Virus diseases of strawberry present several complex problems. More than 25 viruses have been described in the genus thus far. Here, we describe a novel rhabdovirus, tentatively named strawberry virus 1 (StrV-1), that infects and plants. Genomic sequences of three distinct StrV-1 genotypes co-infecting a single host were obtained using combined Illumina and Ion Proton high-throughput sequencing. StrV-1 was transmitted to herbaceous plants via and , further mechanically transmitted to 37B and sub-inoculated to , DCL2/4i, 37B, and plants. Irregular chlorotic sectors on leaf blades and the multiplication of calyx leaves seem to be the diagnostic symptoms for StrV-1 on indexed clones. StrV-1 was detected in asymptomatic grafted plants and in 49 out of 159 field strawberry samples via RT-PCR followed by Sanger sequencing. The bacilliform shape of the virions, which have a cytoplasm-limited distribution, their size, and phylogenetic relationships support the assignment of StrV-1 to a distinct species of the genus . , and were shown to transmit StrV-1 under experimental conditions.