Identification of tomato infecting viruses that co-isolate with nanovesicles using a combined proteomics and electron-microscopic approach
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.
2021
2021-07-27 12:18:31
1033
tomato, plant viruses, tomato brown rugose fruit virus, tomato mosaic virus, tomato spotted wilt virus, extracellular vesicles, capsid protein
dk_c
Ramila
Mammadova
70
Immacolata
Fiume
70
Ramesh
Bokka
70
Veronika
Kralj-Iglič
70
Darja
Božič
70
Matic
Kisovec
70
Marjetka
Podobnik
70
Apolonija
Bedina Zavec
70
Matej
Hočevar
70
Gabriella
Gellén
70
Gitta
Schlosser
70
Gabriella
Pocsfalvi
70
UDK
4
577
ISSN pri članku
9
2079-4991
DOI
15
10.3390/nano11081922
COBISS_ID
3
71549443
RAZ_Mammadova_Ramila_2021.pdf
5783878
Predstavitvena datoteka
2022-03-31 14:20:59
0
Izvorni URL
2021-07-27 12:18:33