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Title:Filling the gaps in diagnostics of Pepino mosaic virus and Potato spindle tuber viroid in water and tomato seeds and leaves
Authors:ID Mehle, Nataša (Author)
ID Kogovšek, Polona (Author)
ID Rački, Nejc (Author)
ID Jakomin, Tjaša (Author)
ID Gutiérrez-Aguirre, Ion (Author)
ID Kramberger, Petra (Author)
ID Ravnikar, Maja (Author)
Files:.pdf PDF - Presentation file, download (312,20 KB)
MD5: 6730A5CAAC1EFBF75A9F9A918923FCB1
 
URL URL - Source URL, visit https://doi.org/10.1111/ppa.12710
 
Language:English
Typology:1.01 - Original Scientific Article
Organization:Logo NIB - National Institute of Biology
Abstract:Waterborne and seedborne Pepino mosaic virus (PepMV) and Potato spindle tuber viroid (PSTVd) pose serious threats to tomato production due to seed transmission and mechanical transmission, coupled with their long-term stability outside the host plant. Therefore, rapid and sensitive diagnostic procedures are needed to prevent the spread of these quarantine pathogens. In particular, water and seed contamination are difficult to detect and confirm without efficient concentration methods. This study presents procedures that improve detection of PSTVd from tomato seeds and leaf tissue, and PepMV from water and tomato leaf tissue. For efficient concentration of PepMV from water samples, a procedure was optimized using convective interaction media monolithic chromatography columns, which provides concentration by three orders of magnitude. For concentration of PSTVd from seed extracts, an easy-to-use and efficient method was developed based on RNA binding to positively charged anion-exchange resin beads that provides up to 100-fold more sensitive detection in comparison with procedures without a concentration step. This thus allows confirmation of RT-qPCR results with sequencing of RT-PCR products in samples with low viroid levels. In addition, reverse-transcription loop-mediated isothermal amplification assays for detection of PSTVd and PepMV were optimized and adapted to both laboratory and on-site testing requirements. This allows rapid detection of these pathogens in crude leaf homogenates, in under 30 min. These procedures of concentration and detection are shown to be efficient and to fill the gaps in diagnostics of PepMV and PSTVd, especially when these pathogens are present at low levels in difficult matrices such as water and seeds.
Keywords:PSTVd, PepMV, seeds, water, concentration, loop-mediated isothermal amplification
Publication status:Published
Publication version:Version of Record
Publication date:01.09.2017
Year of publishing:2017
Number of pages:str. 1191-1201
Numbering:Vol. 66, issue 7
PID:20.500.12556/DiRROS-19712 New window
UDC:577.2
ISSN on article:0032-0862
DOI:10.1111/ppa.12710 New window
COBISS.SI-ID:4302159 New window
Publication date in DiRROS:24.07.2024
Views:540
Downloads:245
Metadata:XML DC-XML DC-RDF
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MEHLE, Nataša, KOGOVŠEK, Polona, RAČKI, Nejc, JAKOMIN, Tjaša, GUTIÉRREZ-AGUIRRE, Ion, KRAMBERGER, Petra and RAVNIKAR, Maja, 2017, Filling the gaps in diagnostics of Pepino mosaic virus and Potato spindle tuber viroid in water and tomato seeds and leaves. Plant Pathology [online]. 2017. Vol. 66, no. 7, p. 1191–1201. [Accessed 3 April 2025]. DOI 10.1111/ppa.12710. Retrieved from: https://dirros.openscience.si/IzpisGradiva.php?lang=eng&id=19712
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Record is a part of a journal

Title:Plant Pathology
Shortened title:Plant Pathol.
Publisher:Her Majesty's Stationery Office
ISSN:0032-0862
COBISS.SI-ID:5661191 New window

Document is financed by a project

Funder:ARIS - Slovenian Research and Innovation Agency
Project number:L4-5525-2013
Name:Študij epidemiologije in raznolikosti mikrobnih povzročiteljev bolezni rastlin
Funder:ARIS - Slovenian Research and Innovation Agency
Project number:P4-0165-2015
Name:Biotehnologija in sistemska biologija rastlin
Funder:EC - European Commission
Funding programme:ArimNet
Name:Emergent viruses and virus vectors in Mediterranean Basin crops
Acronym:Emaramb

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License:CC BY-NC 4.0, Creative Commons Attribution-NonCommercial 4.0 International
Link:http://creativecommons.org/licenses/by-nc/4.0/
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