20.500.12556/DiRROS-19651 High-throughput sequencing facilitates characterisation of a ʺforgottenʺ plant virus the case of a henbane mosaic virus infecting tomato High-throughput sequencing has dramatically broadened the possibilities for plant virus research and diagnostics, enabling discovery of new or obscure viruses, and virus strains and rapid sequencing of their genomes. In this research, we employed high-throughput sequencing to discover a new virus infecting tomato, Henbane mosaic virus (Potyvirus, Potyviridae), which was first discovered at the beginning of 20th century in the United Kingdom in cultivated henbane. A field tomato plant with severe necrotic symptoms of unknown etiology was sampled in Slovenia and high-throughput sequencing analysis using small RNA and ribosomal RNA depleted total RNA approaches revealed a mixed infection with Potato virus M (Carlavirus, Betaflexiviridae), Southern tomato virus (Amalgavirus, Amalgamaviridae) and henbane mosaic virus in the sample. The complete genomic sequence of henbane mosaic virus was assembled from the sequencing reads. By re-inoculation of the infected material on selected test plants, henbane mosaic virus was isolated and a host range analysis was performed, demonstrating the virus was pathogenic on several plant species. Due to limited metadata in public repositories, the taxonomic identification of the virus isolate was initially putative. Thus, in the next step, we used small RNA sequencing to determine genomic sequences of four historic isolates of the virus, obtained from different virus collections. Phylogenetic analyses performed using this new sequence information enabled us to taxonomically position Henbane mosaic virus as a member of the Potyvirus genus within the chili veinal mottle virus phylogenetic cluster and define the relationship of the new tomato isolate with the historic ones, indicating the existence of at least four putative strains of the virus. The first detection of henbane mosaic virus in tomato and demonstration of its pathogenicity on this host is important for plant protection and commercial tomato production. Since the virus was initially present in a mixed infection, and its whole genome was not sequenced, it has probably been overlooked in routine diagnostics. This study confirms the applicability of a combination of high-throughput sequencing and classic plant virus characterization methods for identification and phylogenetic classification of obscure viruses and historical viral isolates, for which no or limited genome sequence data is available. henbane mosaic virus Tomato high-throughput sequencing host range analysis phylogeny Potyvirus true false true Angleški jezik Ni določen Neznano 2024-07-23 03:54:59 2024-07-24 13:34:01 2024-07-25 04:14:52 0000-00-00 00:00:00 2018 0 0 Nasl. z nasl. zaslona; Opis vira z dne 26. 10. 2018; str. 1-11 Vol. 9 2018 0000-00-00 Zaloznikova Objavljeno NiDoloceno 0000-00-00 0000-00-00 2018-11-19 578 1664-302X 10.3389/fmicb.2018.02739 4877391 https://www.frontiersin.org/articles/10.3389/fmicb.2018.02739/abstract 1 ce216432-4885-11ef-b487-001a4af901a5 https://dirros.openscience.si/Dokument.php?lang=slv&id=27143 RAZ_Pecman_Anja_2018.pdf RAZ_Pecman_Anja_2018.pdf 1 003CD9BAD2BD42C0200291C548BA20A6 493a94ed0cc0b79a42cf633443a29ae2eef7000c56ce495c2cad9bb5651b07d0 e79334d8-499f-11ef-b487-001a4af901a5 https://dirros.openscience.si/Dokument.php?lang=slv&id=27330 Nacionalni inštitut za biologijo 0 0 0