2011 Annual Report 1a.Objectives (from AD-416) Develop cost effective and highly sensitive methods to detect graft transmissible pathogens of citrus. 1b.Approach (from AD-416) Establish reliable real-time PCR detection of graft transmissible pathogens of citrus including but not limited to Pospiviroids, Hostuviroids, Spiroplasma citri, Dweet mottle virus, citrus tatter leaf virus. Establish protocols for rapid extraction of pathogen nucleic acid from target tissue. Develop genomic information of graft transmissible pathogens of citrus including pathogens which are presently uncharacterized. 3.Progress Report The project relates to objective 4 of the parent project. Sequences of citrus pathogens including citrus viroids, Citrus psorosis virus, Spiroplasma citri and Citrus tatter leaf virus have been obtained from GenBank and are being used to develop primers for detection of these pathogens. Different protocols for rapid extraction of DNA and RNA and total nucleic acids are being evaluated for their adaptability to 96 well plate format and efficiency of extraction The project is monitored by meeting, site visits, e-mail correspondence, and phone calls.