title: Hibiscus latent Fort Pierce virus in Brazil and synthesis of its biologically active full-length cDNA clone.
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.
authors.affiliation: Department of Biological Sciences, National University of Singapore, Lower Kent Ridge Road, Singapore, 119543, Singapore.
authors.forename: Ruimin
authors.initials: R
authors.lastname: Gao
authors.affiliation: Department of Biological Sciences, National University of Singapore, Lower Kent Ridge Road, Singapore, 119543, Singapore.
authors.forename: Shengniao
authors.initials: S
authors.lastname: Niu
authors.affiliation: Summer Undergraduate Research Internship Program, University of Toronto, Toronto, ON, M5S 1A8, Canada.
authors.forename: Weifang
authors.initials: W
authors.lastname: Dai
authors.affiliation: Departamento de Fitopatologia e Nematologia, Escola Superior e Agricultura Luiz de Queiroz, Universidade de São Paulo, Piracicaba, SP, Brazil.
authors.forename: Elliot
authors.initials: E
authors.lastname: Kitajima
authors.affiliation: Department of Biological Sciences, National University of Singapore, Lower Kent Ridge Road, Singapore, 119543, Singapore. dbswsm@nus.edu.sg.|Temasek Life Sciences Laboratory, Singapore, Singapore. dbswsm@nus.edu.sg.|National University of Singapore Suzhou Research Institute, Suzhou Industrial Park, Jiangsu, China. dbswsm@nus.edu.sg.
authors.forename: Sek-Man
authors.initials: SM
authors.lastname: Wong
mesh_terms: D020413:3' Untranslated Regions / Q000235:genetics; D020121:5' Untranslated Regions / Q000235:genetics; D001483:Base Sequence; D001938:Brazil; D027465:Chenopodium quinoa / Q000821:virology; D003001:Cloning, Molecular / Q000379:methods; D018076:DNA, Complementary / Q000235:genetics*; D012321:DNA-Directed RNA Polymerases / Q000235:genetics; D016679:Genome, Viral / Q000235:genetics; D031584:Hibiscus / Q000821:virology*; D016366:Open Reading Frames / Q000235:genetics; D018515:Plant Leaves / Q000821:virology; D012367:RNA, Viral / Q000235:genetics; D017898:Tobamovirus / Q000235:genetics*; D014764:Viral Proteins / Q000235:genetics
chemical_list: D020413:3' Untranslated Regions; D020121:5' Untranslated Regions; D018076:DNA, Complementary; D012367:RNA, Viral; D014764:Viral Proteins; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases
keywords: Biologically active HLFPV cDNA clone; Brazil; Hibiscus latent Fort Pierce virus; In vitro transcripts