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

Buchs, Natasha; Braga, Sophie Marie-Pierre; Uldry, Anne-Christine; Brodard, Justine; Debonneville, Christophe; Reynard, Jean-Sébastien; Heller, Manfred (2018). Absolute Quantification of in Grapevine Leaf and Petiole Tissues by Proteomics. Frontiers in Plant Science, 9, p. 1735. Frontiers 10.3389/fpls.2018.01735

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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 (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 infection on the transcriptome level. Finally and importantly, we identified some other microorganisms belonging to the grapevine leaf microbiota, two bacterial species ( sp. and ) and one virus, .

Item Type:

Journal Article (Original Article)

Division/Institute:

04 Faculty of Medicine > Pre-clinic Human Medicine > BioMedical Research (DBMR)
04 Faculty of Medicine > Pre-clinic Human Medicine > BioMedical Research (DBMR) > Unit Childrens Hospital > Protein- und Zellbiologie
04 Faculty of Medicine > Pre-clinic Human Medicine > BioMedical Research (DBMR) > DCR Services > Core Facility Massenspektrometrie- und Proteomics-Labor

UniBE Contributor:

Buchs, Natasha; Braga, Sophie Marie-Pierre; Uldry, Anne-Christine and Heller, Manfred

Subjects:

600 Technology > 610 Medicine & health

ISSN:

1664-462X

Publisher:

Frontiers

Language:

English

Submitter:

Marla Rittiner

Date Deposited:

12 Feb 2019 14:50

Last Modified:

17 Feb 2019 02:42

Publisher DOI:

10.3389/fpls.2018.01735

PubMed ID:

30555495

Uncontrolled Keywords:

Grapevine red blotch virus absolute quantification grapevine parallel reaction monitoring proteomics shotgun LC-MS/MS

BORIS DOI:

10.7892/boris.124523

URI:

https://boris.unibe.ch/id/eprint/124523

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