Abstract
A Brazilian isolate of Hibiscus latent Fort Pierce virus (HLFPV-BR) was firstly found in a hibiscus plant in Limeira, SP, Brazil. RACE PCR was carried out to obtain the full-length sequences of HLFPV-BR which is 6453 nucleotides and has more than 99.15 % of complete genomic RNA nucleotide sequence identity with that of HLFPV Japanese isolate. The genomic structure of HLFPV-BR is similar to other tobamoviruses. It includes a 5′ untranslated region (UTR), followed by open reading frames encoding for a 128-kDa protein and a 188-kDa readthrough protein, a 38-kDa movement protein, 18-kDa coat protein, and a 3′ UTR. Interestingly, the unique feature of poly(A) tract is also found within its 3′-UTR. Furthermore, from the total RNA extracted from the local lesions of HLFPV-BR-infected Chenopodium quinoa leaves, a biologically active, full-length cDNA clone encompassing the genome of HLFPV-BR was amplified and placed adjacent to a T7 RNA polymerase promoter. The capped in vitro transcripts from the cloned cDNA were infectious when mechanically inoculated into C. quinoa and Nicotiana benthamiana plants. This is the first report of the presence of an isolate of HLFPV in Brazil and the successful synthesis of a biologically active HLFPV-BR full-length cDNA clone.
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Acknowledgments
This study was funded by the National University of Singapore (NUS) research grant R-154-000-655-112.
Author contributions
Conceived and designed the study: RMG, EK, and SMW. Performed the experiments: RMG, SNN, and WFD. Analyzed the data: RMG, SNN, WFD, EK, and SMW. Wrote and revised the manuscript: RMG, EK, and SMW.
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Edited by Seung-Kook Choi.
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11262_2016_1344_MOESM1_ESM.tif
Supplementary Fig. 1 Viral symptoms of H. rosa sinensis and transmission electron micrographs of HLFPV-BR. A and B The leaves of HLFPV-BR infected H. rosa sinensis showing chlorotic spots and mottling. C and D Rod-shaped HLFPV-BR viruses were purified from infected Nicothana benthaminan leaves.(TIFF 4318 kb)
11262_2016_1344_MOESM2_ESM.tif
Supplementary Fig. 2 Schematic diagram of construction of the full-length cDNA clone of HLFPV-BR isolate. The grey circle represents the backbone of the pBluescript SK (+) vector. The restriction enzyme sites of SalI and SacI are presented as bars in green and purple, respectively. The T7 RNA polymerase promoter is presented as a red arrow.(TIFF 130 kb)
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Gao, R., Niu, S., Dai, W. et al. Hibiscus latent Fort Pierce virus in Brazil and synthesis of its biologically active full-length cDNA clone. Virus Genes 52, 754–757 (2016). https://doi.org/10.1007/s11262-016-1344-8
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DOI: https://doi.org/10.1007/s11262-016-1344-8