ToBRFV Mexican Strain: Seed Transmission Rate, Efficacy of Seed Disinfection Treatment, and Rapid Sensitive Detection in Seed Lots

Abstract

Tomato brown rugose fruit virus (ToBRFV) is a causal agent of severe emergent diseases in Solanaceae hosts with agronomic relevance, such as tomato and pepper. Herein, we analyzed the seed transmission rate and efficacy of different seed disinfection treatments and performed a comparison of molecular biology techniques seeking rapid and sensitive detection in seed lots. We evaluated several total RNA extraction and RT-PCR protocols using a distinct combination of PCR primers to test for the presence of the ToBRFV Mexican strain in tobacco host. Our results showed that the percentage of seed and seedlings transmission was less than 1% and that a 3% sodium hypochlorite solution was effective as a seed disinfection treatment. Finally, the most sensitive molecular method identified for virus detection consisted of a CTAB-Trizol RNA extraction followed by nested PCR using primers reported by Dovas et al. (2004). Additionally, we tested potential natural hosts in selected Cucurbitaceae and Solanaceae species. Our results showed that the ToBRFV Mexican strain is capable of experimentally infecting eggplant (Solanum melongena), tomatillo (Physalis ixocarpa), and tobacco (Nicotiana rustica).