RT Journal Article
SR Electronic
T1 Development and validation of a real-time RT-PCR test for screening pepper and tomato seed lots for the presence of pospiviroids
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2020.04.17.046508
DO 10.1101/2020.04.17.046508
A1 Marleen Botermans
A1 Johanna W. Roenhorst
A1 Marinus Hooftman
A1 Jacobus Th.J. Verhoeven
A1 Eveline Metz
A1 Esther J. van Veen
A1 Bart P.J. Geraats
A1 Mark Kemper
A1 Debora C.M. Beugelsdijk
A1 Harrie Koenraadt
A1 Agata Jodlowska
A1 Marcel Westenberg
YR 2020
UL http://biorxiv.org/content/early/2020/04/17/2020.04.17.046508.abstract
AB 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.