Widely-based full-genome analyses enable development of universal and strain-specific PCR toolkit for wheat dwarf virus detection, revealing new alternative hosts and challenging strain-host specificity

Abstract Background Wheat dwarf virus (WDV) is a destructive cereal virus causing significant yield losses in wheat and barley. It is transmitted by the leafhopper Psammotettix alienus and can persist in wild grasses between growing seasons, making reliable detection and strain differentiation criti...

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Main Authors: Botond Zsombor Pertics, Gergely Tholt, András Kis, Éva Szita, Kornél Gerő, Regina Gerstenbrand, Janka Simon, Ferenc Samu
Format: Article
Language:English
Published: BMC 2025-07-01
Series:Plant Methods
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Online Access:https://doi.org/10.1186/s13007-025-01420-6
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author Botond Zsombor Pertics
Gergely Tholt
András Kis
Éva Szita
Kornél Gerő
Regina Gerstenbrand
Janka Simon
Ferenc Samu
author_facet Botond Zsombor Pertics
Gergely Tholt
András Kis
Éva Szita
Kornél Gerő
Regina Gerstenbrand
Janka Simon
Ferenc Samu
author_sort Botond Zsombor Pertics
collection DOAJ
description Abstract Background Wheat dwarf virus (WDV) is a destructive cereal virus causing significant yield losses in wheat and barley. It is transmitted by the leafhopper Psammotettix alienus and can persist in wild grasses between growing seasons, making reliable detection and strain differentiation critical for disease management. Results We developed a comprehensive PCR toolkit for WDV by analysing 38 complete genome sequences, reviewing, validating, and upgrading existing primers and designing new primers spanning multiple viral genome regions. The primer toolkit achieved high diagnostic and analytical specificity as it consistently detected WDV in plants and insect vectors. This enabled the separation of WDV wheat- and barley-strains through a two-step workflow: screening with universal primers, and strain assignment with strain-specific primer pairs. Field testing across 13 Hungarian sites revealed barley strain dominance in the samples, infecting not only barley but also wheat and multiple grass species. Our surveys identified three previously undocumented reservoir grasses adding to the reviewed host range of 42 species. Complete genome sequencing of one wheat-strain and two barley-strain isolates confirmed > 99% intra-strain nucleotide identity but only ~ 85% between strains. Spatial mapping demonstrated virus concentration in grassy islands with declining titers toward cultivated areas, suggesting these serve as infection reservoirs. Conclusions This validated primer panel provides a robust framework for studying WDV epidemiology and developing targeted management strategies for this economically important pathogen. Understanding this model of virus-vector system and the improvement of the presented methods are key factors to combat other similarly operating plant-vector-pathogen systems.
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institution Kabale University
issn 1746-4811
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publishDate 2025-07-01
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spelling doaj-art-fea2b30435b14332834faa3d771ecfb82025-08-20T04:03:03ZengBMCPlant Methods1746-48112025-07-0121111710.1186/s13007-025-01420-6Widely-based full-genome analyses enable development of universal and strain-specific PCR toolkit for wheat dwarf virus detection, revealing new alternative hosts and challenging strain-host specificityBotond Zsombor Pertics0Gergely Tholt1András Kis2Éva Szita3Kornél Gerő4Regina Gerstenbrand5Janka Simon6Ferenc Samu7National Laboratory for Health Security, HUN-REN Centre for Agricultural Research Plant Protection InstituteNational Laboratory for Health Security, HUN-REN Centre for Agricultural Research Plant Protection InstituteInstitute of Genetics and Biotechnology, Hungarian University of Agriculture and Life SciencesNational Laboratory for Health Security, HUN-REN Centre for Agricultural Research Plant Protection InstituteNational Laboratory for Health Security, HUN-REN Centre for Agricultural Research Plant Protection InstituteNational Laboratory for Health Security, HUN-REN Centre for Agricultural Research Plant Protection InstituteNational Laboratory for Health Security, HUN-REN Centre for Agricultural Research Plant Protection InstituteNational Laboratory for Health Security, HUN-REN Centre for Agricultural Research Plant Protection InstituteAbstract Background Wheat dwarf virus (WDV) is a destructive cereal virus causing significant yield losses in wheat and barley. It is transmitted by the leafhopper Psammotettix alienus and can persist in wild grasses between growing seasons, making reliable detection and strain differentiation critical for disease management. Results We developed a comprehensive PCR toolkit for WDV by analysing 38 complete genome sequences, reviewing, validating, and upgrading existing primers and designing new primers spanning multiple viral genome regions. The primer toolkit achieved high diagnostic and analytical specificity as it consistently detected WDV in plants and insect vectors. This enabled the separation of WDV wheat- and barley-strains through a two-step workflow: screening with universal primers, and strain assignment with strain-specific primer pairs. Field testing across 13 Hungarian sites revealed barley strain dominance in the samples, infecting not only barley but also wheat and multiple grass species. Our surveys identified three previously undocumented reservoir grasses adding to the reviewed host range of 42 species. Complete genome sequencing of one wheat-strain and two barley-strain isolates confirmed > 99% intra-strain nucleotide identity but only ~ 85% between strains. Spatial mapping demonstrated virus concentration in grassy islands with declining titers toward cultivated areas, suggesting these serve as infection reservoirs. Conclusions This validated primer panel provides a robust framework for studying WDV epidemiology and developing targeted management strategies for this economically important pathogen. Understanding this model of virus-vector system and the improvement of the presented methods are key factors to combat other similarly operating plant-vector-pathogen systems.https://doi.org/10.1186/s13007-025-01420-6Wheat dwarf virusPsammotettix alienusWDV host rangeWDV strain specificityWDV wheat strainWDV barley strain
spellingShingle Botond Zsombor Pertics
Gergely Tholt
András Kis
Éva Szita
Kornél Gerő
Regina Gerstenbrand
Janka Simon
Ferenc Samu
Widely-based full-genome analyses enable development of universal and strain-specific PCR toolkit for wheat dwarf virus detection, revealing new alternative hosts and challenging strain-host specificity
Plant Methods
Wheat dwarf virus
Psammotettix alienus
WDV host range
WDV strain specificity
WDV wheat strain
WDV barley strain
title Widely-based full-genome analyses enable development of universal and strain-specific PCR toolkit for wheat dwarf virus detection, revealing new alternative hosts and challenging strain-host specificity
title_full Widely-based full-genome analyses enable development of universal and strain-specific PCR toolkit for wheat dwarf virus detection, revealing new alternative hosts and challenging strain-host specificity
title_fullStr Widely-based full-genome analyses enable development of universal and strain-specific PCR toolkit for wheat dwarf virus detection, revealing new alternative hosts and challenging strain-host specificity
title_full_unstemmed Widely-based full-genome analyses enable development of universal and strain-specific PCR toolkit for wheat dwarf virus detection, revealing new alternative hosts and challenging strain-host specificity
title_short Widely-based full-genome analyses enable development of universal and strain-specific PCR toolkit for wheat dwarf virus detection, revealing new alternative hosts and challenging strain-host specificity
title_sort widely based full genome analyses enable development of universal and strain specific pcr toolkit for wheat dwarf virus detection revealing new alternative hosts and challenging strain host specificity
topic Wheat dwarf virus
Psammotettix alienus
WDV host range
WDV strain specificity
WDV wheat strain
WDV barley strain
url https://doi.org/10.1186/s13007-025-01420-6
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