Grapevine vein clearing virus (GVCV) is associated with a vein-clearing and vine-decline disease. In this study, we surveyed wild Ampelopsis cordata from the Vitaceae family and found that 31% (35 of 113) of native A. cordata plants are infected with GVCV. The full-length genome sequence of one GVCV isolate from A. cordata shared 99.8% identical nucleotides with an isolate from a nearby cultivated 'Chardonel' grapevine, suggesting the occurrence of an insect vector. To identify a vector, we collected Aphis illinoisensis (common name: grape aphids) from wild A. cordata plants and detected GVCV in the aphid populations. We Materials and MethodsCollection of Ampelopsis and Vitis samples. Two mildly symptomatic A. cordata plants were collected, AMP1 from Linn Creek, MO, in the summer of 2015 and AMP2 from Springfield, MO, in †

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Grapevines are frequently infected by multiple viruses. Our previous study showed that ‘Norton’ grapevine (Vitis aestivalis) is resistant to grapevine vein clearing virus, a DNA virus in the family Caulimoviridae. To study the reaction of ‘Norton’ to RNA viruses, we transferred seven RNA viruses to ‘Norton’ from ‘Kishmish Vatkana’ (‘KV’) (Vitis vinifera) via graft-transmission. We profiled viral small RNAs (vsRNAs) of the seven viruses and compared viral titers in ‘Norton’ and ‘KV’. Total vsRNAs of grapevine leafroll-associated virus 1 (GLRaV-1), GLRaV-2, GLRaV-3, grapevine virus A (GVA) and grapevine Pinot gris virus (GPGV) were significantly less abundant in ‘Norton’ than in ‘KV’, but total vsRNAs of grapevine fleck virus (GFkV) were more abundant in ‘Norton’ than in ‘KV’. Total vsRNAs of grapevine rupestris stem pitting-associated virus (GRSPaV) were not different between ‘Norton’ and ‘KV’. Grafting direction of ‘Norton’ to ‘KV’ or ‘KV’ to ‘Norton’ did not affect the quantity of vsRNAs. The genome coverage of GLRaV-1, GLRaV-2, GLRaV-3 and GVA vsRNAs was lower in ‘Norton’ than ‘KV’. The 21-nt and 22-nt classes of vsRNAs were predominant for all seven viruses. Virus quantification by qPCR indicated that GLRaV-1 was undetectable in ‘Norton’, GLRaV-2, GLRaV-3, and GVA were less abundant in ‘Norton’, but GFkV was more abundant in ‘Norton’ than in ‘KV’. These results demonstrated that ‘Norton’ grapevine suppresses GLRaV-1, GLRaV-2, GLRaV-3, and GVA, but supports GFkV in comparison with ‘KV’. This study revealed new facets of complex molecular interactions between grapevines and multiple viruses.

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Grapevines (Vitis spp.) host viruses belonging to 17 families. Virus-associated diseases are a constant challenge to grape production. Genetic resources for breeding virus-resistant grape cultivars are scarce. ‘Norton’ is a hybrid grape of North American Vitis aestivalis and is resistant to powdery mildew and downy mildew. In this study, we assessed resistance of ‘Norton’ to grapevine vein clearing virus (GVCV), which is prevalent in native, wild Vitaceae and in vineyards in the Midwest region of the U.S. We did not detect GVCV in ‘Norton’ as either the scion or the rootstock up to 3 years after it was grafted with a GVCV-infected ‘Chardonel’ grapevine. Upon sequencing of small RNAs, we were able to assemble the GVCV genome from virus small RNAs in GVCV-infected ‘Chardonel’ scion or rootstock, but not from grafted ‘Norton’ scion and rootstock. This study unveils a new trait of ‘Norton’ that can be used in breeding GVCV-resistant grape cultivars, and to investigate genetic mechanisms of ‘Norton’ resistance to GVCV.