High throughput sequencing reveals mixed infections of impatiens necrotic spot virus and tomato brown rugose fruit virus in bell pepper crops in Mexico

Authors: ANGELES MORA-UGALDE, MARIA - Technologico De Monterrey; MIRANDA-CAMPANA, OMAR - Centro De Investigacion En Alimentaction Y Desarollo; RAMIREZ-SOTO, MILAGROS - Instituto Nacional De Investigaciones Forestales Y Agropecuarias (INIFAP); CRUZ-JARAMILLO, JOSE - Non ARS Employee; DE LOS REYES BOLAÑOS, IGNACIO JAVIER - Technologico De Monterrey; TOVAR-PEDRAZA, JUAN - Centro De Investigacion En Alimentaction Y Desarollo; Wintermantel, William; DIAZ-LARA, ALFREDO - Technologico De Monterrey

Submitted to: Journal of Plant Pathology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 10/29/2024
Publication Date: 11/13/2024
Citation: de los Angeles Mora-Ugalde, M., Miranda-Campana, O.A., Ramirez-Soto, M., Cruz-Jaramillo, J.L., de los Reyes Bolaños, I.E., Tovar-Pedraza, J.M., Wintermantel, W.M., Diaz-Lara, A. 2024. High throughput sequencing reveals mixed infections of impatiens necrotic spot virus and tomato brown rugose fruit virus in bell pepper crops in Mexico. Journal of Plant Pathology. 107:711–714. https://doi.org/10.1007/s42161-024-01806-w.
DOI: https://doi.org/10.1007/s42161-024-01806-w

Interpretive Summary: Pepper crops are affected by several viruses worldwide. In September 2022, mosaic, ringspot, and deformation symptoms, were observed on bell pepper leaves and fruits in a commercial greenhouse located in Queretaro, Mexico. Symptoms were indicative of infection by viruses in the genera, orthotospovirus and tobamovirus, but some plants exhibited increased severity compared to what is typically observed for these types of viruses in pepper plants. Testing with antibody-based tests identified infection by tomato brown rugose fruit virus (ToBRFV) in six of 10 plants, but due to the severity of the symptoms, the six ToBRFV-positive samples were additionally tested using high throughput sequencing (HTS) analysis. Results of HTS confirmed infection by both ToBRFV and impatiens necrotic spot virus in two of the plants. Results were further validated using additional molecular biology- and serology-based methods. This is the first report of a natural mixed infection of INSV and ToBRFV in bell pepper in Mexico and potentially around the world, and the increased symptom severity in these plants suggests the possibility of a synergistic interaction between these viruses in pepper.

Technical Abstract: Pepper crops are affected by several viruses worldwide. In September 2022, symptoms of mosaic, deformation and ringspot were observed on bell pepper leaves and fruits in a commercial greenhouse located in Queretaro, Mexico. Symptoms were indicative of infection by an orthotospovirus and/or a tobamovirus, but some plants exhibited increased severity compared to what is typically observed for these viruses in pepper plants. Six out of a total of 10 symptomatic plants tested positive for tomato brown rugose fruit virus (ToBRFV) using ImmunoStrips during initial testing of field plant samples. Due to the severity of the damage, the six ToBRFV-positive samples were independently subjected to high throughput sequencing (HTS) analysis. Sequences of ToBRFV and impatiens necrotic spot virus (INSV) were identified from two of the HTS-analyzed plants, whereas the other four plants were infected with ToBRFV alone. HTS results were validated by reverse transcription-PCR employing specific primers amplifying genomic fragments of those viruses. Finally, using ELISA, INSV presence was confirmed in the two coinfected bell peppers in addition to the four original plants not analyzed by HTS. This is the first report of a natural mixed infection of INSV and ToBRFV in bell pepper in Mexico and potentially around the world, and suggests the possibility of a synergistic interaction between these viruses in pepper.