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A Robust Potato Tuber Tissue Collection Method to Investigate Potato Virus Y, Potato Mop-Top Virus, and Tobacco
Jason Ingram1, Erika Mudrak2, Lisa Tran3
1Emerging Pest and Pathogens Unit, USDA-Agricultural Research Service, Ithaca, NY 14853, U.S.A.
Plant Disease
|April 29, 2025
Summary
Accurate potato virus detection requires molecular assays, not just visual inspection. This study optimized sample collection from tubers using FTA cards and RT-PCR for improved identification of Potato virus Y (PVY), PMTV, TRV, and PVS.
Area of Science:
- Plant Pathology
- Molecular Biology
- Agricultural Science
Background:
- Visual foliar symptoms for Potato virus Y (PVY), potato mop-top virus (PMTV), potato virus S (PVS), and tobacco rattle virus (TRV) are often unreliable for accurate diagnosis.
- Effective detection methods are crucial for managing these significant potato pathogens.
Purpose of the Study:
- To develop and optimize a molecular assay for efficient detection of PVY, PMTV, PVS, and TRV in potato tubers.
- To determine optimal sample collection and nucleic acid extraction methods for improved virus detection rates.
Main Methods:
- A reverse-transcription polymerase chain reaction (RT-PCR) assay was developed using customized Whatman Flinders Technology Associates Plantsaver® Cards (FTA cards) for sample collection and nucleic acid extraction.
- Tuber samples from 12 cultivars across seven locations (2017-2019) were analyzed, assessing virus localization within tubers and optimal tissue depth (surface to 0.5 cm).
- Virus detection rates were evaluated at different time points post-harvest (25 days to 175 days).
Main Results:
- Virus distribution varied within tubers: PMTV and PVY were more frequent at the stem-end, TRV at the rose-end, and PVS was evenly distributed.
- Composite sampling from multiple tuber locations and shallow tissue excision (0-0.5 cm depth) significantly improved detection rates for all four viruses.
- Optimal detection times post-harvest varied, with peak detection for PVY, PMTV, and TRV at 100, 150, and 175 days, respectively. However, PMTV and PVY detection at 25 days was comparable to peak times.
Conclusions:
- Optimized molecular assays using FTA cards and RT-PCR provide efficient and reliable detection of key potato viruses.
- Understanding virus localization within tubers and optimal sampling depths is critical for maximizing diagnostic accuracy.
- The developed method offers a valuable tool for early and accurate detection of potato viruses, aiding in disease management strategies.
