2006 Annual Report 4d.Progress report. This report serves to document the research conducted under a specific cooperative agreement (58-5442-5-342) between ARS and North Dakota State University-Department of Entomology. Additional details of this research area can be found in the report for the parent ARS Project 5442-22000-039-00D, "Insect Genomic Biodiversity and Molecular Regulation of Diapause." The objective of this cooperative research project is to characterize the molecular mechanism(s) regulating diapause initiation in the Colorado potato beetle and to isolate key orthologs from other Chrysomelids. Several closely related species such as the sunflower beetle will be examined to find conserved regions of these genes and their promoters. Conserved regions will be used to design PCR primers to expand to more distantly related pest Chrysomelids. The sequence data generated will be used to identify regulatory elements and describe the evolutionary divergence of diapause specific genes. The research accomplishment of this reporting period was the isolation the DAT-1 orthlog from the sunflower beetle. Comparing the promoter regions of DAT-1 from the Colorado potato beetle and the sunflower beetle revealed a conserved motif for the transcription binding factor broad-complex-3. This indicates that broad-complex-3 is involved in diapause regulation in these two beetle species.