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ARS Home » Pacific West Area » Parlier, California » San Joaquin Valley Agricultural Sciences Center » Crop Diseases, Pests and Genetics Research » Research » Research Project #442343

Research Project: CAP- Advanced Testing and Commercialization of Novel Defensin Peptides and Therapies for HLB Control

Location: Crop Diseases, Pests and Genetics Research

Project Number: 2034-22000-014-009-R
Project Type: Reimbursable Cooperative Agreement

Start Date: Oct 1, 2021
End Date: Sep 14, 2025

Objective 1. Develop genetically-engineered Citrus tristeza virus (GE-CTV) using endogenous California CTV strains. Objective 2. Determine citrus cultivar differences in expression levels of GE-CTV with the hypothesis that the greater expression results in greater control of HLB.

Molecular and recombinant approaches will be used by replacing/inserting foreign genes in restriction sites in the cDNA genome of CTV GE-CTV and transforming clones into Agrobacterium EHA 105, and infiltrating Nicotiana benthamiana to systemically infect and produce infectious virions (rCTV) in tobacco, which will be collected, purified and inoculated in citrus (Alemow) to establish systemic infection. Alemow will be used to inoculate various citrus scions with rCTV. Enzyme-linked immunoassay (ELISA) and real-time polymerase chain reaction (RT-qPCR) will be used to measure virus titer. Test plants will be inoculated with HLB to determine efficacy of control.