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Identification of tomato infecting viruses that co-isolate with nanovesicles using a combined proteomics and electron-microscopic approach
ID
Mammadova, Ramila
(
Author
),
ID
Fiume, Immacolata
(
Author
),
ID
Bokka, Ramesh
(
Author
),
ID
Kralj-Iglič, Veronika
(
Author
),
ID
Božič, Darja
(
Author
),
ID
Kisovec, Matic
(
Author
),
ID
Podobnik, Marjetka
(
Author
),
ID
Bedina Zavec, Apolonija
(
Author
),
ID
Hočevar, Matej
(
Author
),
ID
Gellén, Gabriella
(
Author
),
ID
Schlosser, Gitta
(
Author
),
ID
Pocsfalvi, Gabriella
(
Author
)
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https://www.mdpi.com/2079-4991/11/8/1922
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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
Typology:
1.01 - Original Scientific Article
Organization:
ZF - Faculty of Health Sciences
Publication status:
Published
Publication version:
Version of Record
Year:
2021
Number of pages:
19 str.
Numbering:
Vol. 11, iss. 8, art. 1922
PID:
20.500.12556/RUL-128748
UDC:
577
ISSN on article:
2079-4991
DOI:
10.3390/nano11081922
COBISS.SI-ID:
71549443
Publication date in RUL:
27.07.2021
Views:
1200
Downloads:
162
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Record is a part of a journal
Title:
Nanomaterials
Shortened title:
Nanomaterials
Publisher:
MDPI
ISSN:
2079-4991
COBISS.SI-ID:
523286297
Licences
License:
CC BY 4.0, Creative Commons Attribution 4.0 International
Link:
http://creativecommons.org/licenses/by/4.0/
Description:
This is the standard Creative Commons license that gives others maximum freedom to do what they want with the work as long as they credit the author.
Licensing start date:
01.08.2021
Projects
Funder:
EC - European Commission
Funding programme:
H2020
Project number:
798576
Name:
Conversion of natural plant nanovesicles into nutraceutical delivery system
Acronym:
nanoTOM
Funder:
ARRS - Slovenian Research Agency
Project number:
P3-0388
Name:
Mehanizmi varovanja zdravja
Funder:
ARRS - Slovenian Research Agency
Project number:
P1-0391
Name:
Molekulske interakcije
Funder:
ARRS - Slovenian Research Agency
Project number:
P2-0132
Name:
Fizika in kemija površin kovinskih materialov
Funder:
ARRS - Slovenian Research Agency
Project number:
J1-2467
Name:
Odkrivanje dinamike omrežja v interakciji krompirjev virus Y-rastlinska celica
Funder:
Other - Other funder or multiple funders
Funding programme:
Hungarian Academy of Sciences, Lendület (Momentum) Program
Funder:
Other - Other funder or multiple funders
Funding programme:
Hungary, National Research, Development and Innovation Fund
Project number:
2018-1.2.1-NKP 00005
Funder:
Other - Other funder or multiple funders
Funding programme:
Hungary, Ministry for Innovation and Technology, Thematic Excellence Programme
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