The genome of a novel rhabdovirus was detected in yerba mate (Ilex paraguariensis St. Hil.). The newly identified virus, tentatively named yerba mate virus A (YmVA), has a genome of 14,961 nucleotides.Notably, eight open reading frames were identified in the antigenomic orientation of the negative-sense, single-stranded viral RNA, including two novel accessory genes, in the order 3'-N-P-3-4-M-G-L-8-5'. Sequence identity of the encoded proteins as well as phylogenetic analysis suggest that YmVA is a new member of the genus Cytorhabdovirus, family Rhabdoviridae. YmVA unique genomic organization and phylogenetic relationships indicate that this virus likely represents a distinct evolutionary lineage within the cytorhabdoviruses. Yerba mate (Ilex paraguariensis St. Hil., Aquifoleaceae) is a subtropical tree cultivated in the northeast of Argentina, south of Brazil, and the west of Paraguay. Its leaves and stems are widely utilized in the preparation of an infusion popularly known as "mate." In Argentina, the main producer, the cultivated area has reached 165,327 ha [1]. Although yerba mate is of significant economic, social and traditional importance in the region, studies on plant diseases are still scarce, and only a handful of viruses have been identified in this holly species [2-4] Recently, the complete genome sequence of another cytorhabdovirus was described, associated with chlorotic linear patterns, chlorotic rings, and veins yellowing symptoms in yerba mate, which was named yerba mate chlorosis-associated virus (YmCaV) [3]. The genus Cytorhabdovirus in the family Rhabdoviridae has the largest number of members among the four genera of plant-infecting viruses in the family. Cytorhabdoviruses have a unsegmented, negative-sense, single-stranded RNA genomes with six to ten open reading frames (ORFs) with a canonical genomic organization that includes six proteins in the order 3ʹ-nucleocapsid protein (N) -phosphoprotein (P) -(putative) cell-to-cell movement protein -matrix protein (M) -glycoprotein (G) -polymerase (L) -5ʹ [5]. In this work, we report the molecular characterization of a novel and distinct cytorhabdovirus associated with yerba mate in Argentina, which we tentatively named yerba mate virus A (YmVA).