Haplotype graph analysis of PdR1 uncovers resistance diversity to Pierce's Disease in Vitis arizonica and its hybrids

Previous genetic mapping studies indicate that multiple haplotypes of the Pierces disease (PD) Resistance 1 (PdR1) locus occur in Vitis arizonica and its hybrids. To characterize sequence diversity at this locus, we assembled chromosome-scale diploid genomes for four PD-resistant (PD-R) accessions: b43-17 (PdR1a/PdR1b), the backcross 07744-094 (PdR1c/PdR1-), b46-43 (PdR1e/PdR1f), and b42-26 (PdR1-/PdR1-), which displays quantitative PD resistance not associated with PdR1. Haplotype resolution of PdR1a, PdR1b, PdR1c, and PdR1e revealed substantial variation in intergenic repeat content and gene composition between PdR1 and their alternative haplotype at the PdR1 locus not associated with PD resistance phenotype (PdR1-), as well as among PdR1 haplotypes, demonstrating extensive sequence diversity at the PdR1 locus. Sequence graph analysis uncovered substantial structural divergence concentrated in approximately one quarter of the locus, together with smaller-scale variation across haplotypes. This analysis identified PdR1-specific graph nodes, showing that PdR1a and PdR1b share most of their PdR1-specific features, whereas PdR1c contains the highest number of private nodes, followed by PdR1e. Integration of sequence graph features with gene expression data further refined a set of defense-related candidate genes within PdR1c. Together, these results identify candidate genes for functional validation and indicate that multiple resistance determinants co-localized within the PdR1 locus may contribute to PD resistance, highlighting opportunities for targeted genetic improvement strategies.