2007 Annual Report 1a.Objectives (from AD-416) Development of tightly regulated gene switches for trait improvement and functional genomics applications in forage plants: (1) Development of a EcR gene switch for use in monocot system. (2) Testing of EcR gene switch in functional genomics application using zinc finger genes as a model system. 1b.Approach (from AD-416) Expression of transgenes is becoming a powerful tool in many biotechnological applications in agriculture. Constitutive promoters, those that are expressed all the time in all tissues, are presently the primary means used to express transgenes in plants. Metabolic energy waste, negative pleiotropic effects and potential gene escape are some of the disadvantages associated with the use of constitutive promoters. To counter these problems, gene switches that can regulate the expression of transgenes through hormone analogs and antibiotics have been developed. Ligands that are suitable for regulation of transgenes for large-scale production and field use are the commercially available non-steroidal ecdysone agonists. Ecdysone agonist and ecdysone receptor (EcR) have been shown to regulate gene expression in tobacco, Arabidopsis and corn. In a previous project, highly sensitive EcR gene switches that can support the induction of transgenes with nanomolar concentration of ecdysone agonist in both Arabidopsis and tobacco plants were developed. This work aims to test the use of these gene switches in monocots, ultimately to be used in forage based systems. The first approach will be used to use the gene switch in rice as a model monocot system with routine transformation procedures. A second approach will be to use the gene switches to evaluate the performance of the EcR gene switch system in functional genomics using a zinc finger transcription factors. This will be tested as a model to system to evaluate its use for transcription factors, and will be tested in Arabidopsis, and if successful, will be converted for use in rice and Lolium ssp. 3.Progress Report This report serves to document research conducted under a Non-Funded Cooperative Agreement between ARS and the University of Kentucky. Additional details of research can be found in the report for the in-house project 6440-21310-001-00D, "Enhance Forage-Based Livestock Production Systems." We have tested the use of the EcR gene switch for functional genomic approaches using a zinc finger protein in Arabidopsis. This work has been recently published (reference below). We are in the process of incorporating the gene switch in rice and tall fescue for analysis. Transformation is ongoing. Forage-Animal Production Research (FAPRU) scientist is an intimate component of this research. Meeting regularly with the research team on progress and needs. Further, this scientist keeps FAPRU's Research Leader apprised of progress on a regular basis via face to face meetings.