Influence of adjuvants added to teflubenzuron spray on resistant and susceptible strains of the soybean looper Chrysodeixis includens (Lepidoptera: Noctuidae) / Influência da adição de adjuvantes à calda de pulverização de teflubenzuron em linhagens resistentes e suscetíveis da lagarta falsa-medideira Chrysodeixis includens em soja (Lepidoptera: Noctuidae)

The soybean looper (SBL), Chrysodeixis includens (Walker, [1858]) (Lepidoptera: Noctuidae), is a soybean and cotton pest in South America countries. Field-evolved resistance of SBL to inhibitors of chitin biosynthesis has been reported in Brazil; however, this mode of action is still widely used against SBL. On this basis, we conducted laboratory bioassays to investigate if adjuvants (Nimbus®, TA 35®, Break-Thru® S 240, and Rizospray Extremo®) added to the teflubenzuron spray increase the mortality of SBL strains (resistant, heterozygous, and susceptible to chitin biosynthesis inhibitors). Using chromatography analysis, we also evaluated the amount of teflubenzuron on soybean leaves when applied alone or in combination with adjuvants. In laboratory bioassays, the biological activity of teflubenzuron increased against the susceptible SBL strain when adjuvants were added. In contrast, no relevant effects of adjuvants added to the teflubenzuron spray against heterozygous and resistant SBL larvae were detected. In leaf bioassays, even leaves from the upper third part of the plants containing a significantly higher amount of teflubenzuron (3.4 mg/kg vs 1.7 and 0.6 mg/kg); the mortality of SBL strains was similar when teflubenzuron was applied alone or in mixture with adjuvants. Our findings indicated that adjuvants added to teflubenzuron spray do not provide a substantial increase in the mortality of SBL strains resistant to chitin biosynthesis inhibitors. Therefore, it is necessary to reduce the use of this mode-ofaction insecticide against SBL and to give preference to other insecticides or control tactic.

A lagarta falsa-medideira, Chrysodeixis includens (Walker, [1858]) (Lepidoptera: Noctuidae), é uma praga da soja e do algodão nos países da América do Sul. A resistência de C. includens a inibidores da biossíntese de quitina tem sido relatada no Brasil. Entretanto, esse modo de ação ainda é amplamente utilizado para controle de C. includens. Com base nisso, conduzimos bioensaios em laboratório para investigar se adjuvantes (Nimbus®, TA 35®, Break-Thru® S 240 e Rizospray Extremo®) adicionados à calda inseticida de teflubenzuron aumentam a mortalidade de linhagens de C. includens (resistentes, heterozigotos e suscetíveis a inibidores da biossíntese de quitina). Usando análise cromatográfica, também avaliamos a quantidade de teflubenzuron em folhas de soja quando aplicado isolado ou em combinação com adjuvantes. Em bioensaios de laboratório, a atividade biológica do teflubenzuron aumentou para a linhagem suscetível quando os adjuvantes foram adicionados à calda inseticida. Em contraste, nenhum efeito relevante de adjuvantes adicionados ao teflubenzuron foi detectado para os heterozigotos e resistentes. Em bioensaios de folhas, mesmo naquelas do terço superior das plantas, as quais apresentaram uma maior deposição de teflubenzuron (3,4 mg/kg vs 1,7 e 0,6 mg/kg); a mortalidade das linhagens de C. includens foi semelhante quando o teflubenzuron foi aplicado isolado ou com adjuvantes. Nossos resultados indicam que os adjuvantes adicionados ao teflubenzuron não fornecem um aumento substancial na mortalidade de linhagens de C. includens resistentes aos inibidores da biossíntese de quitina. Portanto, é necessário reduzir o uso desse modo de ação para o manejo de C. includens e dar preferência a outros inseticidas ou tática de controle.

Do fall armyworm's Metaflumizone resistante populations affect the activity of Trichogramma pretiosum? / Populações da lagarta-do-cartucho resistentes à Metaflumizona afetam a atividade de Trichogramma pretiosum?

The possible interference of resistant pest’s populations to insecticides in natural enemies in the action thas not been clarified yet. Thus, this study aimed to evaluate Trichogramma pretiosum Riley (Hymenoptera: Trichogrammatidae) performance on Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae) eggs with resistance frequency to the Metaflumizone over six generations of product exposure. Egg cards (2.0 x 7.0 cm) containing eggs from two populations of S. frugiperda, (resistant to Metaflumizone and the other susceptible), were exposed to T. pretiosum females for 24 hours in free-choice and no-choice testing in three generations (G1, G4, and G6). A completely randomized experimental design was used with 25 replications, each consisting of an egg card (experimental unit) containing 20 eggs. The parameters evaluated were: parasitism (%), emergence (%), sex ratio, number of emerged parasitoids per egg and males/females longevity. ANOVA and Tukey test (P≤ 0.05) were applied on the results. Results showed a reduction in parasitism [41.0% (G1) and 28.4% (G4)], egg emergence (17.5%) and parasitoids/egg [16.2 (G4) and 17.2 (G6)] in eggs originating from the population with resistance frequency. Females emerging from G6 populations eggs without exposure to Metaflumizone had greater longevity (3.5 days more) than the resistant population. The sex ratio and male longevity were not affected. The results indicate a reduction in T. pretiosum activity if S. frugiperda populations have some frequency of resistance to Metaflumizone.

A possível interferência de populações de pragas resistentes na ação de inimigos naturais ainda não foi esclarecida. Assim, este trabalho teve como objetivo avaliar o desempenho de Trichogramma pretiosum Riley (Hymenoptera: Trichogrammatidae) em ovos de Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae) com frequência de resistência à Metaflumizona ao longo de seis gerações de exposição ao produto. Cartelas (2,0 x 7,0 cm) com ovos de duas populações de S. frugiperda, (resistente à Metaflumizona e outra suscetível), foram expostas às fêmeas de T. pretiosum por 24 horas em condições de livre escolha e sem chance de escolha por três gerações (G1, G4 e G6). O delineamento experimental foi inteiramente casualizado com 25 repetições, sendo cada repetição composta por uma cartela (unidade experimental) contendo 20 ovos. Os parâmetros avaliados foram: parasitismo (%), emergência (%), razão sexual, número de parasitoides emergidos por ovo e longevidade de machos e fêmeas. ANOVA e teste de Tukey (P≤ 0,05) foram aplicados aos dados coletados. Os resultados mostraram redução do parasitismo [41,0% (G1) e 28,4% (G4)], emergência de ovos (17,5%) e parasitoides/ovo [16,2 (G4) e 17,2 (G6)] em ovos oriundos da população com frequência de resistência. As fêmeas emergidas de ovos da população G6 sem exposição à Metaflumizona, tiveram maior longevidade (3,5 dias a mais) do que a população exposta ao inseticida. A razão sexual e a longevidade de machos não foram afetadas. Os resultados indicam uma redução na atividade de T. pretiosum se as populações de S. frugiperda apresentarem alguma frequência de resistência à Metaflumizona.

Do fall armyworms Metaflumizone resistante populations affect the activity of Trichogramma pretiosum?

Abstract The possible interference of resistant pests populations to insecticides in natural enemies in the action thas not been clarified yet. Thus, this study aimed to evaluate Trichogramma pretiosum Riley (Hymenoptera: Trichogrammatidae) performance on Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae) eggs with resistance frequency to the Metaflumizone over six generations of product exposure. Egg cards (2.0 x 7.0 cm) containing eggs from two populations of S. frugiperda, (resistant to Metaflumizone and the other susceptible), were exposed to T. pretiosum females for 24 hours in free-choice and no-choice testing in three generations (G1, G4, and G6). A completely randomized experimental design was used with 25 replications, each consisting of an egg card (experimental unit) containing 20 eggs. The parameters evaluated were: parasitism (%), emergence (%), sex ratio, number of emerged parasitoids per egg and males/females longevity. ANOVA and Tukey test (P 0.05) were applied on the results. Results showed a reduction in parasitism [41.0% (G1) and 28.4% (G4)], egg emergence (17.5%) and parasitoids/egg [16.2 (G4) and 17.2 (G6)] in eggs originating from the population with resistance frequency. Females emerging from G6 populations eggs without exposure to Metaflumizone had greater longevity (3.5 days more) than the resistant population. The sex ratio and male longevity were not affected. The results indicate a reduction in T. pretiosum activity if S. frugiperda populations have some frequency of resistance to Metaflumizone.

Resumo A possível interferência de populações de pragas resistentes na ação de inimigos naturais ainda não foi esclarecida. Assim, este trabalho teve como objetivo avaliar o desempenho de Trichogramma pretiosum Riley (Hymenoptera: Trichogrammatidae) em ovos de Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae) com frequência de resistência à Metaflumizona ao longo de seis gerações de exposição ao produto. Cartelas (2,0 x 7,0 cm) com ovos de duas populações de S. frugiperda, (resistente à Metaflumizona e outra suscetível), foram expostas às fêmeas de T. pretiosum por 24 horas em condições de livre escolha e sem chance de escolha por três gerações (G1, G4 e G6). O delineamento experimental foi inteiramente casualizado com 25 repetições, sendo cada repetição composta por uma cartela (unidade experimental) contendo 20 ovos. Os parâmetros avaliados foram: parasitismo (%), emergência (%), razão sexual, número de parasitoides emergidos por ovo e longevidade de machos e fêmeas. ANOVA e teste de Tukey (P 0,05) foram aplicados aos dados coletados. Os resultados mostraram redução do parasitismo [41,0% (G1) e 28,4% (G4)], emergência de ovos (17,5%) e parasitoides/ovo [16,2 (G4) e 17,2 (G6)] em ovos oriundos da população com frequência de resistência. As fêmeas emergidas de ovos da população G6 sem exposição à Metaflumizona, tiveram maior longevidade (3,5 dias a mais) do que a população exposta ao inseticida. A razão sexual e a longevidade de machos não foram afetadas. Os resultados indicam uma redução na atividade de T. pretiosum se as populações de S. frugiperda apresentarem alguma frequência de resistência à Metaflumizona.

Do fall armyworm's Metaflumizone resistante populations affect the activity of Trichogramma pretiosum? / Populações da lagarta-do-cartucho resistentes à Metaflumizona afetam a atividade de Trichogramma pretiosum?

Abstract The possible interference of resistant pest's populations to insecticides in natural enemies in the action thas not been clarified yet. Thus, this study aimed to evaluate Trichogramma pretiosum Riley (Hymenoptera: Trichogrammatidae) performance on Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae) eggs with resistance frequency to the Metaflumizone over six generations of product exposure. Egg cards (2.0 x 7.0 cm) containing eggs from two populations of S. frugiperda, (resistant to Metaflumizone and the other susceptible), were exposed to T. pretiosum females for 24 hours in free-choice and no-choice testing in three generations (G1, G4, and G6). A completely randomized experimental design was used with 25 replications, each consisting of an egg card (experimental unit) containing 20 eggs. The parameters evaluated were: parasitism (%), emergence (%), sex ratio, number of emerged parasitoids per egg and males/females longevity. ANOVA and Tukey test (P≤ 0.05) were applied on the results. Results showed a reduction in parasitism [41.0% (G1) and 28.4% (G4)], egg emergence (17.5%) and parasitoids/egg [16.2 (G4) and 17.2 (G6)] in eggs originating from the population with resistance frequency. Females emerging from G6 populations eggs without exposure to Metaflumizone had greater longevity (3.5 days more) than the resistant population. The sex ratio and male longevity were not affected. The results indicate a reduction in T. pretiosum activity if S. frugiperda populations have some frequency of resistance to Metaflumizone.

Resumo A possível interferência de populações de pragas resistentes na ação de inimigos naturais ainda não foi esclarecida. Assim, este trabalho teve como objetivo avaliar o desempenho de Trichogramma pretiosum Riley (Hymenoptera: Trichogrammatidae) em ovos de Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae) com frequência de resistência à Metaflumizona ao longo de seis gerações de exposição ao produto. Cartelas (2,0 x 7,0 cm) com ovos de duas populações de S. frugiperda, (resistente à Metaflumizona e outra suscetível), foram expostas às fêmeas de T. pretiosum por 24 horas em condições de livre escolha e sem chance de escolha por três gerações (G1, G4 e G6). O delineamento experimental foi inteiramente casualizado com 25 repetições, sendo cada repetição composta por uma cartela (unidade experimental) contendo 20 ovos. Os parâmetros avaliados foram: parasitismo (%), emergência (%), razão sexual, número de parasitoides emergidos por ovo e longevidade de machos e fêmeas. ANOVA e teste de Tukey (P≤ 0,05) foram aplicados aos dados coletados. Os resultados mostraram redução do parasitismo [41,0% (G1) e 28,4% (G4)], emergência de ovos (17,5%) e parasitoides/ovo [16,2 (G4) e 17,2 (G6)] em ovos oriundos da população com frequência de resistência. As fêmeas emergidas de ovos da população G6 sem exposição à Metaflumizona, tiveram maior longevidade (3,5 dias a mais) do que a população exposta ao inseticida. A razão sexual e a longevidade de machos não foram afetadas. Os resultados indicam uma redução na atividade de T. pretiosum se as populações de S. frugiperda apresentarem alguma frequência de resistência à Metaflumizona.

RNA silencing of hormonal biosynthetic genes impairs larval growth and development in cotton bollworm, Helicoverpa armigera

The cotton bollworm, Helicoverpa armigera, is a highly polyphagous pest, causing enormous losses to variouseconomically important crops. The identification and in vitro functional validation of target genes of a pest is aprerequisite to combat pest via host-mediated RNA interference (RNAi). In the present study, six hormonalbiosynthesis genes of H. armigera were chosen and evaluated by feeding insect larvae with dsRNAs corresponding to each target gene, viz., juvenile hormone acid methyltransferase (HaJHAMT), prothoracicotropichormone (HaPTTH), pheromone biosynthesis-activating peptide (HaPBAP), molt regulating transcription factor(HaHR3), activated protein 4 (HaAP-4) and eclosion hormone precursor (HaEHP). The loss of function phenotypes for these hormonal genes were observed by releasing second instar larvae on to artificial diet containingtarget gene-specific dsRNAs. Ingestion of dsRNAs resulted in mortality ranging from 60% to 90%, reduced larvalweight, phenotypic deformities and delayed pupation. The quantitative real-time PCR (qRT-PCR) analysisshowed that the target gene transcript levels were decreased drastically (31% to 77%) as compared to control orunrelated control (GFP-dsRNA), and correlated well with the mortality and developmental defects of larvae.Also, a comparison of the silencing efficacy of un-diced long HaPTTH-dsRNA with RNase III diced HaPTTHdsRNA (siRNAs) revealed that long dsRNAs were more efficient in silencing the target gene. These resultsindicated that the hormonal biosynthesis genes have varied sensitivity towards RNAi and could be the vital targetsfor insect resistance in crop plants like cotton which are infested by H. armigera

Genetic engineering of crops for insect resistance: An overview

Phytophagous insect incidence is a serious threat for reduction of crop productivity globally. There is anestimation of one fourth of crop is being destroyed by insects annually. Indeed, the development of insectresistant crops is a great milestone in agriculture to increase crop yield and reduce pesticide dependency.Genetic engineering facilitates development of insect resistant crops by expressing bacterial d-endotoxins andvegetative insecticidal proteins and other plant genes like lectins, protease inhibitors, etc. In addition, RNAinterference and genome editing through CRISPR Cas9 also provides new solutions for the development ofinsect-resistant crops. The resultant genetically modified crops showed resistance against lepidopteran, dipteran, homopteran and coleopteran insects. The insect-resistant crops have made a significant economic impactworldwide in terms of higher yield and low pesticide usage. In this review, we focus on different strategies fordeveloping transgenics against insect pest control by expressing different insecticidal proteins in crops

Expression of Pinellia ternata leaf agglutinin under rolC promoter confers resistance against a phytophagous sap sucking aphid, Myzus persicae

BACKGROUND: Piercing/sucking insect pests in the order Hemiptera causes substantial crop losses by removing photoassimilates and transmitting viruses to their host plants. Cloning and heterologous expression of plantderived insect resistance genes is a promising approach to control aphids and other sap-sucking insect pests. While expression from the constitutive 35S promoter provides broad protection, the phloem-specific rolC promoter provides better defense against sap sucking insects. The selection of plant-derived insect resistance genes for expression in crop species will minimize bio-safety concerns. RESULTS: Pinellia ternata leaf agglutinin gene (pta), encodes an insecticidal lectin, was isolated and cloned under the 35S and rolC promoters in the pGA482 plant transformation vector for Agrobacterium-mediated tobacco transformation. Integration and expression of the transgene was validated by Southern blotting and qRT-PCR, respectively. Insect bioassays data of transgenic tobacco plants showed that expression of pta under rolC promoter caused 100% aphid mortality and reduced aphid fecundity up to 70% in transgenic tobacco line LRP9. These results highlight the better effectivity of pta under rolC promoter to control phloem feeders, aphids. CONCLUSIONS: These findings suggested the potential of PTA against aphids and other sap sucking insect pests. Evaluation of gene in tobacco under two different promoters; 35S constitutive promoter and rolC phloemspecific promoter could be successfully use for other crop plants particularly in cotton. Development of transgenic cotton plants using plant-derived insecticidal, PTA, would be key step towards commercialization of environmentally safe insect-resistant crops.

Development of Helicoverpa armigera (Hübner, 1805) and Spodoptera frugiperda (SMITH, 1797) in winter forages / Desenvolvimento de Helicoverpa armigera (Hübner, 1805) e Spodoptera frugiperda (SMITH, 1797) em forragens de inverno

Helicoverpa armigera Hübner, 1805 and Spodoptera frugiperda Smith, 1797 are polyphagous pests of great agricultural importance in subtropical and temperate climate regions. The usual management of production areas in the southern region of Brazil occurs after the harvesting of summer crops, and the areas are sown with pasture. Thus, forages recommended for grazing are azevém (Lolium multiflorumLam.) and cornichão (Lotus corniculatus L.) due to their nutritional benefits, good palatability, regrowth, and hardiness. Considering the high degree of polyphagia of H. armigera and S. frugiperda, and the impact of maintaining continuous feeding areas (green bridges) in the management of these species, this work aimed to evaluate the development of H. armigera and S. frugiperda fed leaves of azevém and cornichão under laboratory conditions. The insects were collected in the city of Capão do Leão/RS in corn and soybean plantations. For each forage species, 130 newly hatched caterpillars were each placed in autoclaved glass tubes, with one-third of the tube length containing forage. The tubes were capped with waterproof cotton and placed in a climate-controlled room at 25 ± 1°C, 70 ± 10% relative humidity, and a 12-h photophase. The food was replenished daily until the caterpillars entered the pupae stage. The pupae were sexed and weighed, and the newly emerged adults were placed in pairs in PVC cages, lined with white A4 sulfite paper. The papers used as a laying substrate were removed and changed daily, and eggs were counted. The number of instars was determined by the linearized Dyar rule model. The complete randomization design was employed for the variables biological cycle length and viability of egg, caterpillar, pre-pupa, pupa, adult, and pre-oviposition phases and weight of caterpillars on the 14th day and pupae after 24 h. Based on the results obtained, a fertility life table was prepared. H. armigera did not complete the cycle, with only three instars and a duration of 22.1 and 24.6 days in azevém and cornichão, respectively. When evaluated in S. frugiperda caterpillars, development in forage species was observed, with five and six instars and duration of 51.7 and 45.1 days in azevém and cornichão, respectively. The azevém was distinguished by interference in the development cycle of the species, reducing the effect of the green bridge. In addition, surviving insects were susceptible to the effects of the agroecosystem due to the low-quality food source.

Helicoverpa armigera Hübner, 1805 e Spodoptera frugiperda Smith, 1797 são pragas polífagas de grande importância agrícola, em regiões de clima subtropical e temperado. Após a colheita das culturas de verão no sul do Brasil, as áreas são utilizadas como pastagem. Com isso, as forrageiras recomendadas ao pastoreio são cornichão (Lotuscorniculatus L.) e azevém (Loliummultiflorum Lam.) por apresentarem benefícios nutricional, palatabilidade, rebrota e rusticidade. Devido ao alto grau de polifagia de H. armigera e S. frugiperda e o impacto de áreas continuas de alimento (pontes verdes) no manejo destas espécies, este trabalho objetivou-se avaliar o desenvolvimento de H. armigera e S. frugiperda alimentadas com azevém e cornichão em condições de laboratório. Os insetos foram coletados no município de Capão do Leão/RS em plantios de milho e soja. Para cada espécie forrageira foram individualizadas 130 lagartas recém-eclodidas em tubos de vidro autoclavados, com um terço (1/3) do seu comprimento contendo a respectiva forrageira. Os tubos foram tampados com algodão hidrófugo e levados para a sala climatizada a 25 ± 1°C, UR de 70 ± 10% e 12 horas de fotofase. Diariamente, o alimento foi reposto até que as lagartas entrarem em estágio de pupa. As pupas foram sexadas e pesadas e os adultos recém-emergidos foram individualizados em casais em gaiolas de PVC, forradas com papel sulfite A4 branco no seu interior. Os papéis utilizados como substrato de postura foram retirados e trocados diariamente e os ovos foram contabilizados. O número de instares foi determinado pelo modelo linearizado da regra de Dyar. O delineamento utilizado foi inteiramente casualizado (DIC) para as variáveis duração do ciclo biológico e viabilidade das fases de ovo, lagarta, pré-pupa, pupa, adultos e pré-oviposição, peso de lagartas ao decimo quarto (14°) dia e pupas após 24 horas. Com base nos resultados obtidos, foi elaborada tabela de vida de fertilidade. H. armigera não completou o ciclo, com apenas três instares e duração de 22,1 e 24,6 dias em azevém e cornichão, respectivamente. Quando avaliado em lagartas de S. frugiperda observou o desenvolvimento nas espécies forrageiras, com 5 e 6 instares e duração de 51,7 e 45,1 dias em azevém e cornichão, respectivamente. Contudo, considerando o critério que evite a formação de ponte verde, azevém destacou-se por interferir no ciclo de desenvolvimento das espécies. Onde, mesmos os que sobrevivem, estão suscetíveis aos efeitos do agroecossitema devido a fonte alimentar ser de baixa qualidade.

Preliminary Environmental Risk Assessment of Insect Resistant Genetically Modified Maize MON 89034 for Cultivation (EFSA/GMO/BE/2011/90)

The environmental risk assessment of the insect resistant genetically modified maize MON 89034 (Reference EFSA/GMO/BE/2011/90) has been performed by the Panel on Genetically Modified Organisms (GMO) of the Norwegian Scientific Committee for Food Safety (VKM). VKM has been requested by the Norwegian Directorate for Nature Management and the Norwegian Food Safety Authority to issue a preliminary scientific opinion on the safety of the genetically modified maize MON 89034 (Unique identifier MON-89Ø34-3) for cultivation, and submit relevant scientific comments or questions to EFSA on the application EFSA/GMOBE/2011/90. The current submission is intended to complement application EFSA-GMO-NL-2007-37, which was approved by Commission Decision 2009/813/EC of 30 October 2009, authorising the placing on the market of products containing, consisting of, or produced from genetically modified maize MON 89034 (scope import, processing, food and feed). Maize MON89034 has previously been assessed by the VKM GMO Panel in connection with EFSA´s public hearing of the application EFSA/GMO/NL/2007/37 (VKM 2008a). Preliminary health- and environmental risk assessments of several stacked events, with MON 89034 as one of the parental lines, have also been performed by the VKM GMO Panel (VKM 2009a, b, c; VKM 2010a,b). The environmental risk assessment of the maize MON 89034 is based on information provided by the applicant in the application EFSA/GMO/BE/2011/90, and scientific comments from EFSA and other member states made available on the EFSA website GMO Extranet. The risk assessment also considered peer-reviewed scientific literature as relevant. The VKM GMO Panel has evaluated MON 89034 with reference to its intended uses in the European Economic Area (EEA), and according to the principles described in the Norwegian Food Act, the Norwegian Gene Technology Act and regulations relating to impact assessment pursuant to the Gene Technology Act, Directive 2001/18/EC on the deliberate release into the environment of genetically modified organisms, and Regulation (EC) No 1829/2003 on genetically modified food and feed. The Norwegian Scientific Committee for Food Safety has also decided to take account of the appropriate principles described in the EFSA guidelines for the risk assessment of GM plants and derived food and feed (EFSA 2006, 2011a), the environmental risk assessment of GM plants (EFSA 2010), the selection of comparators for the risk assessment of GM plants (EFSA 2011b), and for the post-market environmental monitoring of GM plants (EFSA 2006, 2011c). The scientific risk assessment of maize MON 89034 include molecular characterisation of the inserted DNA and expression of target proteins, comparative assessment of agronomic and phenotypic characteristics, unintended effects on plant fitness, potential for gene transfer, interactions between the GM plant and target and non-target organisms, effects on biogeochemical processes and evaluations of the post-market environmental plan. In line with its mandate, VKM emphasised that assessments of sustainable development, societal utility and ethical considerations, according to the Norwegian Gene Technology Act and Regulations relating to impact assessment pursuant to the Gene Technology Act, shall not be carried out by the Panel on Genetically Modified Organisms. The genetically modified maize MON 89034 was developed to provide protection against certain lepidopteran target pest, including European corn borer (Ostrinia nubilalis) and Mediterranean corn borer (Sesamia nonagrioides). Protection is achieved through expression in the plant of two insecticidal Cry proteins, Cry1A.105 and Cry2Ab2, derived from Bacillus thuringiensis, a common soil bacterium. Cry1A.105, encoded by the cry1A.105 gene, is a chimeric protein made up of different functional domains derived from three wild-type Cry proteins from B. thuringiensis subspecies kurstaki and aizawai. The Cry2Ab2 protein is encoded by the cry2Ab2 gene derived from B. thuringiensis subspecies kurstaki. Molecular characterization: Appropriate analysis of the integration site, including flanking sequence and bioinformatics analysis, has been performed to characterise the transformation event MON 89034. The results of the segregation analysis are consistent with a single site of insertion for the cry1A.105 and cry2Ab2 gene expression cassettes and confirm the results of the molecular characterisation. Molecular analysis of both self-pollinated and cross-fertilised lines, representing a total of seven different generations, indicates that the inserted DNA is stably transformed and inherited from one generation to the next. No genes that encode resistance to antibiotics are present in the genome of MON 89034 maize. The molecular characterisation confirmed the absence of both the aad and nptII genes, which were used in the cloning and transformation process. Event MON 89034 and the physical, chemical and functional characteristics of the proteins have previously been evaluated by The VKM Panel on Genetically Modified Organisms, and considered satisfactory (VKM 2008a). Comparative assessment: The field trials for comparative assessment of agronomic and phenotypic characteristics of maize MON 89034 in the USA (2004-2005) and Europe (2007), have been performed in accordance with the EFSAs guidelines for risk assessment of genetically modified plants and derived food and feed (EFSA 2010, 2011a). Based on results from the comparative analyses, it is concluded that maize MON 89034 is agronomically and phenotypically equivalent to the conventional counterpart and commercial available reference varieties, with the exception of the lepidopteran-protection trait. The field evaluations support a conclusion of no phenotypic changes indicative of increased plant weed/pest potential of MON 89034 compared to conventional maize. Evaluations of ecological interactions between maize MON 89034 and the biotic and abiotic environment indicate no unintended effects of the introduced trait on agronomic and phenotypic characteristics. Environmental risk: There are no reports of the target Lepidopteran species attaining pest status on maize in Norway. Since there are no Bt-based insecticides approved for use in Norway, and lepidopteran pests have not been registered in maize, issues related to resistance evolution in target pests are not relevant at present for Norwegian agriculture. Published scientific studies show no or negligible adverse effects of Cry1A.105 and Cry2Ab2 proteins on non-target arthropods that live on or in the vicinity of maize plants. Cultivation of maize MON 89034 is not considered to represent a threat to the prevalence of red-listed species in Norway. Few studies have been published examining potential effects of Cry1A.105 and Cry2Ab toxin on ecosystems in soil, mineralization, nutrient turnover and soil communities. Some field studies have indicated that root exudates and decaying plant material containing Cry proteins may affect population size and activity of rhizosphere organisms (soil protozoa and microorganisms). However, data are only available from short term experiments and predictions of potential long term effects are difficult to deduce. Most studies conclude that effects on soil microorganisms and microbial communities are transient and minor compared to effects caused by agronomic and environmental factors. Few studies have assessed the impact of Cry proteins on non-target aquatic arthropods and the fate of these proteins in senescent and decaying maize detritus in aquatic environments. Further studies with better experimental design are needed for the assessment of the potential effects of Bt crops on aquatic organisms. However, exposure of non-target organisms to Cry proteins in aquatic ecosystems is likely to be very low, and potential exposure of Bt toxins to non-target organisms in stream ecosystems in Norway is considered to be negligible. Maize is the only representative of the genus Zea in Europe, and there are no cross-compatible wild or weedy relatives outside cultivation with which maize can hybridise and form backcross progeny. Vertical gene transfer in maize therefore depends on cross-pollination with other conventional or organic maize varieties. In addition, unintended admixture of genetically modified material in seeds represents a possible way for gene flow between different crop cultivations. The risk of pollen flow from maize volunteers is negligible under Norwegian growing conditions. In addition to the data presented by the applicant, the VKM GMO Panel is not aware of any scientific report of increased establishment and spread of maize MON 89034 and any change in survival (including over-wintering), persistence and invasiveness capacity. Because the general characteristics of maize MON 89034 are unchanged, insect resistance are not likely to provide a selective advantage outside cultivation in Norway. Since MON 89034 has no altered agronomic and phenotypic characteristics, except for the specific target pest resistance, the VKM GMO Panel is of the opinion that the likelihood of unintended environmental effects due to the establishment and survival of maize MON 89034 will be no different to that of conventional maize varieties in Norway The environmental risk assessment will be completed and finalized by the VKM Panel on Genetically Modified Organisms when requested additional information from the applicant is available.

Environmental Risk Assessment of Insect Resistant Genetically Modified Maize MON810 for Cultivation, Seed Production, Import, Processing and Feed Uses under Directive 2001/18/EC (Notification C/F/95/12/02)

In preparation for a legal implementation of EU-regulation 1829/2003, the Norwegian Scientific Committee for Food Safety (VKM) has been requested by the Norwegian Directorate for Nature Management to conduct final environmental risk assessments for all genetically modified organisms (GMOs) and products containing or consisting of GMOs that are authorized in the European Union under Directive 2001/18/EC or Regulation 1829/2003/EC. The request covers scope(s) relevant to the Gene Technology Act. The request does not cover GMOs that VKM already has conducted its final risk assessments on. However, the Directorate requests VKM to consider whether updates or other changes to earlier submitted assessments are necessary. MON810 notification C/F/95/12-02 is approved under Directive 90/220/EEC for cultivation, seed production, import and processing into feeding stuffs and industrial purposes since 22 April 1998 (Commission Decision 98/294/EC). In December 1997, food and food ingredients derived from the progeny of maize line MON810 were notified under Article 5 of Regulation (EC) No 258/97 on novel foods and novel food ingredients. In addition, existing food and feed products containing, consisting of or produced from MON810 were notified according to Articles 8 and 20 of Regulation (EC) No 1829/2003 and were placed in the Community Register in 2005. Three applications for renewal of the authorisation for continued marketing of (1) existing food and food ingredients produced from MON810; (2) feed consisting of and/or containing maize MON810, and MON810 for feed use (including cultivation); and (3) food and feed additives, and feed materials produced from maize MON810 within the framework of Regulation (EC) No 1829/2003 were submitted in 2007. Maize MON810 has previously been assessed by the VKM GMO Panel commissioned by the Norwegian Directorate for Nature Management in connection with the national finalisation of the procedure of the notification C/F/95/12/02 (VKM 2007a,b). In addition, MON810 has been evaluated by the VKM GMO Panel as a component of several stacked GM maize events (VKM 2005a,b,c, VKM 2007c, VKM 2008, VKM 2009, VKM 2012). Due to the publication of updated guidelines for environmental risk assessments of genetically modified plants and new scientific literature, the VKM GMO Panel has decided to deliver an updated environmental risk assessment of MON810. The environmental risk assessment of the maize MON810 is based on information provided by the applicant in the notification C/F/95/12/02 and application EFSA/GMO/RX/MON810, and scientific comments from EFSA and other member states made available on the EFSA website GMO Extranet. The risk assessment also considered other peer-reviewed scientific literature as relevant. The VKM GMO Panel has evaluated MON810 with reference to its intended uses in the European Economic Area (EEA), and according to the principles described in the Norwegian Food Act, the Norwegian Gene Technology Act and regulations relating to impact assessment pursuant to the Gene Technology Act, Directive 2001/18/EC on the deliberate release into the environment of genetically modified organisms, and Regulation (EC) No 1829/2003 on genetically modified food and feed. The Norwegian Scientific Committee for Food Safety has also decided to take account of the appropriate principles described in the EFSA guidelines for the risk assessment of GM plants and derived food and feed (EFSA 2006, 2011a), the environmental risk assessment of GM plants (EFSA 2010), the selection of comparators for the risk assessment of GM plants (EFSA 2011b), and for the post-market environmental monitoring of GM plants (EFSA 2006, 2011c). The scientific risk assessment of maize MON810 include molecular characterisation of the inserted DNA and expression of the target protein, comparative assessment of agronomic and phenotypic characteristics, unintended effects on plant fitness, potential for gene transfer, interactions between the GM plant and target and non-target organisms, effects on biogeochemical processes and evaluations of the post-market environmental plan. In line with its mandate, VKM emphasised that assessments of sustainable development, societal utility and ethical considerations, according to the Norwegian Gene Technology Act and Regulations relating to impact assessment pursuant to the Gene Technology Act, shall not be carried out by the Panel on Genetically Modified Organisms. The genetically modified maize MON810 was developed to provide protection against certain lepidopteran target pests, including European corn borer (Ostrinia nubilalis) and species belonging to the genus Sesamia. Protection is achieved through expression in the plant of the insecticidal Cry protein, Cry1Ab, derived from Bacillus thuringiensis ssp. kurstaki, a common soil bacterium. Molecular characterisation Appropriate analysis of the integration site including flanking sequences and bioinformatics analyses have been performed to analyse the construct integrated in the GM plant. Updated bioinformatics analyses revealed that one ORF shared sequence similarity to a putative HECT-ubiquitin ligase protein. The VKM GMO Panel found no safety implications from the interruption of this gene sequence. Analyses of leaf, grains, whole plant tissue and pollen from the maize MON 810 demonstrated that the Cry1Ab protein is expressed at very low levels in all tissues tested and constitutes less than 0.001% of the fresh weight in each tissue. The cry1Ab gene is the only transgene expressed in line MON 810 and was expressed highest in the leaves. The stability of the genetic modification has been demonstrated over several generations. Event MON810 and the physical, chemical and functional characteristics of the proteins have previously been evaluated by The VKM Panel on Genetically Modified Organisms, and considered satisfactory (VKM 2007a,b). Comparative assessment: Comparative analyses of data from field trials located at representative sites and environments in the USA and Europe indicate that maize MON810 is agronomically and phenotypically equivalent to the conventional counterpart and commercially available reference varieties, with the exception of the lepidopteran-protection trait, conferred by the expression of the Cry1Ab protein. The field evaluations support a conclusion of no phenotypic changes indicative of increased plant weed/pest potential of MON810 compared to conventional maize. Evaluations of ecological interactions between maize MON810 and the biotic and abiotic environment indicate no unintended effects of the introduced trait on agronomic and phenotypic characteristics. Environmental risk: There are no reports of the target lepidopteran species attaining pest status on maize in Norway. Since there are no Bt-based insecticides approved for use in Norway, and lepidopteran pests have not been registered in maize, issues related to resistance evolution in target pests are not relevant at present for Norwegian agriculture. Published scientific studies show no or negligible adverse effects of Cry1Ab protein on non-target arthropods that live on or in the vicinity of maize plants. Cultivation of maize MON810 is not considered to represent a threat to the prevalence of red-listed species in Norway. Few studies have been published examining potential effects of Cry1Ab toxin on ecosystems in soil, mineralization, nutrient turnover and soil communities. Some field studies have indicated that root exudates and decaying plant material containing Cry proteins may affect population size and activity of rhizosphere organisms (soil protozoa and microorganisms). Most studies conclude that effects on soil microorganisms and microbial communities are transient and minor compared to effects caused by agronomic and environmental factors. However, data are only available from short term experiments and predictions of potential long term effects are difficult to deduce. Few studies have assessed the impact of Cry proteins on non-target aquatic arthropods and the fate of these proteins in senescent and decaying maize detritus in aquatic environments. However, exposure of non-target organisms to Cry proteins in aquatic ecosystems is likely to be very low, and potential exposure of Bt toxins to non-target organisms in aquatic ecosystems in Norway is considered to be negligible. Maize is the only representative of the genus Zea in Europe, and there are no cross-compatible wild or weedy relatives outside cultivation with which maize can hybridise and form backcross progeny. Vertical gene transfer in maize therefore depends on cross-pollination with other conventional or organic maize varieties. In addition, unintended admixture of genetically modified material in seeds represents a possible way for gene flow between different crop cultivations. The risk of pollen flow from maize volunteers is negligible under Norwegian growing conditions. In addition to the data presented by the applicant, the VKM GMO Panel is not aware of any scientific report of increased establishment and spread of maize MON810 and any change in survival (including over-wintering), persistence and invasiveness capacity. Because the general characteristics of maize MON810 are unchanged, insect resistance are not likely to provide a selective advantage outside cultivation in Norway. Since MON810 has no altered agronomic and phenotypic characteristics, except for the specific target pest resistance, the VKM GMO Panel is of the opinion that the likelihood of unintended environmental effects due to the establishment and survival of maize MON810 will be no different to that of conventional maize varieties in Norway. Overall conclusion: The VKM GMO Panel concludes that cultivation of maize MON810 is unlikely to have any adverse effect on the environment in Norway.

Genetic parameters of growth, and resistance to the shoot borer, in young clones of the tree Cedrela odorata (Meliaceae)

Damage by Hypsipyla grandella Zeller restricts the success of plantations of Cedrela odorata. The most critical and vulnerable infection period by H. grandella is during the first three years. The aim was to estimate the expected genetic gain for the selection of clones of fast growth and resistant to the attack by this insect. A trial with 40 clones produced by grafting was planted in Veracruz, Mexico. Heritability, genetic correlations and genetic gain of clones were estimated with data at two years-old. The survival rate of the trial was of 97 % (466 living ramets in total), 2.84 m in total high and 2.32 cm of average normal diameter. Aditionally, 9.9 % of the ramets did not present attacks, indication of evasion, and 0.6 percent without response to attack. With response to attack 89.5 % of the ramets (several degrees of tolerance): 29 % with a single shoot, 14.2 % with several shoots and 46.3 % with a dominant shoot of several shoots formed. The clonal heritability of normal diameter, height, volume, stem taper index, number of attacks and response to the attack was H2c = 0.81, 0.80, 0.81, 0.61, 0.34, and 0.26. The genetic correlations between height, diameter and volume were ≥ 0.95, and the correlation of the volume with the incidence and the response to the attack was rg = -0.31 and rg = 0.62, that is a decrease in number of attacks and moderate increase in tolerance. With a selection intensity of 10 %, selecting the four clones with higher volume will produce a genetic gain of 82 % for volume, a decrease of 10.9 % of number of attacks and 6.3 % of better recovery from damage; this is more tolerance to the insect attack. The genetic gain justifies the use of the best clones in commercial plantations in Veracruz, Mexico.

El daño por Hypsipyla grandella limita el éxito de las plantaciones de Cedrela odorata. El periodo más crítico y vulnerable es durante los primeros tres años. El objetivo del estudio fue estimar la ganancia genética esperada en la selección de clones de mayor crecimiento y resistentes al ataque del insecto. Un ensayo de 40 clones producidos por injertos fue plantado en Veracruz, México. Con datos a la edad de dos años se estimaron: la heredabilidad, correlaciones genéticas y ganancia genética de los clones. El ensayo presentó una supervivencia del 97 % (466 rametos vivos en total), con 2.84 m de altura y 2.32 cm de diámetro normal promedio. El 9.9 % de los rametos no presentaron ataques, indicando evasión, y se estimó un 0.6 % sin respuesta al ataque. Con respuesta al ataque 89.5 % de los rametos (diferentes niveles de tolerancia): 29 % con un solo brote, 14.2 % con varios brotes, y 46.3 % con un brote dominante de varios brotes formados. La heredabilidad clonal del diámetro normal, altura, volumen, índice de conicidad, número de ataques y respuesta al ataque fue de H2 c = 0.81, 0.80, 0.81, 0.61, 0.34 y 0.26, respectivamente. Las correlaciones genéticas entre la altura, diámetro y volumen fueron ≥ 0.95, y del volumen con la incidencia y la respuesta al ataque de rg = -0.31 y rg = 0.62, es decir se presentó una disminución en el número de ataques y el aumento moderado en la tolerancia. Con una intensidad de selección del 10 %, seleccionando los cuatro clones de mayor volumen se obtuvo una ganancia genética de 82 % para el volumen, una disminución de 10.9 % de ataques y 6.3 % mejor recuperación del daño, esto es más tolerancia al ataque del insecto. La ganancia genética justifica la utilización de los mejores clones en plantaciones comerciales para Veracruz.

CALIDAD MOLINERA DE LAS VARIEDADES DE ARROZ SD20A Y PAYARA 1FL, Y SU RESISTENCIA A Sitophilus oryzae (L.) Y Rhyzopertha dominica (F.) / MILLING QUALITY OF RICE VARIETIES SD20A AND PAYARA 1FL, AND THEIR RESISTANCE TO Sitophilus oryzae (L.) AND Rhyzopertha dominica (F.)

RESUMEN La agroindustria requiere de variedades de arroz, que permitan obtener granos enteros durante su procesamiento y que resistan el ataque de insectos. En este estudio, se determinó la calidad molinera de las variedades SD20A y Payara 1FL y su resistencia a Sitophilus oryzae (L.) y Rhyzopertha dominica (F.). En la primera fase, se calculó el índice de blancura (IB) y rendimiento en molino (RM) de ambas variedades, mientras que, en la segunda, fueron sometidas a los ataques de S. oryzae y R. dominica, en su condición de arroz paddy y pulido, durante tres meses. La resistencia a los insectos, se interpretó con base al número de adultos emergidos al finalizar el periodo de evaluación. Hubo diferencias estadísticas (p< 0,01) entre el IB y el RM de las variedades de arroz SD20A (32,40 ± 1,87°Kett y 44,40 ± 2,55%) y Payara 1FL (35,90 ± 0,63°Kett y 48,20 ± 1,69%), respectivamente. El número de adultos de S. oryzae y R. dominica fue diferente entre SD20A y Payara 1FL (p< 0,01), siendo mayores los valores registrados para las variedades en condición de arroz pulido que sobre arroz paddy. La cantidad de adultos de R. dominica fue siempre superior a la de S. oryzae, excepto para el arroz pulido de Payara 1FL, donde no hubo diferencias significativas (p> 0,01). Los resultados muestran que Payara 1FL presentó mayor RM que SD20A y mostró mayor resistencia a los ataques de S. oryzae que a los de R. dominica.

ABSTRACT Agroindustry requires rice varieties that allow obtaining whole grains during its processing and also to resist insect attacks. In this study was determined milling quality of varieties SD20A and Payara 1FL, and their resistance to Sitophilus oryzae (L.) and Rhyzopertha dominica (F.). In the first phase, the whiteness ratio (WR) and grain yield in a mill (GY) were calculated for both varieties, while in the second phase, those were through the attack of S. oryzae and R. dominica, in paddy and milled rice condition, during three months. The resistance was interpreted based on the number of adults emerged at the end of the experiment. There were statistical differences (p< 0.01) between WR and GY of rice varieties SD20A (32.40 ± 1.87°Kett and 44.40 ± 2.55%) and Payara 1FL (35.90 ± 0.63°Kett and 48.20 ± 1.69%). Adult emerged numbers of S. oryzae and R. dominica was different between SD20A and Payara 1FL (p< 0.01), being higher in milled rice than those registered on paddy rice. The amount of adult of R. dominica was always higher than S. oryzae, except for Payara 1FL milled rice in where no significative difference (p> 0.01). Results showed that Payara 1FL had a higher yield in the mill than SD20A, and higher resistance to S. oryzae attacks than to R. dominica.

Risk Assessment of Insect-resistant and Herbicide Tolerant Genetically Modified Maize Bt11 for Cultivation, Import, Processing, Food and Feed Uses under Directive 2001/18/EC and Regulation (EC) Opinion

The Norwegian Environment Agency (NEA) and the Norwegian Food Safety Authority (NFSA) requested the Norwegian Scientific Committee for Food Safety (Vitenskapskomiteen for mattrygghet, VKM) for an opinion of potential risks to biodiversity and agriculture in Norway associated with import of seeds for sowing and cultivation of insect-resistant and herbicide tolerant genetically modified maize Bt11 under Directive 2001/18/EC (Notification C/F/96.05.10). The notification is still pending for authorisation in the European Union. VKM is also requested to assess the applicant´s post-market environmental monitoring plan, and the management measures suggested in the draft implementing decision of the European Commission. As the scope of the notification does not cover food and feed uses of maize Bt11, VKM was not asked for a health risk assessment of maize Bt11. However, VKM has decided to update a previous safety evaluation of the food and feed uses of maize Bt11 and derived products (VKM, 2014). VKM appointed a working group consisting of members from the Panel on Genetically Modified Organisms, the Panel on Alien Organisms and trade in Endangered Species (CITES) and the VKM staff to answer the requests. The Panel on Genetically Modified Organisms assessed and approved the final report. The genetically modified maize Bt11 was developed to provide protection against certain lepidopteran target pests, such as the European corn borer (ECB, Ostrinia nubilalis), and some species belonging to the genus Sesamia . The insect resistence is achieved by the expression of a truncated form of a Cry1Ab protein encoded by a modified cry1Ab gene derived from the soil microorganism Bacillus thuringiensis subsp kurstaki HD-1. Maize Bt11 also expresses the phosphinothricin - N - ace tyltransferase (pat) gene, derived from the soil microorganism Streptomyces viridochromogenes strain Tu494, which encodes the enzyme: phosphinothricin acetyl transferase (PAT). PAT protein confers tolerance to the herbicidal active substance glufosinate-ammonium. The PAT protein expressed in Bt11 was used as a selectable marker to facilitate the selection process of transformed plant cells and is not intended for weed management purposes. Since the scope of the notification C/F/96.05.10 does not cover the use of glufosinate-ammonium-containing herbicides on maize Bt11, potential effects due to the use of such herbicides on maize Bt11 are not considered by VKM. In delivering its scientific opinion, VKM considered relevant peer-reviewed scientific publications and information provided by the applicant in the notification C/F/96.05.10, the renewal application EFSA/GMO/RX/Bt11, and scientific opinions and comments from EFSA and other EU-member states. VKM has evaluated maize Bt11 with reference to its intended uses in the European Economic Area (EEA), and according to the principles described in the Norwegian Food Act, the Norwegian Gene Technology Act and regulations relating to impact assessment pursuant to the Gene Technology Act, Directive 2001/18/EC on the deliberate release into the environment of genetically modified organisms, and Regulation (EC) No 1829/2003 on genetically modified food and feed. The Norwegian Scientific Committee for Food Safety has also decided to take account of the appropriate principles described in the EFSA guidelines for the risk assessment of GM plants and derived food and feed (EFSA 2011a), the environmental risk assessment of GM plants (EFSA 2010a), selection of comparators for the risk assessment of GM plants (EFSA, 2011b) and for the post-market environmental monitoring of GM plants (EFSA, 2011c). The scientific risk assessment of maize Bt11 includes molecular characterisation of the inserted DNA and expression of novel proteins, comparative assessment of agronomic and phenotypic characteristics, nutritional assessments, toxicology and allergenicity. An evaluation of unintended effects on plant fitness, potential for gene transfer, interactions between the GM plant and target and non-target organisms, effects on biogeochemical processes, the post-market environmental monitoring plan and coexistence measures at the farm level has also been undertaken. It is emphasised that the VKM mandate does not include assessments of contribution to sustainable development, societal utility and ethical considerations, according to the Norwegian Gene Technology Act and Regulations relating to impact assessment pursuant to the Gene Technology Act. These considerations are therefore not part of the risk assessment provided by the VKM. Molecular Characterization: Appropriate analyses of the integration site, inserted DNA sequence, flanking regions, and bioinformatics have been performed. The molecular characterisation reported by the applicant shows that the DNA-fragment containing the cry1Ab and pat genes, is integrated as a single copy at a single locus in the nuclear genome of maize Bt11 and that it is stably inherited as a dominant trait. VKM considers the molecular characterisation of maize Bt11 satisfactory. Comparative Assessment: Comparative analyses of data from field trials located at representative sites and environments in North America and Europe indicates that maize Bt11 is compositionally equivalent to its conventional counterpart, with the exception of the herbicide tolerance and insect resistance traits, conferred by the expression of the PAT and Cry1Ab proteins. However, data on the amino acid tryptophan, is only given in one out of six studies. Based on current knowledge, VKM concludes that maize Bt11 is compositionally equivalent to conventional maize. The data provided by the applicant are not sufficient to show that Bt11 maize is phenotypically and agronomically equivalent to conventional near-isogenic maize lines. The agronomic assessment data are provided from one growing season in the North America and one growing season in France. This is not considered to be sufficient for representative testing of agricultural environments. Food and Feed Risk Assessment: Whole food feeding studies have not indicated any adverse health effects of maize Bt11. These studies further support that maize Bt11 is nutritionally equivalent to conventional maize. The Cry1Ab and PAT proteins do not show sequence resemblance to other known toxins or IgE allergens, nor have they been reported to cause IgE mediated allergic reactions. Some studies have however indicated a potential role of Cry-proteins as adjuvants in allergic reactions. Based on current knowledge, the VKM concludes that maize Bt11 is nutritionally equivalent to conventional maize varieties. It is unlikely that the Cry1Ab and PAT proteins will introduce a toxic or allergenic potential in food or feed based on maize Bt11 compared to conventional maize. Environmental Risk Assessment: Maize is the only representative of the genus Zea in Europe, and there are no cross-compatible wild or weedy relatives outside cultivated maize with which maize can hybridise and form backcross progeny. Vertical gene transfer in maize therefore depends on cross-pollination with other conventional or organic maize varieties. In addition, unintended admixture of genetically modified material in seeds represents a possible way for gene flow between different crop cultivations. The risk of pollen flow from maize volunteers is negligible under Norwegian growing conditions. Since maize Bt11 has no altered agronomic and phenotypic characteristics, except for the specific target insect resistance and herbicide tolerance, the likelihood of unintended environmental effects as a consequence of spread of genes from maize Bt11 is considered to be extremely low. There are no reports of the target lepidopteran species attaining pest status on maize in Norway. Since there are no Bt-based insecticides approved for use in Norway, and lepidopteran pests have not been registered in maize, issues related to resistance evolution in target pests are not relevant at present for Norwegian agriculture. Published scientific studies showed that the likelihood of negative effects of Cry1Ab protein on non-target arthropods that live on or in the vicinity of maize plants is low. In Norway, the maize cultivation is marginal. The total crop area of forage maize is estimated to 2000-2800 decares, equivalent to less than 0.1% of the areas with cereal crops. The area of individual fields is limited by the topography such that the quantity of maize pollen produced under flowering is also limited. The potential exposure of Cry1Ab-containing maize pollen on non-target lepidopteran species in Norway is therefore negligible. Cultivation of maize Bt11 is not considered to represent a threat to the prevalence of red-listed species in Norway. Exposure of nontarget organisms to Cry proteins in aquatic ecosystems is likely to be very low, and potential exposure of Cry proteins to non-target organisms in aquatic ecosystems in Norway is considered to be negligible. VKM concludes that, although the data on the fate of the Cry1Ab protein and its potential interactions in soil are limited, the relevant scientific publications analysing the Cry1Ab protein, together with the relatively broad knowledge about the environmental fate of other Cry1 proteins, do not indicate significant direct effects on the soil environment. Despite limited number of studies, most studies conclude that effects on soil microorganisms and microbial communities are transient and minor compared to effects caused by agronomic and environmental factors. However, data are only available from short-term experiments and predictions of potential long-term effects are difficult to deduce. Coexistence: VKM concludes that separation distances of 200 meters most likely will ensure coexistence between genetically modified maize and conventional and organic maize varieties in Norway. Overall Conclusion: Based on current knowledge, VKM concludes that maize Bt11 is nutritionally equivalent t

Final Health and Environmental Risk Assessment of Genetically Modified Maize MON 89034 x NK 603

In preparation for a legal implementation of EU-regulation 1829/2003, the Norwegian Scientific Committee for Food Safety (VKM) has been requested by the Norwegian Environment Agency (former Norwegian Directorate for Nature Management) and the Norwegian Food Safety Authority (NFSA) to conduct final food/feed and environmental risk assessments for all genetically modified organisms (GMOs) and products containing or consisting of GMOs that are authorized in the European Union under Directive 2001/18/EC or Regulation 1829/2003/EC. The request covers scope(s) relevant to the Gene Technology Act. The request does not cover GMOs that VKM already has conducted its final risk assessments on. However, the Agency and NFSA requests VKM to consider whether updates or other changes to earlier submitted assessments are necessary. The insect-resistant and glyphosate-tolerant genetically modified maize MON 89034 x NK 603 from Monsanto (Unique Identifier MON-89Ø34-3 × MON-ØØ6Ø3-6) was approved under Regulation (EC) No 1829/2003 in the EU for food and feed uses, import and processing on 28 July 2010 (Commission Decision 2010/420/EC). Genetically modified maize MON 890314 x NK 603 has previously been risk assessed by the VKM Panel on Genetically Modified Organisms (GMO), commissioned by the Norwegian Food Safety Authority and the Norwegian Environment Agency related and to the EFSA public hearing of the applications EFSA/GMO/NL/2007/38 and EFSA/GMO/NL/2009/72 in 2007 and 2009/2010 (VKM 2008a, VKM 2010a). In addition, the parental lines MON 89034 and NK 603 have been evaluated by the VKM GMO Panel as single events and as a component of several stacked GM maize events (VKM 2005a,b,c,d,e, VKM 2007a,b, VKM 2008b,c,d, VKM 2009a,b, VKM 2010 a,b, VKM 2011, VKM 2012a,b, VKM 2013 a,b, VKM 2014). The food/feed and environmental risk assessment of the maize MON 89034 x NK 603 is based on information provided by the applicant in the applications EFSA/GMO/NL/2007/38 EFSA/GMO/NL/2009/72 and scientific comments from EFSA and other member states made available on the EFSA website GMO Extranet. The risk assessment also considered other peer-reviewed scientific literature as relevant. The VKM GMO Panel has evaluated MON 89034 x NK 603 with reference to its intended uses in the European Economic Area (EEA), and according to the principles described in the Norwegian Food Act, the Norwegian Gene Technology Act and regulations relating to impact assessment pursuant to the Gene Technology Act, Directive 2001/18/EC on the deliberate release into the environment of genetically modified organisms, and Regulation (EC) No 1829/2003 on genetically modified food and feed. The Norwegian Scientific Committee for Food Safety has also decided to take account of the appropriate principles described in the EFSA guidelines for the risk assessment of GM plants and derived food and feed (EFSA 2011a), the environmental risk assessment of GM plants (EFSA 2010a), selection of comparators for the risk assessment of GM plants (EFSA 2011b) and for the post-market environmental monitoring of GM plants (EFSA 2011c). The scientific risk assessment of maize MON 89034 x NK 603 include molecular characterisation of the inserted DNA and expression of novel proteins, comparative assessment of agronomic and phenotypic characteristics, nutritional assessments, toxicology and allergenicity, unintended effects on plant fitness, potential for gene transfer, effects on biogeochemical processes and interactions between the GM plant and target and non-target organisms. It is emphasized that the VKM mandate does not include assessments of contribution to sustainable development, societal utility and ethical considerations, according to the Norwegian Gene Technology Act and Regulations relating to impact assessment pursuant to the Gene Technology Act. These considerations are therefore not part of the risk assessment provided by the VKM Panel on Genetically Modified Organisms. Likewise, the VKM mandate does not include evaluations of herbicide residues in food and feed from genetically modified plants. The hybrid maize MON 89034 x NK 603 has been produced by conventional crosses between inbred lines containing MON 89034 and NK 603 events to combine resistance to certain lepidopteran pests and to confer tolerance towards glyphosate-containing herbicides. Maize MON 89034 was developed to provide protection against specific lepidopteran target pest, including Ostrinia nubilalis, S podoptera spp. and Agrotis ipsilon. Protection is achieved through expression in the plant of two insecticidal Cry proteins, Cry1A.105 and Cry2Ab2, derived from Baci llus thuringiensis subsp. a izawai and kurstaki. Maize NK 603 has been developed to provide tolerance to glyphosate by the introduction, of a gene coding for 5enolpyruvylshikimate-3-phosphate synthase (EPSPS) from Agrobacterium sp. strain CP4 (CP4 EPSPS). Molecular Characterisation: Southern and PCR analyses indicate that the recombinant inserts in the single maize events MON 89034 and NK 603 are retained in maize stack MON 89034 x NK603. Genetic stability of the inserts has previously been demonstrated in the parental lines MON 89034and NK603. The level of Cry1A.105, Cry2Ab2 and CP4 EPSPS proteins in grain and forage from the stacked event are comparable to the levels in the corresponding single events. Phenotypic analyses also indicate stability of the insect resistance and herbicide tolerance traits of the stacked event. Based on current knowledge and the previous assessments of the parental maize events, the VKM GMO Panel considers the molecular characterisation of maize MON 89034 x NK 603 satisfactory. 6 VKM Report 2016: 17. Comparative Assessment: The applicant has performed comparative analyses of data from field trials located at representative sites and environments in Argentina in 2004/2005 and Europe in 2007. With the exception of small intermittent variations and the insect resistance and herbicide tolerance conferred by the Cry1A.105, Cry2Ab2 and CP4 EPSPS proteins, the results showed no biologically relevant differences between maize stack MON 89034 x NK 603 and conventional control. Based on the assessment of available data, the VKM GMO Panel concludes that maize MON 89034 x NK 603 is compositionally, agronomical and phenotypically equivalent to its conventional counterpart, except for the new proteins. Food/feed Safety Assessment: A whole food feeding study on broilers has not indicated any adverse health effects of maize MON 89034 x NK 603, and shows that it is nutritionally equivalent to conventional maize varieties. The Cry1A.105, Cry2Ab2, and CP4 EPSPS proteins do not show sequence resemblance to other known toxins or IgE allergens, nor have they been reported to cause IgE mediated allergic reactions. However, some studies have indicated a potential role of Cry-proteins as adjuvants in allergic reactions. Based on current knowledge, the VKM GMO Panel concludes that maize MON 89034 x NK 603 is nutritionally equivalent to conventional maize varieties. It is unlikely that the Cry1A.105, Cry2Ab2, and CP4 EPSPS proteins will cause toxic or IgE-mediated allergic reactions to food or feed based on maize MON 89034 x NK 603 compared to conventional maize. Environmental Risk: Considering the intended uses of maize MON 89034 x NK603, excluding cultivation, the environmental risk assessment is concerned with accidental release into the environment of viable grains during transportation and processing, and indirect exposure, mainly through manure and faeces from animals fed grains from maize MON 89034 x NK603. Maize MON 89034 x NK 603 has no altered survival, multiplication or dissemination characteristics, and there are no indications of an increased likelihood of spread and establishment of feral maize plants in the case of accidental release into the environment of seeds from maize MON 89034 x NK603. Maize is the only representative of the genus Zea in Europe, and there are no cross-compatible wild or weedy relatives outside cultivation. The VKM GMO Panel considers the risk of gene flow from occasional feral GM maize plants to conventional maize varieties to be negligible in Norway. Considering the intended use as food and feed, interactions with the biotic and abiotic environment are not considered by the GMO Panel to be an issue. 7 VKM Report 2016: 17. Overall Conclusion: Based on current knowledge, the VKM GMO Panel concludes that maize MON 89034 x NK 603 is compositionally, nutritionally, agronomically and phenotypically equivalent to its conventional counterpart except for the new proteins. It is unlikely that the Cry1A.105, Cry2Ab2 and CP4 EPSPS proteins will cause an increased risk of toxic or IgE-mediated allergic reactions to food or feed based on maize MON 89034 x NK 603 compared to conventional maize varieties. The VKM GMO Panel concludes that maize MON 89034 x NK603, based on current knowledge, is comparable to conventional maize varieties concerning environmental risk in Norway with the intended usage.

Final Health- and Environmental Risk Assessment of Genetically Modified Maize MON 89034 x MON 88017

In preparation for a legal implementation of EU-regulation 1829/2003, the Norwegian Scientific Committee for Food Safety (VKM) has been requested by the Norwegian Environment Agency and the Norwegian Food Safety Authority (NFSA) to conduct final food/feed and environmental risk assessments for all genetically modified organisms (GMOs) and products containing or consisting of GMOs that are authorized in the European Union under Directive 2001/18/EC or Regulation 1829/2003/EC. The request covers scope(s) relevant to the Gene Technology Act. The request does not cover GMOs that VKM already has conducted its final risk assessments on. However, the Agency and NFSA requests VKM to consider whether updates or other changes to earlier submitted assessments are necessary. The insect-resistant and glyphosate-tolerant genetically modified maize MON 89034 x MON 88017 from Monsanto (Unique Identifier MON-89Ø34-3 × MON-88Ø17-3) was approved under Regulation (EC) No 1829/2003 in the EU for food and feed uses, import and processing on 17th of June 2011 (Commission Decision 2011/366/EC). Genetically modified maize MON 890314 x MON 88017 has previously been risk assessed by the VKM Panel on Genetically Modified Organisms (GMO), commissioned by the Norwegian Food Safety Authority and the Norwegian Environment Agency related and to the EFSA public hearing of the applications EFSA/GMO/NL/2007/39 and EFSA/GMO/BE/2009/71 in 2007 and 2009/2010 (VKM 2008a, VKM 2010a). In addition, the parental lines MON 89034 and MON 88017 have been evaluated by the VKM GMO Panel as single events and as a component of several stacked GM maize events (VKM 2007a,b, VKM 2008b, VKM 2009a,b,c, VKM 2010b,c, VKM 2012, VKM 2013, VKM 2014). The food/feed and environmental risk assessment of the maize MON 89034 x MON 88017 is based on information provided by the applicant in the applications EFSA/GMO/NL/2007/39 EFSA/GMO/BE/2009/71 and scientific comments from EFSA and other member states made available on the EFSA website GMO Extranet. The risk assessment also considered other peer-reviewed scientific literature when relevant. The VKM GMO Panel has evaluated MON 89034 x MON 88017 with reference to its intended uses in the European Economic Area (EEA), and according to the principles described in the Norwegian Food Act, the Norwegian Gene Technology Act and regulations relating to impact assessment pursuant to the Gene Technology Act, Directive 2001/18/EC on the deliberate release into the environment of genetically modified organisms, and Regulation (EC) No 1829/2003 on genetically modified food and feed. The Norwegian Scientific Committee for Food Safety has also decided to take account of the appropriate principles described in the EFSA guidelines for the risk assessment of GM plants and derived food and feed (EFSA 2011a), the environmental risk assessment of GM plants (EFSA 2010), selection of comparators for the risk assessment of GM plants (EFSA 2011b) and for the post-market environmental monitoring of GM plants (EFSA 2011c). The scientific risk assessment of maize MON 89034 x MON 88017 include molecular characterisation of the inserted DNA and expression of novel proteins, comparative assessment of agronomic and phenotypic characteristics, nutritional assessments, toxicology and allergenicity, unintended effects on plant fitness, potential for gene transfer, effects on biogeochemical processes and interactions between the GM plant and target and non-target organisms. It is emphasised that the VKM mandate does not include assessments of contribution to sustainable development, societal utility and ethical considerations, according to the Norwegian Gene Technology Act and Regulations relating to impact assessment pursuant to the Gene Technology Act. These considerations are therefore not part of the risk assessment provided by the VKM Panel on Genetically Modified Organisms. Likewise, the VKM mandate does not include evaluations of herbicide residues in food and feed from genetically modified plants.The hybrid maize MON 89034 x MON 88017 has been produced by conventional crosses between inbred lines containing MON 89034 and MON 88017 events to combine resistance to certain coleopteran and lepidopteran pests, and to confer tolerance towards glyphosate-containing herbicides. Maize MON 89034 was developed to provide protection against specific lepidopteran target pest, including Ostrinia nubilalis , S podoptera spp. and Agrotis ipsilon. Protection is achieved through expression in the plant of two insecticidal Cry proteins, Cry1A.105 and Cry2Ab2, derived from Bacillus thuringiensis subsp. a izawai and kurstaki. Maize MON 88017 was developed to express a modified Cry3Bb1 insecticidal protein, derived from B. thuringiensis subsp. kumamotoensis , which confers protection against coleopteran target pests belonging to the genus Diabrotica such as Western corn rootworm ( D . virgifera virgifera ). MON 88017 is also developed to provide tolerance to the herbicidal active substance glyphosate by the introduction of a gene coding for the enzyme 5enolpyruvylshikimate-3-phosphate synthase (EPSPS), from Agrobacterium tumefaciens strain CP4 (CP4 EPSPS). Molecular Characterisation: Southern and PCR analyses indicate that the recombinant inserts in the single maize events MON 89034 and MON 88017 are retained in the stacked event MON 89034 x MON 88017. Genetic stability of the inserts has previously been demonstrated in the single events. The levels of Cry1A.105, Cry2Ab2, CP4 EPSPS and Cry3Bb1 proteins in grain and forage from the stacked event are comparable to the levels in the corresponding single events. Phenotypic analyses also indicate stability of the insect resistance and herbicide tolerance traits of the stacked event. Based on current knowledge and the previous assessments of the parental maize events, the VKM GMO Panel considers the molecular characterisation of maize MON 89034 x MON 88017 satisfactory. Comparative Assessment: Comparative analyses of maize MON 89034 x MON 88017 and its conventional counterpart have been performed by the applicant during field trials located at representative sites and environments in USA during 2004, and in Europe in 2007. Several different conventional maize varieties were included in the field trials and used as references. With the exception of small variations, and the insect resistance and herbicide tolerance conferred by the Cry3Bb1, Cry1A105, Cry2Ab2, and CP4 EPSPS proteins, the results from these studies showed no biologically relevant differences between the maize stack MON 89034 x MON 88017 and its conventional counterpart. Based on the assessment of available data, the VKM GMO Panel concludes that maize MON 89034 x MON 88017 is compositionally, agronomically and phenotypically equivalent to its conventional counterpart, except for the new proteins. Food and Feed Safety Assessment: A whole food feeding study performed on broilers indicates no adverse health effects of maize MON 89034 x MON 88017, and shows that it is nutritionally equivalent to conventional maize varieties. The Cry1A.105, Cry2Ab2, Cry3Bb1 and CP4 EPSPS proteins do not show relevant sequence resemblance to other known toxins or IgE-allergens, nor have they been reported to cause IgE-mediated allergic reactions. However, some studies have indicated a potential role of Cry-proteins as adjuvants in allergic reactions. Based on current knowledge, the VKM GMO Panel concludes that maize MON 89034 x MON 88017 is nutritionally equivalent to conventional maize varieties. It is unlikely that the Cry1A.105, Cry2Ab2, Cry3Bb1 and CP4 EPSPS proteins will cause toxic or IgE-mediated allergic reactions to food or feed derived from maize MON 89034 x MON 88017 compared to conventional maize. Environmental Risk: Considering the intended uses of maize MON 89034 x MON 88017, excluding cultivation, the environmental risk assessment is concerned with accidental release into the environment of viable grains during transportation and processing, and indirect exposure, mainly through manure and faeces from animals fed grains from maize MON 89034 x MON 88017. Maize MON 89034 x MON 88017 has no altered survival, multiplication or dissemination characteristics, and there are no indications of an increased likelihood of spread and establishment of feral maize plants in the case of accidental release into the environment of seeds from maize MON 89034 x MON 88017. Maize is the only representative of the genus Zea in Europe, and there are no cross-compatible wild or weedy relatives outside cultivation. The VKM GMO Panel considers the risk of gene flow from occasional feral GM maize plants to conventional maize varieties to be negligible in Norway. Considering the intended use as food and feed, interactions with the biotic and abiotic environment are not considered by the GMO Panel to be an issue. Overall Conclusion: Based on current knowledge, the VKM GMO Panel concludes that maize MON 89034 x MON 88017 is compositionally, nutritionally, agronomically and phenotypically equivalent to its conventional counterpart except for the new proteins. It is unlikely that the Cry1A.105, Cry2Ab2, CryBb1 and CP4 EPSPS proteins will cause an increased risk of toxic or IgE-mediated allergic reactions to food or feed based on maize MON 89034 x MON 88017 compared to conventional maize varieties. The VKM GMO Panel concludes that maize MON 89034 x MON 88017, based on current knowledge, is comparable to conventional maize varieties concerning environmental risk in Norway with the intended usage.

Final Health- and Environmental Risk Assessment of Genetically Modified Maize MON 88017 x MON 810

In preparation for a legal implementation of EU-regulation 1829/2003, the Norwegian Scientific Committee for Food Safety (VKM) has been requested by the Norwegian Environment Agency and the Norwegian Food Safety Authority (NFSA) to conduct final food/feed and environmental risk assessments for all genetically modified organisms (GMOs) and products containing or consisting of GMOs that are authorized in the European Union under Directive 2001/18/EC or Regulation 1829/2003/EC. The request covers scope(s) relevant to the Gene Technology Act. The request does not cover GMOs that VKM already has conducted its final risk assessments on. However, the Agency and NFSA requests VKM to consider whether updates or other changes to earlier submitted assessments are necessary. The insect-resistant and glyphosate-tolerant genetically modified maize MON 88017 x MON 810 from Monsanto (Unique Identifier DAS-MON 88017-3 x MON-ØØ81Ø-6) was approved under Regulation (EC) No 1829/2003 in the EU for food and feed uses, import and processing on 28th of July 2010 (Commission Decision 2010/429/EC). Genetically modified maize MON 88017 x MON 810 has previously been risk assessed by the VKM Panel on Genetically Modified Organisms (GMO), commissioned by the Norwegian Food Safety Authority related to the EFSA public hearing of the application in 2007 (VKM 2007a). In addition, MON 88017 and MON 810 has been evaluated by the VKM GMO Panel as single events and as a component of several stacked GM maize events and Regulation (EC) 1829/2003 and Directive 2001/18/EC (VKM 2005a,b,c, VKM 2007b,c,d, VKM 2008, VKM 2009, VKM 2010 a,b,c, VKM 2012, VKM 2013, VKM 2016). The food/feed and environmental risk assessment of the maize MON 88017 x MON 810 is based on information provided by the applicant in the application EFSA/GMO/CZ/2006/33 and scientific comments from EFSA and other member states made available on the EFSA website GMO Extranet. The risk assessment also considered other peer-reviewed scientific literature as relevant. The VKM GMO Panel has evaluated MON 88017 x MON 810 with reference to its intended uses in the European Economic Area (EEA), and according to the principles described in the Norwegian Food Act, the Norwegian Gene Technology Act and regulations relating to impact assessment pursuant to the Gene Technology Act, Directive 2001/18/EC on the deliberate release into the environment of genetically modified organisms, and Regulation (EC) No 1829/2003 on genetically modified food and feed. The Norwegian Scientific Committee for Food Safety has also decided to take account of the appropriate principles described in the EFSA guidelines for the risk assessment of GM plants and derived food and feed (EFSA 2011a), the environmental risk assessment of GM plants (EFSA 2010), selection of comparators for the risk assessment of GM plants (EFSA 2011b) and for the post-market environmental monitoring of GM plants (EFSA 2011c). The scientific risk assessment of maize MON 88017 x MON 810 include molecular characterisation of the inserted DNA and expression of novel proteins, comparative assessment of agronomic and phenotypic characteristics, nutritional assessments, toxicology and allergenicity, unintended effects on plant fitness, potential for gene transfer, effects on biogeochemical processes and interactions between the GM plant and target and non-target organisms. It is emphasized that the VKM mandate does not include assessments of contribution to sustainable development, societal utility and ethical considerations, according to the Norwegian Gene Technology Act and Regulations relating to impact assessment pursuant to the Gene Technology Act. These considerations are therefore not part of the risk assessment provided by the VKM Panel on Genetically Modified Organisms. Likewise, the VKM mandate does not include evaluations of herbicide residues in food and feed from genetically modified plants. The hybrid maize MON 88017 x MON 810 was produced by conventional crosses between inbred lines containing MON 88017 and MON 810 events to combine resistance to certain coleopteran and lepidopteran pests, and to confer tolerance towards glyphosate-containing herbicides. Maize MON 88017 was developed to express a modified Cry3Bb1 insecticidal protein, derived from Bacillus thuringiensis subsp. kumamotoensis , which confers protection against coleopteran target pests belonging to the genus Diabrotica such as Western corn rootworm ( Diabrotica virgifera virgifera ). MON 88017 is also developed to provide tolerance to the herbicidal active substance glyphosate by the introduction of a gene coding for the enzyme 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS), from Agrobacteri um tumefaciens strain CP4 (CP4 EPSPS). Maize MON 810 expresses the Cry1Ab insecticidal protein, derived from Bacillus thuringiensis subsp. k u rstaki, which confers protection against lepidopteran pests such as Ostrinia nubilaris and species belonging to the genus Sesamia. Molecular characterisation Southern and PCR analyses indicate that the recombinant inserts in the single maize events MON 88017 and MON 810 are retained in the stacked event MON 88017 x MON 810. Genetic stability of the inserts has previously been demonstrated in the single events. The levels of CP4 EPSPS, Cry3Bb1 and Cry1Ab proteins in grain and forage from the stacked event are comparable to the levels in the corresponding single events. Phenotypic analyses also indicate stability of the insect resistance and herbicide tolerance traits of the stacked event. Based on current knowledge and the previous assessments of the parental maize events, the VKM GMO Panel considers the molecular characterisation of maize MON 88017 x MON 810 satisfactory. Comparative assessment The applicant has performed comparative analyses of data from field trials located at representative sites and environments in USA during the 2002 growing season. With the exception of small intermittent variations and the insect resistance and herbicide tolerance conferred by the CP4 EPSPS, Cry3Bb1 and Cry1Ab proteins, the results showed no biologically relevant differences between maize stack MON 88017 x MON 810 and its conventional counterpart. Based on the assessment of available data, the VKM GMO Panel concludes that maize MON 88017 x MON 810 is compositionally, agronomically and phenotypically equivalent to its conventional counterpart, except for the new proteins. Food and feed safety assessment A whole food feeding study on broilers indicates no adverse health effects of maize MON 88017 x MON 810, and shows that it is nutritionally equivalent to conventional maize varieties. The Cry3Bb1, Cry1Ab and CP4 EPSPS proteins do not show relevant sequence resemblance to other known toxins or IgE-allergens, nor have they been reported to cause IgE-mediated allergic reactions. However, some studies have indicated a potential role of Cry-proteins as adjuvants in allergic reactions. Based on current knowledge, the VKM GMO Panel concludes that maize MON 88017 x MON 810 is nutritionally equivalent to conventional maize varieties. It is unlikely that the Cry3Bb1, Cry1Ab and CP4 EPSPS proteins will cause toxic or IgE-mediated allergic reactions to food or feed based on maize MON 88017 x MON 810 compared to conventional maize. Environmental risk assessment Considering the intended uses of maize MON 88017 x MON 810, excluding cultivation, the environmental risk assessment is concerned with accidental release into the environment of viable grains during transportation and processing, and indirect exposure, mainly through manure and faeces from animals fed grains from maize MON 88017 x MON 810. Maize MON 88017 x MON 810 has no altered survival, multiplication or dissemination characteristics, and there are no indications of an increased likelihood of spread and establishment of feral maize plants in the case of accidental release into the environment of seeds from maize MON 88017 x MON 810. Maize is the only representative of the genus Zea in Europe, and there are no cross-compatible wild or weedy relatives outside cultivation. The VKM GMO Panel considers the risk of gene flow from occasional feral GM maize plants to conventional maize varieties to be negligible in Norway. Considering the intended use as food and feed, interactions with the biotic and abiotic environment are not considered by the GMO Panel to be an issue. Overall conclusion Based on current knowledge, the VKM GMO Panel concludes that maize MON 88017 x MON 810 is compositionally, nutritionally, agronomically and phenotypically equivalent to its conventional counterpart except for the new proteins. It is unlikely that the Cry3Bb1, Cry1Ab and CP4 EPSPS proteins will cause an increased risk of toxic or IgE-mediated allergic reactions to food or feed based on maize MON 88017 x MON 810 compared to conventional maize varieties. The VKM GMO Panel concludes that maize MON 88017 x MON 810, based on current knowledge, is comparable to conventional maize varieties concerning environmental risk in Norway with the intended usage.

Risk Assessment of Insect-resistant and Herbicide Tolerant Genetically Modified Maize 1507 for Cultivation, Import, Processing, Food and Feed Uses under Directive 2001/18/EC and Regulation (EC) No 1829/2003

The Norwegian Environment Agency (NEA) and the Norwegian Food Safety Authority (NFSA) requested the Norwegian Scientific Committee for Food Safety (Vitenskapskomiteen for mattrygghet, VKM) for an opinion of potential risks to biodiversity and agriculture in Norway associated with import of seeds for sowing, and cultivation of insect-resistant and herbicide-tolerant genetically modified maize 1507 under Directive 2001/18/EC (Notification C/ES/01/01). The notification is still pending for authorisation in the European Union. VKM is also requested to assess the applicant´s post-market environmental monitoring plan, and the management measures suggested in the draft implementing decision of the European Commission. As VKM delivered a scientific opinion on this application including cultivation in 2014 (VKM, 2014), VKM is asked to assess whether the previous risk assessment is still valid concerning cultivation, and to update the opinion after current knowledge. The assessment shall specifically consider Norwegian conditions. Furthermore, as the notification does not cover food and feed uses of maize 1507, VKM was not asked for a health risk assessment of maize 1507. However, VKM has decided to update the previous safety evaluation of the food and feed uses of maize 1507 and derived products from 2014. VKM appointed a working group consisting of members from the Panel on Genetically Modified Organisms, the Panel on Alien Organisms and trade in Endangered Species (CITES) and the VKM staff to answer the requests. The Panel on Genetically Modified Organisms has assessed and approved the final report. The genetically modified maize 1507 was developed to provide protection against certain lepidopteran target pests, such as the European corn borer (ECB, Ostrinia nubilalis), and some species belonging to the genus Sesamia. The insect resistence is achieved by the expression of a synthetic version of the truncated c ry1F gene derived from Bacillus thuringiensis subsp. aizawai, a common soil bacterium. Maize 1507 also expresses the phosphinothricin - N - acetyltransferase (pat) gene, derived from the soil bacterium Streptomyces viridochromogenes. PAT protein confers tolerance to the herbicidal active substance glufosinate-ammonium. The PAT protein expressed in maize 1507 was used as a selectable marker to facilitate the selection process of transformed plant cells and is not intended for weed management purposes. Since the scope of the notification C/ES/01/01 does not cover the use of glufosinate-ammonium-containing herbicides on maize 1507, potential effects due to the use of such herbicides on maize 1507 are not considered by VKM. In delivering its scientific opinion, VKM considered relevant peer-reviewed scientific publications and information provided by the applicant in the notifications C/ES/01/01, C/NL/00/10, the applications EFSA/GMO/NL/2004/02 and EFSA/GMO/RX/1507, and scientific opinions and comments from EFSA and other EU member states. VKM has evaluated maize 1507 with reference to its intended uses in the European Economic Area (EEA), and according to the principles described in the Norwegian Food Act, the Norwegian Gene Technology Act and regulations relating to impact assessment pursuant to the Gene Technology Act, Directive 2001/18/EC on the deliberate release into the environment of genetically modified organisms, and Regulation (EC) No 1829/2003 on genetically modified food and feed. VKM has also decided to take into account, the appropriate principles described in the EFSA guidelines for the risk assessment of GM plants and derived food and feed (EFSA, 2011a), the environmental risk assessment of GM plants (EFSA, 2010a), selection of comparators for the risk assessment of GM plants (EFSA, 2011b) and for the post-market environmental monitoring of GM plants (EFSA, 2011c). The scientific opinion of maize 1507 include molecular characterisation of the inserted DNA and expression of novel proteins, comparative assessment of agronomic and phenotypic characteristics, nutritional assessments, toxicology and allergenicity. An evaluation of unintended effects on plant fitness, potential for gene transfer, interactions between the GM plant and target and non-target organisms, effects on biogeochemical processes, the postmarket environmental monitoring plan and coexistence measures at the farm level has also been undertaken. It is emphasised that VKM’s mandate does not include assessments of contribution to sustainable development, societal utility and ethical considerations, according to the Norwegian Gene Technology Act and Regulations relating to impact assessment pursuant to the Gene Technology Act. These considerations are therefore not part of the risk assessment provided by VKM. Molecular Characterization: Appropriate analyses of the transgenic DNA insert, its integration site, number of inserts and flanking sequences in the maize genome, have been performed. The results show that one copy only of the insert is present in maize 1507. Homology searches with databases of known toxins and allergens have not indicated any potential production of harmful proteins or polypeptides caused by the genetic modification in maize 1507. Southern blot analyses and segregation studies show that the introduced genes cry1F and pat are stably inherited and expressed over several generations along with the phenotypic characteristics of maize 1507. VKM considers the molecular characterisation of maize 1507 satisfactory. Comparative Assessment: Comparative analyses of maize 1507 to its non-GM conventional counterpart have been performed during multiple field trials located at representative sites and environments in Chile (1998/99), USA (1999) and in Europe (1999, 2000 and 2002). With the exception of small intermittent variations, no biologically significant differences were found between maize 1507 and the conventional maize. Based on the assessment of available data, VKM concludes that maize 1507 is compositionally, agronomically and phenotypically equivalent to its conventional counterpart, except for the introduced characteristics, and that its composition fell within the normal ranges of variation observed among non-GM varieties. The field evaluations support a conclusion of no phenotypic changes indicative of increased plant weed/pest potential of 1507 compared to conventional maize. Food and Feed Safety Assessment: Whole food feeding studies on rats, broilers, pullets, pigs and cattle have not indicated any adverse health effects of maize 1507. These studies further indicate that maize 1507 is nutritionally equivalent to conventional maize. The PAT and Cry1F proteins do not show sequence resemblance to other known toxins or IgE allergens, nor have they been reported to cause IgE mediated allergic reactions. Some studies have however, indicated a potential role of Cry-proteins as adjuvants in allergic reactions. Based on current knowledge, VKM concludes that maize 1507 is nutritionally equivalent to conventional maize varieties. It is unlikely that the PAT and Cry1F proteins will introduce a toxic or allergenic potential in food or feed based on maize 1507 compared to conventional maize. Environmental Risk Assessment: Maize is the only representative of the genus Zea in Europe, and there are no cross-compatible wild or weedy relatives outside cultivated maize with which maize can hybridise and form backcross progeny. Vertical gene transfer in maize therefore depends on cross-pollination with other conventional or organic maize varieties. In addition, unintended admixture of genetically modified material in seeds represents a possible way for gene flow between different crop cultivations. The risk of pollen flow from maize volunteers is negligible under Norwegian growing conditions. Since maize 1507 has no altered agronomic and phenotypic characteristics, except for the specific target insect resistance and herbicide tolerance, the likelihood of unintended environmental effects as a consequence of spread of genes from maize 1507 is considered to be extremely low. There are no reports of the target lepidopteran species attaining pest status on maize in Norway. Since there are no Bt-based insecticides approved for use in Norway, and lepidopteran pests have not been registered in maize, issues related to resistance evolution in target pests are not relevant at present for Norwegian agriculture. There are a limited number of published scientific studies on environmental effects of Cry1F protein. Published scientific studies show that the likelihood of negative effects of Cry1F protein on non-target arthropods that live on or in the vicinity of maize plants is low. In Norway, the maize cultivation is marginal. The total crop area of forage maize is estimated to 2000-2800 decares, equivalent to less than 0.1 % of the areas with cereal crops. The area of individual fields is limited by the topography such that the quantity of maize pollen produced under flowering is also limited. The potential exposure of Cry1F-containing maize pollen on non-target lepidopteran species in Norway is therefore negligible. Cultivation of maize 1507 is not considered to represent a threat to the prevalence of red-listed species in Norway. Exposure of non-target organisms to Cry proteins in aquatic ecosystems is likely to be very low, and potential exposure of Cry proteins to non-target organisms in aquatic ecosystems in Norway is considered to be negligible. VKM concludes that, although the data on the fate of the Cry1F protein and its potential interactions in soil are limited, the relevant scientific publications analysing the Cry1F protein, together with the relatively broad knowledge about the environmental fate of other Cry1 proteins, do not indicate significant direct effects on the soil environment. Despite limited number of studies, most studies conclude that effects on soil microorganisms and microbial communities are transient and minor c

Susceptibility of Grapholita molesta to insecticides in Brazil / Suscetibilidade de Grapholita molesta a inseticidas no Brasil

ABSTRACT: The use of insecticides has been the main tool for Grapholita molesta (Busck) control in Brazil, which is considered one of the most important pests in apple and peach orchards. In order to implement an Insect Resistance Management (IRM) program, studies were conducted to characterize the baseline susceptibility of G. molesta to major insecticides for its control. Then, we conducted an insecticide susceptibility monitoring in thirteen field-collected populations of the pest. Neonates (0-24h old) were exposed to insecticides applied on surface of artificial diet. A high susceptibility was verified when neonates of the Laboratory population of G. molesta were exposed to insecticides with LC50 values (µg a.i./cm2) of 0.1 (spinetoram), 1.0 (metaflumizone), 1.2 (chlorantraniliprole), 4.8 (novaluron), 5.1 (tebufenozide), 11.3 (phosmet) and 222.5 (pyriproxyfen). Based on the LC99 (µg a.i./cm2), the diagnostic concentrations of 0.6 (spinetoram), 5.5 (metaflumizone), 5.6 (chlorantraniliprole), 19.6 (tebufenozide), 37.4 (phosmet), 37.8 (novaluron) and 2011 pyriproxyfen) caused high mortality (>95%) of neonates from field populations. These diagnostic concentrations will be used in resistance monitoring programs of G. molesta in Brazil.

RESUMO: O uso de inseticidas tem sido a principal ferramenta para o controle da Grapholita molesta (Busck) no Brasil, considerada uma das mais importantes pragas em pomares de macieira e pessegueiro. Para implementar um programa de Manejo de Resistência a Insetos (MRI), estudos foram conduzidos para estabelecer uma linha básica de suscetibilidade de G. molesta a inseticidas utilizados para o seu controle. Posteriormente, foi realizado o monitoramento da suscetibilidade a inseticidas em treze populações da praga provenientes do campo. Lagartas (0-24 horas de idade) foram expostas a inseticidas aplicados na superfície da dieta artificial. Verificou-se alta suscetibilidade de lagartas neonatas de G. molesta (população de laboratório) quando foram expostas aos inseticidas, com valores de CL50 (µg i.a./cm2) de 0,1 (espinetoram), 1,0 (metoxifenozida), 1,2 (clorantraniliprole), 4,8 (novaluron), 5,1 (tebufenozida), 11,3 (fosmete) e 222,5 (piriproxifem). Com base na CL99 (µg i.a./cm2), as concentrações diagnósticas de 0,6 (espinetoram), 5,5 (metaflumizona), 5,6 (clorantraniliprole), 19,6 (tebufenozida), 37,4 (fosmete), 37,8 (novaluron) e 2.011 (piriproxifem) ocasionaram alta mortalidade (> 95%) de neonatas provenientes de populações de campo. Essas concentrações diagnósticas poderão ser utilizadas em programas de monitoramento da resistência de G. molesta no Brasil.

Progress on Function and Biosynthesis of Benzoxazinoids / 中国生物工程杂志

Benzoxazinoids (BXs) are important secondary metabolites in plants.There has been a wide range of attention and research of them because of their role in defensive and allelopathy.With the development of genomics and molecular biology,the BXs biosynthesis and other molecular areas research has made great progress.The BXs profile,the function of BXs,the genetic basis of BXs biosynthesis and expression regulation were briefly introduced.

Biological activity of Bt proteins expressed in different structures of transgenic corn against Spodoptera frugiperda / Atividade biológica de proteínas Bt expressas em diferentes estruturas de milho transgênico em Spodoptera frugiperda

ABSTRACT: Spodoptera frugiperda (J. E. Smith) is the main target pest of Bt corn technologies, such as YieldGard VT PRO(tm) (Cry1A.105/Cry2Ab2) and PowerCore(tm) (Cry1A.105/Cry2Ab2/Cry1F). In this study, it was evaluated the biological activity of Bt proteins expressed in different plant structures of YieldGard VT PRO(tm) and PowerCore(tm) corn against S. frugiperda . Complete mortality of S. frugiperda neonates was observed on leaf-disc of both Bt corn technologies. However, the mortality in silks and grains was lower than 50 and 6%, respectively. In addition, more than 49% of the surviving larvae in silks and grains completed the biological cycle. However, all life table parameters were negatively affected in insects that developed in silks and grains of both Bt corn events. In summary, the low biological activity of Bt proteins expressed on silks and grains of YieldGard VT PRO(tm) and PowerCore(tm) corn can contribute to the resistance evolution in S. frugiperda populations.

RESUMO: Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae ) é a principal praga-alvo dos eventos de milho YieldGard VT PRO(tm) (Cry1A.105/Cry2Ab2) e PowerCore(tm) (Cry1A.105/Cry2Ab2/Cry1F). Para subsidiar o manejo de resistência, avaliou-se a atividade biológica das proteínas de Bt expressas em diferentes estruturas dessas tecnologias de milho Bt contra S. frugiperda . Em discos de folha de milho YieldGard VT PRO(tm) e PowerCore(tm), houve mortalidade completa de neonatas de S. frugiperda . No entanto, em estilo-estigmas e grãos, a mortalidade foi inferior a 50% e 6%, respectivamente. Em adição, mais de 49% das larvas sobreviventes em estilos-estigmas e grãos completaram o ciclo biológico. No entanto, todos os parâmetros de tabela de vida de fertilidade foram negativamente afetados nos insetos que se desenvolveram em estilos-estigmas e grãos de milho Bt. A baixa atividade biológica das proteínas Bt expressas em estilos-estigmas e grãos de milho YieldGard VT PRO(tm) e PowerCore(tm) pode contribuir para a evolução de resistência em populações de S. frugiperda .