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Lepidoptera are the phytophagous group most at risk of impact by insect-resistant genetically modified (GM) plants since most of these crops have been developed to express toxins to specifically target lepidopteran pests. In the case of Bt-maize cultivation, pollen grains released by GM plants could dust larval host plants growing inside or nearby maize fields, thus leading to the exposure of caterpillars to the Cry toxins. This way of exposure has been studied mainly for butterflies, whereas moths, in spite of their great diversity and abundance, were rarely considered. To evaluate the potential exposure of moth species to maize pollen in absence of Cry-expressing plants, macro-moths were sampled by means of light traps in three protected areas of Northern, Central and Southern Italy where conventional maize is among the main crops. Light traps were activated in two consecutive years (2010- 11) from the beginning of June to the end of July, a period that encompasses the anthesis of most maize cultivars grown in Italy. Overall, 11446 individuals belonging to 201 species and nine families were collected. Given that larvae of several sampled species are likely to match the criteria of potential exposure to maize pollen and susceptibility to Cry toxins, macro-moths might be considered among the potential assessment endpoints for environmental risk assessment of GM plants, especially in the vicinity of protected areas. © 2017, Bulletin of Insectology. All rights reserved.

Genetically modified (GM) plants may show unintended differences compared to the original varieties, due to the modification process. Such differences might in some cases affect non-target organisms linked to the crop into an agro-ecosystem. In this paper, we aimed to study interactions of two blight-resistant GM potato clones with the aphid species Macrosiphum euphorbiae Thomas, a non-target arthropod frequently feeding on potato plants and one of the major pests of that crop. One of the potato events used in our experiments caused an increased fertility of the aphids in the first generation, and consequently, a positive effect on the growth of the aphid population was estimated. When a second generation of the aphid was reared on potato leaves of the same GM event, differences in aphid fertility were no longer observed. Behavioural studies conducted in a wind tunnel using the aphid parasitoid Aphidius ervi Haliday indicated that neither of the two tested GM varieties had a significant effect on the attractiveness of potato plants towards the parasitoid. In planta tests proved to be sensitive protocols to detect unintended effects on a non-target arthropod; experimental results, however, indicate that these effects are not expected to be biologically relevant in this tritrophic system, if these GM events become available for commercial use in the future.

Ladybird beetles (Coleoptera Coccinellidae) are important predators of aphids and other crop pests, and there is great interest in their conservation in agroecosystems. Bt-maize, genetically engineered to express insecticidal Cry proteins, is regarded as a taxon-specific way of controlling pests, but some concerns have been raised about the possibility of unintended negative effects on non-target organisms, including coccinellids. One of the possible routes of exposure of ladybird beetles to Cry toxins is by feeding on maize pollen, as for many species pollen is an important integrative food source. In this study, coccinellid adults were sampled by sweep netting in Ecological Compensation Areas (ECAs) in three sites of Northern and Central Italy, where conventional maize cultivars are grown (Bt-maize is currently forbidden for commercial cultivation in Italy). The coccinellid communities were sampled during and around the typical flowering periods of maize in order to check their diversity and their overlap with pollen shedding. A total of 11 species were recorded. Hippodamia variegata (Goeze), Coccinella septempunctata L. and the exotic Harmonia axyridis (Pallas) were the most abundant species in Northern Italy, whereas Tytthaspis sedecimpunctata (L.) and Coccinula quatuordecimpustulata (L.) were dominant in Central Italy. The potential exposure to maize pollen was different in the two areas, since in Northern Italy the maize flowering coincided with a period of high coccinellid activity in the field, whereas in Central Italy the ladybird population peaks occurred roughly one month earlier than the anthesis. The collected data might be useful for exposure characterization of ladybird beetles in a possible future scenario of Bt-maize cropping in the studied areas. © 2016, Universita degli Studi di Bologna, Department of Agroenvironmental Sciences and Technologies. All rights reserved.

Sicily (Italy) is one of the richest European regions in animal biodiversity. Butterflies offer good opportunities for studies on biodiversity, population and community ecology. Many species are strictly seasonal, preferring only a particular set of habitats. Five typical Mediterranean habitats: olive grove (conducted following organic farm management), mixed wood, ampelodesmos prairie (with dominance of Ampelodesmos mauritanicus), Mediterranean shrub and shrub at the coastal areas, were monitored for diversity and seasonal patterns in butterfly communities for 2 years. All habitats were inside protected areas indicated as Sites of Community Importance. Butterfly species richness was highest in ampelodesmos prairie and in olive grove. Pieris brassicae, Lasiommata megera, Maniola jurtina, Pararge aegeria have been reported in all habitats, although with different abundance whilst other species were observed only in one particular habitat. Melanargia pherusa, endemic species, and Hipparchia statilinus, listed in the IUCN Red List, respectively as “Least Concern” and “Near Threatened”, were observed in the ampelodesmos prairie. The morphotype Coenonympha pamphilus f. lyllus was reported in olive grove and ampelodesmos prairie. The majority of species showed abundance peaks in June and July, however many species showed a longer flight period than reported in literature with detection of specimens until November or December. The analysis of the similarity of communities among habitats shows a similarity between ampelodesmos prairie and olive grove and between Mediterranean shrub and Mediterranean shrub at the coastal areas, while the butterfly community in mixed wood is substantially different. The results of several diversity indexes suggest that ampelodesmos prairie has an important ecological role, as it supports butterfly abundance and species diversity. Copyright 2015, Universita degli Studi di Bologna, Department of Agroenvironmental Sciences and Technologies, All Rights Reserved.

Editorial on the Research Topic: Advances and Challenges of RNAi Based Technologies for Plants—Volume 2

Bee pollinators are an important guild delivering a fundamental input to European agriculture due to the ecological service they provide to crops in addition to the direct economic revenues from apiculture. Bee populations are declining in Europe as a result of the effects of several environmental stressors, both natural and of anthropic origin. Efforts are ongoing in the European Union (EU) to improve monitoring and management of pollinator populations to arrest further declines. Genetically modified (GM) crops are currently cultivated in a limited area in Europe, and an environmental risk assessment (ERA) is required prior to their authorization for cultivation. The possible impacts of GM crops on pollinators are deemed relevant for the ERA. Existing ecotoxicological studies indicate that traits currently expressed in insect‐resistant GM plants are unlikely to represent a risk for pollinators. However, new mechanisms of insect resistance are being introduced into GM plants, including novel combinations of Cry toxins and double strand RNA (dsRNA), and an ERA is required to consider lethal and sublethal effects of these new products on nontarget species, including insect pollinators. The evaluation of indirect effects linked to the changes in management practices (e.g. for herbicide‐tolerant GM crops) is an important component of EU regulations and a requirement for ERA. This paper reviews current approaches used to test the sensitivity of pollinators to GM plants and their products to determine whether sufficient data are being provided on novel GM plants to satisfy EU risk assessment requirements. © 2021 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry., There is concern that genetically modified (GM) plants might induce adverse effects on pollinators either directly or indirectly via reduced food availability. Biosafety regulation in the EU aims to assess these potential risks to ensure the biosafety of GM crops.

The use of RNA interference (RNAi) enables the silencing of target genes in plants or plant-dwelling organisms, through the production of double stranded RNA (dsRNA) resulting in altered plant characteristics. Expression of properly synthesized dsRNAs in plants can lead to improved crop quality characteristics or exploit new mechanisms with activity against plant pests and pathogens. Genetically modified (GM) crops exhibiting resistance to viruses or insects via expression of dsRNA have received authorization for cultivation outside Europe. Some products derived from RNAi plants have received a favourable opinion from the European Food Safety Authority (EFSA) for import and processing in the European Union (EU). The authorization process in the EU requires applicants to produce a risk assessment considering food/feed and environmental safety aspects of living organisms or their derived food and feed products. The present paper discusses the main aspects of the safety assessment (comparative assessment, molecular characterization, toxicological assessment, nutritional assessment, gene transfer, interaction with target and non-target organisms) for GM plants expressing dsRNA, according to the guidelines of EFSA. Food/feed safety assessment of products from RNAi plants is expected to be simplified, in the light of the consideration that no novel proteins are produced. Therefore, some of the data requirements for risk assessment do not apply to these cases, and the comparative compositional analysis becomes the main source of evidence for food/feed safety of RNAi plants. During environmental risk assessment, the analysis of dsRNA expression levels of the GM trait, and the data concerning the observable effects on non-target organisms (NTO) will provide the necessary evidence for ensuring safety of species exposed to RNAi plants. Bioinformatics may provide support to risk assessment by selecting target gene sequences with low similarity to the genome of NTOs possibly exposed to dsRNA. The analysis of these topics in risk assessment indicates that the science-based regulatory process in Europe is considered to be applicable to GM RNAi plants, therefore the evaluation of their safety can be effectively conducted without further modifications. Outcomes from the present paper offer suggestions for consideration in future updates of the EFSA Guidance documents on risk assessment of GM organisms.

Facing current climate challenges and drastically reduced chemical options for plant protection, the exploitation of RNA interference (RNAi) as an agricultural biotechnology tool has unveiled possible new solutions to the global problems of agricultural losses caused by pests and other biotic and abiotic stresses. While the use of RNAi as a tool in agriculture is still limited to a few transgenic crops, and only adopted in restricted parts of the world, scientists and industry are already seeking innovations in leveraging and exploiting the potential of RNAi in the form of RNA-based biocontrol compounds for external applications. Here, we highlight the expanding research and development pipeline, commercial landscape and regulatory environment surrounding the pursuit of RNA-based biocontrol compounds with improved environmental profiles. The commitments of well-established agrochemical companies to invest in research endeavours and the role of start-up companies are crucial for the successful development of practical applications for these compounds. Additionally, the availability of standardized guidelines to tackle regulatory ambiguities surrounding RNA-based biocontrol compounds will help to facilitate the entire commercialization process. Finally, communication to create awareness and public acceptance will be key to the deployment of these compounds. © 2019 Society of Chemical Industry.

RNA interference (RNAi) is being developed and exploited to improve plants by modifying endogenous gene expression as well as to target pest and pathogen genes both within plants (i.e. host-induced gene silencing) and/or as topical applications (e.g. spray-induced gene silencing). RNAi is a natural mechanism which can be exploited to make a major contribution towards integrated pest management and sustainable agricultural strategies needed worldwide to secure current and future food production. RNAi plants are being assessed and regulated using existing regulatory frameworks for GMO. However, there is an urgent need to develop appropriate science-based risk assessment procedures for topical RNAi applications within existing plant protection products legislation.

peer-reviewed This paper considers the statistical analysis of entomological count data from field experiments with genetically modified (GM) plants. Such trials are carried out to assess environmental safety. Potential effects on nontarget organisms (NTOs), as indicators of biodiversity, are investigated. The European Food Safety Authority (EFSA) gives broad guidance on the environmental risk assessment (ERA) of GM plants. Field experiments must contain suitable comparator crops as a benchmark for the assessment of designated endpoints. In this paper, a detailed protocol is proposed to perform data analysis for the purpose of assessing environmental safety. The protocol includes the specification of a list of endpoints and their hierarchical relations, the specification of intended levels of data analysis, and the specification of provisional limits of concern to decide on the need for further investigation. The protocol emphasizes a graphical representation of estimates and confidence intervals for the ratio of mean abundances for the GM plant and its comparator crop. Interpretation relies mainly on equivalence testing in which confidence intervals are compared with the limits of concern. The proposed methodology is illustrated with entomological count data resulting from multiyear, multilocation field trials. A cisgenically modified potato line (with enhanced resistance to late blight disease) was compared to the original conventional potato variety in the Netherlands and Ireland in two successive years (2013, 2014). It is shown that the protocol encompasses alternative schemes for safety assessment resulting from different research questions and/or expert choices. Graphical displays of equivalence testing at several hierarchical levels and their interpretation are presented for one of these schemes. The proposed approaches should be of help in the ERA of GM or other novel plants.

By means of a literature survey, earthworm species of significant relevance for soil functions in different biogeographical regions of Europe (Atlantic, Boreal, Mediterranean) were identified. These focal earthworm species, defined here according to the EFSA Guidance Document on the environmental risk assessment (ERA) of genetically modified plants, are typical for arable soils under crop rotations with maize and/or potatoes within the three regions represented by Ireland, Sweden and Spain, respectively. Focal earthworm species were selected following a matrix of four steps: Identification of functional groups, categorization of non-target species, ranking species on ecological criteria, and final selection of focal species. They are recommended as appropriate non-target organisms to assess environmental risks of genetically modified (GM) crops; in this case maize and potatoes. In total, 44 literature sources on earthworms in arable cropping systems including maize or potato from Ireland, Sweden and Spain were collected, which present information on species diversity, individual density and specific relevance for soil functions. By means of condensed literature data, those species were identified which (i) play an important functional role in respective soil systems, (ii) are well adapted to the biogeographical regions, (iii) are expected to occur in high abundances under cultivation of maize or potato and (iv) fulfill the requirements for an ERA test system based on life-history traits. First, primary and secondary decomposers were identified as functional groups being exposed to the GM crops. In a second step, anecic and endogeic species were categorized as potential species. In step three, eight anecic and endogeic earthworm species belonging to the family Lumbricidae were ranked as relevant species: Aporrectodea caliginosa, Aporrectodea rosea, Aporrectodea longa, Allolobophora chlorotica, Lumbricus terrestris, Lumbricus friendi, Octodrilus complanatus and Octolasion cyaneum. Five out of these eight species are relevant for each biogeographical region with an overlap in the species. Finally, the earthworm species Ap. caliginosa (endogeic, secondary decomposer) and L. terrestris (anecic, primary decomposer) were selected as focal species. In the Mediterranean region L. terrestris may be substituted by the more relevant anecic species L. friendi. The selected focal species are recommended to be included in a standardized laboratory ERA test system based on life-history traits. © 2016 Elsevier B.V.

Genetically modified (GM) plants may show unintended differences compared to the original varieties, due to the modification process. Such differences might in some cases affect non-target organisms linked to the crop into an agro-ecosystem. In this paper, we aimed to study interactions of two blight-resistant GM potato clones with the aphid species Macrosiphum euphorbiae Thomas, a non-target arthropod frequently feeding on potato plants and one of the major pests of that crop. One of the potato events used in our experiments caused an increased fertility of the aphids in the first generation, and consequently, a positive effect on the growth of the aphid population was estimated. When a second generation of the aphid was reared on potato leaves of the same GM event, differences in aphid fertility were no longer observed. Behavioural studies conducted in a wind tunnel using the aphid parasitoid Aphidius ervi Haliday indicated that neither of the two tested GM varieties had a significant effect on the attractiveness of potato plants towards the parasitoid. In planta tests proved to be sensitive protocols to detect unintended effects on a non-target arthropod; experimental results, however, indicate that these effects are not expected to be biologically relevant in this tritrophic system, if these GM events become available for commercial use in the future.

In legal frameworks worldwide, genetically modified plants (GMPs) are subjected to pre-market environmental risk assessment (ERA) with the aim of identifying potential effects on the environment. In the European Union, the EFSA Guidance Docuinent introduces the rationale that GMPs, as well as their newly produced metabolites, represent the potential stressor to be evaluated during ERA. As a consequence, during several phases of ERA for cultivation purposes, it is considered necessary to use whole plants or plant parts in experimental protocols. The importance of in planta studies as a strategy to address impacts of GMPs tin non-target organislns is demonstrated, to evaluate both effects due to the intended modification in plant phenotype (e.g. expression of Cry proteins) and effects due to unintended modifications in plant phenotype resultihg from the transformation process (e.g. due to somaclonal variations or pleiotropic effects). In planta tests are also necessary for GMPs in which newly expressed metabolites cannot easily be studied in vitro. This paper reviews the scientific literature supporting the choice of in planta studies as a fundamental tool in ERA of GMPs in cultivation dossiers; the evidence indicates they can realistically mimic the ecological relationships occurring in their receiving environments and provide important insights into the biology and sustainable management of GMPs.

Insect–plant interactions may be unintentionally affected when introducing genetically modified (GM) crops into an agro-ecosystem. Our aim was to test the non-target effects of a late blight-resistant GM potato on Myzus persicae in greenhouse and climate room experiments and understand how position and number of R gene insertions can affect non-targets in GM events. We also aimed to compare results to baseline differences among three conventional potato varieties varying in resistance to late blight. Aphid development and survival were affected by some GM events in the first generation, though effects disappeared in the second generation. Effects were not dependent on the presence of a marker gene or the insertion of a second resistance gene. Positional effects of gene insertion influenced aphid performance on certain GM events. However, aphid fitness varied considerably more between conventional potato varieties than between Désirée and the GM events. Comparing different GM events to the non-transformed variety is relevant, since unintended effects of insertion can occur. Our protocols can be recommended for in planta risk assessments with aphids. Ecological perspective is gained by selecting several measured endpoints and by comparing the results with a baseline of conventional cultivars.

The yield of several commodity crops is provided in large part by genetically modified crops in North and South America. However, reservations exist in Europe due to possible negative effects on human health or environment. This paper aims to analyse the current research priorities identified in EU countries and to engage European stakeholders into the formulation of future common research needs regarding the effects of the possible adoption of commercially available and forthcoming genetically modified organisms (GMOs) in the areas of socio-economics, human and animal health, and environment. Additionally, it aims to identify the requirements for sharing available research capacities and existing infrastructures. First a mapping exercise of existing research activities in Europe was performed. A questionnaire was developed on a web-based platform and submitted to national focal points to collect information from EU Member States. Information was collected from 320 research projects conducted in the last 10 years in Europe. To refine results of the surveys, twenty invited experts and stakeholders from the public funding agencies of different EU Member States participated in an international workshop. This paper reports the main findings of these activities., Bio-based and Applied Economics, Vol 6 No 1 (2017)

Sicily (Italy) is one of the richest European regions in animal biodiversity. Butterflies offer good opportunities for studies on biodiversity , population and community ecology. Many species are strictly seasonal, preferring only a particular set of habitats. Five typical Mediterranean habitats: olive grove (conducted following organic farm management), mixed wood, ampelodesmos prairie (with dominance of Ampelodesmos mauritanicus), Mediterranean shrub and shrub at the coastal areas, were monitored for diversity and seasonal patterns in butterfly communities for 2 years. All habitats were inside protected areas indicated as Sites of Community Importance. Butterfly species richness was highest in ampelodesmos prairie and in olive grove. Pieris brassicae, Lasiommata megera, Maniola jurtina, Pararge aegeria have been reported in all habitats, although with different abundance whilst other species were observed only in one particular habitat. Melanargia pherusa, endemic species, and Hipparchia statil-inus, listed in the IUCN Red List, respectively as " Least Concern " and " Near Threatened " , were observed in the ampelodesmos prairie. The morphotype Coenonympha pamphilus f. lyllus was reported in olive grove and ampelodesmos prairie. The majority of species showed abundance peaks in June and July, however many species showed a longer flight period than reported in literature with detection of specimens until November or December. The analysis of the similarity of communities among habitats shows a similarity between ampelodesmos prairie and olive grove and between Mediterranean shrub and Mediterranean shrub at the coastal areas, while the butterfly community in mixed wood is substantially different. The results of several diversity indexes suggest that ampelodesmos prairie has an important ecological role, as it supports butterfly abundance and species diversity.