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This document provides supplementary guidance on specific topics for the allergenicity risk assessment of genetically modified plants. In particular, it supplements general recommendations outlined in previous EFSA GMO Panel guidelines and Implementing Regulation (EU) No 503/2013. The topics addressed are non-IgE-mediated adverse immune reactions to foods, in vitro protein digestibility tests and endogenous allergenicity. New scientific and regulatory developments regarding these three topics are described in this document. Considerations on the practical implementation of those developments in the risk assessment of genetically modified plants are discussed and recommended, where appropriate. (C) 2017 European Food Safety Authority. EFSA Journal published by John Wiley and Sons Ltd on behalf of European Food Safety Authority

Field trials were established at three European sites (Denmark, Eastern France, South-West France) of genetically modified maize (Zea mays L.) expressing the CryIAb Bacillus thuringiensis toxin (Bt), the nearisogenic non-Bt cultivar, another conventional maize cultivar and grass. Soil from Denmark was sampled at sowing (May) and harvest (October) over two years (2002, 2003); from E France at harvest 2002, sowing and harvest 2003; and from SW France at sowing and harvest 2003. Samples were analysed for microbial community structure (2003 samples only) by community-level physiological-profiling (CLPP) and phospholipid fatty acid analysis (PLFA), and protozoa and nematodes in all samples. Individual differences within a site resulted from: greater nematode numbers under grass than maize on three occasions; different nematode populations under the conventional maize cultivars once; and two occasions when there was a reduced protozoan population under Bt maize compared to non-Bt maize. Microbial community structure within the sites only varied with grass compared to maize, with one occurrence of CLPP varying between maize cultivars (Bt versus a conventional cultivar). An overall comparison of Bt versus non-Bt maize across all three sites only revealed differences for nematodes, with a smaller population under the Bt maize. Nematode community structure was different at each site and the Bt effect was not confined to specific nematode taxa. The effect of the Bt maize was small and within the normal variation expected in these agricultural systems.

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In Brassica crops differences in susceptibility to root fly attack can be largely attributed to antixenotic resistance. Plants of four genotypes (two swedes and two kales) with widely differing resistance in field trials, were compared in laboratory choice assays for their susceptibility to oviposition by the root flies Delia radicum (L.) and D. floralis (Fallen) (Diptera, Anthomyiidae). For both species the preference among the genotypes corresponded to the susceptibility of the genotypes in the field. The preference ranking in response to surrogate leaves treated with methanolic surface extracts of the four genotypes was identical to the preference among potted plants, demonstrating that chemical factors on the leaf surface mediate host preference for oviposition in these species. For both species of fly, glucosinolates are major oviposition stimulants and for D. radicum an additional, non‐glucosinolate oviposition stimulant, presently called CIF, is known. We describe a procedure for chromatographic separation of glucosinolates from CIF in leaf surface extracts. In oviposition‐choice assays with D. radicum, the CIF‐fractions of the two swede genotypes applied to surrogate leaves received a 1.8 and 4.6 times higher proportion of eggs than the respective glucosinolate‐fractions, confirming the major importance of CIF as an oviposition stimulant. The genotype of swede that was preferred by both fly species in tests with plants and methanolic leaf surface extracts, also stimulated oviposition more in tests with the glucosinolate‐fractions or the CIF‐fractions derived from the surface extracts, respectively. Thus, glucosinolates and CIF together account for the observed preference among the genotypes and may also be responsible for their susceptibility under field conditions. In the two kale genotypes the preference for plants or surface extracts differed from the preference among the corresponding glucosinolate‐ and CIF‐fractions, indicating that additional, as yet unknown chemical factors may also be involved. For both groups of stimulants tarsal chemoreceptors allow electrophysiological monitoring of glucosinolate‐ and CIF‐activity in fractionated surface extracts. For D. radicum the chemosensory activity of both glucosinolate‐ and CIF‐fractions corresponded to the respective behavioural activity in the oviposition preference tests, suggesting that preference for oviposition among genotypes can be predicted from the electrophysiological activity of their fractions. The chemosensory response of D. floralis, in particular to the CIF‐fractions, was less pronounced than the response of D. radicum, indicating interspecific differences in the perception of the major oviposition stimulants. We discuss the potential application of electrophysiological techniques in support of other screening methods used in breeding for root fly resistance in Brassica crops.

Two genotypes of kale (Brassica olerucea L). two of rape (one forage and one winter oilseed type, B riapus L) and two of swede ( B rtupus L) grown in a glasshouse were inoculated with turnip root fly (Deliafloralis Fall) eggs at 5 weeks and their roots, stems and leaves sampled after a further 8 weeks of larval feeding. Changes in total and individual glucosinolates were examined in roots, stems and leaves after attack. Damage by Dflorulis larvae to roots caused tissue weight losses in roots ranging from 26 to 48 Yo, relative to controls. The resultant plant stress also caused smaller weight losses in leaf tissue (10-27 Yo) and stems (6-20 YO). On susceptible brassica genotypes, D florulis percent pupal development and mean pupal weights were a factor of x 1.5 greater than on resistant genotypes, resulting in a difference of x 2.4 in total pupal biomass per root. There was no apparent link between D floralis susceptibility and total or individual glucosinolate content of undamaged control roots. Major differences in glucosinolate composition were found between control roots of the six brassicas. Five of the 15 compounds detected had aromatic side chains and were present in all samples. Only three of the 10 aliphatic glucosinolates were common to all six brassicas. The predominant compound in the two kale roots was prop-2-enyl glucosinolate, in the rapes 2hydroxy-but-3-enyl (forage rape cv Hobson) and 2-phenylethyl (oilseed rape cv Ariana) glucosinolates, and in the swedes 2-hydroxy-but-3-enyl glucosinolate. All brassicas showed a similar response in glucosinolate metabolism after root damage. Total glucosinolate content in roots increased due to a two to four fold increase in indole-based compounds. The largest increase for an individual compound after attack was found for I -methoxy-3-indolylmethyl glucosinolate, which increased four to 17 fold. Root damage did not significantly affect stem glucosinolate composition but in leaves caused a 1.2 fold increase in aliphatic glucosinolates with a corresponding decrease in indole-based compounds. The possible ecological and nutritional significance of glucosinolatc responses to root fly darnage is discussed.

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This document provides guidance for the agronomic and phenotypic characterisation of genetically modified (GM) plants and clarifies the EFSA GMO Panel?s view on how agronomic and phenotypic data support the risk assessment of GM plants. Specific recommendations are given on (1) the selection of sites and test materials; (2) the quality and design of field trials; (3) the selection of relevant agronomic and phenotypic endpoints; and (4) data analysis. The guidance proposes a comprehensive and harmonised approach for the agronomic and phenotypic characterisation of GM plants, which should ensure the best use of agronomic and phenotypic data for the comparative analysis of GM plants and derived food and feed products, and for their food and feed and environmental risk assessment.publishersversionNon peer reviewe