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ORIGINAL RESEARCH article

Front. Plant Sci.
Sec. Plant Pathogen Interactions
Volume 16 - 2025 | doi: 10.3389/fpls.2025.1535175

Validation of a One-Step Reverse Transcription PCR detection method for Tobamovirus maculatessellati, in tomato (Solanum lycopersicum L.) and pepper (Capsicum annuum L.)

Provisionally accepted
Chellappan Padmanabhan Chellappan Padmanabhan 1*Andrea C Gilliard Andrea C Gilliard 2Kai-Shu Ling Kai-Shu Ling 2Yazmin Rivera Yazmin Rivera 1*
  • 1 Plant Pathogen Confirmatory Diagnostics Laboratory, USDA APHIS PPQ Science and Technology, Laurel, Maryland, United States
  • 2 United States Department of Agriculture, Agricultural Research Service, U. S. Vegetable Laboratory, Charleston, SC 29414, United States

The final, formatted version of the article will be published soon.

    The solanaceous-infecting tobamoviruses are closely related and hence it can be challenging to detect them using serological or molecular methods, particularly when present in a mixed infection. Tomato mottle mosaic virus (ToMMV) is a newly identified tobamovirus that poses serious risk to tomato (Solanum lycopersicum L.) and pepper (Capsicum annuum L.) production worldwide. Species-specific identification is crucial to prevent the entry and establishment of plant pathogens and protect the billiondollar tomato industry. In this study, we report the validation of a previously described reverse transcription polymerase chain reaction (RT-PCR) assay that amplifies a 289 bp fragment of the coat protein coding region of ToMMV genome. This assay has 100% specificity for ToMMV. Inclusivity tests were performed against a diverse collection of six ToMMV isolates in North America. Exclusivity tests showed no cross reaction with eleven non-target viruses and seven viroids commonly found on tomato and pepper host plants. The detection limit of the one-step RT-PCR was determined to be at 10 -5 (or 0.25pg/μl) dilution in plant samples, with its amplicon sequence confirmed by Sanger sequencing. The RT-PCR can detect ToMMV consistently on contaminated seed or leaf tissues. This validated assay could serve as a standard method for detecting ToMMV in seed health testing and for plant disease diagnosis, thus to prevent inadvertent introduction and spread of this emerging and economically important tobamovirus in tomato and pepper fields.

    Keywords: Tobamovirus maculatessellati, Tomato mottle mosaic virus, RT-PCR, Method validation, Tomato, pepper

    Received: 27 Nov 2024; Accepted: 13 Jan 2025.

    Copyright: © 2025 Padmanabhan, Gilliard, Ling and Rivera. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

    * Correspondence:
    Chellappan Padmanabhan, Plant Pathogen Confirmatory Diagnostics Laboratory, USDA APHIS PPQ Science and Technology, Laurel, Maryland, United States
    Yazmin Rivera, Plant Pathogen Confirmatory Diagnostics Laboratory, USDA APHIS PPQ Science and Technology, Laurel, Maryland, United States

    Disclaimer: All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher.