Gao Ruimin, Niu Shengniao, Dai Weifang, Kitajima Elliot, Wong Sek-Man
Department of Biological Sciences, National University of Singapore, Lower Kent Ridge Road, Singapore, 119543, Singapore.
Summer Undergraduate Research Internship Program, University of Toronto, Toronto, ON, M5S 1A8, Canada.
Virus Genes. 2016 Oct;52(5):754-7. doi: 10.1007/s11262-016-1344-8. Epub 2016 May 2.
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.
巴西木槿潜隐福特皮尔斯病毒分离株(HLFPV-BR)首次在巴西圣保罗州利梅拉的一株木槿植物中被发现。进行了RACE PCR以获得HLFPV-BR的全长序列,该序列为6453个核苷酸,与HLFPV日本分离株的完整基因组RNA核苷酸序列同一性超过99.15%。HLFPV-BR的基因组结构与其他烟草花叶病毒相似。它包括一个5'非翻译区(UTR),随后是编码128 kDa蛋白和188 kDa通读蛋白的开放阅读框、一个38 kDa运动蛋白、18 kDa外壳蛋白以及一个3'UTR。有趣的是,在其3'-UTR内也发现了poly(A)尾的独特特征。此外,从HLFPV-BR感染的藜麦叶片局部病斑中提取的总RNA中,扩增出一个包含HLFPV-BR基因组的具有生物活性的全长cDNA克隆,并将其置于T7 RNA聚合酶启动子附近。克隆cDNA的加帽体外转录本在机械接种到藜麦和本氏烟草植株中时具有感染性。这是巴西存在HLFPV分离株以及成功合成具有生物活性的HLFPV-BR全长cDNA克隆的首次报道。