Molecular analysis of potato virus X isolates in relation to the potato hypersensitivity gene Nx

The coat protein gene of potato virus X is known to affect the outcome of interactions between different strains of the virus and potato plants carrying the Nx resistance gene. To analyze the role of the coat protein in interactions with Nx hosts, we used the potato virus X strain PVXDX, which induces a hypersensitive response on potato cultivars carrying the Nx resistance gene and the strain PVXDX4, which was originally derived from PVXDX and which overcomes Nx-mediated resistance. Sequencing of cloned coat protein genes representing the strains PVXDX and PVXDX4 showed that they differed at a single nucleotide. This change results in the substitution of glutamine-78 in the PVXDX coat protein for proline in PVXDX4. We constructed hybrid viral genomes by replacing the coat protein gene of a full-length clone of isolate PVXUK3 with the corresponding sequence from either PVXDX or PVXDX4. Progeny virus, derived from in vitro transcripts of these hybrid clones, showed that the single nucleotide difference between the coat protein genes of isolates PVXDX and PVXDX4 was sufficient to alter the outcome of the interaction between the virus and potato plants carrying the resistance gene Nx. Additional coat protein mutants generated in planta from transcript-derived inocula induced an intermediate host response on Nx potato cultivars which is influenced by the presence of a second, PVX-specific, resistance gene in the host plant genome.