Location: Crop Diseases, Pests and Genetics Research
Project Number: 2034-22000-015-441-T
Project Type: Trust Fund Cooperative Agreement
Start Date: Apr 15, 2024
End Date: Apr 15, 2025
Objective:
The overall goal of this project is to develop enhanced grape rootstock varieties with broad-spectrum resistance to common grapevine diseases. CRSPR/Gene-editing techniques will be used to target cytidine diphosphate diacylglycerol (CDP-DAG) genes (CDSs) in grapevine.
Approach:
Cytidine diphosphate diacylglycerol (CDP-DAG) is a critical intermediate for glycerolipid (GL) biosynthesis in both prokaryotic and eukaryotic cells. CDP-DAG synthase (CDS) is a membrane-bound enzyme that catalyzes the transfer of a cytidyl group from cytidine triphosphate (CTP) to phosphatidic acid (PA) to form CDP-DAG. CDS is the gateway to phosphatidylinositol (PI) and cardiolipin (CL) biosynthesis in eukaryotes. Deletion in a rice CDS gene named RBL1 can generate lesion mimic mutants (LMMs) exhibiting broad-spectrum disease resistance to bacterial and fungal pathogens. RBL1 catalyzes the biosynthesis of PI and its derivative phosphatidylinositol 4,5-bisphosphate (PtdIns(4,5)P2), which is associated with effector secretion and fungal infection. As a disease-susceptibility factor, RBL1 can be genome-edited to confer broad-spectrum disease resistance with no growth penalty or yield decrease. Protein BLAST (BLASTP) search of rice RBL1 against grapevine annotated genome PN40024.v4 found its two orthologs Vitvi10g00174 and Vitvi19g00570. We propose to perform genome editing on the two putative CDS synthase genes and screen edited grapevine plants for those exhibiting normal fruit production and increased broad-spectrum resistance to common grapevine diseases, such as Pierce’s disease caused by Xylella fastidiosa and powdery mildew caused by Uncinula necator.
