Dataset Information

Absolute Quantification of Grapevine Red Blotch Virus in Grapevine Leaf and Petiole Tissues by Proteomics.

ABSTRACT: Grapevine red blotch is a recently identified viral disease that was first recognized in the Napa Valley of California. Infected plants showed foliar symptoms similar to leafroll, another grapevine viral disease, on vines testing negative for known grapevine leafroll-associated virus. Later, the Grapevine red blotch virus (GRBV) was independently discovered in the US states of California and New York and was demonstrated to be the causal agent of red blotch disease. Due to its wide occurrence in the United States, vector transmission, and impacts on grape industry, this virus has the potential to cause serious economic losses. Despite numerous attempts, it has yet not been possible to isolate or visualize viral particles from GRBV-infected plants, thereby hampering the development of a serological assay that would facilitate GRBV detection in grapevine. In this work, mass spectrometry approaches were applied in order to quantify GRBV in infected plants and identify potential biomarkers for viral infection. We present for the first time the physical detection on the protein level of the two GRBV genes V1 (coat protein) and V2 in grapevine tissue lysates. The GRBV coat protein load in petioles was determined to be in the range of 100-900 million copies per milligram wet weight by using three heavy isotope labeled reference peptides as internal standards. In leaves on the other hand, the V1 copy number per unit wet tissue weight appeared to be about six times lower than in petioles, and about 300 times lower in terms of protein concentration in the extractable protein mass, albeit these estimations could only be made with one reference peptide detectable in leaf extracts. Moreover, we found in leaf and petiole extracts of GRBV-infected plants a consistent upregulation of several enzymes involved in flavonoid biosynthesis by label-free shotgun proteomics, indicating the activation of a defense mechanism against GRBV, a plant response already described for Grapevine leafroll-associated virus infection on the transcriptome level. Finally and importantly, we identified some other microorganisms belonging to the grapevine leaf microbiota, two bacterial species (Novosphingobium sp. Rr 2-17 and Methylobacterium) and one virus, Grapevine rupestris stem pitting-associated virus.

SUBMITTER: Buchs N

PROVIDER: S-EPMC6281998 | biostudies-literature | 2018

REPOSITORIES: biostudies-literature

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Publications

Absolute Quantification of <i>Grapevine Red Blotch Virus</i> in Grapevine Leaf and Petiole Tissues by Proteomics.

Buchs Natasha N Braga-Lagache Sophie S Uldry Anne-Christine AC Brodard Justine J Debonneville Christophe C Reynard Jean-Sébastien JS Heller Manfred M

Frontiers in plant science 20181129

Grapevine red blotch is a recently identified viral disease that was first recognized in the Napa Valley of California. Infected plants showed foliar symptoms similar to leafroll, another grapevine viral disease, on vines testing negative for known grapevine leafroll-associated virus. Later, the <i>Grapevine red blotch virus</i> (GRBV) was independently discovered in the US states of California and New York and was demonstrated to be the causal agent of red blotch disease. Due to its wide occurr ...[more]

PMID: 30555495

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Project description:Grapevine red blotch is a recently identified viral disease that was first recognized in the Napa Valley of California. Infected plants showed foliar symptoms similar to leafroll, another grapevine viral disease, on vines testing negative for known grapevine leafroll-associated virus. Later, the Grapevine red blotch virus (GRBV) was independently discovered in the US states of California and New York and was demonstrated to be the causal agent of red blotch disease. Due to its wide occurrence in the US, vector transmission and impacts on grape industry, this virus has the potential to cause serious economic losses. Despite numerous attempts, it was not possible to isolate or visualize viral particles from GRBV infected plants. Consequently, this has hampered the development of a serological assay that would facilitate GRBV detection in grapevine. We therefore decided to explore mass spectrometry approaches in order to quantify GRBV in infected plants and to identify potential biomarkers for viral infection. We present for the first time the physical detection on the protein level of the two GRBV genes V1 (coat protein) and V2 in grapevine tissue lysates. The GRBV coat protein load in leaf petioles was determined to be in the range of 100 to 900 million copies per milligram wet weight by using three heavy isotope labeled reference peptides as internal standards. The V1 copy number per unit wet tissue weight in leaves appeared to be about six times lower, and about 200-times lower in terms of protein concentration in the extractable protein mass than in petioles. We found a consistent upregulation of several enzymes involved in flavonoid biosynthesis in leaf and petiole extracts of GRBV-infected plants by label-free shotgun proteomics, indicating the activation of a defense mechanism against GRBV, a plant response already described for grapevine leafroll associated virus infection on the transcriptome level. Last but not least, we identified some other microorganisms belonging to the grapevine leaf microbiota, two bacterial species (Novosphingobium sp. Rr 2-17 and Methylobacterium) and one virus, Grapevine rupestris stem pitting associated virus.

Project description:Although deficit irrigation is used to improve fruit quality in healthy grapevines, it can potentially amplify negative effects of viral disease and reduce fruit quality in Grapevine Red Blotch Virus (GRBV) infected grapevines. Therefore, a 2-year field experiment was conducted to understand the interaction between GRBV infection and water deficits on disease development and vine physiology. Well-watered (WW) vines were irrigated at 100% of estimated crop evapotranspiration (ETc), while water deficit (WD) vines received water at 66 and 50% ETc in 2017 and 2018, respectively. Healthy (GRBV-) and infected (GRBV+) vines were confirmed by PCR assays. There were no significant effects of water deficits on foliar symptom onset in either year, but more severe water deficits in 2018 resulted in a more rapid symptom progression. GRBV+ vines had a higher ?stem compared to GRBV- vines, but the effects of virus only appeared post-veraison and corresponded to decreased leaf gas exchange. In general, vine vegetative and reproductive growth were not reduced in GRBV+ vines. Yields were highest in WW/GRBV+ vines due to larger clusters containing larger berries. Consistent treatment effects on berry primary chemistry were limited to sugars, with no interactions between factors. Water deficits were able to somewhat increase berry anthocyanin concentration in GRBV+ fruit, but the effects were dependent on year. By comparison, virus status and water deficits interacted on skin tannins concentration such that they were decreased in WD/GRBV+ vines, but increased in WD/GRBV- vines. Water deficits had no effect on seed phenolics, with only virus status having a significant diminution. Although keeping GRBV+ vines well-watered may mitigate some of the negative effects of GRBD, these results suggest that water deficits will not improve overall fruit quality in GRBV+ vines. Ultimately, the control of fruit ripening imparted by GRBV infection seems to be stronger than abiotic control imparted by water deficits.

Dataset Information

Absolute Quantification of Grapevine Red Blotch Virus in Grapevine Leaf and Petiole Tissues by Proteomics.

Publications

Absolute Quantification of <i>Grapevine Red Blotch Virus</i> in Grapevine Leaf and Petiole Tissues by Proteomics.

Similar Datasets

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