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Evaluating the Negative-Strand Coding Potential in Plum Pox Virus.

Beatriz García¹, José Vicente Del Álamo¹, Mario Rincón¹

¹Departamento de Genética Molecular de Plantas, Centro Nacional de Biotecnología (CNB-CSIC), Campus Universidad Autónoma de Madrid, Madrid, Spain.

Molecular Plant Pathology

|December 9, 2025

Summary

This summary is machine-generated.

Reverse open reading frames (rORFs) in plum pox virus (PPV) do not appear to be functional. Studies found no evidence of rORF-encoded peptides or impact on PPV infection, suggesting they are not produced or are irrelevant.

Keywords:

Potyviridae alternative proteins negative strand plum pox virus rORFs

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Area of Science:

Virology
Molecular Biology
Genomics

Background:

Single-stranded, positive-sense RNA viruses can possess reverse open reading frames (rORFs) in their negative strands, encoding functional peptides.
Highly conserved rORFs are present in the negative strand of Potyviridae family viruses, such as turnip mosaic virus (TuMV), where rORF2 is crucial for infection.

Purpose of the Study:

To investigate the existence and potential functionality of rORFs in plum pox virus (PPV).
To determine if rORFs play a role in PPV infection and survival in experimental and natural hosts.

Main Methods:

Utilized mass spectrometry for unbiased identification of rORF-encoded peptides in PPV.
Employed mutagenesis to create PPV mutants lacking individual rORF-derived peptides.
Conducted comparative infection experiments in Nicotiana benthamiana and Prunus persica.
Performed competitive infection assays between wild-type PPV and rORF knockout mutants.

Main Results:

Mass spectrometry analyses did not detect any peptides encoded by PPV rORFs.
PPV mutants with abolished rORF expression showed no significant difference in infection rates compared to wild-type.
Competitive infection experiments revealed no advantage for either wild-type PPV or rORF2 knockout mutants.

Conclusions:

rORFs in plum pox virus are likely not produced or are functionally insignificant for infection.
The role of rORFs in potyviruses requires individual assessment, as functionality is not conserved across the family.
These findings contrast with the essential role of rORF2 in turnip mosaic virus.