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Background: Phthorimaea absoluta, a highly destructive invasive pest, poses a significant threat to tomato production globally. Exploring alternative control methods, such as host plant resistance can contribute to diminish reliance on insecticides and promote sustainable integrated pest management (IPM) practices. Thus, the identification of new P. absoluta-resistant tomato cultivars and potential wild sources for breeding programmes remains imperative. We evaluated the effect of 19 tomato genotypes, comprising 16 domesticated varieties and three wild tomato species, on oviposition output of female P. absoluta, as well as on larval performance under no-choice conditions using detached leaves. We also characterized and quantified glandular and nonglandular trichomes, exploring their potential correlation with the response of P. absoluta to the tomato plants., Results: Generally, fewer eggs were oviposited on domesticated plants, whereas the wild tomatoes Solanum arcanum and S. neorickii and the domesticated tomato Corona F1 impaired larval development. The pest larvae consumed a limited area of leaflets from S. arcanum and S. neorickii compared to other genotypes, leading to the lowest weights in both male and female pupae. All tomato plants exhibited a prevalence of nonglandular over glandular trichomes, except for S. arcanum, which exhibited a higher abundance of glandular trichomes. Although higher trichome density correlated with longer larval settlement on the leaflets, it did not influence female oviposition., Conclusion: Our findings demonstrate that the wild tomatoes S. arcanum and S. neorickii could be considered as potential sources for breeding programmes, and the domesticated Corona F1 could offer IPM options against P. absoluta. © 2024 The Author(s). Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry., (© 2024 The Author(s). Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.)

Which option for regulating plants derived from new genomic techniques in European Union law is feasible and justifiable scientifically? The European Commission has proposed a new regulation on plants obtained by specific new genomic techniques, which is now subject to discussion in the legislative process. From the perspective of the European Commission's envisaged legal reforms of European Union law towards the integration of greater sustainability, we conclude that the option focusing on plant traits delivering sustainability benefits should be chosen, which is most fitting to facilitate a contribution to climate action, the transition towards climate neutrality, and promptly integrate sustainability into all food-related policies. To assist the decision-making in the legislative process, we outline six regulatory options resulting from regulatory research involving interdisciplinary teams., (© 2023. Springer Nature Limited.)

Genetically engineered gene drives (geGD) are potentially powerful tools for suppressing or even eradicating populations of pest insects. Before living geGD insects can be released into the environment, they must pass an environmental risk assessment to ensure that their release will not cause unacceptable harm to non-targeted entities of the environment. A key research question concerns the likelihood that nontarget species will acquire the functional GD elements; such acquisition could lead to reduced abundance or loss of those species and to a disruption of the ecosystem services they provide. The main route for gene flow is through hybridization between the geGD insect strain and closely related species that co-occur in the area of release and its expected dispersal. Using the invasive spotted-wing drosophila, Drosophila suzukii, as a case study, we provide a generally applicable strategy on how a combination of interspecific hybridization experiments, behavioral observations, and molecular genetic analyses can be used to assess the potential for hybridization., (© 2023 The Authors. Risk Analysis published by Wiley Periodicals LLC on behalf of Society for Risk Analysis.)

Genetically engineered (GE) cotton, MON 88702, is protected against certain sucking pests, such as plant bugs and thrips, by producing mCry51Aa2, a modified protein from Bacillus thuringiensis (Bt). Predatory pirate bugs (Orius spp.), natural enemies contributing to biological pest control, are also sensitive to the insecticidal protein when exposed continuously to high concentrations. We evaluated effects of MON 88702 on Orius majusculus when fed prey types with different mCry51Aa2 concentrations. When neonates were provided exclusively Tetranychus urticae spider mites reared on MON 88702 (high mCry51Aa2 content), adverse effects on predator survival and development were confirmed, compared with specimens fed prey from near-isogenic non-Bt cotton. When fed a mixture of T. urticae and Ephestia kuehniella eggs (mCry51Aa2-free), predator life table parameters were similar to the treatment where eggs were fed exclusively. When mCry51Aa2-containing spider mites were provided for a limited time at the beginning or the end of juvenile development, effects were less pronounced. While pirate bug nymphs showed similar consumption rates for prey from Bt and non-Bt cotton, choice experiments revealed a preference for E. kuehniella eggs over spider mites. Lepidopteran larvae (Spodoptera littoralis, high mCry51Aa2 content) or cotton aphids (Aphis gossypii, mCry51Aa2-free) reared on MON 88702 as alternative prey did not result in adverse effects on O. majusculus. Our study suggests limited risk of mCry51Aa2-producing cotton for O. majusculus, because its sensitivity for the Bt protein is relatively low and its natural food consists of diverse prey species with varying concentrations of Bt protein., (© 2022 The Authors. Insect Science published by John Wiley & Sons Australia, Ltd on behalf of Institute of Zoology, Chinese Academy of Sciences.)

Background: Combining different biocontrol agents, particularly micro- and macroorganisms, can contribute to new and sustainable pest control approaches. Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae) is one of the most destructive pests of solanaceous crops. An emerging management strategy consists of biological control using microbial insecticides such as baculoviruses, but with limited efficacy. Thanks to their high target specificity, baculoviruses can be used simultaneously with natural enemies such as parasitoids for improved control of T. absoluta. However, potential indirect nontarget effects of baculoviruses on parasitoids can result from overlapping resource requirements. We assessed whether ovipositing parasitoid females discriminated against virus-treated hosts and examined the outcome of within-host competition between the hymenopteran parasitoids Necremnus tutae (Reuter) (Eulophidae) and Dolichogenidea gelechiidivoris Marsch (Braconidae), and the Phthorimaea operculella granulovirus (PhopGV, Baculoviridae) that infects T. absoluta larvae., Results: Female D. gelechiidivoris discriminated against virus-treated hosts, whereas N. tutae did not. We found few indirect virus-related effects depending on the species, the sex, and the time of virus treatment. Effects were ambivalent for D. gelechiidivoris offspring and ranged from increased male longevity when infection occurred before parasitization to reduced emergence and male longevity when infection occurred after parasitization. N. tutae offspring showed a longer development time and shorter male longevity when they developed in virus-treated hosts., Conclusion: The virus had a low impact on parasitoid offspring. In rare cases, adverse effects were detected; however, the low magnitude of these effects is unlikely to reduce the fitness of parasitoid offspring, therefore both parasitoids seem compatible with the baculovirus for control of T. absoluta. © 2022 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry., (© 2022 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.)

Engineered gene drives, which bias their own inheritance to increase in frequency in target populations, are being developed to control mosquito malaria vectors. Such mosquitoes can belong to complexes of both vector and nonvector species that can produce fertile interspecific hybrids, making vertical gene drive transfer (VGDT) to sibling species biologically plausible. While VGDT to other vectors could positively impact human health protection goals, VGDT to nonvectors might challenge biodiversity ones. Therefore, environmental risk assessment of gene drive use in species complexes invites more nuanced considerations of target organisms and nontarget organisms than for transgenes not intended to increase in frequency in target populations. Incorporating the concept of target species complexes offers more flexibility when assessing potential impacts from VGDT., Competing Interests: Declaration of interests J.B.C. and G.T.: Employment by Imperial College London in a position which is solely funded through a grant from the Bill & Melinda Gates Foundation, Member of Target Malaria not-for-profit research consortium; travel grants from Foundation for the National Institutes of Health. M.B.C.: Member of Target Malaria not-for-profit research consortium and supported by research grants from Bill & Melinda Gates Foundation., (Copyright © 2022 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Novel genetically modified biological control products (referred to as "GM biocontrol products") are being considered to address a range of complex problems in public health, conservation, and agriculture, including preventing the transmission of vector-borne parasitic and viral diseases as well as the spread of invasive plant and animal species. These interventions involve release of genetically modified organisms (GMOs) into the environment, sometimes with intentional dissemination of the modification within the local population of the targeted species, which presents new challenges and opportunities for regulatory review and decision-making. Practices developed for GMOs, primarily applied to date for GM crops may need to be adapted to accommodate different types of organisms, such as insects, and different technologies, such as gene drive. Developers of new GM biocontrol products would benefit from an early understanding of safety data and information that are likely to be required within the regulatory dossier for regulatory evaluation and decision making. Here a generalizable tool drawing from existing GM crop dossier requirements, forms, and relevant experience is proposed to assist researchers and developers organize and plan their research and trialing. This tool requires considering specifics of each investigational product, their intended use, and country specific requirements at various phases of potential product development, from laboratory research through contained field testing and experimental release into the environment. This may also be helpful to risk assessors and regulators in supporting their systematic and rigorous evaluation of new biocontrol products., (© 2022. The Author(s).)

Genetic modification of apple cultivars through cisgenesis can introduce traits, such as disease resistance from wild relatives, quickly and without crossing. This approach was used to generate the cisgenic apple line C44.4.146, a 'Gala Galaxy' carrying the fire blight resistance gene FB_MR5. In contrast to traditionally bred apple cultivars, genetically modified (GM) plants need to undergo a regulatory risk assessment considering unintended effects before approval for commercial release. To determine potential unintended effects of C44.4.146, we assessed major leaf components and effects on the fitness of the decomposers Drosophila melanogaster (fruit fly) and Folsomia candida (collembolan), which were fed a diet amended with powdered apple leaf material. Leaf material of 'Gala Galaxy', several natural 'Gala' mutants, and the unrelated apple cultivar 'Ladina' were used for comparison. The genetic modification did not alter major leaf components and did not adversely affect survival, growth, or fecundity of the two decomposers. Consistent with previous studies with other GM crops, the differences between conventionally bred cultivars were greater than between the GM line and its non-GM wild type. These data provide a baseline for future risk assessments., (Copyright © 2022. Published by Elsevier Inc.)

Background: Hundreds of studies on environmental effects of genetically modified (GM) crops became available over the past 25 years. For maize producing insecticidal proteins from Bacillus thuringiensis (Bt), potential adverse effects on non-target organisms are a major area of concern and addressed in risk assessments. Reviews and meta-analyses have helped various stakeholders to address uncertainties regarding environmental impacts of the technology. Many field studies from Europe and other parts of the world have been published in the last decade, and those data are often not covered by previous meta-analyses. Therefore, we conducted a systematic review to answer the question: "Does the growing of Bt maize change abundance or ecological function of non-target animals compared to the growing of non-GM maize?", Methods: Literature published until August 2019 was searched systematically in 12 bibliographic databases, 17 specialized webpages, and reference sections of 78 review articles. Defined eligibility criteria were applied to screen titles, abstracts, and full texts of the retrieved references. A custom-made database was developed with quantitative data on invertebrate abundance, activity density, or predation/parasitism rates. Eligible data that did not fit the quantitative database were captured in detailed tables and summarized narratively. For the first time, a critical appraisal scheme for field studies on non-targets in GM crops was developed to estimate the risk of bias (internal validity) and the suitability to answer the review question (external validity) of all primary data. Meta-analyses on different taxonomic levels, functional groups, and types of Bt maize were conducted. Untreated Bt maize was either compared with untreated non-Bt maize, or with insecticide-treated non-Bt maize. The influence of contributions by private sector product developers on reported effects was investigated., Review Findings: The database on non-target effects of Bt maize field trials contains more than 7200 records from 233 experiments and 120 articles. Meta-analyses on different taxonomic levels revealed only few and often non-robust significant effect sizes when both Bt maize and non-Bt maize were untreated. Bt maize harboured fewer parasitoids (Braconidae, Tachinidae) of the European corn borer, the main target pest of Lepidoptera-active Bt maize, compared with non-Bt maize. Similarly, sap beetles (Nitidulidae), that are associated with Lepidoptera damage, were recorded less in Bt maize. In some analyses, a negative effect of Bt maize was observed for rove beetles (Staphylinidae) and hoverflies (Syrphidae) and a positive effect for ladybeetles (Coccinellidae), flower bugs (Anthocoridae), and lacewings (Neuroptera). However, those effects were not consistent for different analyses and often related to individual articles. When untreated Bt maize was compared with pyrethroid-treated non-Bt maize, more effect sizes were significant. In particular, populations of predators were reduced after pyrethroid treatment, while few data were available for other insecticides. Funnel plots showed no evidence for publication bias and the analyses of private sector contribution revealed no evidence for influence of vested interests. Conclusions about potential effects of Bt maize on vertebrates or on animals inhabiting off-crop habitats were not possible, because only few such studies fitting the format of direct Bt/non-Bt comparisons on plot or field level were identified., Conclusions: The current work largely confirmed previously published results. The effects of Bt maize on the community of non-target invertebrates inhabiting maize fields were small and mostly neutral, especially when compared with the effects of broad-spectrum pyrethroid insecticide treatments., (© 2022. The Author(s).)

Objectives: To assess potential non-target effects of genetically engineered/modified (GM) maize that produces insecticidal proteins from Bacillus thuringiensis (Bt), numerous field experiments have been conducted worldwide. Field data are often variable and influenced by uncontrolled factors and meta-analyses can recognize general effects with increased statistical power compared to individual studies. This database represents a comprehensive collection of experimental field data on non-target invertebrates in Bt and non-Bt maize. It was created for a systematic review with the question if growing Bt maize changes abundance or ecological function of non-target animals compared to growing of non-GM maize. Systematic literature searches identified relevant data. Authors were contacted for additional information or raw data if needed and a critical appraisal scheme was developed and applied to each data record., Data Description: The database contains 7279 records of non-target invertebrate abundance, activity density, or predation or parasitism extracted from 120 articles. Records for individual species and life stages, but also aggregated data are available. Each record represents a comparison of invertebrates in Bt and non-Bt maize and includes means, standard deviations and sample sizes. Additional variables characterize publication details, experimental setup, cultivars, Bt proteins, geographic location, field management, insecticide treatments, sampling details, and taxonomy., (© 2022. The Author(s).)

Building on an exercise that identified potential harms from simulated investigational releases of a population suppression gene drive for malaria vector control, a series of online workshops identified nine recommendations to advance future environmental risk assessment of gene drive applications., (© 2022. The Author(s).)

The overwintering capacity of biocontrol agents is of fundamental relevance for biological control of pests in temperate regions. In this study we tested the cold tolerance of the indigenous Drosophila pupal parasitoids Pachycrepoideus vindemmiae and Trichopria drosophilae at constant low temperature in the laboratory and exposed different preimaginal parasitoid stages in the field during winter. We evaluated whether semi-natural habitats promote overwintering via more favorable microclimatic conditions as well as higher host availability compared to orchards. Further, we studied the parasitoids' phenology in a semi-field experiment during autumn. We found that P. vindemmiae larvae and pupae were most cold tolerant under laboratory and field conditions, while all preimaginal stages of T. drosophilae displayed similar cold tolerance. Semi-natural habitats buffered temperature extremes, yet overwintering survival was not enhanced compared to orchards. Suitable overwintering hosts were present in all habitats at times when parasitoids were still active parasitizing. These results demonstrate that P. vindemmiae overwinters most likely as larva or pupa and that T. drosophilae can overwinter in a preimaginal life stage. Further, we provide evidence that both parasitoids can overwinter in a wide range of habitats and that the availability of hosts for overwintering is unlikely a limiting factor for the parasitoids during fall., (Copyright © 2022 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Material from genetically engineered maize producing insecticidal Cry proteins from Bacillus thuringiensis (Bt) may enter aquatic ecosystems and expose nontarget organisms. We investigated the effects on life table parameters of the midge Chironomus riparius (Diptera: Chironomidae) of SmartStax maize leaves, which contain six different Cry proteins targeting Lepidoptera and Coleoptera pests, in two plant backgrounds. For midge development and emergence, 95% confidence intervals for the means of six conventional maize lines (Rheintaler, Tasty Sweet, ES-Eurojet, Planoxx, EXP 258, and EXP 262), were used to capture the natural range of variation. For reproduction, lowest and highest means were used. The natural range of variation allows one to judge whether observed effects between Bt maize and the closest non-Bt comparator are likely to be of biological relevance. No adverse effects on C. riparius were observed with any Bt maize line compared with the respective non-Bt counterpart. Development time was shorter when females were fed Bt maize than when they were fed non-Bt maize, but this effect was not considered adverse. Development time, emergence ratio, sex ratio, and larvae/egg rope measured for Bt maize were within the natural range of variation. Fecundity for the Bt lines was equal to or higher than that for the conventional lines. Future risk assessment studies may consider plant background effects and the natural range of variation to judge the relevance of observed differences between particular genetically engineered and non-genetically engineered plants. Environ Toxicol Chem 2022;41:1078-1088. © 2022 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC., (© 2022 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.)

Previous studies reported adverse effects of genetically engineered maize that produces insecticidal Cry proteins from Bacillus thuringiensis (Bt) on the water flea Daphnia magna. In the current study, effects of flour, leaves, or pollen from stacked Bt maize that contains six Bt proteins (SmartStax) in two plant backgrounds on life table parameters of D. magna were investigated. Adverse effects were observed for Bt maize flour, originating from different production fields and years, but not for leaves or pollen, produced from plants grown concurrently in a glasshouse. Because leaves contained eight to ten times more Cry protein than flour, the effects of the flour were probably not caused by the Cry proteins, but by compositional differences between the plant backgrounds. Furthermore, considering the natural range of variation in the response of D. magna to conventional maize lines, the observed effects of Bt maize flour were unlikely to be of biological relevance. Our study demonstrates how Cry protein effects can be separated from plant background effects in non-target studies using Bt plant material as the test substance and how detected effects can be judged for their biological relevance., (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)

A possible way to alleviate the public skepticism toward regulatory science is to increase transparency by making all data and value judgments used in regulatory decision making accessible for public interpretation, ideally early on in the process, and following the concepts of Open Science. This paper discusses the opportunities and challenges in strengthening Open Science initiatives in regulatory environmental risk assessment (ERA). In this discussion paper, we argue that the benefits associated with Open Science in regulatory ERA far outweigh its perceived risks. All stakeholders involved in regulatory ERA (e.g., governmental regulatory authorities, private sector, academia, and nongovernmental organizations), as well as professional organizations like the Society of Environmental Toxicology and Chemistry, can play a key role in supporting the Open Science initiative, by promoting the use of recommended reporting criteria for reliability and relevance of data and tools used in ERA, and by developing a communication strategy for both professionals and nonprofessionals to transparently explain the socioeconomic value judgments and scientific principles underlying regulatory ERA. Integr Environ Assess Manag 2021;17:1229-1242. © 2021 The Authors. Integrated Environmental Assessment and Management published by Wiley Periodicals LLC on behalf of Society of Environmental Toxicology & Chemistry (SETAC)., (© 2021 The Authors. Integrated Environmental Assessment and Management published by Wiley Periodicals LLC on behalf of Society of Environmental Toxicology & Chemistry (SETAC).)

Use of genetically engineered plants that express insecticidal Cry proteins derived from Bacillus thuringiensis (Bt) have been proven efficacious for managing lepidopteran pests. However, in some cases herbivores that are not targeted by the Bt trait have increased in importance. It has been suggested that reduced caterpillar damage to Bt crops could lead to decreased levels of induced plant defensive compounds which might benefit other non-target herbivores. Here we investigated the potential effect of reduced damage by larvae of Mythimna separata on aphid populations in Bt corn. We compared the performance of Rhopalosiphum maidis feeding on non-Bt corn plants that had been infested by M. separata larvae or were uninfested. The results showed that caterpillar-infested corn plants significantly reduced the fitness of R. maidis leading to a prolonged nymphal development time, reduced adult longevity and fecundity compared to uninfested plants. Consequently, the population growth rate of corn aphids feeding on caterpillar-infested corn plants was significantly lower than on uninfested plants. As expected, the aphids performed significantly better on Lepidoptera-resistant Bt corn than on non-Bt corn when plants were infested with M. separata, since the caterpillars caused very little damage to the Bt plants. The current findings indicate that reduced M. separata infestation could benefit aphid development in Bt corn. Bt corn has the potential to be commercialized in China in the near future and aphids and other non-target pests should be monitored in the farming fields., (© 2020 Institute of Zoology, Chinese Academy of Sciences.)

Non-target effects of genetically engineered (GE) plants on aquatic Daphnia magna have been studied by feeding the species with different maize materials containing insecticidal Cry proteins from Bacillus thuringiensis (Bt). The results of those studies were often difficult to interpret, because only one GE plant was compared to one related non-GE control. In such a setting, effects of the Cry proteins cannot be distinguished from plant background effects, in particular when the test species is nutritionally stressed. In the present study, we tested the suitability of three different maize materials, i.e., flour, leaves and pollen, from five diverse non-GE maize lines (including EXP 258, a breeding line that is closely related to a SmartStax Bt maize) as exclusive food sources for D. magna. The parameters recorded included survival, sublethal endpoints such as body size, number of moltings to first offspring, time to first offspring, number of individuals in first clutch, total number of clutches, total number of offspring, average number of offspring per clutch, and population measures such as net reproductive rate R 0 , generation time T and intrinsic rate of increase r m . The results showed that D. magna can survive, grow and reproduce when fed only maize materials, although the performance was poorer than when fed algae, which indicates nutritional stress. Large differences in life table and population parameters of D. magna were observed among the different maize lines. Our results suggest that confounding effects caused by nutritional stress and plant background might explain some of the conflicting results previously published on the effects of Bt crops on D. magna. Using 95% confidence intervals for the means of the five maize lines for all measured parameters of D. magna performance in our study, we captured the natural range of variation. This information is useful for the interpretation of observed differences in D. magna performance between a GE plant and its non-GE comparator as it helps judging whether observed effects are of biological relevance. If differences between a GE and comparator line are observed and their biological relevance needs to be assessed in future risk assessments of GE maize, 1) the data on natural variation of the different parameters generated by previous studies can be informative (e.g. data from our study for maize fed D. magna); 2) for additional experiments the inclusion of multiple unrelated non-GE comparators should be considered; In addition, it should be taken into account that nutritional stress can affect the outcome of the study., (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)

The complete mitochondrial genome of a predominant parasitoid, Necremnus tutae (Hymenoptera: Eulophidae) (GenBank accession number MT916846) is 15,252 bp in length, and contains 13 protein-coding genes (PCGs), 22 transfer RNA genes (tRNAs), 2 ribosomal RNA genes, and an A + T-rich region. The overall base composition is 38.86% for A, 7.14% for C, 8.57% for G, and 45.43% for T, with a high AT bias of 84.29%. ATA, ATT, ATG were initiation codons and TAA and T were termination codons. All the 22 tRNAs displayed a typical cloverleaf secondary structure, except for trnS 1 and trnR which lacked the dihydrouracil (DHU) arm. Phylogenetic analyses were performed using 13 PCGs showed that N. tutae is closely related to Tenthredo tienmushana , which in accordance with the traditional classification., Competing Interests: No potential conflict of interest was reported by the author(s)., (© 2021 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group.)

In late 2018, the highly destructive and polyphagous fall armyworm was first detected in China. It is now a major economic threat to corn production. In this article, the main control strategies that are available are reviewed and prospects to manage this pest with Bacillus thuringiensis (Bt) corn in China are discussed., (Copyright © 2020 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Insecticidal Cry proteins from Bacillus thuringiensis (Bt) can be transferred from genetically engineered crops to herbivores to natural enemies. For the lady beetle Harmonia axyridis, we investigated potential uptake of Cry proteins from the gut to the body and intergenerational transfer. Third and fourth instar H. axyridis fed with pollen or spider mites from SmartStax maize contained substantial amounts of Cry1A.105, Cry1F, Cry2Ab2, Cry3Bb1, and Cry34Ab1. Cry protein concentrations in lady beetle larvae were typically one order of magnitude lower than in the food. When H. axyridis larvae were fed Bt maize pollen, median amounts of Cry protein in the non-feeding pupae were below the limit of detection except for small amounts of Cry34Ab1. No Cry protein was detected in pupae when spider mites were used as food. Cry protein concentrations decreased quickly after H. axyridis larvae were transferred from pollen or spider mites to Bt-free food. Aphids contained very low or no detectable Cry protein, and no Cry protein was found in H. axyridis larvae fed with aphids, and in pupae. When H. axyridis adults were fed with Bt maize pollen (mixed with Ephestia kuehniella eggs), the median concentrations of Cry proteins in lady beetle eggs were below the limit of detection except for Cry34Ab1 in eggs laid later in adult life. No Bt protein was detected in eggs laid by H. axyridis females fed with aphids from Bt maize. Our results confirm previous observations that Cry proteins are degraded and excreted quickly in the arthropod food web without evidence for bioaccumulation. Despite the fact that small amounts of Cry proteins were detected in some samples of the non-feeding pupal stage of H. axyridis as well as in eggs, we conclude that this route of exposure is unlikely to be significant for predators or parasitoids in a Bt maize field., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Background: In its invaded regions, Drosophila suzukii (Matsumura) is a novel host for the community of resident parasitoids of Drosophila. To attain a high parasitization rate on the novel host, the parasitoids have to locate it and accept it in the presence of other Drosophila hosts. We conducted a laboratory choice experiment and a semifield trial to investigate host searching and host preference of the three pupal parasitoid species Trichopria drosophilae (Perkins), Pachycrepoideus vindemmiae (Rondani) and Spalangia erythromera Förster., Results: All three parasitoid species preferred D. suzukii over two common native hosts in the choice experiment. In field cages, most parasitoid offspring emerged from D. suzukii hosts. While P. vindemmiae mainly parasitized hosts in the foliage, most T. drosophilae offspring emerged from pupae presented on the ground., Conclusions: Both P. vindemmiae and T. drosophilae have the potential to find and parasitize D. suzukii in the field. If released early in the season, possible nontarget effects on native Drosophila should be minimal., (© 2020 Society of Chemical Industry.)

Background: Most recently, major federal regulatory agencies deregulated an in planta RNA interference (RNAi) trait against a devastating corn pest, the western corn rootworm Diabrotica virgifera virgifera, in the United States and Canada. The impact of double-stranded RNA (dsRNA) plant-incorporated protectants (PIPs) and dietary RNAi to non-target organisms, however, still needs further investigation. In this study, we assessed the potential risks of a Diabrotica virgifera virgifera active dsRNA to a group of predatory biological control agents, including Hippodamia convergens, Harmonia axyridis, Coleomegilla maculata, and Coccinella septempunctata. The overarching hypothesis is that the insecticidal dsRNA targeting Diabrotica virgifera virgifera has no or negligible adverse effect on lady beetles., Results: A 400-bp fragment with the highest sequence similarity between target and tested species was selected as the template for dsRNA synthesis. For the dietary RNAi toxicity assay, newly hatched first instar larvae were administered with v-ATPase A dsRNAs designed from Diabrotica virgifera virgifera and the four lady beetles, respectively. A dsRNA from β-glucuronidase (GUS), a plant gene, and H 2 O were served as the negative controls. The endpoint included both sub-organismal (gene expression), and organismal (survival rate, development time, pupa and adult weight) measurements. The results from dietary RNAi toxicity assay demonstrate significantly impacts of Diabrotica virgifera virgifera-active dsRNAs on lady beetles under the worst-case scenario at both transcriptional and phenotypic level. Interestingly, substantial differences among the four lady beetle species were observed toward the ingested exogenous dsRNAs., Conclusion: Such differential response to dietary RNAi may shed light on the mechanisms underlying the mode-of-action of RNAi-based biopesticides. © 2020 Society of Chemical Industry., (© 2020 Society of Chemical Industry.)

Advancements in -omics techniques provide powerful tools to assess the potential effects in composition of a plant at the RNA, protein and metabolite levels. These technologies can thus be deployed to assess whether genetic engineering (GE) causes changes in plants that go beyond the changes introduced by conventional plant breeding. Here, we compare the extent of transcriptome and metabolome modification occurring in leaves of four GE rice lines expressing Bacillus thuringiensis genes developed by GE and seven rice lines developed by conventional cross-breeding. The results showed that both types of crop breeding methods can bring changes at transcriptomic and metabolic levels, but the differences were comparable between the two methods, and were less than those between conventional non-GE lines were. Metabolome profiling analysis found several new metabolites in GE rice lines when compared with the closest non-GE parental lines, but these compounds were also found in several of the conventionally bred rice lines. Functional analyses suggest that the differentially expressed genes and metabolites caused by both GE and conventional cross-breeding do not involve detrimental metabolic pathways. The study successfully employed RNA-sequencing and high-performance liquid chromatography mass spectrometry technology to assess the unintended changes in new rice varieties, and the results suggest that GE does not cause unintended effects that go beyond conventional cross-breeding in rice., (© 2020 The Authors. The Plant Journal published by Society for Experimental Biology and John Wiley & Sons Ltd.)

Trichopria drosophilae (Perkins) (Hymenoptera: Diapriidae) is a pupal parasitoid of drosophila flies recorded from several parts of the world. It is currently considered for augmentative biological programs to control the severe agricultural pest Drosophila suzukii (Matsumura) (Diptera: Drosophilidae). Since D. suzukii has invaded regions that experience zero and sub-zero winter temperatures, cold tolerance of the parasitoid is an important aspect to consider. We characterized low temperature tolerance and overwintering capacity of a colony of T. drosophilae collected in Northern Switzerland. We used copper-constantan thermocouples to determine the supercooling point and pre-freeze mortality. Moreover, we subjected honey-fed and unfed adult T. drosophilae as well as developing stages within their drosophila host to short- and long-term acclimation conditions and assessed the duration of their survival at low temperatures. Finally, we exposed adult and sub-adult stages to winter conditions in a semi-field experiment and evaluated their survival. We found that T. drosophilae is chill susceptible like D. suzukii, but adults froze and survived at colder temperatures than those reported for D. suzukii. Adult parasitoids could tolerate several days of exposure to sub-zero temperatures and could reproduce afterwards, whereas sub-adult stages could survive longer periods under these conditions. The provision of honey and water enhanced the survival of adults and long-term acclimation led to longer survival in all stages. The semi-field experiment supported the results of the laboratory tests. Based on these results we suggest that in Central Europe, T. drosophilae survives winters mainly in developing stages but adults are likely able to tolerate short periods of low spring temperatures., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Historically, genetically engineered (GE) plants that have incorporated genes conferring insect protection have primarily used Cry proteins derived from Bacillus thuringiensis ( Bt ) to achieve their insecticidal phenotype. As a result, regulators have developed a level of familiarity and confidence in reviewing plants incorporating these insecticidal proteins. However, new technologies have been developed that produce GE plants that incorporate pest protection by triggering an RNA interference (RNAi) response or proteins other than Bt Cry proteins. These technologies have new modes of action. Although the overall assessment paradigm for GE plants is robust, there are ongoing discussions about the appropriate tests and measurement endpoints needed to inform non-target arthropod assessment for technologies that have a different mode of action than the Bt Cry proteins. As a result, increasing attention is being paid to the use of sublethal endpoints and their value for environmental risk assessment (ERA). This review focuses on the current status and history of sublethal endpoint use in insect-active GE crops, and evaluates the future use of sublethal endpoints for new and emerging technologies. It builds upon presentations made at the Workshop on Sublethal Endpoints for Non-target Organism Testing for Non- Bt GE Crops (Washington DC, USA, 4-5 March 2019), and the discussions of government, academic and industry scientists convened for the purpose of reviewing the progress and status of sublethal endpoint testing in non-target organisms., (Copyright © 2020 Roberts, Boeckman, Mühl, Romeis, Teem, Valicente, Brown, Edwards, Levine, Melnick, Rodrigues, Vélez, Zhou and Hellmich.)

RNA interference (RNAi) is a powerful technology that offers new opportunities for pest control through silencing of genes that are essential for the survival of arthropod pests. The approach relies on sequence-specificity of applied double-stranded (ds) RNA that can be designed to have a very narrow spectrum of both the target gene product (RNA) as well as the target organism, and thus allowing highly targeted pest control. Successful RNAi has been reported from a number of arthropod species belonging to various orders. Pest control may be achieved by applying dsRNA as foliar sprays. One of the main concerns related to the use of dsRNA is adverse environmental effects particularly on valued non-target species. Arthropods form an important part of the biodiversity in agricultural landscapes and contribute important ecosystem services. Consequently, environmental risk assessment (ERA) for potential impacts that plant protection products may have on valued non-target arthropods is legally required prior to their placement on the market. We describe how problem formulation can be used to set the context and to develop plausible pathways on how the application of dsRNA-based products could harm valued non-target arthropod species, such as those contributing to biological pest control. The current knowledge regarding the exposure to and the hazard posed by dsRNA in spray products for non-target arthropods is reviewed and suggestions are provided on how to select the most suitable test species and to conduct laboratory-based toxicity studies that provide robust, reliable and interpretable results to support the ERA., (Copyright © 2020 Romeis and Widmer.)

We discuss options to reform the EU genetically modified organism (GMO) regulatory framework, make risk assessment and decision-making more consistent with scientific principles, and lay the groundwork for international coherence. In this third of three articles, we focus on labeling and coexistence as well as discuss the political reality and potential ways forward., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Here, we discuss options to reform the EU genetically modified organism (GMO) regulatory framework, to make risk assessment and decision-making more consistent with scientific principles, and to lay the groundwork for international coherence. We discussed the scope and definitions in a previous article and, thus, here we focus on the procedures for risk assessment and risk management., (Copyright © 2019. Published by Elsevier Ltd.)

Concerns have been raised that multiple insecticidal proteins produced by genetically engineered (GE) crops may interact unexpectedly and pose new threats to biodiversity and nontarget organisms. We reviewed the literature to assess whether this concern is justified and whether the current regulatory framework needs to be adapted to address this concern., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

We discuss options to reform the EU genetically modified organisms (GMO) regulatory framework, make risk assessment and decision-making more consistent with scientific principles, and lay the groundwork for international coherence. The first in a three-part series, this article focuses on reform options related to the scope of the legislation and the GMO definition., (Copyright © 2020 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Winter wheat expressing the sucrose transporter HvSUT1 from barley (HOSUT) has an increased yield potential. Genetic engineering should improve cultivars without increasing susceptibility to biotic stresses or causing negative impacts on ecosystem services. We studied the effects of HOSUT wheat on cereal aphids that feed on the sugar-rich phloem sap. Three HOSUT winter wheat lines, their conventional parental cultivar Certo, and three conventional cultivars were used. Clip cage experiments in the greenhouse showed no differences in life-table parameters of Rhopalosiphum padi and Sitobion avenae (Hemiptera: Aphididae) on transgenic lines compared to Certo, except higher fecundity of S. avenae on one HOSUT line. Population development of both aphid species over three weeks on caged flowering tillers did not reveal differences between the HOSUT lines and Certo. When aphids were monitored in a Swiss field study over two years, no differences between HOSUT lines and Certo were observed. We conclude that HOSUT wheat did not have consistent effects on aphids compared to the parental cultivar and measured parameters were generally in the range observed for the conventional winter wheat cultivars. Thus, HOSUT wheat is unlikely to suffer from increased aphid damage., Competing Interests: The authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, or in the decision to publish the results.

Background: One concern with the adoption of RNAi-based genetically engineered (GE) crops is the potential harm to valued non-target organisms. Species of Coccinellidae (Coleoptera) are important natural enemies and might be exposed to the insecticidal dsRNA produced by the plant. To assess their susceptibility to dietary RNAi, we fed Adalia bipunctata and Coccinella septempunctata with a dsRNA designed to target the vATPase A of the western corn rootworm, Diabrotica virgifera virgifera (Dvv dsRNA). Specific dsRNAs designed to target the vATPase A of the two ladybird beetle species served as positive controls., Results: Our results revealed that both species were sensitive to dietary RNAi when ingesting their own dsRNAs, with C. septempunctata being more sensitive than A. bipunctata. Dvv dsRNA also adversely affected the two ladybird beetles as indicated by a significantly (but marginally) prolonged developmental time for A. bipunctata and a significantly reduced survival rate for C. septempunctata. These results, however, were obtained at Dvv dsRNA concentrations that were orders of magnitude higher than expected to occur in the field. Gene expression analyses confirmed the bioactivity of the dsRNA treatments and the results from the feeding bioassays. These results are consistent with the bioinformatics analyses, which revealed a higher number of 21-nucleotide-long matches, a requirement for effective RNAi, of the Dvv dsRNA with the vATPase A of C. septempunctata (34 matches) than with that of A. bipunctata (six matches)., Conclusion: Feeding bioassays revealed that two ladybird species are responsive to dietary RNAi. The two species, however, differed in their sensitivity. © 2019 Society of Chemical Industry., (© 2019 Society of Chemical Industry.)

Background: Besides fibers, cotton plants also produce a large amount of seeds with a high oil and protein content. The use of these seeds is restricted by their high contents of the terpenoid gossypol, which is harmful to humans and livestock. Using a genetic engineering approach, "Ultra-low gossypol cottonseed" (ULGCS) plants were produced by knocking down an enzyme that catalyzes the formation of a precursor of gossypol. This was accomplished via RNAi-mediated silencing of the target gene using a seed-specific α-globulin promotor. Since gossypol is also a crucial defense mechanism against leaf-feeding herbivores, ULGCS plants might possess lower herbivore resistance than non-engineered plants. Therefore, we tested the constitutive and inducible direct insect resistance of two ULGCS cotton lines against the African cotton leafworm, Spodoptera littoralis., Result: The herbivore was equally affected by both ULGCS lines and the control (Coker 312) line when feeding on fully expanded true leaves from undamaged plants and plants induced by jasmonic acid. When plants were induced by caterpillar-damage, however, S. littoralis larvae performed better on the ULGCS plants. Terpenoid analyses revealed that the ULGCS lines were equally inducible as the control plants. Levels of terpenoids were always lower in one of the two lines. In the case of cotyledons, caterpillars performed better on ULGCS cotton than on conventional cotton. This was likely caused by reduced levels of gossypol in ULGCS cotyledons., Conclusion: Despite those effects, the insect resistance of ULGSC cotton can be considered as largely intact and the plants may, therefore, be an interesting alternative to conventional cotton varieties.

Pre-market/prospective environmental risk assessments (ERAs) contribute to risk analyses performed to facilitate decisions about the market introduction of regulated stressors. Robust ERAs begin with an explicit problem formulation, which involves among other steps: (1) formally devising plausible pathways to harm that describe how the deployment of a regulated stressor could be harmful; (2) formulating risk hypotheses about the likelihood and severity of such events; (3) identifying the information that will be useful to test the risk hypotheses; and (4) developing a plan to acquire new data for hypothesis testing should tests with existing information be insufficient for decision-making. Here, we apply problem formulation to the assessment of possible adverse effects of RNA interference-based insecticidal genetically modified (GM) plants, GM growth hormone coho salmon, gene drive-modified mosquitoes and classical biological weed control agents on non-target organisms in a prospective manner, and of neonicotinoid insecticides on bees in a retrospective manner. In addition, specific considerations for the problem formulation for the ERA of nanomaterials and for landscape-scale population-level ERAs are given. We argue that applying problem formulation to ERA maximises the usefulness of ERA studies for decision-making, through an iterative process, because: (1) harm is defined explicitly from the start; (2) the construction of risk hypotheses is guided by policy rather than an exhaustive attempt to address any possible differences; (3) existing information is used effectively; (4) new data are collected with a clear purpose; (5) risk is characterised against well-defined criteria of hypothesis corroboration or falsification; and (6) risk assessment conclusions can be communicated clearly. However, problem formulation is still often hindered by the absence of clear policy goals and decision-making criteria (e.g. definition of protection goals and what constitutes harm) that are needed to guide the interpretation of scientific information. We therefore advocate further dialogue between risk assessors and risk managers to clarify how ERAs can address policy goals and decision-making criteria. Ideally, this dialogue should take place for all classes of regulated stressors, as this can promote alignment and consistency on the desired level of protection and maximum tolerable impacts across regulated stressors., (© 2019 European Food Safety Authority. EFSA Journal published by John Wiley and Sons Ltd on behalf of European Food Safety Authority.)

Trichopria drosophilae is a pupal parasitoid that can develop in a large number of drosophilid host species including the invasive pest Drosophila suzukii , and is considered a biological control agent. We investigated the influence of the rearing host on the preference and performance of the parasitoid, using two different strains of T. drosophilae , reared on D. melanogaster or D. suzukii for approximately 30 generations. Host switching was employed to assess the impact of host adaptation on T. drosophilae performance. In a no-choice experimental setup, T. drosophilae produced more and larger offspring on the D. suzukii host. When given a choice, T. drosophilae showed a preference towards D. suzukii , and an increased female ratio on this host compared to D. melanogaster and D. immigrans . The preference was independent from the rearing host and was confirmed in behavioral assays. However, the preference towards D. suzukii increased further after a host switch from D. melanogaster to D. suzukii in just one generation. Our data indicate that rearing T. drosophilae for several years on D. melanogaster does not compromise its performance on D. suzukii in the laboratory. However, producing a final generation on D. suzukii prior to release could increase its efficacy towards the pest.

This review describes the current status and future challenges of risk assessment and regulation of plants modified by modern biotechniques, namely genetic engineering and genome editing. It provides a general overview of the biosafety and regulation of genetically modified plants and details different regulatory frameworks with a focus on the European situation. The environmental risk and safety assessment of genetically modified plants is explained, and aspects of toxicological assessments are discussed, especially the controversial debate in Europe on the added scientific value of untargeted animal feeding studies. Because RNA interference (RNAi) is increasingly explored for commercial applications, the risk and safety assessment of RNAi-based genetically modified plants is also elucidated. The production, detection, and identification of genome-edited plants are described. Recent applications of modern biotechniques, namely synthetic biology and gene drives, are discussed, and a short outlook on the future follows.

Bt cotton was genetically modified to produce insecticidal proteins targeting Lepidopteran pests and is therefore only minimally affected by caterpillar damage. This could lead to reduced levels of inherent, systemically inducible defensive compounds in Bt cotton which might benefit other important cotton herbivores such as plant bugs. We studied the effects of plant defense induction on the performance of the plant bug Lygus hesperus by caging nymphs on different food sources (bolls/squares) of Bt and non-Bt cotton which were either undamaged, damaged by Bt tolerant caterpillars, or treated with jasmonic acid (JA). Terpenoid induction patterns of JA-treated and L. hesperus-damaged plants were characterized for different plant structures and artificial diet assays using purified terpenoids (gossypol/heliocide H1/4) were conducted. Nymphs were negatively affected if kept on plants damaged by caterpillars or sprayed with JA. Performance of nymphs was increased if they fed on squares and by the Bt-trait which had a positive effect on boll quality as food. In general, JA-sprayed plants (but not L. hesperus infested plants) showed increased levels of terpenoids in the plant structures analyzed, which was especially pronounced in Bt cotton. Nymphs were not negatively affected by terpenoids in artificial diet assays indicating that other inducible cotton responses are responsible for the found negative effects on L. hesperus. Overall, genetically engineered plant defenses can benefit plant bugs by releasing them from plant-mediated indirect competition with lepidopterans which might contribute to increasing numbers of hemipterans in Bt cotton.

The ladybird Propylea japonica , adults of the green lacewing Chrysoperla nipponensis and the honey bee Apis mellifera are common pollen feeders in many crop systems. They could therefore be directly exposed to Cry proteins in Bacillus thuringiensis ( Bt )-transgenic crop fields by ingestion of pollen. They, or closely related species, are therefore often selected as surrogate test species in non-target risk assessment of Bt plants. In the current study, we evaluated the potential effects of the ingestion of Bt maize pollen containing the Cry1Ab/Cry1Ac fusion protein on various life-table parameters of the three pollen-feeding non-target species in laboratory feeding assays. The results showed that pupation rate and male adult fresh weight of P. japonica were significantly increased when fed pollen from Bt maize compared to control maize pollen, but other test life-table parameters were not affected. For the other two species, none of the tested life-table parameters (survival, pre-oviposition period, fecundity and adult fresh weight for C. nipponensis; survival and mean acinus diameter of hypopharyngeal glands for A. mellifera ) differed between non- Bt and Bt maize pollen treatments. ELISA measurements confirmed the stability and uptake of the Cry protein by all three species during the feeding bioassays. In addition, a sensitive insect bioassay confirmed the bioactivity of the Cry1Ab/Cry1Ac protein in the Bt maize pollen used. Overall, the results suggested that the three pollen feeders are not sensitive to the Cry1Ab/Cry1Ac protein, and planting of the Bt maize variety will pose a negligible risk to P. japonica , adult C. nipponensis and adult A. mellifera .

The transgenic rice line T1C-19 provides high resistance to lepidopteran pests because of the synthesis of the Bacillus thuringiensis (Bt) insecticidal protein Cry1C. It thus shows good prospect for commercial planting in China. Species of Cladocera, an order of aquatic arthropods commonly found in aquatic ecosystems such as rice paddies, might be exposed to the insecticidal protein released from Bt-transgenic rice-straw residues. For the study reported herein, we used Daphnia magna (water flea) as a representative of Cladocera to evaluate whether aquatic arthropods are adversely affected when exposed to Bt rice-straw leachates. We exposed D. magna to M4 medium containing various volume percentages of medium that had been incubated with T1C-19 rice straw or rice straw from its non-transformed near-isoline Minghui 63 (MH63) for 21 days. Compared with pure M4 medium (control), the fitness and developmental and reproduction parameters of D. magna decreased significantly when exposed to rice-straw leachates; conversely, no significant differences between the T1C-19 and MH63 rice-straw leachate treatments were observed, indicating that the Bt rice straw leachate did not adversely affect this non-target species., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Phytochemical defense responses of plants are often herbivore-specific and can be affected by a herbivore's feeding mode. However, comprehensive studies documenting the impact of multiple herbivores from different feeding guilds on induced phytochemical responses in distal leaves and its consequences for plant-mediated herbivore interactions are limited and findings are inconsistent. We investigated how herbivory by leaf-chewing caterpillars, cell-content feeding spider mites and phloem-feeding aphids and whiteflies affect secondary metabolomes and phytohormone levels in youngest, non-damaged cotton leaves (distal leaves). Furthermore, bioassays with caterpillars were conducted to assess their performance on distal leaves of plants infested with different herbivores. Caterpillars, and to a lesser degree spider mites, led to a systemic induction of terpenoids with negative consequences for caterpillar performance in the bioassays. Both herbivores reduced levels of various nutrients and potentially antioxidative compounds. Caterpillar damage increased levels of jasmonoyl-L-isoleucine and abscisic acid (ABA), whereas spider mite infestation had no effect on phytohormone levels. Aphid and whitefly infestation did not systemically affect secondary metabolites. Aphids decreased salicylic acid levels while whitefly-infested plants contained increased ABA levels. Neither aphid nor whitefly infestation affected caterpillar performance. In general, feeding mode of a herbivore can affect systemically induced changes in phytochemistry and plant-mediated indirect interactions even though the two phloem-feeding herbivores triggered different phytohormonal responses. The observed reduction of nutrients and potentially antioxidative compounds upon caterpillar and spider mite herbivory underlines the importance of further elucidating the role of resource sequestration as a potential systemic defensive response following herbivory by chewers and cell-content feeding herbivores.

Background: Legumes have been genetically engineered to express α-amylase inhibitor 1 (αAI-1) from common bean in their seeds. Whereas the genetically modified (GM) seeds are immune to multiple bruchid pest species, the cosmopolitan bruchid Acanthoscelides obtectus is tolerant to αAI-1 and their larvae develop normally inside the seeds. Hymenopteran bruchid parasitoids, the most important natural enemies of bruchids, might thus be exposed to αAI-1 when attacking A. obtectus larvae developing inside GM seeds. Exposure might reduce parasitoid fitness, resulting in a decline in the natural control of A. obtectus, and thus promote the spread of this pest. We investigated the impact of the presence of αAI-1 in legume seeds on parasitoid fitness in tritrophic experiments with αAI-1 GM or non-GM chickpea seeds, A. obtectus, and three parasitoid species. Additionally, we investigated the exposure of parasitoids to αAI-1 using a fourth, highly sensitive parasitoid species., Results: Parasitoid fitness was not affected when A. obtectus was used in GM chickpea seeds as hosts, and this lack of effects was probably attributable to the fact that exposure of the parasitoids to αAI-1 was negligible., Conclusion: We conclude that the release of GM chickpeas containing αAI-1 should not harm this important group of non-target insects. © 2018 Society of Chemical Industry., (© 2018 Society of Chemical Industry.)

With the cultivation of Bt cotton, the produced insecticidal Cry proteins are ingested by herbivores and potentially transferred along the food chain to natural enemies, such as predators. In laboratory experiments with Bollgard II cotton, concentrations of Cry1Ac and Cry2Ab were measured in Lepidoptera larvae (Spodoptera littoralis, Heliothis virescens), plant bugs (Euschistus heros), aphids (Aphis gossypii), whiteflies (Bemisia tabaci), thrips (Thrips tabaci, Frankliniella occidentalis), and spider mites (Tetranychus urticae). Tritrophic experiments were conducted with caterpillars of S. littoralis as prey and larvae of ladybird beetles (Harmonia axyridis, Adalia bipunctata) and lacewings (Chrysoperla carnea) as predators. Immunological measurements (ELISA) indicated that herbivores feeding on Bt cotton contained 5%-50% of the Bt protein concentrations in leaves except whiteflies and aphids, which contained no or only traces of Bt protein, and spider mites, which contained 7 times more Cry1Ac than leaves. Similarly, predators contained 1%-30% of the Cry protein concentration in prey. For the nontarget risk assessment, this indicates that Bt protein concentrations decrease considerably from one trophic level to the next in the food web, except for spider mites that contain Bt protein concentrations higher than those measured in the leaves. Exposure of phloem sucking hemipterans is negligible., (© 2017 Institute of Zoology, Chinese Academy of Sciences.)

Genetically engineered (GE) rice lines expressing Lepidoptera-active insecticidal cry genes from the bacterium Bacillus thuringiensis (Bt) have been developed in China. Field surveys indicated that Bt rice harbours fewer rice planthoppers than non-Bt rice although planthoppers are not sensitive to the produced Bt Cry proteins. The mechanisms underlying this phenomenon remain unknown. Here, we show that the low numbers of planthoppers on Bt rice are associated with reduced caterpillar damage. In laboratory and field-cage experiments, the rice planthopper Nilapavata lugens had no feeding preference for undamaged Bt or non-Bt plants but exhibited a strong preference for caterpillar-damaged plants whether Bt or non-Bt. Under open-field conditions, rice planthoppers were more abundant on caterpillar-damaged non-Bt rice than on neighbouring healthy Bt rice. GC-MS analyses showed that caterpillar damage induced the release of rice plant volatiles known to be attractive to planthoppers, and metabolome analyses revealed increased amino acid contents and reduced sterol contents known to benefit planthopper development. That Lepidoptera-resistant Bt rice is less attractive to this important nontarget pest in the field is therefore a first example of ecological resistance of Bt plants to nontarget pests. Our findings suggest that non-Bt rice refuges established for delaying the development of Bt resistance may also act as a trap crop for N. lugens and possibly other planthoppers., (© 2018 The Authors. Plant Biotechnology Journal published by Society for Experimental Biology and The Association of Applied Biologists and John Wiley & Sons Ltd.)

Background: Honeydew is a sugar-rich excretion produced by sap-feeding Sternorrhyncha and is an important source of carbohydrates for natural enemies, especially for parasitoids. Honeydew derived from genetically modified (GM) crops can contain amounts of the transgene product. Thus, it is a possible route of exposure for natural enemies feeding on honeydew. In the present study, the potential effects of Nilaparvata lugens honeydew derived from Cry1C and Cry2A rice on different life-table parameters and parasitism dynamics of the egg parasitoid Anagrus nilaparvatae were evaluated under laboratory and field conditions. Furthermore, the Bacillus thuringiensis (Bt) levels and the sugar and amino acid composition of honeydew were analyzed., Results: Results indicated that A. nilaparvatae was exposed to Bt proteins by feeding on N. lugens honeydew produced from Bt rice. However, honeydew derived from the tested Cry1C and Cry2A rice lines did not affect the development, longevity, emergence rate and fecundity of A. nilaparvatae. Also, the parasitism dynamics in the field remained unaffected. In addition, the sugar and amino acid composition of N. lugens honeydew was not significantly altered for the tested Bt rice lines compared with the parental non-Bt plant., Conclusion: The quality of honeydew derived from the tested Bt rice lines as a food resource for natural enemies was maintained. © 2018 Society of Chemical Industry., (© 2018 Society of Chemical Industry.)

The area planted with insect-resistant genetically engineered crops expressing Bacillus thuringiensis ( Bt ) genes has greatly increased in many areas of the world. Given the nearby presence of non- Bt crops (including those planted as refuges) and non-crop habitats, pests targeted by the Bt trait have a choice between Bt and non- Bt crops or weeds, and their host preference may greatly affect insect management and management of pest resistance to Bt proteins. In this study, we examined the oviposition preference of the target pest of Bt rice, Chilo suppressalis , for Bt versus non- Bt rice plants as influenced by previous damage caused by C. suppressalis larvae. The results showed that C. suppressalis females had no oviposition preference for undamaged Bt or non- Bt plants but were repelled by conspecific-damaged plants whether Bt or non- Bt Consequently, C. suppressalis egg masses were more numerous on Bt plants than on neighbouring non- Bt plants both in greenhouse and in field experiments due to the significantly greater caterpillar damage on non- Bt plants. We also found evidence of poorer performance of C. suppressalis larvae on conspecific-damaged rice plants when compared with undamaged plants. GC-MS analyses showed that larval damage induced the release of volatiles that repelled mated C. suppressalis females in wind tunnel experiments . These findings suggest that Bt rice could act as a dead-end trap crop for C. suppressalis and thereby protect adjacent non- Bt rice plants. The results also indicate that the oviposition behaviour of target pest females should be considered in the development of Bt resistance management strategies., (© 2018 The Authors.)

Assessing the potential effects of insect-resistant genetically engineered (GE) plants on collembolans is important because these common soil arthropods may be exposed to insecticidal proteins produced in GE plants by ingestion of plant residues, crop pollen, or root exudates. Laboratory studies were conducted to evaluate the potential effects of two Bacillus thuringiensis ( Bt )-rice lines expressing Cry1C and Cry2A in pollen and leaves and of their non- Bt conventional isolines on the fitness of the collembolan Folsomia candida and on the activities of its antioxidant-related enzymes, superoxide dismutase and peroxidase, and of its detoxification-related enzymes, glutathione reductase and glutathione S -transferase. Survival, development, reproduction, and the intrinsic rate of increase ( r m ) were not significantly reduced when F. candida fed on the Bt rice pollen or leaf powder than on the non- Bt rice materials; these parameters, however, were significantly reduced when F. candida fed on non- Bt rice pollen or non- Bt leaf-based diets containing the protease inhibitor E-64 at 75 μg/g. The activities of the antioxidant-related and detoxification-related enzymes in F. candida were not significantly affected when F. candida fed on the Bt rice materials, but were significantly increased when F. candida fed on the non- Bt rice materials containing E-64. The results demonstrate that Cry1C and Cry2A are not toxic to F. candida , and also indicate the absence of unintended effects on the collembolan caused by any change in plant tissue nutritional composition due to foreign gene transformation.

In the agroecosystem, genetically engineered plants producing insecticidal Cry proteins from Bacillus thuringiensis (Bt) interact with non-target herbivores and other elements of the food web. Stacked Bt crops expose herbivores to multiple Cry proteins simultaneously. In this study, the direct interactions between SmartStax ® Bt maize producing six different Cry proteins and two herbivores with different feeding modes were investigated. Feeding on leaves of Bt maize had no effects on development time, fecundity, or longevity of the aphid Rhopalosiphum padi (Hemiptera: Aphididae), and no effects on the egg hatching time, development time, sex ratio, fecundity, and survival of the spider mite Tetranychus urticae (Acari: Tetranychidae). The results thus confirm the lack of effects on those species reported previously for some of the individual Cry proteins. In the Bt maize leaves, herbivore infestation did not result in a consistent change of Cry protein concentrations. However, occasional statistical differences between infested and non-infested leaves were observed for some Cry proteins and experimental repetitions. Overall, the study provides evidence that the Cry proteins in stacked Bt maize do not interact with two common non-target herbivores.

Cabbage looper, Trichoplusia ni (Hübner) is an important lepidopteran pest on many vegetable and greenhouse crops, and some field crops. Although there are no commercial transgenic Bt vegetable or greenhouse crops, T. ni is a target of Bollgard II cotton, which produces Cry1Ac and Cry2Ab. We expand on previous work that examined the effect of Bt crops on parasitoids using Bt-resistant lepidopteran populations as hosts. Cry1Ac/Cry2Ab-resistant T. ni larvae were used to eliminate host quality effects and to evaluate the direct effects of Bt cotton on the parasitoids Copidosoma floridanum (Ashmead) and Cotesia marginiventris (Cresson). These tri-trophic studies confirm that Bt cotton had no significant impact on development, success of parasitism, survival and adult longevity of C. marginiventris when using Bt-resistant T. ni fed on Bt cotton. Similarly, this Bt cotton had no significant impact on the development, mummy weight and the number of progeny produced by C. floridanum. Our studies verified that lyophilized Bt crop tissue maintained its insecticidal bioactivity when incorporated into an artificial diet, demonstrating that hosts and parasitoids were exposed to active Cry proteins. The egg-larval parasitoid C. floridanum, or similar species that consume their entire host, should be considered useful surrogates in risk assessment of Bt crops to non-target arthropods.