A proteômica expandiu-se dentro da área biológica a partir da década de 90 com o desenvolvimento de técnicas de ionização branda que permitem analisar macromoléculas por espectrometria de massas (MS) e é uma nova estratégia na busca de genes de interesse e de informações sobre o controle da expressão gênica para a manipulação genética de plantas. Na busca por genes diferencialmente expressos durante o desenvolvimento da madeira, foi estabelecida uma plataforma 2D-LC-MS/MS (eletroforese bidimensional seguida de cromatografia líquida e espectrometria de massas em tandem) e identificadas 72 proteínas da região cambial de Eucalyptus grandis de árvores de 2 anos e 8 meses. A identificação foi feita por homologia com o banco de proteínas SwissProt, com um banco de ESTs específico de Eucalyptus spp. (GENOLYPTUS) e com o NCBI. Os dados de proteínas também foram confrontados com a expressão dos transcritos correspondentes obtidos a partir do mesmo tecido por SAGE (Serial Analysis of Gene Expression). Dentre as proteínas identificadas, a maior parcela (24,1 %) tem função relacionada com resposta a estresse, 20,5 % pertence a metabolismo de energia, 17,8 % são componentes estruturais, 14,3 % participam em metabolismo de macromoléculas e 15,2 % em metabolismos básicos como de nitrogênio, aminoácidos, nucleotídeos, lipídeos e metabolismo secundário. / Proteomics impacted the biological sciences since the 90´s after the development of soft ionization techniques that allow to analyse macromolecules by mass spectrometry (MS). It became a new strategy to identify genes and obtain information about the molecular control of gene expression, of importance to genetic manipulation of plants. In order to search for genes involved in wood quality and to understand gene expression during wood development, a 2D-LC-MS/MS system (bidimensional electrophoresis followed by liquid cromatography coupled to mass spectrometry in tandem) was stablished. 72 proteins from the cambial region of Eucalyptus grandis trees at the age of 2 years and 8 months were identified. The MS/MS spectra were processed using the SwissProt databank, an EST (Expressed Sequence Tags) bank of Eucalyptus spp. (GENOLYPTUS) and the NCBI. A comparation of gene and protein expression was carried out using a databank constructed in our laboratory using SAGE (Serial Analysis of Gene Expression) obtained from the same cambial tissue. Most identified proteins are related to stress response (24.1 %), 20.5 % participate in energy metabolism, 17.8 % are important to the cell as structural components, 14,3 % are related to macromolecular metabolism and 15,2 % to basic metabolism of nitrogen, amino acids, nucleotides, lipids and secondary metabolism.

Plants possess a complex and poorly understood network of defence mechanisms that enable them to counteract the effects of abiotic and biotic stress. Aphid phloem feeding is source of biotic stress in plants. Russian wheat aphid and Bird Cherry-Oat aphid feeding cause significant losses in the annual wheat crop, and control by conventional methods such as pesticide application, has proved to be ineffective. Infestation by the Russian wheat aphid has a particularly devastating effect in South Africa. Aphid-resistant wheat cultivars have been identified but an incomplete understanding of the mechanism of the plant’s resistance thwarts the development of improved cultivars. A two-dimensional gel electrophoresis method was developed, partially optimised and validated in order to determine the effect of Russian wheat aphid and Bird Cherry-Oat aphid phloem feeding on the Betta and Betta DN wheat proteome. Differentially expressed proteins that were up or down regulated more than two fold were identified using PDQuest™ Basic software and matched to known wheat proteins stored in the SwissProt protein database on the basis of their molecular mass and isolectric point. Initial analysis of the differential protein expression of Betta and Betta DN wheat in response to Russian wheat aphid and Bird Cherry-Oat aphid phloem feeding at different growth stages revealed that younger plants display higher levels of resistance than older plants. Feeding by the Bird-Cherry Oat aphid does not result in the upregulation of proteins implicated in a defence response, which indicates that the damage incurred by the plant due to feeding by this aphid is not enough to trigger a classic defence response. Feeding by the more damaging Russian wheat aphid resulted in a stress response in susceptible wheat cultivar Betta, and a defence response in resistant wheat cultivar Betta DN. The infestation of Betta DN resulted in the upregulation of putative thaumatins and amylase trypsin inhibitors, indicating that the Betta DN resistance response could be due to the combined effect of protease inhibitors that discourage aphid phloem feeding and the activation of the salicylic acid and jasmonic acid plant defence pathways.

Prof. I.A. Dubery

There are apprehensions that genetic modification of maize with Bacillus thuringiensis (Bt) may have negative effects on soil biodiversity, ecosystem processes and functions. This study aimed at determining the effect of Bt maize crop, Bt maize residues and its genetic modification on microbial biomass carbon (MBC), selected enzyme activities, vesicular arbuscular mycorrhizal (VAM) fungi and N and P release patterns. The study was conducted under field, glasshouse and laboratory conditions. In 2010/2011 season, four maize cultivars; DKC 61-25B (Bt), DKC 61-24 (non-Bt), PAN 6Q-321B (Bt) and PAN6777 (non-Bt) were planted. Determination of MBC, enzyme activities and fungal spore count was done at 42, 70, and 105 days after planting (DAP). A loam soil amended with Bt or non-Bt maize leaf residues from a study of 2009/2010 season was incubated to investigate effects of Bt maize residues on MBC and soil enzyme activities. Leaf residues of Bt and non-Bt maize cultivars (DKC 61-25B, DKC 61-24, PAN 6Q-321B and PAN6777) were used and soil without residues was used as a control. Samples were collected at 7, 28 and 56 days of incubation (DOI). An incubation study was also carried out in the laboratory to determine the effect of Bt maize residues (i.e. leaf, stem and root) and its genetic modification on N and P release patterns. Residues of DKC 61-25B, DKC 61-24, PAN 6Q-321B and PAN6777and soil without residues as a control were incubated in the laboratory. After destructive sampling at 0, 7, 14, 28, and 56 DOI, N in the form of NH4-N and NO3-N and P mineralisation were determined. Amendment of soil with residues enhanced MBC (p < 0.05) at all the sampling dates. For example MBC increased from 95 in the control to 146.3 mg/kg in the DKC 61-25B treatment at the end of the glasshouse trial. In the field DKC 61-25B had 9.1 mg/kg greater MBC than DKC 61-24, while PAN 6Q-321B had 23.9 mg/kg more MBC than PAN6777 at the end of the trial. However, no differences (p < 0.05) were observed in enzyme activities under field and glasshouse conditions except for dehydrogenase that had greater activity where DKC 61-25B and PAN 6777 were grown. There were no differences between the type of residues (Bt and non-Bt) on enzyme activities tested. However, differences were observed among the sampling dates. No effects of Bt maize crop on fungal spore count were observed. Similarly no differences were observed in leaf, stem and root tissues composition between Bt and non-Bt maize cultivars. Net N and P mineralisation from Bt maize cultivars did not differ from that of non-Bt maize cultivars. However, differences were observed among the cultivars. The results of this study suggested that Bt maize with Bt MON810 event can be grown in the central region of the Eastern Cape (EC), South Africa without affecting MBC, soil enzyme activities, VAM, and release of N and P nutrients from its residues.

There are concerns that genetic modification of maize with Bacillus thuringiensis may influence its structural and chemical composition which, together with the Cry1Ab proteins, may affect agro-ecosystem processes and feed quality. This study was aimed at investigating the expression of Cry1Ab protein in leaves, stems and roots of Bt maize and the effect of genetic modification (MON810) on activity of earthworms, silage quality and decomposition of residues in the field. In 2009/10 four maize cultivars; DKC61-25B, DKC61-24, PAN6Q-321B and PAN6777 were planted. Expression of Cry1Ab in leaves, stems and roots was analyzed at 6, 8, 10 and 12 weeks after planting (WAP). Earthworms were also sampled from the same treatments at 6, 9 and 18 WAP. Two silage experiments were conducted using maize cultivars, DKC80-12B and DKC80-10 produced in the 2008/09 season and DKC61-25B, DKC61-24, PAN6Q-321B and PAN6777, produced in 2009/10. The silage was sampled at 0, 2, 4, 8, 15 and 42 days in 2008/09 and 0, 8 and 42 days in 2009/10 and analyzed for Ash Free Dry Matter, Crude Fiber, Neutral Detergent Fiber, Acid Detergent Fiber, Acid Detergent Lignin, Crude Protein and Total Digestible Nutrients. Two litter-bag decomposition studies were also carried out (i) in 2008 (surface applied) using maize cultivars DKC80-12B, DKC80-10 and DKC6-125 residues and (ii) in 2009 (soil incorporated) using DKC75-15B, CRN3505, PAN6Q-321B v and PAN6Q-121. Ash-free dry matter and Cry1Ab protein were measured throughout the incubation time. There were no differences between DKC61-25B and PAN6Q-321B in terms of expression of Cry1Ab in leaves, stems and roots over time. The Cry1Ab expression levels were in decreasing order: leaves > stems > roots. No effects of Bt maize on earthworm numbers and biomass were observed. There were no differences in all silages parameters except NDF and ADF, which were higher in the Bt maize silage than that of the non-Bt maize from the 2008/09 season. The Cry1Ab levels were essentially not reduced during ensiling. The maize residues (both Bt and non-Bt maize) degraded to similar levels, either when surface-applied or incorporated into soil but soil-incorporated residues decomposed faster than surface-applied ones. Cry1Ab degraded as the plant matrix decomposed. The findings suggested that maize genetically modified with the Bt MON810 event can be grown in the Central Eastern Cape without affecting earthworm numbers and biomass, silage quality and decomposition of maize residues.

Grain protein content is an essential component to producing a profitable Hard Red Spring Wheat (HRSW) (Triticum aestivum L.) crop in the northern Great Plains. Growers can increase grain protein content through in-season N fertilization; however, the cost of these applications may outweigh the benefits. Predicting the grain protein content of early-season HRSW would give growers crucial information as they decide whether to apply in-season fertilizer to boost grain protein content. This research encompasses three studies; two of which aim to predict grain protein content with hand-held and aerial sensors respectively, and a third, which investigates the optimal rate, timing, and source of N fertilizer to boost grain protein content. Results of these experiments seemed to be greatly influenced by environmental factors. Findings of this research suggest that an in-season N application should be used for ameliorative purposes and not as a regular practice. / Minnesota Research and Promotion Council / Minnesota Association of Wheat Growers

Les systèmes alimentaires occidentaux ne sont pas durables en termes d‘impacts environnementaux et d‘effet sur la santé. Une solution est de revisiter les modes de consommation en favorisant les produits à base de protéines végétales. Cependant, un des verrous à l‘introduction des protéines végétales dans notre alimentation est leurs défauts sensoriels qui sont un frein à l‘acceptabilité des produits par les consommateurs. La fermentation est un procédé ancien qui pourrait être un levier à ce verrou sensoriel. Dans ce contexte, l‘objectif de ce projet de thèse est d‘étudier les bénéfices sensoriels apportés par la fermentation de gels enrichis en protéines végétales à l‘aide de consortia microbiens sélectionnés. Pour cela, une stratégie alliant analyses sensorielles, microbiologie et physicochimie a été mise en œuvre. Deux types de matrices contenant 10% de protéines et 10% d‘huile de colza ont été étudiés : la première est constituée de 100% de protéines de pois et la deuxième est constituée d‘un mélange de protéines de lait (50%) et de pois (50%). Sur la base des connaissances de la matrice et des propriétés fonctionnelles des micro-organismes, 56 souches microbiennes ont été sélectionnées. Une stratégie raisonnée d‘assemblage de ces souches a été mise en place, basée sur la répartition équilibrée selon leur groupe phylogénétique, mais aussi sur la connaissance experte des fonctions cibles aromatiques recherchées. En parallèle, plusieurs procédés de gélification ont été étudiés pour structurer les gels. Dans un premier temps, la fermentation a été étudiée sur des émulsions non gélifiées pour permettre la sélection des communautés microbiennes spécifiques pour chaque matrice. Dans un second temps, le potentiel d‘adaptation et de fonctionnement des écosystèmes sélectionnés ont été étudiés sur les émulsions gélifiées. La croissance et l‘abondance des microorganismes dans les gels après trois et sept jours de fermentation ont été évaluées sur milieu spécifique et le potentiel aromatique des consortia a été cartographié par un panel sensoriel.Dans une dernière partie du travail, les défauts/bénéfices sensoriels de la fermentation ont été étudiés de point de vue sensoriel et analytique. Les résultats montrent un fort potentiel d‘implantation des bactéries lactiques (principalement Lactobacillus plantarum, Lactobacillus rhamnosus, Lactococcus lactis et Lactobacillus casei) et de la majorité des eucaryotes (en particulier Mucor et Geotrichum) pour l‘émulsion mixte comme pour l‘émulsion végétale. Même si les souches appartenant aux groupes Actinobacteria et Proteobacteria ne soient pas compétitives vis-à-vis de la flore endogène (Bacillus), certaines espèces comme Hafnia alvei, Acinetobacter johnsonii et Glutamicibacter arilaitensis, ont montré une forte croissance quand elles sont inoculées en associations. Des notes aromatiques spécifiques pour chaque émulsion ont été générées permettant de masquer la note végétale « verte » caractéristique du pois. Ainsi, deux consortia ont pu être sélectionnés sur la base de notes lactiques et fruitées pour l‘émulsion végétale (VEGAN), et de notes fruitées pour l‘émulsion mixte (MEGAN). Le potentiel d‘adaptation de ces deux consortia dépendait de la composition et de la structure des gels. Ainsi, le consortium VEGAN semble bien adapté pour le gel végétal et génère des notes torréfiées/grillées, alors qu‘il génère des notes laitières (crème fraiche/caillé frais) dans le gel mixte. Le consortium MEGAN s‘implante très bien dans les deux types de gels, générant des notes fromagères et fruitées dans le gel mixte, mais ne permet pas de masquer les notes vertes dans le gel végétal. Les défauts sensoriels attribués aux notes vertes et à l‘amertume sont liés principalement à la présence des aldéhydes et des acides aminés hydrophobes respectivement, mais restent encore à approfondir. (suite et fin du résumé dans la thèse) / Western food systems are not sustainable in terms of environmental impacts and health effects. One solution is to revisit consumption patterns by favoring products based on plant proteins. However, one of the barriers to the introduction plant proteins in our diet is their sensory defects which can be obstacles for the acceptability of products by consumers. Fermentation is an ancient process that could be a solution to this sensory issue. In this context, the aim of this PhD thesis is to study the sensory benefits provided by the fermentation of plant protein-based gels using selected microbial consortia. In order to accomplish this, a strategy combining sensory analyzes, microbiology and physicochemistry was implemented. Two types of matrices containing 10% protein and 10% rapeseed oil were studied: the first consists of 100% pea protein and the second consists of a mixture of milk proteins (50%) and pea proteins (50%).Based on knowledge of the matrix and the functional properties of microorganisms, 56 microbial strains were selected. A reasoned strategy of assembly of these strains was put in place, based on the balanced distribution according to their phylogenetic group, but also on the expert knowledge of the desired aromatic functions. In parallel, several gelling processes were studied to structure the gels. In the first step, the fermentation was studied on non-gelled emulsions to allow the selection of specific microbial communities for each matrix. In the second step, the adaptation and functioning potential of the selected ecosystems were studied on gelled emulsions. The growth and abundance of microorganisms in the gels after three and seven days of fermentation were evaluated on a specific growing medium and the aromatic potential of the consortia was mapped by a sensory panel. In the final section of this study, the sensory defects / benefits of fermentation were studied from a sensorial and analytical point of view. The results show a highpotential for implantation of lactic acid bacteria and the majority of eukaryotes (in particular Mucor and Geotrichum) for the mixed emulsion and for the vegetable emulsion. Although strains belonging to the Actinobacteria and Proteobacteria groups are not competitive with endogenous flora (Bacillus), certain species such as Hafnia alvei, Acinetobacter johnsonii and Glutamicibacter arilaitensis, have shown strong growth when inoculated into associations. Specific aromatic notes for each emulsion were generated to mask the green note characteristic of peas. Thus, two consortia were selected on the basis of lactic and fruity notes for the vegetable emulsion (VEGAN), and fruity notes for the mixed emulsion (MEGAN). The adaptation potential of these two consortia depended on the composition and structure of the gel. Thus, the VEGAN consortium seems well suited for vegetable gels and generates roasted / grilled notes, while it generates dairy notes (fresh cream / fresh curd) in the mixed gels. The MEGAN consortium implements itself very well in both types of gels, generating cheesy and fruity notes in the mixed gel, but does not mask the green notes in the vegetable gels. The sensory defects attributed to green notes and bitterness are mainly related to the presence of aldehydes and hydrophobic amino acids respectively, but still need to be deepened. This study validated a proof of concept of formulation of fermented food products and will create opportunities for innovation.

There has been growing interest in the utilization of plant-derived proteins as functional ingredients in many food and beverage applications because they are perceived as being more sustainable, healthy, and ethical than animal-derived proteins by many consumers. Traditionally, soy proteins have been the most widely employed plant protein in the food industry. However, a number of alternative plant-based protein sources have recently become available, with pulse proteins being one of the most popular. In this study, the physicochemical properties and functional attributes of various commercially available pulse protein isolates were compared with those of soy protein isolate to evaluate their potential application in foods and beverages. The water holding capacity, oil holding capacity, gelation properties, emulsifying properties, and color of faba bean (FPI), pea (PPI), lentil (LPI), and soy (SPI) protein isolates were therefore measured. SPI had a significantly higher water holding capacity (7.6 g/g) than the pulse protein isolates (2.2-5.1 g/g). Among the plant protein isolates, PPI had a significantly lower oil holding capacity and gelling property. LPI was more effective at producing small oil droplet sizes during homogenization than the other protein isolates. Nevertheless, all of the plant proteins were capable of forming relatively small oil droplets (D32 = 1-3 mm) at a protein-to-oil ratio of 1:10. As expected, droplet size decreased with increasing protein concentration for all plant protein isolates, which increased their resistance to creaming. These results suggest that pulse proteins may have similar or better techno-functional properties than soy proteins for certain applications. In particular, lentil proteins were more effective emulsifiers, whereas faba bean proteins were more effective gelling agents. These proteins may therefore be suitable for application in plant-based milks, eggs, cheese, or meats where emulsifying or gelling properties are required.

The purpose of this study was to explore the influence of external product attributes on consumers’ perceived acceptance of pea protein as a protein substitute in food products. The interest in pea protein as a possible protein source was investigated and the various advantages of consuming pea protein were highlighted. In previous studies, pea protein was identified as a possible substitute for red meat (Davies & Lightowler, 1998), but also as protein containing the amino acid profile which corresponds with the profile recommended for adults by the Food and Agriculture Organisation (FAO) (Swendseid, 1981). Additionally the nutritional and health benefits, the environmental benefits of growing pea protein as well as the non-allergen declaration accompanying pea protein were discussed. The various factors that will influence the consumer’s decision making process when introduced to a new and unfamiliar protein source such as pea protein were investigated in terms of the external product attributes as well as the factors indicated in the Theory of Reasoned Action (TRA) (Fishbein & Ajzen, 1975). The external product attributes were identified as the influence of price, marketing of the product, packaging attributes of new food products, branding importance of a product and new product education and knowledge perceived necessary for acceptability of a new food product. The additional factors which were investigated, as indicated by the TRA, included the influence of the subjective norms or significant others, as well as the consumers’ personal attitude. attitude. Due to the limited insight and understanding of the influences that external product attributes have on a consumer’s perceived acceptance of pea protein as a protein substitute in food products, a qualitative exploratory research design was initiated. An exploratory research design was incorporated because this afforded the researcher the opportunity to gather thorough and comprehensive data or information and it also increased the richness and depth of explorations that could be researched (Key, 2002). The fact that an exploratory research design was incorporated, a non-probability sampling strategy was implemented. Additionally a convenient sampling strategy was also implemented because this type of sampling strategy represents sites or individuals from which data can easily be collected and where access is convenient (Cresswell, 2007). The unit of analysis for this research was female shoppers, being the main grocery and food shoppers in the household, who were living in suburban areas. The participants of the research, therefore, resided mainly in the Johannesburg and Pretoria suburban areas, within the Gauteng province and were chosen because these are two of the major suburban cities in South Africa. The data were collected through means of a multi-method approach whereby different data collection instruments were applied. In this regard focus group interviews, sentence completion exercise and a vignette were used which addressed the objectives set out in the study. From the findings of the study on the influence of external product attributes on consumers’ perceived acceptance of new products in general and new products containing pea protein, it was found that there were not any noticeable differences between a new food product and a new food product containing pea protein. The consumer did not indicate a greater willingness to accept general new products in comparison to a pea protein product. On the contrary, the fact that a new product contained pea protein did not make any difference. For consumers external product attributes in general played a major role in their purchasing decision making process. The most important requirement that the consumers had was that additional information regarding the pea protein should be provided. It was indicated by the participants that they would be interested in the pea protein product and consider purchasing it and incorporating it into their daily diet, providing they knew more about the product and had sufficient background information about the new product. Furthermore, the importance of marketing of a new product was highlighted and the fact that the packaging of the new product should be attractive and eye-catching. Price competiveness was mentioned, however, should a new and unfamiliar product be introduced by a well-known brand, this would lower the initial resistance to the new product. One of the main limitations of this study is the relatively small sample size from a particular area in South Africa and, therefore, resulting in a very selective sample. The sample, therefore, remains one of convenience which limits the ability to generalize or transfer findings to a bigger group of consumers. Furthermore the study location was within a limited area, namely Gauteng. For this reason no assumptions were made nor were findings considered to be transferable to other population groups or consumer groups. Another limitation is that the topic of this study is very new and unfamiliar; there are, therefore, no previous studies with which to compare the findings. This study was exploratory in nature and the instruments were chosen specifically to explore a new and unfamiliar topic. The transferability of this study might be limited, but care was taken to describe the research context in as much detail as possible. In doing so, an attempt was made to enhance the transferability, should another researcher wish to duplicate the study. From the findings of this study certain recommendation can be made as most participants were of the opinion that they would be interested in the pea protein product and consider purchasing it and incorporating it into their daily diet provided they knew more about the product and sufficient background information had been supplied. Furthermore, it was noted from the findings that these external attributes would influence the new product acceptability and, therefore, giving the research and developers as well as the marketing team of a manufacturing company the information necessary to understand the requirements from the consumers regarding the product containing pea protein better and how this product should be marketed as a healthy alternative to other vegetable protein sources. Further recommendations are that should a food manufacturing company wish to develop a pea protein product, they would need to promote the product beforehand and create an awareness of the advantages and benefits of pea proteins. This can be done by editorials in magazines as well as promotions in printed media. Additionally, the competition between products and brands is fierce and, therefore, manufacturing companies need to differentiate their products to keep consumers interested in their product or brand. Pea protein is a new and innovative product and can, therefore, add that differentiation and benefit to food products. Future research that can be considered is that of a sensory study, to determine the actual viability of the pea protein products based on sensory evaluation. One of the main criteria indicated for the acceptability of a food product containing pea protein has been the taste. The present research was theoretical and exploratory in nature and, therefore, there were no actual products which the participants could taste. Additionally, the inclusion criteria for this study was identified as female shoppers who are in charge of household grocery and food purchases and were, therefore, specifically recruited to participate in the research. However, based on a study done by Jensen (2011), it is recommended that further studies specifically include males because men are doing more of the household grocery shopping and they are not as brand loyal as females and would, therefore, be more willing to try and experiment with new food products. A study based on the gender differences of the acceptability of new products, could highlight different extrinsic attributes that would influence the purchasing decision. / Business Management / M. Consumer Sc.

A soja geneticamente modificada MON 87701 × MON 89788, Glycine max (L.) Merrill, que expressa genes que codificam a proteína Cry1Ac de Bacillus thuringiensis var. kurstaki Berliner e a proteína 5-enolpiruvilchiquimato-3-fosfato sintase de Agrobacterium sp., foi liberada para uso comercial no Brasil em 2010. Para subsidiar um programa de Manejo da Resistência de Insetos (MRI) foi realizada a avaliação de risco de evolução de resistência a Cry1Ac para as pragas-alvo primárias, Anticarsia gemmatalis Hübner e Pseudoplusia includens (Walker), a praga-alvo secundária Heliothis virescens (Fabricius) e as pragas nãoalvo Spodoptera cosmioides (Walker), Spodoptera eridania (Stoll) e Spodoptera frugiperda (J. E. Smith). Em bioensaios com a proteína Cry1Ac purificada incorporada em dieta artificial, verificou-se que Cry1Ac foi extremamente ativa para A. gemmatalis [CL50 (IC 95%) = 0,23 (0,15 - 0,34) g Cry1Ac/mL de dieta], P. includens [CL50 (IC 95%) = 3,72 (2,65 - 4,86) g Cry1Ac/mL de dieta] e H. virescens [CL50 (IC 95%) = 0,026 (0,021 - 0,033) g de Cry1Ac/mL de dieta]. Em contraste, na máxima concentração testada de 100 g Cry1Ac/mL de dieta, S. cosmioides e S. eridania apresentaram mortalidade < 13%, e S. frugiperda de » 50%. Em bioensaios com tecido liofilizado de soja MON 87701 × MON 89788, diluído 25 vezes em dieta artificial, houve 100% de mortalidade para A. gemmatalis e H. virescens e até 96% para P. includens. Entretanto, as lagartas sobreviventes de P. includens apresentaram enfezamento larval > 95%. Em bioensaios com discos de folha em laboratório (para A. gemmatalis, P. includens e H. virescens), vagens (para H. virescens) e elevada infestação em condições de casa de vegetação e de infestação natural em campo (para A. gemmatalis e P. includens) foram obtidas alta eficácia da soja MON 87701 × MON 89788 no controle das pragas-alvo primárias e secundária. Por outro lado, a soja MON 87701 × MON 89788 apresentou baixa eficácia para S. cosmioides e S. eridania (mortalidade < 7%) e atividade intermediária para S. frugiperda (32 a 47% mortalidade). A soja MON 87701 × MON 89788 não afetou os parâmetros biológicos de S. cosmoides e S. eridania. Para S. frugiperda houve prolongamento da fase larval (» 5 dias), menor viabilidade larval e total, aumento no intervalo entre gerações (» 8 dias) e redução na taxa intrínseca de crescimento (» 41%). No contexto do MRI, a soja MON 87701 × MON 89788 expressa a proteína Cry1Ac em níveis que constituem alta dose para A. gemmatalis e H. virescens, e muito próximo a alta dose para P. includens. Por outro lado, para as espécies de Spodoptera, a soja MON 87701 × MON 89788 é um evento de baixa dose, pois permite que indivíduos suscetíveis completem o ciclo biológico. Em termos de MIP, a soja MON 87701 × MON 89788 apresenta um elevado nível de controle de A. gemmatalis, P. includens e H. virescens. No entanto, a soja MON 87701 × MON 89788 não ocasiona controle efetivo de espécies de Spodoptera. As informações obtidas no presente trabalho contribuirão para subsidiar programas de MRI e preservar a durabilidade dessa tecnologia para o MIP-Soja no Brasil. / Genetically modified MON 87701 × MON 89788 soybean, Glycine max (L.) Merrill, expressing genes that code for the Cry1Ac protein of Bacillus thuringiensis var. kurstaki Berliner and the 5-enolpyruvylshikimate-3-phosphate synthase protein of Agrobacterium sp., has been registered for commercial use in Brazil in 2010. To develop a program of Insect Resistance Management (IRM) for this event, we conducted resistance risk assessment to the primary target pests, Anticarsia gemmatalis Hübner and Pseudoplusia includens (Walker), the secondary target pest Heliothis virescens (Fabricius) and the non-target pests Spodoptera cosmioides (Walker), Spodoptera eridania (Stoll) and Spodoptera frugiperda (J. E. Smith) to the Cry1Ac protein. In bioassays with purified Cry1Ac protein incorporated into artificial diet, Cry1Ac was highly active for A. gemmatalis [LC50 (95% FL) = 0.23 (0.15 - 0.34) g Cry1Ac/mL of diet], P. includens [LC50 (95% FL) = 3.72 (2.65 - 4.86) g Cry1Ac/mL of diet] and H. virescens [LC50 (95% FL) = 0.026 (0.021 - 0.033) g Cry1Ac/mL of diet]. In contrast, even at the highest concentration tested of 100 g Cry1Ac/mL of diet, the mortality observed to S. cosmioides and S. eridania was < 13% and to S. frugiperda was » 50%. In bioassays with freeze-dried MON 87701 × MON 89788 soybean tissue, diluted 25 times in artificial diet, there was 100% mortality to A. gemmatalis and H.virescens and up to 96% for P. includens. However, the surviving P. includens larvae showed > 95% larval stunting. In bioassays with leaf discs in the laboratory to (A. gemmatalis, P. includens and H. virescens), pods (H. virescens), high pest infestation studies under greenhouse conditions and in natural infestation in the field (to A. gemmatalis and to P. includens) showed a very high level of efficacy against these primary and secondary target pests. On the other hand, soybean MON 87701 × MON 89788 showed low efficacy to S. cosmioides and S.eridania (mortality < 7%), and intermediate activity to S. frugiperda (32 to 47% of mortality). MON 87701 × MON 89788 soybean did not affect the biological parameters of S. cosmoides and S. eridania. For S. frugiperda, the larval stage was prolonged (» 5 days), with reduction on larval and total viability, increase in generation time (» 8 days) and reduction in the intrinsic rate of increase (» 41%). In the context of IRM, MON 87701 × MON 89788 soybean expresses the Cry1Ac protein at levels that are high-dose for A. gemmatalis and H. virescens, and near to the highdose for P. includens. On the other hand, for the Spodoptera species, MON 87701 × MON 89788 soybean is a low-dose event, because it allows susceptible individuals to complete larval developement. In terms of IPM, MON 87701 × MON 89788 soybean provided a high level of control of A. gemmatalis, P. includens and H. virescens. However, MON 87701 × MON 89788 soybean was not effective to control Spodoptera species. The information obtained in this research will contribute to develop IRM programs and to preserve the durability of this technology for soybean IPM in Brazil.