Pakistan-US Science and Technology Cooperation Program Phase 6 (2015 Deadline) Application of Molecular Methods for the Development of a Pathogen-Free Certification Program for Grapevine in Pakistan US partner: Adib Rowhani, University of California, Davis, and Maher Al Rwahnih, University of California, Davis Pak partner: Shagufta Naz, Lahore College for Women University Progress Reports 2016: Pakistan does not currently have the highly technical tools that could vastly improve the national harvest of perennial crops, such as grapes. Microbial diseases greatly decrease the yield and quality of those harvests. During the first year of this project, we have made progress towards bringing expertise in plant disease detection and management from the US to Pakistan. This past year, the Pakistani collaborator was trained in the US in the use of molecular techniques to detect and identify viruses in commercially valuable grape varieties in Pakistan. These techniques have now been used to detect viral diseases in different grapevine varieties sampled from various ecological zones in Pakistan. This is the foundation for further work of conducting a survey of grapevine viruses, which will also tell us the extent of virus infection in Pakistan, as an important first step in the development of a management program for their control. 2017: Pakistan does not currently have the highly technical tools that could vastly improve the national harvest of perennial crops, such as grapes. Microbial diseases greatly decrease the yield and quality of those harvests. During the second year of this project, we have made progress towards bringing expertise in plant disease detection and management from the US to Pakistan. This past year, the Pakistani collaborators were trained in the US in the use of molecular techniques to detect and identify viruses in commercially valuable grape varieties in Pakistan. These techniques were used to detect viral diseases in 249 grapevine samples collected from different grapevine varieties from 13 vineyards in Pakistan. These results confirmed the presence of grapevine viruses in Pakistan for the first time. Furthermore, it shows that economically important grapevine viruses are present and pose a threat to the grapevine industry. These results are critical in providing basic information about the sanitary status of vineyards in Pakistan and will aid growers and nurseries in their vineyard management decisions. In addition, this research demonstrates the use of diagnostic tests for the detection of viruses of grapevines which can continue to be used for routine virus screening. Implementation of screening programs to produce virus tested grapevines and maintain healthy vineyards would be a significant benefit to the grape industry in Pakistan. 2018: This project has also provided training in the establishment and maintenance of clean plant material and the use of tissue culture techniques for a micro shoot-tip virus elimination program. The training, tools and resources provided by this project will be tremendous assets in developing a grapevine clean stock program which will lead to the improvement of the quality and quantity of grape production in Pakistan.