Development of Bacteriophage Plaque Assay for Liberibacter

Development of Bacteriophage Plaque Assay for Liberibacter

Report Date: 07/15/2016
Project: 726-Liberibacter   Year: 2016
Category: Horticultural & Management
Author: Carlos Gonzalez
Sponsor: Citrus Research and Development Foundation

�Candidatus Liberibacter asiaticus� has not been cultured. However, Liberibacter crescens, has been cultured under laboratory conditions. The focus of our project has been to develop a detection system for bacteriophages (phages) and/or phage components (tailocins) using L. crescens strain BT-1 as a model system. We have accomplished the development of the assay system, which we have used to conduct screening of phages and broad host tailocins. Liberibacter is a member of the Rhizobiaceae. It is our experience that phylogenetically related microorganisms can share common surface components, such as phage receptor sites. Bioinformatic and structural analyses indicates that there is high homology in the surface structures of Rhizobium spp., Agrobacterium spp. and Liberibacter spp. Therefore, one of our strategies has been to search for naturally occurring phages active against Rhizobium spp. or Agrobacterium spp. that may also show activity against Liberibacter spp. We are continuing our screening of both Rhizobium spp. and Agrobacterium spp. phages against BT-1 using modifications of the assay, since growth condition can affect phage susceptibility. Strain BT-1 harbors two prophages (LC1 and LC2) that we have determined are not inducible by UV or oxidative stress. This indicates that the prophages are defective. The phages are predicted to be podophages, because their genomes exhibit no separate tail structure gene. We will continue to construct fusions between N-terminal tail fiber region of a broad host tailocin and the putative C-terminal portions of tail spike from BT-1 prophages to obtain active tailocins against Liberibacter.


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