Abstract
An orthotospovirus distinct from all other orthotospoviruses was isolated from naturally infected alstroemeria plants. Disease symptoms caused by this virus mainly consisted of yellow spots on the leaves based on which the name alstroemeria yellow spot virus (AYSV) was coined. A host range analysis was performed and a polyclonal antiserum was produced against purified AYSV ribonucleoproteins which only reacted with the homologous antigen and not with any other (established or tentative) orthotospovirus from a selection of American and Asian species. Upon thrips transmission assays the virus was successfully transmitted by a population of Thrips tabaci. The entire nucleotide sequence of the M and S RNA segments was elucidated by a conventional cloning and sequencing strategy, and contained 4797 respectively 2734 nucleotides (nt). Simultaneously, a next generation sequencing (NGS) approach (RNAseq) was employed and generated contigs covering the entire viral tripartite RNA genome. In addition to the M and S RNA nucleotide sequences, the L RNA (8865 nt) was obtained. The nucleocapsid (N) gene encoded by the S RNA of this virus consisted of 819 nucleotides with a deduced N protein of 272 amino acids and by comparative sequence alignments to other established orthotospovirus species showed highest homology (69.5% identity) to the N protein of polygonum ringspot virus. The data altogether support the proposal of AYSV as a new orthotospovirus species within a growing clade of orthotospoviruses that seem to share the Middle East basin as a region of origin.
Highlights
Members of the Tospoviridae continuously cause significant losses in both mono- and dicot crops worldwide [1, 2]
From 31 plant species mechanically inoculated with Als2000, from here onwards referred to as alstroemeria yellow spot virus (AYSV), 17 species did not show any sign of infection, 9 species only showed local symptoms while 5 species displayed local and systemic symptoms (Table 1)
A polyclonal antiserum has been raised against AYSV Ribonucleocapsid protein (RNP) and only reacts positively with the homologous virus and not with any other established orthotospovirus species tested
Summary
Members of the Tospoviridae continuously cause significant losses in both mono- and dicot crops worldwide [1, 2]. Ever since TSWV re-emerged in the 1980’s and started its world wide spread by the introduction and spread of Frankliniella occidentalis, new viral species have been discovered. This has led to the identification of a current total of 29 orthotospovirus species of which 11 are officially recognized by the ICTV while the remaining, listed as tentative species [2, 20,21,22,23,24,25,26]. Preliminary serological tests with available antisera to a wide range of different orthotospoviruses were negative and indicated the possible emergence of a new orthotospovirus species. The nucleotide sequence of the entire genome was elucidated using a simultaneous conventional and NGS cloning/ sequencing approach
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