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ARS Home » Southeast Area » Charleston, South Carolina » Vegetable Research » Research » Publications at this Location » Publication #373380
Research Project: Biological, Genetic and Genomic Based Disease Management for Vegetable Crops

Location: Vegetable Research

Title: Development of a triplex AmplifyRP molecular assay for a reliable detection of Tomato brown rugose fruit virus

Author
item LI, RUGANG - Agdia
item DAVENPORT, BRYANT - Agdia
item ZHANG, SHULU - Agdia
item SCHUETZ, KEITH - Agdia
item Ling, Kai-Shu

Submitted to: Phytopathology
Publication Type: Abstract Only
Publication Acceptance Date: 5/15/2020
Publication Date: 12/1/2020
Citation: Li, R., Davenport, B., Zhang, S., Schuetz, K., Ling, K. 2020. Development of a triplex AmplifyRP molecular assay for a reliable detection of Tomato brown rugose fruit virus. Phytopathology Phytopathology 110:12S2.138-S2.139
DOI: https://doi.org/10.1094/PHYTO-110-12-S2.1

Interpretive Summary:

Technical Abstract: Tomato brown rugose fruit virus (ToBRFV) is an emerging Tobamovirus initially identified infecting greenhouse tomatoes in Israel and Jordan in 2014-2015. ToBRFV has since been reported in many countries in Asia, North America, and Europe indicating it has become widely distributed throughout the world. The serious damage to tomato and pepper crop productions, the Tm-2^2 resistance-breaking ability, and the seedborne nature make ToBRFV a serious concern to vegetable growers, seed production industries, and governmental organizations responsible for plant biosecurity. Serological detection using tomato mosaic virus (TMV) or tomato mosaic virus (ToMV) antibodies can make a general detection of a Tobamovirus infection with cross serological reactivity. However, in order to facilitate reliable detection of ToBRFV, we developed a ToBRFV-specific isothermal AmplifyRP assay. The AmplifyRP assay targets two virus-specific genes of ToBRFV and a host-specific gene as an internal control. Screening of primer and probe combinations led to the discovery that one set of primer/probe from the RNA dependent RNA polymerase (RdRp) gene and another set from the coat protein (CP) gene exhibited specific and sensitive detection for ToBRFV, plus one set of primer/probe specific for the plant ribosomal RNA as internal control. The preliminary experiments showed that both the RdRp-specific and the CP-specific assays can detect ToBRFV from infected leaves using a simple diluted crude tissue extract. No cross reactions were observed to other Tobamoviruses. Data from the optimization of this assay will be reported in this meeting.