Using Improved Diagnostics to Determine Citrus Blight Prevalence

Using Improved Diagnostics to Determine Citrus Blight Prevalence

Report Date: 12/20/2023
Project: 23-021   Year: 2023
Category: Other
Author: Ronald Brlansky
Sponsor: Citrus Research and Development Foundation

1. Please state project objectives and what work was done this quarter to address them:

Objective 1 – The development of diagnostic assays. We have focused our efforts on this objective. Antibody testing is ongoing, but we have had some positive results. The antibody binds to the movement protein antigen as expected. This has been determined using dot blot and western assays. Because blight has not been demonstrated in tobacco, we tested tobacco as a negative control. The dot blot and western have both shown that there is no reaction between the antibody and protein extracted from the tobacco control. Therefore, we conclude that the antibody is likely specific to the detection of citrus blight moment protein. Further testing is needed to continue antibody validation. We have completed water-uptake tests, canopy scores, root collection and protein extractions for 30 field trees. Roots have been processed and protein has been extracted. We have positive and negative results using extracted protein from these trees, completing from our first set of field tests. This needs to be repeated two more times before data can be analyzed. We are currently developing an ELISA assay to using the antibody to determine concentration curves of known movement protein antigen dilutions.

Objective two – We have processed 30 nursery trees for germplasm analysis.

2. Please state what work is anticipated for next quarter:

Objective 1: Next quarter we plan to continue developing the antibody assay and complete its validation using dot blot, westerns, and ELISAs. We will continue testing field trees and generating data to assess the diagnostic capacity of this method compared to other existing methods (i.e. water uptake and PCR). We expect to also process at least 20 more field samples for analysis using the developed antibody-based diagnostic assays.

Objective 2: Germ plasm studies will begin using the tissue that have processed. We expect to have at least 30 samples total from 5-10 different rootstocks tested in the next quarter.

3. Please state budget status (underspend or overspend, and why):
We have purchased the necessary start-up supplies needed and managing our resources well. We are on track with spending for supplies this quarter. We are underspent on salary money because of hiring delays with UF resulting in the onboarding of OPS positions a month after the grant began.

4. Please show all potential commercialization products resulting from this research, and the status of each:

At this time there is no potential commercialization of any products associated with this research.


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