2011 Annual Report 1a.Objectives (from AD-416) The objectives of this project will be to: . 1)collect data on siRNA expression needed for the successful development, submission, and shepherding of a deregulation dossier for 'HoneySweet' plum in Europe;. 2)organize a workshop entitled, "The deregulation of genetically engineered Plum pox virus resistant 'HoneySweet' plum in the U.S. and the potential impacts on regulatory approval and plum cultivation in Europe"; and. 3)support the exchange of information necessary for the successful development, submission, and shepherding of a deregulation dossier for 'HoneySweet' plum in Europe. 1b.Approach (from AD-416) 'HoneySweet' siRNA will be purified and sequenced using next generation sequencing technologies to identify the specific species of siRNAs produced. Candidate siRNAs will be confirmed via conventional molecular approaches and assessed for potential animal toxicity via bioinformatic analyses. A visiting scientist from the Crop Research Institute, Drnovska, Czech Republic, will conduct the research at the Appalachian Fruit Research Station. This person will be educated and trained in 'HoneySweet' molecular characterization and U.S. regulatory policy to serve as a technical liaison between the U.S. and EU partner laboratories. Meetings will be organized with the specific purpose of developing a dossier on 'HoneySweet' plum for the submission to the European Food Safety Association by the Crop Research Institute, Drnovska, Czez Republic (a European public institution), and meetings at regular intervals to monitor the progress of that dossier and to address roadblocks that may develop in the progress of the dossier. 3.Progress Report Leaf tissues were collected from Plum pox virus (PPV) infected and uninfected plum trees grown in a containment greenhouse (Fort Detrick, MD). These have been stored at -80 C for RNA and siRNA extraction. Dr. Ralph Scorza has coordinated with European partners in Czech Republic, Poland, and Spain to obtain field samples of fruit and leaves for mRNA and siRNA experiments. A visiting scientist from Poland has agreed to come to the Appalachian Fruit Research Station (AFRS) to conduct experiments and gain regulatory expertise. An international meeting was held in Prague, Czech Republic, to facilitate the submission of a deregulation petition to the EU. A large group of 'HoneySweet' researchers, regulatory experts, government representatives, and advisors participated in the meeting. Outcomes were as follows: . 1)the Crop Research Institute (CRI) in the Czech Republic agreed to submit the deregulation petition and. 2)dossier writing tasks were assigned and deadlines were set (Nov 2010-completion of all data and Dec 2010-Dossier submission).