Development and validation of a real-time RT-PCR test for screening pepper and tomato seed lots for the presence of pospiviroids

Potato spindle tuber viroid and other pospiviroids can cause serious diseases in potato and tomato crops. Consequently, pospiviroids are regulated in several countries. Since seed transmission is considered as a pathway for the introduction and spread of pospiviroids, some countries demand for the t...

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Bibliographic Details
Published in:PloS one 2020-09, Vol.15 (9), p.e0232502-e0232502
Main Authors: Botermans, Marleen, Roenhorst, Johanna W, Hooftman, Marinus, Verhoeven, Jacobus Th. J, Metz, Eveline, van Veen, Esther J, Geraats, Bart P.J, Kemper, Mark, Beugelsdijk, Debora C.M, Koenraadt, Harrie, Jodlowska, Agata, Westenberg, Marcel
Format: Article
Language:English
Subjects:
Biology and Life Sciences
Capsicum annuum
Crops
Design
Diagnosis
Diseases and pests
Fruits
Peppers
Plant diseases
Polymerase chain reaction
Potatoes
Real time
Research and Analysis Methods
Screening
Seeds
Solanum lycopersicum
Stunt
Testing
Testing laboratories
Tomatoes
Viroids
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Summary:Potato spindle tuber viroid and other pospiviroids can cause serious diseases in potato and tomato crops. Consequently, pospiviroids are regulated in several countries. Since seed transmission is considered as a pathway for the introduction and spread of pospiviroids, some countries demand for the testing of seed lots of solanaceous crops for the presence of pospiviroids. A real-time RT-PCR test, named PospiSense, was developed for testing pepper (Capsicum annuum) and tomato (Solanum lycopersicum) seeds for seven pospiviroid species known to occur naturally in these crops. The test consists of two multiplex reactions running in parallel, PospiSense 1 and PospiSense 2, that target Citrus exocortis viroid (CEVd), Columnea latent viroid (CLVd), pepper chat fruit viroid (PCFVd), potato spindle tuber viroid (PSTVd), tomato apical stunt viroid (TASVd), tomato chlorotic dwarf viroid (TCDVd) and tomato planta macho viroid (TPMVd, including the former Mexican papita viroid). Dahlia latent viroid (DLVd) is used as an internal isolation control. Validation of the test showed that for both pepper and tomato seeds the current requirements of a routine screening test are fulfilled, i.e. the ability to detect one infested seed in a sample of c.1000 seeds for each of these seven pospiviroids. Additionally, the PospiSense test performed well in an inter-laboratory comparison, which included two routine seed-testing laboratories, and as such provides a relatively easy alternative to the currently used tests.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0232502