1. Please state project objectives and what work was done this quarter to address them:
Objective 1: Develop an antibody based CB diagnostic assay. We have successfully completed this objective and concluded that the antibody based diagnostic assay allows for earlier detection of CB than the water uptake test.
Objective 2: Determine how prevalent inserted copies of viral DNA are in commercial citrus. We have successfully completed this objective and have concluded that the CBaPRV retrovirus is in every cultivar, suggesting that the viral insertion of CB predates the conventional breeding of citrus. We are currently working on publishing this work. a rough draft will be circulated across collaborators in May and we hope to submit that manuscript this summer.
Objective 3: Development a CRISPR solution to inactvate CBaPRV. A total of 37 lines with the MP construct and 23 with the RT construct have been micro grafted on rootstocks. Graft success rate was 90% and these lines are being sized up to obtain lead materials for molecular analysis to identify positive lines with the DNA edits. This objective has been completed, but we are waiting to test these pants in objective 4.
Objective 4: Determine the efficacy of CRISPR as a way to eradicate CB in plantlets. Because there has been a delay in the generating CRISPR plant to test. We are expecting to start testing plants in May. We expect to have all plants PCR tested to validate edits by the end of June.
2. Please state what work is anticipated for next quarter:
In the 10th quarter we will also finalize the germplasm publication and test CRISPR-edited plants to validate that CRISPR was effective.
Completion of objectives 85%
The progress of our production during the grant has improved now that CRISPR-edited plants are ready to test and determine if the edits were successful and we are expecting to start testing thses plants in the next quarter.
3. Please state budget status (underspend or overspend, and why):
We are on track with spending. Our supply budget has been spent out and we will use our remainder travel, publication, and salary budget to continue to work on the project until the end of the grant.
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.