Comparison of molecular protocols to detect Tomato brown rugose fruit virus in solanaceae hosts

Authors: ZAMORA-MACORRA, ERIKA - Universidad Nacianal Autonoma De Mexico; AVINA-PADILLA, KATIA - Langebio Cinvestav; Hammond, Rosemarie; OCHOA-MARTINEZ, DANIEL - Colegio De Postgraduados

Submitted to: Mexican Journal of Phytopathology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 11/24/2023
Publication Date: 12/23/2023
Citation: Zamora-Macorra, E.J., Avina-Padilla, K., Hammond, R., Ochoa-Martinez, D.L. 2023. Comparison of molecular protocols to detect Tomato brown rugose fruit virus in solanaceae hosts. Mexican Journal of Phytopathology. 41(4):2. https://doi.org/10.18781/R.MEX.FIT.2023-5.
DOI: https://doi.org/10.18781/R.MEX.FIT.2023-5

Interpretive Summary: Tomato brown rugose fruit virus (ToBRFV) is an emerging and rapidly spreading RNA virus that infects tomato and pepper, with tomato as the primary host. The virus is stable and mechanically transmitted, with standard horticultural practices and contaminated seeds, that are untreated or insufficiently treated with disinfectants leading to infection of the growing seedling, as the primary modes of transmission. Our work examined the relevance of emergent diseases associated with potential seed dispersion of ToBRFV Mexican strains to agronomically important Solanaceous hosts, focusing on comparison of molecular biology techniques for rapid and sensitive virus detection in seed lots. Herein we are contributing findings for improved diagnostic methods with potential application in the routine detection of seeds lots and plants, reducing the risk of dispersion. The results of this study will be of interest to the vegetable seed industry and growers, regulatory agencies, and researchers worldwide.

Technical Abstract: Objective/Background: The Tomato brown rugose fruit virus (ToBRFV) has emerged as a significant threat to Solanaceae crops, including tomato and pepper. Its presence in Mexico since 2018 has raised concerns about its impact on agricultural production. Early and accurate detection of this pathogen is essential to prevent its spread and mitigate its effects. In Mexico, several molecular techniques are employed for its diagnosis, including endpoint RT-PCR, RT-qPCR, and multiplex RT-qPCR. Materials and Methods: This research aimed to assess the efficiency of different RNA extraction methods in combination with specific PCR primers for detecting ToBRFV. Results: Among the methods tested, the CTAB-Trizol RNA extraction protocol combined with nested PCR using primers reported by Dovas et al. (2004) was identified as the most sensitive molecular method for detecting the virus. Conclusion: This finding highlights the importance of selecting the appropriate combination of extraction and amplification protocols to achieve optimal sensitivity and accuracy in ToBRFV detection.