Box 1: Crucial aspects for a two-pronged assessment strategy to address trait-related effects and method-related modifications, respectively.

(1) The assessment of effects associated with the newly developed trait(s) in GE plants should consider, among others: The level of knowledge and familiarity with the particular crop and trait combination needs to be considered. As indicated in Section 4.1, only limited scientific knowledge is available for some GE applications. Some applications may lead to changes in agricultural management; possible indirect effects resulting from their use need to be addressed during the ERA. Complex GE modifications should be thoroughly scrutinized regarding adverse environmental effects resulting from these changes. A robust assessment should be provided for physiological effects of multiple simultaneous changes (multiplexed GE) and for regulatory effects of the introduced modifications on morphology, development and reproduction of the GE plant. The ERA conducted for GE plants should also address secondary effects associated with the intended trait(s). This should encompass pleiotropic effects of the intended trait(s). (2) The assessment of method-related unintended changes associated with the intended trait(s) or with other modifications in the GE plant should take into account the following aspects: The available body of evidence with regard to off-target-effects, their occurrence and their identification as indicated in Section 4.2. The likelihood that off-target modifications are still present in the final breeding product. This likelihood may be higher with fast-tracked breeding applications, i.e., aimed at modification of elite lines, modification of vegetatively propagated crops, and modification of plant species with longer generation cycles such as trees. The available information on unintended secondary modifications introduced by GE systems in the vicinity of the intended genomic target site. Such modifications are tightly linked to the intended traits and are not easily lost during subsequent breeding steps. The available recommendations on how an assessment of unintended and off-target effects may be conducted and which kind of aspects should be considered in the framework of the assessment.