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. The fungus was readily detectable, using V. dahliae-expressing green fluorescent protein and confocal microscopy in the xylem of spinach plants, in root cross sections, and in the fruit walls of spinach seed. The two fungicidal seed treatments did not kill all of the fungus within the seed although the fungus was not detectable in the embryo and centralized seed tissues.