Location: Subtropical Insects and Horticulture Research
Project Number: 6034-22320-007-019-A
Project Type: Cooperative Agreement
Start Date: Oct 1, 2020
End Date: Jun 30, 2024
Objective:
Funds provided to cooperator are to support the following objectives:
1. Further develop PHACT technology by refining export and systemic movement, peptide production, cell culture, transplantation, and vector design as in Objective 1 and 3a.
2. Assist with bend-to-field pipeline assay design using PHACT.
3. Conduct field trials to test candidate therapeutics and delivery systems identified in Objective 2.
4. Collaborate to design direct delivery device in Objective 3C.
5. Conduct preliminary economic assessment of lead candidate therapeutics.
Approach:
Objective 1&2. Optimize use of PHACT as peptide production method. Produce AMPs, fungal entomopathogen metabolites and Ribonucleic acid (RNA) aptamers, using PHACT and compare with Vmax to determine best production method for each type of peptide. Use PHACT to produce peptides for use in objective 2 pipeline assays, as well as 3a and 3c delivery methods. Determine whether PHACT can be used as a screening method for therapeutics. Assist with field testing of molecules that show anti-ACP or anti-CLas activity in greenhouse bioassays.
Objective 3a. PHACT Delivery. Optimize biofactory morphological structure, recombinant gene expression and defense gene product export as well as cell culture and delivery strategies and demonstrate the use of PHACT for HLB control. Optimize Cell activation genes. Optimize protein and dsRNA molecule export and systemic movement in the plant. Optimize cell culture of activated citrus and tomato biofactory cell lines for transplantation. Determine whether PHACT can function as an HLB control strategy for curing infected trees and/or preventing infection of healthy trees.
Objective 3c. Direct Plant Infusion. A collaborative effort between agricultural engineers and biologists will use current knowledge from laboratory device development to construct rugged attachment devices that can remain attached to the tree and allow rapid refilling over multiple years. Agricultural industry key personnel will support commercial production pathway evaluations.
Objective 5. Economics analysis. Conduct preliminary economic evaluations of cost and commercial viability on lead candidates in field trials, including Objective 5A cost estimates and 5B a detailed outline of EPA registration costs.
