First report of <i>Tomato brown rugose fruit virus</i> infecting tomato in Florida, USA

Florida is a leading producer of fresh-market tomatoes (Solanum lycopersicum) in the US, $262 million industry for state. In October 2019, eight tomato plants showing chlorotic, cupped leaves and stunted foliage (Fig. 1) were collected from c. 1.6 ha community garden Alachua County, Florida. The symptoms observed characteristic Tomato yellow leaf curl virus (TYLCV). Samples tested by PCR using TYLCV-specific primers Navot et al. (1992). Three samples positive TYLCV. Lateral flow immunoassay tests (“immunostrip assays”; Agdia, USA) Pepino mosaic Cucumber negative. However, an immunostrip Tobacco (TMV) gave faint result one three TYLCV-positive sample, suggesting mixed infection. Tobamovirus-like rigid rod-shaped particles ?300 nm length, also dips this plant LVEM5 transmission electron microscope (DeLong America, USA). Tests common tobamoviruses, i.e. TMV, virus, mottle (Sui al., 2017), mild green (Cohen 2006), brown rugose fruit (ToBRFV) (Ling 2019) conducted specific RT-PCR with total RNA extracted RNeasy® Plant Mini kits (Qiagen, Germany). Additionally, conventional primers, DPI-782(5859nt-5879nt) = 5?-TAGCGAAGTGTGGAAACCTG-3? DPI-783(6154nt-6183nt) 5?-GGTGCAGAGGACCATTGTAA-3? (amplicon size 342 bp), probes real-time Taqman assay DPI-784(6029nt-6049nt) 5?-CCGAAACGTTAGACGCTACTC-3?, DPI-785(6106nt -6127nt) ACCTGTTCCTTTGACCAA TTCT-3?, DPI-787(6060nt-6083nt) TaqMan probe 5?-ATG CAACGGTGGCTATAAGGA-3’) coat protein ?5800-6130 nt region developed ToBRFV. Six samples, including sequenced products (GenBank Accession No. MW322766) 98 to 99 % identical ToBRFV isolates Jordan Mexico (KT383474, MK319944). identification was further confirmed USDA-APHIS-PPQ-S&T laboratory (Laurel, Maryland, sequencing. Ct values range 27 31. weak immunostrip, relatively high co-infection TYLCV suggest that present low titre these samples. detected imported sale grocery stores Naples Gainesville University Department Agriculture Consumer Services (FDACS) around same time (Batuman 2020), Immediately following survey, all solanaceous organic destroyed. FDACS taking precautionary measures test produced Further studies are needed better understand risk factors authors would like thank Industry (DPI), assistance project.