Following the submission of application EFSA-GMO-NL-2009-68 under Regulation (EC) No 1829/20031 from Dow Agrosciences LLC, the Panel on Genetically Modified Organisms of the European Food Safety Authority (GMO Panel) was asked to deliver a scientific opinion on the safety of insect-resistant and herbicide-tolerant genetically modified (GM) cotton 281-24-236 × 3006-210-23 × MON 88913 (hereafter referred to as ‘three-event stack cotton’). The scope of application EFSA-GMO-NL-2009-68 is for food and feed uses, import and processing, but excludes cultivation within the European Union (EU).

The applicant restricted the scope of application EFSA-GMO-NL-2009-68 to the three-event stack cotton only. As cotton (Gossypium hirsutum) is predominantly a self-pollinator and the three-event stack cotton 281-24-236 × 3006-210-23 × MON 88913 is homozygous for all traits,2 the produced and imported cottonseed of this GM cotton will contain all traits, and segregants are expected only at very low frequency.

In accordance with the GMO Panel guidance document applicable to this application (EFSA, 2007), where all single events have been assessed, the risk assessment of stacked events should focus mainly on issues related to (a) stability, (b) expression of the events and (c) potential interactions between the events. For application EFSA-GMO-NL-2009-68, previous assessments of the three single events (281-24- 236, 3006-210-23 and MON 88913) provided a basis to evaluate the three-event stack cotton.

The three-event stack cotton 281-24-236 × 3006-210-23 × MON 88913 was produced by conventional crossing of the two-event stack cotton 281-24-236 9 3006-210-23, containing the previously assessed single cotton events 281-24-236, 3006-210-23 and MON 88913. The single events 281-24-236 (expressing Cry1F and phosphinothricin acetyltransferase (PAT) proteins) and 3006-210-23 (expressing Cry1Ac and PAT proteins), in the frame of the two-event stack cotton 281-24-236 9 3006-210-23, and MON 88913 (expressing two copies of the CP4 5-enolpyruvylshikimate-3-phosphate synthase (CP4 EPSPS) protein) were assessed previously and no concerns were identified. No safety issue was identified by updated bioinformatic analyses, nor reported by the applicant concerning the three single cotton events, since the publication of the respective scientific opinions. Consequently, the GMO Panel considers that its previous conclusions on the safety of the single cotton events remain valid (Section 3.2).

For the three-event stack cotton, the risk assessment included the molecular characterisation of the inserted DNA and the analysis of the protein expression. An evaluation of the comparative analyses of compositional, agronomic and phenotypic characteristics was undertaken, and the safety of the newly expressed proteins and the whole food/feed was evaluated with respect to potential toxicity, allergenicity and nutritional characteristics. Evaluation of environmental impacts and of the post-market environmental monitoring (PMEM) plan was also undertaken.

The molecular data establish that the events stacked in cotton 281-24-236 × 3006-210-23 × MON 88913 have retained their integrity. Protein expression analyses showed that the levels of the newly expressed proteins are similar in the three-event stack cotton, and the two GM parental lines, i.e. the two-event stack (281-24-236 9 3006-210-23) and single event (MON 88913). There is no indication of an interaction that may affect the integrity of the events and the levels of the newly expressed proteins in this stack.

The combination of the newly expressed proteins in the three-event stack cotton did not raise concerns for human and animal health. The combination of cotton events 281-24-236, 3006-210-23 and MON 88913 in the three-event stack cotton did not give rise to issues – based on the agronomic, phenotypic or compositional characteristics – regarding food and feed safety and nutrition.

Considering the introduced traits, the outcome of the comparative analysis, the routes of exposure and the limited exposure levels, the GMO Panel concluded that this three-event stack cotton would not raise safety concerns in the event of accidental release of viable GM cottonseeds into the environment.

The GMO Panel is of the opinion that the PMEM plans provided by the applicant are in line with the scope of the three-event stack cotton and that post-market monitoring of food/feed derived from the three-event stack cotton is not considered necessary.

In conclusion, the GMO Panel concludes that the three-event stack cotton is as safe and as nutritious as its conventional counterpart in the context of its scope.