Visualization and Detection of Proteins Produced by ‘ca. Liberibacter Asiaticus’ in Infected Sweet Orange Plants and Vector Psyllids
Molecular Plant Pathology
Project Number: 1245-22000-281-13
Start Date: Apr 01, 2012
End Date: Mar 31, 2015
With previous funding from the FL Citrus Research and Development Foundation (CRDF), ARS' Molecular Plant Pathology Laboratory (MPPL) has developed a library of scFv antibody fragments that bind specific targets on the HLB pathogen that are believed to be exposed on the surface of the pathogen and therefore useful for novel ultrasensitive detection assays. The University of Florida has developed expertise in the use of antibodies to enhance visualization of bacteria for detection and characterization of host/pathogen interactions. The objectives of this proposal are to transfer recently developed scFv antibodies to commercial partners for the development of novel diagnostic tests, as well as to use them to study host/pathogen interactions at the ultrastructural level.
ARS' MPPL, Beltsville, will convert scFv genes from phagemid to plasmid inserts, ensure the sequence is free of unwanted stop codons and send both purified scFv and the genes encoding them to commercial partners. ARS MPPL, Beltsville, will also purify and transfer scFv to the University of Florida to develop serologically enhanced methods to detect and visualize ‘Ca. Liberibacter asiaticus’ in infected plants and insects. The University of Florida and ARS will both maintain HLB infected plants, and the University of Florida will maintain colonies of infected citrus psyllids. Agdia will carry out serological characterization of scFv received from ARS and test them in serological assays for detection of ‘Ca. Liberibacter asiaticus’.