Comparative analyses of three grapevine Pinot gris virus cDNA clones reveal insights into the pathological properties of different phylogroups

Grapevine Pinot gris virus (GPGV) is an emerging grapevine virus associated with grapevine leaf mottling and deformation (GLMD) disease. Being a recently identified virus, the molecular biology, pathological properties, and etiological complexity of GPGV remain poorly studied. Previous research revealed that GPGV comprises genetically different variants, some encoding a larger movement protein (MP) and others a shorter MP due to a C/T polymorphic site in ORF2 encoding MP. Variants that encode the shorter MP are associated with severe disease, whereas variants encoding the longer MP are associated with mild or no symptoms. However, this has yet to be demonstrated experimentally. Here, we report the construction of a wildtype cDNA clone, pGPGV-SY, based on ON93-12, a local isolate from Syrah closely related to the variants encoding the larger MP. Surprisingly, our clone exhibited significantly faster replication and caused more severe disease symptoms than pRI::GPGV-lat, an Italian GPGV clone, with a longer MP and demonstrated similar efficacies with that of pRI::GPGV-vir, another Italian clone with a shorter MP. A single C to T mutation at the polymorphic site of pGPGV-SY resulted in a two-fold higher RNA accumulation in the grapevine. Findings from this work constitute a leap toward the long-standing and complex question pertaining to the relationship between GPGV variant groups and GLMD. Integrating findings from this work and those by others, we propose an updated model to explain the complex relationship between GPGV variants and GLMD.Grapevine Pinot gris virus (GPGV) is an emerging grapevine virus associated with grapevine leaf mottling and deformation (GLMD) disease. Being a recently identified virus, the molecular biology, pathological properties, and etiological complexity of GPGV remain poorly studied. Previous research revealed that GPGV comprises genetically different variants, some encoding a larger movement protein (MP) and others a shorter MP due to a C/T polymorphic site in ORF2 encoding MP. Variants that encode the shorter MP are associated with severe disease, whereas variants encoding the longer MP are associated with mild or no symptoms. However, this has yet to be demonstrated experimentally. Here, we report the construction of a wildtype cDNA clone, pGPGV-SY, based on ON93-12, a local isolate from Syrah closely related to the variants encoding the larger MP. Surprisingly, our clone exhibited significantly faster replication and caused m