Skip to main content
APS Plant Health 2025 Main banner
Icon Legend
This session is not in your schedule.
This session is in your schedule. Click again to remove it.

POSTERS: Analytical and Theoretical Plant Pathology II

P-004 - Effects of Tagging Potyvirus rapae NIb Protein on Virus Pathogenicity

Tuesday, August 5, 2025
9:30 AM - 10:15 AM HST
Location: Exhibit Hall 3

    Presenting Author(s)

  • AG

    Amany E. Gomaa

    PhD Student
    Department of Plant Pathology and Nebraska Center for Virology, University of Nebraska-Lincoln, Lincoln, NE 68583, USA
    Lincoln, Nebraska, United States

    • Author(s)

  • EP

    Eric Parperides

    Department of Plant Pathology and Nebraska Center for Virology, University of Nebraska-Lincoln, Lincoln, NE 68583, USA
    Lincoln, NE, USA

  • HG

    Hernan Garcia-Ruiz

    Principal investigator
    Department of Plant Pathology and Nebraska Center for Virology, University of Nebraska-Lincoln, Lincoln, NE 68583, USA
    Lincoln, Nebraska, United States

Abstract Text:
Turnip mosaic virus (TuMV, Potyvirus rapae), a member of the Potyvirus genus, is widely used in plant-virus interaction studies. Potyviruses cause significant crop losses. Their positive-sense single-stranded RNA genome encodes 11 genes, some of which remain poorly characterized. We aimed to construct a TuMV infectious clone with a tagged nuclear inclusion protein b (NIb) for protein tracking and interaction studies. However, tagging can interfere with protein structure, function, or expression. We tested 6×His and 3×Flag tags at the N- and C-termini of NIb in Nicotiana benthamiana and Arabidopsis thaliana. Our results showed that tagged clones were temperature-sensitive for symptom formation, requiring 22°C instead of 24°C, and exhibited reduced infectivity compared to the wild-type virus. Co-infection with a silencing suppressor did not restore infectivity, suggesting that tagging disrupts viral function rather than being affected by RNA silencing. Western blot confirmed tagged NIb stability. We used AlphaFold to predict the structure of NIb before and after tagging to assess stability and identify an optimal tag that minimally disrupts NIb function. We also predicted the structural impact of tagging other viral proteins, providing insights into potential interference. This study provides a framework for selecting suitable tags for viral proteins, saving time and effort in future tagging experiments, contributing to improving the methodology for studying virus-host interactions, and may facilitate similar research in other viral systems.