture and other economic sectors. This is due to the large number of potential benefits and risks, which are quite often insufficiently scientifically researched, exacerbating the controversy associated with their use. It is a technology that has been used for a relatively long time, but its legal regulation is constantly challenged due to the dynamic development of the related scientific advances. Despite the aforementioned, some issues of the topic receive only marginal attention, which makes it difficult to create and apply a fully functioning legal regulation. The diploma thesis itself deals with selected issues of legal regulation of genetically modified organisms, focusing on the readiness of legal regulation, functionality of auxiliary instruments, as well as de lege ferenda considerations. These topics are formulated into three basic research questions, which the author tries to answer by analyzing relevant parts of the legal regulation. The aim of this thesis is to expand this vital part of environmental law with new findings and thus supplement other academic works that deal with legal regulation of genetically modified organisms....

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Genetically modified (GM) crops gained attention in southern Africa as countries are struggling with food insecurity and poverty to achieve sustainable development. The controversy around GM crops have provoked heated debates. GM crops are often perceived as a global risk to human health and the environment. The research question is what are the perceptions of environmental educators regarding the use of GM crops toward sustainable development. In the Decade of Education for Sustainable Development environmental educators will need to be key role players in addressing crucial issues such as GM crops. Their perceptions hold many implications for educational programmes. Environmental educators interviewed perceive GM crops as a serious issue. As mediators in a multidisciplinary setting between science and society, environmental educators can play a functional role. Open processes that require greater participation, criticality and reflexivity need to be facilitated in a complex biophysical and social context in southern Africa. / Educational Studies / M. Ed.(Environmental Education)

Genetically modified crops are submitted to strict regulation to ensure the safety of consumers and the environment. To complement the comparison between GM plants and their counterparts, in the present Thesis, we evaluated the possible unexpected effects of the transgene on the host plant, by means of transcriptomic technologies. More exactly, we studied three pathogen-resistant GM rice lines: S-afp, expressing constitutively the antifungal protein AFP; and S-bp217 and S-bp213, expressing undecapeptide BP100 derivatives, which were developed in the UdG in the context of this Thesis. Although the high phytotoxicity of the BP100 derivatives on the host plant the transcriptional changes observed in S-afp, S-bp217 and S-bp213 compared to the conventional line Senia were similar that those observed in other GM crops, of other species and with different transgenes, and only the half of them was attributed to the insertion and/or expression of the transgene. / Les plantes modificades genèticament (MG) destinades a comercialització estan sotmeses a estricta legislació per garantir la seguretat del consumidor i del medi ambient. Per complementar la comparativa entre plantes MG i convencionals, en aquesta tesi s’ha abordat l’avaluació dels possibles efectes no esperats del transgèn sobre la planta hoste, mitjançant tècniques de transcriptòmica. Concretament s’han estudiat línies d'arròs MG que presenten fenotips de resistència a patògens: S-afp, que expressa constitutivament la proteïna antifúngica AFP, i S-bp213 i S-bp217, que expressen derivats de l’undecapèptid BP100, desenvolupat a la UdG, que s’han obtingut també en el marc d’aquesta tesi. Malgrat l’elevada fitotoxicitat dels derivats de BP100 enfront la planta hoste, els canvis transcripcionals de S-afp, S-bp213 i S-bp217 respecte la línia convencional Senia són similars als observats en altres events MG, de diferents espècies i amb diferents transgens; i només la meitat d’ells s’ha atribuit a la presència o expressió del transgèn.

Genetically modified (GM) crops gained attention in southern Africa as countries are struggling with food insecurity and poverty to achieve sustainable development. The controversy around GM crops have provoked heated debates. GM crops are often perceived as a global risk to human health and the environment. The research question is what are the perceptions of environmental educators regarding the use of GM crops toward sustainable development. In the Decade of Education for Sustainable Development environmental educators will need to be key role players in addressing crucial issues such as GM crops. Their perceptions hold many implications for educational programmes. Environmental educators interviewed perceive GM crops as a serious issue. As mediators in a multidisciplinary setting between science and society, environmental educators can play a functional role. Open processes that require greater participation, criticality and reflexivity need to be facilitated in a complex biophysical and social context in southern Africa. / Educational Studies / M. Ed.(Environmental Education)

Soil is an important ecosystem that supports a wide variety of organisms such as bacteria, fungi, arthropods and nematodes. This sensitive ecosystem may be influenced by various factors, including agricultural management practices. With the introduction of genetically modified (GM) glyphosate-tolerant (RoundUp ® Ready: RR) crops, herbicides such as glyphosate have been increasingly used. However, little is known about the effect of glyphosate on the biological communities in these herbicide-sprayed soils. With the intimate proximity that microorganisms and nematodes have with the roots of plants, these organisms can be used to assess changes that may occur in the soil surrounding roots of RR crops. The aim of this study was to determine microbial community structure and nematode diversity, with emphasis on that of non-parasitic nematodes, in soil samples from conventional soybean (CS) - and RR- soybean fields compared to that in adjacent natural veld (NV) areas. Samples were collected from twenty three sites at six localities that are situated within the soybean-production areas of South Africa. These sites represented fields where RR and CS soybean grew, as well as surrounding NV. All RR fields have been treated with glyphosate for no less than five years. Microbial community structures of the twenty three sites in the RR, CS and NV ecosystems were determined by phospholipid fatty acid (PLFA) analyses. Nematode diversity was determined by extracting the nematodes from soil samples and conducting a faunal analysis. Soil physical and chemical properties were determined by an independent laboratory, Eco-Analytica (North West University, Potchefstroom) according to standard procedures. Results from this study indicated differences in microbial community structure between the various localities. However, there were no significant (p ≤ 0.05) differences in microbial community structures between RR- and CS ecosystems. Soils of both RR- and CS crops were primarily dominated by bacteria. Nematode identification and faunal analysis also indicated no significant (p ≤ 0.05) differences between the different non-parasitic/beneficial nematodes that were present in soils of these two ecosystems during the time of sampling. Non-parasitic nematode communities were primarily dominated by bacterivores. A faunal analysis indicated that most of the sites contained enriched, but unstructured soil food-webs. However, four of the sites showed enriched and structured food webs due to the presence of non-parasitic nematodes with high coloniser-persister (cp) values. Relationships between non-parasitic nematode – and microbial communities showed that there was a positive relationship between nematode functional groups and their corresponding microbial prey. From the results obtained in this study, it can be concluded that the community structures of both non-parasitic nematodes and microorganisms shared similarities. These community structures showed no long-term detrimental effects of glyphosate application in the soils surrounding roots of RR soybean crops. Relationships existed between non-parasitic nematode and microbial communities in the rhizosphere of soybean crops and natural veld. For example, bacterivore nematodes had a strong positive relationship with gram-negative bacteria. Similar but weaker relationships also existed between carnivores, omnivores, plantparasitic nematodes and gram-negative bacteria. A positive relationship also existed between fungivores and fungal fatty acids. This emphasises the value of these organisms as indicators of soil health and also the impact that agricultural practices can have on soils. / MSc (Environmental Sciences), North-West University, Potchefstroom Campus, 2014

Soil is an important ecosystem that supports a wide variety of organisms such as bacteria, fungi, arthropods and nematodes. This sensitive ecosystem may be influenced by various factors, including agricultural management practices. With the introduction of genetically modified (GM) glyphosate-tolerant (RoundUp ® Ready: RR) crops, herbicides such as glyphosate have been increasingly used. However, little is known about the effect of glyphosate on the biological communities in these herbicide-sprayed soils. With the intimate proximity that microorganisms and nematodes have with the roots of plants, these organisms can be used to assess changes that may occur in the soil surrounding roots of RR crops. The aim of this study was to determine microbial community structure and nematode diversity, with emphasis on that of non-parasitic nematodes, in soil samples from conventional soybean (CS) - and RR- soybean fields compared to that in adjacent natural veld (NV) areas. Samples were collected from twenty three sites at six localities that are situated within the soybean-production areas of South Africa. These sites represented fields where RR and CS soybean grew, as well as surrounding NV. All RR fields have been treated with glyphosate for no less than five years. Microbial community structures of the twenty three sites in the RR, CS and NV ecosystems were determined by phospholipid fatty acid (PLFA) analyses. Nematode diversity was determined by extracting the nematodes from soil samples and conducting a faunal analysis. Soil physical and chemical properties were determined by an independent laboratory, Eco-Analytica (North West University, Potchefstroom) according to standard procedures. Results from this study indicated differences in microbial community structure between the various localities. However, there were no significant (p ≤ 0.05) differences in microbial community structures between RR- and CS ecosystems. Soils of both RR- and CS crops were primarily dominated by bacteria. Nematode identification and faunal analysis also indicated no significant (p ≤ 0.05) differences between the different non-parasitic/beneficial nematodes that were present in soils of these two ecosystems during the time of sampling. Non-parasitic nematode communities were primarily dominated by bacterivores. A faunal analysis indicated that most of the sites contained enriched, but unstructured soil food-webs. However, four of the sites showed enriched and structured food webs due to the presence of non-parasitic nematodes with high coloniser-persister (cp) values. Relationships between non-parasitic nematode – and microbial communities showed that there was a positive relationship between nematode functional groups and their corresponding microbial prey. From the results obtained in this study, it can be concluded that the community structures of both non-parasitic nematodes and microorganisms shared similarities. These community structures showed no long-term detrimental effects of glyphosate application in the soils surrounding roots of RR soybean crops. Relationships existed between non-parasitic nematode and microbial communities in the rhizosphere of soybean crops and natural veld. For example, bacterivore nematodes had a strong positive relationship with gram-negative bacteria. Similar but weaker relationships also existed between carnivores, omnivores, plantparasitic nematodes and gram-negative bacteria. A positive relationship also existed between fungivores and fungal fatty acids. This emphasises the value of these organisms as indicators of soil health and also the impact that agricultural practices can have on soils. / MSc (Environmental Sciences), North-West University, Potchefstroom Campus, 2014

"Mémoire présenté à la Faculté des études supérieures en vue de l'obtention du grade de Maître en droit (LL.M)" / Depuis quelques années, le recours à la biotechnologie dans le secteur de l'alimentation est omniprésent. Les techniques, évoluant au gré de l'avancement de la science, permettent aux gens faisant partie de plusieurs sphères de la société, tels les scientifiques, producteurs, agriculteurs, consommateurs etc., de profiter de ces percées technologiques. Toutefois, afin d'encadrer la commercialisation de ces nouveaux produits alimentaires découlant de cette technologie, un cadre réglementaire devait, il va sans dire, être élaboré. Ce fut le cas de part et d'autre de l'Atlantique. Toutefois, bien qu'appréciée des scientifiques, cette technologie est loin de faire l'assentiment de tous. D'ailleurs, différentes perceptions se reflètent dans les différents cadres réglementaires mis de l'avant, tout aussi bien canadien que français, quant à l'utilisation et la mise en vente des nouveaux aliments. Ce travail décrit les systèmes réglementaires visant la commercialisation des aliments nouveaux tant canadien que français. Par ailleurs, il relate les différents débats, sociologique, éthique, et techniques, dont ces organismes génétiquement modifiés (OGM) sont la cible. / Since the past decades, the use of biotechnology in the industry of novel food was very considerable. This new technique is very useful, and can be easily important for an appreciable quantity of people, like productors, agricultors and consumers. Since it was used in the production of novel food, a new regulatory enforcement was needed. It was done either in France and Canada, but differently. As we know, the different way of life in both countries play an important role in the acceptance of the genetically modified organisms (GMO's). In this thesis, we will give a description of the different regulatory frameworks for food products of biotechnology. Then, we will see what was important, according to the different sociologie and ethics point of view, in the elaboration of these relevant legislation.

by Ng Wang Kit. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2001. / Includes bibliographical references (leaves 131-139). / Abstracts in English and Chinese. / Acknowledgements --- p.iii / Abstract --- p.v / List of Tables --- p.ix / List of Figures --- p.x / List of Abbreviations --- p.xiii / Table of Contents --- p.xv / Chapter Chapter 1: --- General Introduction --- p.1 / Chapter Chapter 2: --- Literature Review --- p.3 / Chapter 2.1 --- Malaria --- p.3 / Chapter 2.1.1 --- Global picture --- p.3 / Chapter 2.1.2 --- Malaria mechanics --- p.4 / Chapter 2.1.3 --- Life cycle of malaria parasite --- p.4 / Chapter 2.2 --- Treatment of malaria ´ؤ malaria drugs --- p.5 / Chapter 2.2.1 --- Antimalarial drugs --- p.5 / Chapter 2.2.2 --- Drug resistance --- p.6 / Chapter 2.3 --- Treatment of malaria - malarial vaccines --- p.7 / Chapter 2.3.1 --- Malarial vaccine developments --- p.7 / Chapter 2.3.2 --- Transmission blocking vaccines --- p.7 / Chapter 2.3.3 --- Pre-erythrocytic vaccines --- p.9 / Chapter 2.3.4 --- Blood stage vaccines --- p.10 / Chapter 2.4 --- The major merozoite protein - gpl95 --- p.11 / Chapter 2.5 --- Plants as bioreactors --- p.12 / Chapter 2.5.1 --- Products of transgenic plants --- p.13 / Chapter 2.6 --- Transgenic plants for production of subunit vaccines --- p.14 / Chapter 2.6.1 --- Norwalk virus capsid protein production --- p.15 / Chapter 2.6.2 --- Hepatitis B surface antigen production --- p.15 / Chapter 2.7 --- Tobacco and Arabidopsis as model plants --- p.16 / Chapter 2.7.1 --- Arabidopsis --- p.16 / Chapter 2.7.2 --- Tobacco --- p.17 / Chapter 2.8 --- Transformation methods --- p.17 / Chapter 2.8.1 --- Direct DNA uptake --- p.17 / Chapter 2.8.1.1 --- Plant protoplast transformation --- p.17 / Chapter 2.8.1.2 --- Biolistic transformation --- p.18 / Chapter 2.8.2 --- Agrobacterium-mediated transformation --- p.18 / Chapter 2.8.2.1 --- Leaf-disc technique --- p.18 / Chapter 2.8.2.2 --- In planta transformation --- p.19 / Chapter 2.9 --- Phaseolin --- p.20 / Chapter 2.10 --- Detection and purification of recombinant products - Histidine tag --- p.21 / Chapter 2.11 --- Aims of study and hypotheses --- p.22 / Chapter Chapter 3: --- Materials and Methods --- p.24 / Chapter 3.1 --- Introduction --- p.24 / Chapter 3.2 --- Chemicals --- p.24 / Chapter 3.3 --- Expression in tobacco system --- p.24 / Chapter 3.3.1 --- Plant materials --- p.24 / Chapter 3.3.2 --- Bacterial strains --- p.25 / Chapter 3.3.3 --- Chimeric gene construction for tobacco transformation --- p.25 / Chapter 3.3.3.1 --- The cloning of pTZPhasp/flgp42-His/Phast (F1) --- p.26 / Chapter 3.3.3.2 --- The cloning of pBKPhasp-sp/flgp42-His/Phast (P9) --- p.30 / Chapter 3.3.3.3 --- The cloning of pHM2Ubip/flgp42-His/Nost (C2) --- p.30 / Chapter 3.3.4 --- Confirmation of sequence fidelity of chimeric gene by DNA sequencing --- p.33 / Chapter 3.3.5 --- Cloning of chimeric gene into binary vector --- p.34 / Chapter 3.3.6 --- Triparental mating of Agrobacterium tumefaciens LBA4404/pAL4404 --- p.35 / Chapter 3.3.7 --- Tobacco transformation and regeneration --- p.36 / Chapter 3.3.8 --- GUS assay --- p.37 / Chapter 3.3.9 --- Genomic DNA isolation --- p.37 / Chapter 3.3.10 --- PCR amplification and detection of transgene --- p.38 / Chapter 3.3.11 --- Southern blot analysis --- p.38 / Chapter 3.3.12 --- Total seeds RNA isolation --- p.39 / Chapter 3.3.13 --- RT-PCR --- p.39 / Chapter 3.3.14 --- Northern blot analysis --- p.40 / Chapter 3.3.15 --- Protein extraction and SDS-PAGE --- p.40 / Chapter 3.3.16 --- Western blot analysis --- p.41 / Chapter 3.4 --- Expression in Arabidopsis system --- p.42 / Chapter 3.4.1 --- Plant materials --- p.42 / Chapter 3.4.2 --- Bacterial strains --- p.42 / Chapter 3.4.3 --- Chimeric gene construction --- p.42 / Chapter 3.4.3.1 --- The cloning of pBKPhasp-sp/His/EK/p42/Phast (DH) --- p.43 / Chapter 3.4.3.2 --- The cloning of pTZPhaSp/His/EK/p42/Phast (EH) --- p.45 / Chapter 3.4.3.3 --- The cloning of pBKPhasp-sp/His/EK/flgp42/Phast (DHF) and pTZPhasp/His/EK/flgp42/Phast (EHF) --- p.45 / Chapter 3.4.4 --- Confirmation of sequence fidelity of chimeric genes --- p.45 / Chapter 3.4.5 --- Cloning of chimeric gene into Agrobacterium binary vector --- p.49 / Chapter 3.4.6 --- Transformation of Agrobacterium tumefaciens GV3101/pMP90 with chimeric gene constructs --- p.49 / Chapter 3.4.7 --- Arabidopsis Transformation --- p.49 / Chapter 3.4.8 --- Vacuum infiltration transformation --- p.50 / Chapter 3.4.9 --- Selection of successful transformants --- p.51 / Chapter 3.4.10 --- Selection for homozygous plants with single gene insertion --- p.51 / Chapter 3.4.11 --- GUS assay --- p.52 / Chapter 3.4.12 --- Genomic DNA isolation --- p.52 / Chapter 3.4.13 --- PCR amplification and detection of transgenes --- p.52 / Chapter 3.4.14 --- Southern Blot analysis --- p.52 / Chapter 3.4.15 --- Total siliques RNA isolation --- p.53 / Chapter 3.4.16 --- RT-PCR --- p.53 / Chapter 3.4.17 --- Northern blot analysis --- p.53 / Chapter 3.4.17 --- Protein extraction and SDS-PAGE --- p.54 / Chapter 3.4.18 --- Western blot analysis --- p.54 / Chapter 3.5 --- In vitro transcription and translation --- p.54 / Chapter 3.5.1 --- In vitro transcription --- p.54 / Chapter 3.5.2 --- In vitro translation --- p.55 / Chapter 3.6 --- Particle bombardment of GUS fusion gene --- p.56 / Chapter 3.6.1 --- Chimeric gene constructs --- p.56 / Chapter 3.6.2 --- Particle bombardment using snow bean cotyledon --- p.61 / Chapter Chapter 4: --- Results --- p.63 / Chapter 4.1 --- Tobacco system --- p.63 / Chapter 4.1.1 --- Chimeric gene constructs --- p.63 / Chapter 4.1.2 --- Tobacco transformation and regeneration --- p.65 / Chapter 4.1.3 --- GUS activity assay --- p.67 / Chapter 4.1.4 --- Molecular analysis of transgene integration --- p.68 / Chapter 4.1.4.1 --- Genomic DNA extraction and PCR --- p.68 / Chapter 4.1.4.2 --- Southern blot analysis --- p.70 / Chapter 4.1.5 --- Molecular analysis of transgene expression --- p.72 / Chapter 4.1.5.1 --- Total RNA isolation and RT-PCR --- p.72 / Chapter 4.1.5.2 --- Northern blot analysis --- p.75 / Chapter 4.1.6 --- Genomic PCR to confirm whole gene transfer --- p.76 / Chapter 4.1.7 --- Biochemical analysis of transgene expression --- p.78 / Chapter 4.1.7.1 --- Protein extraction and SDS-PAGE --- p.78 / Chapter 4.1.7.2 --- Western blot analysis --- p.78 / Chapter 4.2 --- Arabidopsis system --- p.83 / Chapter 4.2.1 --- Chimeric gene constructs --- p.83 / Chapter 4.2.2 --- Arabidopsis transformation and selection --- p.85 / Chapter 4.2.3 --- Selection of transgenic plants --- p.87 / Chapter 4.2.4 --- Assay of GUS activity --- p.91 / Chapter 4.2.5 --- Molecular analysis of transgene integration --- p.92 / Chapter 4.2.5.1 --- Genomic DNA extraction and PCR --- p.92 / Chapter 4.2.5.2 --- Southern blot analysis --- p.96 / Chapter 4.2.6 --- Molecular analysis of transgene expression --- p.99 / Chapter 4.2.6.1 --- Total RNA isolation and RT-PCR --- p.99 / Chapter 4.2.6.2 --- Northern blot analysis --- p.106 / Chapter 4.2.7 --- Genomic PCR for confirmation of whole gene transfer --- p.107 / Chapter 4.2.8 --- Biochemical analysis of transgene expression --- p.108 / Chapter 4.2.8.1 --- Protein extraction and SDS-PAGE --- p.108 / Chapter 4.2.8.2 --- Western blot analysis --- p.108 / Chapter 4.3 --- In vitro transcription and translation --- p.112 / Chapter 4.4 --- Particle bombardment of p42/ GUS fusion gene --- p.115 / Chapter Chapter 5: --- Discussion and Future perspectives --- p.117 / Chapter 5.1 --- Failure in detecting transgene expression --- p.117 / Chapter 5.2 --- Poor transgene expression --- p.120 / Chapter 5.2.1 --- Bacillus thuringiensis toxin and green fluorescent protein --- p.120 / Chapter 5.2.2 --- AT-richness --- p.121 / Chapter 5.2.3 --- Deleterious sequence - AUUUA --- p.123 / Chapter 5.2.4 --- Presence of AAUAAA or AAUAAA-like motifs --- p.125 / Chapter 5.2.5 --- Codon usage --- p.126 / Chapter 5.3 --- Future perspectives --- p.127 / Chapter Chapter 6: --- Conclusion --- p.129 / References --- p.131

Made available in DSpace on 2017-07-25T19:30:58Z (GMT). No. of bitstreams: 1 Lucas N Wisch.pdf: 1238431 bytes, checksum: fab61da00fd601e632d7feb86156e3da (MD5) Previous issue date: 2016-02-26 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Soybean is the main crop of economic importance and commercially produced on a large scale in Brazil. Nevertheless many are the factors that affect the productive potential of this oilseed, among them stand out the insects of the order Lepidoptera. As an alternative to control these pests, many growers have adopted the use of Bt soybean, which expresses the Cry1Ac toxin. However, the large area with this technology can generates a main concern that is Cry1Ac exposure to target and non-target insects and its entomopathogens, such Metarhizium rileyi and nuclear polyhedrosis virus. The aim of this study were to elucidate the HD-73 impacts, that expresses Cry1Ac, i) in the immune system of S. frugiperda in order to investigate whether Bt challenge parental induce "immune priming or transgenerational effects"; ii) on virulence and occlusion body yield in Spodoptera frugiperda iii) level of larval mortality of the noctuid species when applied simultaneously or sequentially with NPV and M. rileyi; and iv) the biological parameters of the noctuid species, when exposed in conjunction with entomopathogens. In the immune assay, neonate larvae were exposed ad libitum to different concentrations of HD-73 (0, 15, 33, 72.6 μg.ml-1), in the fifth instar were extracted the haemolymph and measured the number of haemocytes, total protein and phenoloxidase activity (PO). Neonate offspring of those individuals were inoculated with 0 and 40 μg.ml-1 of HD-73 and again was quantified the same immune components in fifth instar larvae, including antibacteriana activity. In addition, biological parameters were observed in these tests. In the evaluation of Cry1Ac impacts on virus infectivity in S. frugiperda, neonate were exposed to HD-73 (0, 10, 40 and 80 μg.ml-1) until the end of the third instar and early of the fourth instar was inoculated with viruses (0, 103 and 104 OB.larvae-1). For this assay was evaluated larval weight (4 DAE), mortality, number of OB/caterpillar and subsequently, virulence test (LC50). Spodoptera cosmioides, Spodoptera eridania, S. frugiperda, Chrysodeixis includens and Rachiplusia nu caterpillars were simultaneously or sequentially exposed to entomopathogens, HD-73, M. rileyi and their viruses, and record mortality and biological parameters of these interactions. The sublethal concentrations of HD-73 did not cause changes in the immune system of the parental, as well as there was no evidence for acquired immunity in the offspring. Sublethal concentration of HD-73 affected the biological components of S. frugiperda, providing lower larval and pupal weight, reduced fecundity and egg viability. HD-73 proved lower OB yield/caterpillar and lower virulence of the virus multiplied in caterpillars prior exposed to 10 μg.ml-1. The interactions did not affect the biological parameters of these noctuids. In most cases was observed negative effect in the interactions between HD-73 and viruses, but may be promising simultaneous inoculation of HD-73 and M. rileyi to control S. eridania and S. frugiperda. Sublethal concentrations of HD-73 did not provide larger mortality of Plusiinae in interactions with M. rileyi and viruses, except for HD-73+ M. rileyi that had positive and negative effects on C. includens. / A soja é a principal cultura de importância econômica e comercialmente produzida em larga escala no Brasil. Porém, diversos são os fatores que afetam o potencial produtivo desta oleaginosa, entre eles destacam-se os insetos da ordem Lepidoptera. Como alternativa para controle destas pragas, diversos produtores têm adotado o uso da soja Bt, que expressa a toxina Cry1Ac. Contudo, a expansão da área com essa tecnologia gera dúvidas quanto ao impacto da exposição de Cry1Ac aos insetos-alvo e não-alvo e seus entomopatógenos, Metarhizium rileyi e os vírus de poliedrose nuclear. Assim, os objetivos desse trabalho foram: elucidar o impacto de concentrações subletais da cepa HD-73, que expressa Cry1Ac, i) no sistema imunológico de Spodoptera frugiperda, a fim de investigar se o desafio com Bt induz a “memória imunológica”; ii) na virulência e produção de corpos de oclusão do vírus em S. frugiperda; iii) na mortalidade larval das espécies de noctuídeos, quando aplicada de forma simultânea ou sequencial com VPN e M. rileyi; e iv) nos parâmetros biológicos de noctuídeos-alvo e não-alvo, quando aplicada em conjunto com os entomopatógenos. No ensaio imunológico, lagartas neonatas foram expostas ad libitum a diferentes concentrações de HD-73 (0, 15, 33, 72,6 μg.ml-1), no quinto instar foi extraída a hemolinfa e avaliado o número de hemócitos, proteína total e atividade da fenoloxidase (PO). Neonatas descendentes destes indivíduos foram inoculadas com 0 e 40 μg.ml-1 de HD-73 e no quinto instar quantificou-se os mesmos componentes de resposta imune observados nos parentais, incluindo a atividade antibacteriana. Os parâmetros biológicos também foram observados nestes ensaios. Nas avaliações do impacto de Cry1Ac na infectividade de vírus sobre S. frugiperda, neonatas foram inoculadas com HD-73 (0, 10, 40 e 80 μg.ml-1) e mantidas até o final do terceiro instar, e no início do quarto instar ocorreu a infecção com vírus (0, 103 e 104 CO.lagarta-1), após foi avaliado o peso larval (4 DAE), a mortalidade, o número de CO.cadáver-1 e, posteriormente, teste de virulência (CL50). Lagartas de Spodoptera cosmioides, Spodoptera eridania, S. frugiperda, Chrysodeixis includens e Rachiplusia nu, foram expostas de forma simultânea ou sequencial aos entomopatógenos, HD-73, M. rileyi e os respectivos vírus, para registrar a mortalidade e os parâmetros biológicos destas interações. As concentrações subletais de HD-73 não provocaram alterações no sistema imune dos parentais, como também não houve evidências de imunidade adquirida nas progênies. HD-73 afetou subletalmente os componentes biológicos de S. frugiperda, proporcionando menor peso larval e pupal, redução na fecundidade e viabilidade dos ovos. HD-73 proporcionou menor rendimento de CO.cadáver-1 e menor virulência dos vírus multiplicados em lagartas expostas a 10 μg.ml-1. As interações não afetaram os parâmetros biológicos dos noctuídeos em estudo. Na maioria dos casos observou-se efeito negativo nas interações entre HD-73 e vírus, mas podendo ser promissor a inoculação simultânea de HD-73 e M. rileyi no controle de S. eridania e S. frugiperda. Concentrações subletais de HD-73 não proporcionaram maior mortalidade das Plusiinae nas interações com M. rileyi e vírus, exceto na associação HD-73+M. rileyi que apresentou efeitos positivos e negativos sobre C. includens.