Tomato spotted wilt virus infects spider lily plants in Australia
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Tomato spotted wilt virus (TSWV) was identified in green red-rimmed ringspots on spider lily (Hymenocallis spp.) leaves in Brisbane, Australia. Tospovirus-like particles were seen in thin sections of those lesions. RT-PCR using tospovirus-specific primers amplified a DNA fragment whose sequence matched TSWV S RNA. The virus caused symptoms in Nicotiana benthamiana following mechanical transmission and was confirmed as TSWV by RT-PCR.
KeywordsOrthotospovirus Ornamental plant Ringspots Hymenocallis spp. TSWV
Total RNA was extracted from ringspot lesions using RNeasy Plant Mini kit (Qiagen). Superscript One-Step RT-PCR system with Platinum Taq DNA polymerase (Thermo Fisher Scientific) was used following the manufacturer’s protocol and primer pair AM1-F and AM1-R using an annealing temperature of 50 °C (Hassani-Mehraban et al. 2016). A single amplicon of 763 bp was obtained, similar in size to the product obtained with TSWV RNA control (data not shown). Amplified DNA was purified using Wizard SV Gel and PCR clean up kit (Promega), cloned into pGEM-T easy vector (Promega) and transformed into Omnimax E. coli competent cells following the manufacturer’s protocols. Following colony PCR, four positive recombinant clones were grown overnight and plasmid DNA was extracted for Sanger sequencing using M13 forward and reverse primers at the Australian Genome Research Facility (Brisbane). The tospovirus infecting spider lily was identified as TSWV based on PCR amplification of a fragment of S RNA using generic American clade tospovirus-specific primers (Hassani-Mehraban et al. 2016) and on the nucleotide sequence of this fragment that was deposited as GenBank accession MG81238. In a Blast N search on NCBI, the TSWV S RNA fragment amplified from spider lily was 99% identical to S RNA sequences from 20 different TSWV accessions.
We conclude that TSWV is associated with the striking red-green ringspots on spider lily and that this isolate was mechanically transmissible to the known experimental host N. benthamiana where TSWV replicated, moved systemically and caused symptoms. To the best of our knowledge this is the first Australian report of TSWV infecting Hymenocallis species.
This research was jointly supported by the Queensland Department of Agriculture and Fisheries and the University of Queensland through the Queensland Alliance for Agriculture and Food Innovation. Reciprocal travel and collaboration was supported by FAPESP-UQ SPRINT grant 2017/50039-1. We thank the friendly volunteers at the Gardens for their interest and assistance in identifying the genus of spider lily.
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The authors have no conflicts of interest.
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This article does not contain any studies with human participants or animals performed by any of the authors.
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