During this reporting period (April, May and June, 2016), we analyzed over two dozen newly transformed plants from Dr. Janice Zale’s program (University of Florida Mature Citrus Transformation Facility, Lake Alfred, FL). These plants are ‘Hamlin’ sweet orange and ‘Carrizo’ rootstock lines. However, we were not able to detect the FLT-antiNodT fusion protein expressed in any of these lines by protein immunoblotting. This contrasts with our results in ‘Duncan’ grapefruit, where nearly all the independently transformed lines expressed detectable levels of full-length FLT-antiNodT fusion protein. Further molecular analyses during the present funding period indicated that the transgene encoding the FLT-antiNodT fusion protein was not detectable in the ‘Hamlin’ and ‘Carrizo’ lines, although the markers for transformation (kanamycin resistance and green fluorescent protein) were detectable. Unfortunately, these results suggest that these plants are not actually transformed with the FLT-antiNodT fusion protein transgene. However, our original and proposed activity, transforming ‘Duncan’ grapefruit and testing for HLB resistance, was successful and is still in progress. Plants are continuing to be propagated for testing for HLB resistance at in Dr. Tim Gottwalds’ lab at Ft. Pierce. When sufficient plants are ready, these will be transferred into an HLB transmission greenhouse and exposed to Asian citrus psyllids carrying Candidatus Liberibacter asiaticus. During the present funding period, protein samples from the Ft. Pierce trees were analyzed by protein immunoblotting and these trees were found to be expressing moderate to high levels of FLT-antiNodT fusion protein, which is very promising. Although Dr. McNellis had planned to send rooted clones of the seven ‘Duncan’ lines at Penn State to Dr. Gottwald’s lab at Ft. Pierce, the permit for such transfer was not approved. Transporting citrus back to Florida is unlikely to ever be approved. Dr. McNellis then pursued an alternate plan to transfer plants to Ft. Detrick, Maryland, to Dr. Bill Schneider’s lab, for testing for HLB resistance. Dr. Schneider has agreed to participate in this collaboration, and Dr. McNellis has applied for a USDA APHIS permit for this plant transfer. This application is still under review by USDA APHIS.