2012 Annual Report 1a.Objectives (from AD-416): The specific objectives of this project are to. 1)develop a rapid detection assay for Verticillium dahliae in spinach seed;. 2)determine where the fungus is located in spinach seed to develop effective seed treatments; and. 3)identify lettuce and spinach resistant to V. dahliae to limit spread via seed. 1b.Approach (from AD-416): Detection of the fungus in vegetable and spinach seeds will be carried out by PCR techniques; localization of the fungus in spinach seed will be determined by confocal microscopy; and spinach and lettuce will be screened for resistance to Verticillium wilt in greenhouses. 3.Progress Report: The agreement was established in support of objective 2, the goal being to develop molecular diagnostic tools for the identification of emerging diseases of vegetables and strawberries, and use these tools in the development of management strategies as alternatives to methyl bromide. Fifteen commercial spinach seed lots were analyzed for percentage of seeds infected with Verticillium dahliae. Seeds obtained from seed producers in the US Pacific Northwest and Europe were disinfested and plated on culture medium to determine whether or not the pathogen was present in each seed. The percentage of seeds infected per lot ranged from 0.3 to 85 percent.