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Grapevine red blotch virus (GRBV) is a recently identified virus. Previous research indicates primarily a substantial impact on berry ripening in all varieties studied. The current study analyzed grapes' primary and secondary metabolism across grapevine genotypes and seasons to reveal both conserved and variable impacts to GRBV infection. Vitis vinifera cv. Cabernet Sauvignon (CS) grapevines grafted on two different rootstocks (110R and 420A) were analyzed in 2016 and 2017. Metabolite profiling revealed a considerable impact on amino acid and malate acid levels, volatile aroma compounds derived from the lipoxygenase pathway, and anthocyanins synthesized in the phenylpropanoid pathway. Conserved transcriptional responses to GRBV showed induction of auxin-mediated pathways and photosynthesis with inhibition of transcription and translation processes mainly at harvest. There was an induction of plant-pathogen interactions at pre-veraison, for all genotypes and seasons, except for CS 110R in 2017. Lastly, differential co-expression analysis revealed a transcriptional shift from metabolic synthesis and energy metabolism to transcription and translation processes associated with a virus-induced gene silencing transcript. This plant-derived defense response transcript was only significantly upregulated at veraison for all genotypes and seasons, suggesting a phenological association with disease expression and plant immune responses.

Feeding by the three-cornered alfalfa hopper, Spissistilus festinus (Say) (Hemiptera: Membracidae) results in girdling of grapevine petioles and shoots. Its significance as an economic pest of grape has increased since it was shown to transmit Grapevine red blotch virus (GRBV) in a greenhouse study. However, the status of grapevines as a reproductive host for S. festinus remained undetermined. Adult S. festinus were caged onto three regions of the grapevines: apical shoot, green shoot, and dormant cane. Their ability to reproduce was determined by weekly destructive sampling for 7 wk. Successful oviposition and nymphal emergence were observed on apical and green shoots, but not on dormant canes. However, insect development beyond the second nymphal instar did not occur. Knowledge of S. festinus reproduction on grapevines will be an important consideration in designing management guidelines to minimize the spread of GRBV in vineyards.

Grapevine leafroll-associated virus 3 (GLRaV-3) is the most widely prevalent and economically important of the complex of RNA viruses associated with grapevine leafroll disease (GLD). Phylogenetic studies have grouped GLRaV-3 isolates into nine different monophyletic groups and four supergroups, making GLRaV-3 a genetically highly diverse virus species. In addition, new divergent variants have been discovered recently around the world. Accurate identification of the virus is an essential component in the management and control of GLRaV-3; however, the diversity of GLRaV-3, coupled with the limited sequence information, have complicated the development of a reliable detection assay. In this study, GLRaV-3 sequence data available in GenBank and those generated at Foundation Plant Services, University of California-Davis, was used to develop a new RT-qPCR assay with the capacity to detect all known GLRaV-3 variants. The new assay, referred to as FPST, was challenged against samples that included plants infected with different GLRaV-3 variants and originating from 46 countries. The FPST assay detected all known GLRaV-3 variants, including the highly divergent variants, by amplifying a small highly conserved region in the 3' untranslated terminal region (UTR) of the virus genome. The reliability of the new RT-qPCR assay was confirmed by an enzyme linked immunosorbent assay (ELISA) that can detect all known GLRaV-3 variants characterized to date. Additionally, three new GLRaV-3 divergent variants, represented by four isolates, were identified using a hierarchical testing process involving the FPST assay, GLRaV-3 variant-specific assays and high-throughput sequencing analysis. These variants were distantly related to groups I, II, III, V, VI, VII and IX, but much similar to GLRaV-3 variants with no assigned group; thus, they may represent new clades. Finally, based on the phylogenetic analysis, a new GLRaV-3 subclade is proposed and named as group X.

Grapevine red blotch-associated virus (GRBaV) is a single-stranded DNA vims, proposed to be a member of the family Geminiviridae, and is associated with grapevines showing red blotch symptoms in North America. The existence of the vims was reported in 2012, and subsequently detected in grapevines in major grape production regions. We investigated if a vector exists that can transmit GRBaV in vineyards. Phylogenetic analysis of the predicted amino acid sequence of coat protein (CP) of GRBaV with the CP of 23 geminivimses representing all seven genera of the family Geminiviridae revealed that GRBaV-CP was most similar to that of Tomato pseudo-curly top virus, a geminivirus known to be transmitted by a treehopper (Membracidae), a family that is closely related to leafhoppers (Cicadellidae). To identify vectors of GRBaV. hemipteran species within and nearby wine grape vineyards where virus spread was suspected were collected and transmission assays were conducted. Among the species tested, the three-cornered alfalfa hopper Spissistilus festinus (Hemiptera: Membracidae) was able to both acquire the virus from a grapevine infected with GRBaV and transmit the virus to healthy grapevines in the laboratory. In commercial vineyards, lateral shoots of grapevines girdled due to feeding injury by the adult three-cornered alfalfa hopper also tested positive for the virus using digital PCR. These findings represent an important step in understanding the biology of GRBaV and develop management guidelines. [ABSTRACT FROM AUTHOR]

Grapevine red blotch virus (GRBV) has become widespread in the United States since its identification in 2012. GRBV is the causative agent of grapevine red blotch disease (GRBD), which has caused detrimental economic impacts to the grape and wine industry. Understanding viral function, plant–pathogen interactions, and the effects of GRBV on grapevine performance remains essential to developing potential mitigation strategies. This comprehensive review examines the current body of knowledge regarding GRBV, to highlight gaps in the knowledge and potential mitigation strategies for grape growers and winemakers. [ABSTRACT FROM AUTHOR]

In the Napa Valley of California, vineyards of 'Cabernet Franc' (CF) clone 214, 'Cabernet Sauvignon' clone 337, and 'Zinfandel' clone 1A (Z1A) with grapevines exhibiting foliar symptoms of red blotches, marginal reddening, and red veins that were accompanied by reduced sugar accumulation in fruit at harvest were initially suspected to be infected with leafroll-associated viruses. However, reverse-transcription polymerase chain reaction (PCR) tests were negative for all known leafroll-associated viruses, with the exception of Grapevine leafrollassociated virus 2 in Z1A. Metagenomic analysis of cDNA libraries obtained from double-stranded RNA enriched nucleic acid (NA) preparations from bark scrapings of dormant canes on an Illumina platform revealed sequences having a distant relationship with members of the family Geminiviridae. Sequencing of products obtained by PCR assays using overlapping primers and rolling circle amplification (RCA) confirmed the presence of a single circular genome of 3,206 nucleotides which was nearly identical to the genome of a recently reported Grapevine cabernet franc-associated virus found in declining grapevines in New York. We propose to call this virus "Grapevine red blotch-associated virus" (GRBaV) to describe its association with grapevine red blotch disease. Primers specific to GRBaV amplified a product of expected size (557 bp) from NA preparations obtained from petioles of several diseased source vines. Chip bud inoculations successfully transmitted GRBaV to test plants of CF, as confirmed by PCR analysis. This is the first report of a DNA virus associated with red blotch disease of grapevines in California. [ABSTRACT FROM AUTHOR]

We have identified the genome of a novel viral satellite in deep sequence analysis of double-stranded RNA from grapevine. The genome was 1,060 bases in length, and encoded two open reading frames. Neither frame was related to any known plant virus gene. But translation of the longer frame showed a protein sequence similar to those of other plant virus satellites. Other than in commonalities they shared in this gene sequence, members of that group were extensively divergent. The reading frame in this gene from the novel satellite could be translationally coupled to an adjacent reading frame in the -1 register, through overlapping start/stop codons. These overlapping AUGA start/stop codons were adjacent to a sequence that could be folded into a pseudoknot structure. Field surveys with PCR probes specific for the novel satellite revealed its presence in 3% of the grapevines (n = 346) sampled.

The development of genetically engineered resistance to plant viruses is a result of efforts to understand the plant-virus interactions involved in "crossprotection," a phenomenon observed with several plant virus diseases. Historically, expression of the coat protein gene of Tobacco mosaic virus in transgenic tobacco (Nicotiana tabacum) plants is the first example of transgene-mediated resistance to a plant virus. Subsequently, virus-derived sequences of several plant viruses were shown to confer virus resistance in experimental and/or natural hosts. For plant RNA viruses, virus complementary DNA sequences shown to confer resistance include wild-type genes, mutated genes that produced truncated protein products, and nontranslatable sense or antisense transcripts to various regions of the virus genome. Resistance also has been demonstrated for some viruses by mutant trans-dominant gene products, derived from the movement protein and replication-associated protein genes. In addition to virus-derived sequences, gene sequences of plant origin have also been used for transgenic resistance, and such resistance can be virus-specific, for instance, R genes isolated from resistant plant genotypes, or nonspecific, for example, ribosome inactivating proteins and proteinase inhibitors. Plantibodies and 2-5A synthetase, a class of proteins of mammalian origin, have also been useful in engineering plant virus resistance. In the case of transgenic resistance mediated by viral coat protein, the mechanism of resistance was suggested to operate during the early events of virus infection. However, transgene-mediated RNA silencing and generation of small interfering RNAs appears to be the primary mechanism that confers resistance to plant viruses. Despite the advantages of transgene-mediated resistance, current interest in the development and use of transgenic virus resistant plants is low in most parts of the world. However, because of its real potential, we believe that this technology will have more widespread and renewed interest in the near future.