2007 Annual Report 1a.Objectives (from AD-416) To discover gene sequences that can be used to distinguish Planococcus citri and P. minor. Preliminary results suggest that segments of the mitochondrial gene COI and the nuclear gene EF1-' will be useful in separating them. We will examine Rapid PCR and other technologies to develop a diagnostic tool that will be useful for an efficient and effective method for separation of the species. We also will examine morphological features to determine if structural characters exist that could be used to make accurate determinations. 1b.Approach (from AD-416) Gather specimens from as many localities as possible. Extract DNA and mount exoskeleton as a voucher and representative for morphological analysis. PCR and sequence extracted DNA. Align sequences using computer algorithms and use data to formulate phylogenetic trees. Preliminary data suggest that segments of EF1-' and COI are useful in distinguishing these species. Utilize unique base pairs from each of Planococcus citri and P. minor to develop a diagnostic tool that will quickly distinguish these species. Examine exoskeletons of molecularly identified specimens to determine if structural characters can be utilized to distinguish these mealybugs. 3.Progress Report This report serves to document research conducted under a Reimbursable agreement between ARS and APHIS/PPQ to discover molecular differences between the mealybugs Planococcus citri and P. minor. Additional details of research can be found in the report for the parent project 1275-22000-230-00D Systematic analysis of aphids, mites, scales, thrips, and termites with emphasis on invasive species. Sequence data from the COI gene seem to discriminate these apparent species, and data from EF-1a confirm these differences. Discovery of a third gene that could be used to sort out rare conflicts between EF-1a and COI will be the next thrust in this research area. Quarterly reports were provided during the reporting period and there were numerous telephone conversations.