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Identification of tomato infecting viruses that co-isolate with nanovesicles using a combined proteomics and electron-microscopic approach
Mammadova, Ramila (Author), Fiume, Immacolata (Author), Bokka, Ramesh (Author), Kralj-Iglič, Veronika (Author), Božič, Darja (Author), Kisovec, Matic (Author), Podobnik, Marjetka (Author), Bedina Zavec, Apolonija (Author), Hočevar, Matej (Author), Gellén, Gabriella (Author), Schlosser, Gitta (Author), Pocsfalvi, Gabriella (Author)

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Abstract
Plant-derived nanovesicles (NVs) have attracted interest due to their anti-inflammatory, anticancer and antioxidative properties and their efficient uptake by human intestinal epithelial cells. Previously we showed that tomato (Solanum lycopersicum L.) fruit is one of the interesting plant resources from which NVs can be obtained at a high yield. In the course of the isolation of NVs from different batches of tomatoes, using the established differential ultracentrifugation or size-exclusion chromatography methods, we occasionally observed the co-isolation of viral particles. Density gradient ultracentrifugation (gUC), using sucrose or iodixanol gradient materials, turned out to be efficient in the separation of NVs from the viral particles. We applied cryogenic transmission electron microscopy (cryo-TEM), scanning electron microscopy (SEM) for the morphological assessment and LC–MS/MS-based proteomics for the protein identification of the gradient fractions. Cryo-TEM showed that a low-density gUC fraction was enriched in membrane-enclosed NVs, while the high-density fractions were rich in rod-shaped objects. Mass spectrometry–based proteomic analysis identified capsid proteins of tomato brown rugose fruit virus, tomato mosaic virus and tomato mottle mosaic virus. In another batch of tomatoes, we isolated tomato spotted wilt virus, potato virus Y and southern tomato virus in the vesicle sample. Our results show the frequent co-isolation of plant viruses with NVs and the utility of the combination of cryo-TEM, SEM and proteomics in the detection of possible viral contamination

Language:English
Keywords:tomato, plant viruses, tomato brown rugose fruit virus, tomato mosaic virus, tomato spotted wilt virus, extracellular vesicles, capsid protein
Work type:Article (dk_c)
Tipology:1.01 - Original Scientific Article
Organization:ZF - University College of Health Studies
Year:2021
Submitted for review:20.07.2021
Article acceptance date:21.07.2021
Article publication date:26.07.2021
Number of pages:1-19
Numbering:[article] 1922, iss. 8
UDC:577
ISSN on article:2079-4991
DOI:10.3390/nano11081922 This link opens in a new window
COBISS.SI-ID:71549443 This link opens in a new window
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Downloads:17
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Record is a part of a journal

Title:Nanomaterials
Shortened title:Nanomaterials
Publisher:MDPI AG
ISSN:2079-4991
COBISS.SI-ID:523286297 This link opens in a new window

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