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First Report of Chilli veinal mottle virus Infecting Lycopersicon esculentum in Jiangsu Province in China

Zhu, F., Che, Y.-P., Qian, K., Zhou, Y.-K., Xu, Y.-J., Zhao, Y.-Q., Ji, Z.-L.
Plant disease 2018 v.102 no.2 pp. 462
Capsicum, Chilli veinal mottle virus, Nicotiana tabacum, RNA, Solanum lycopersicum var. lycopersicum, Western blotting, coat proteins, complementary DNA, computer software, consensus sequence, leaves, nucleotides, plant diseases and disorders, plant viruses, polyclonal antibodies, protocols, reverse transcriptase polymerase chain reaction, reverse transcription, sap, sequence alignment, tobacco, tomatoes, viruses, China
Chilli veinal mottle virus (ChiVMV) belongs to the genus Potyvirus, family Potyviridae, infects mostly Capsicum sp., and is aphid-transmitted in a nonpersistent manner in the field. This virus is widespread and has been reported in Hainan Province (Wang et al. 2006), Yunnan Province (Ding et al. 2011), and Sichuan Province (Zhao et al. 2014) in China. In June 2016, virus-like symptoms such as mottling, mosaic, and narrowing were observed on tomato (Lycopersicon esculentum) plants in Yangzhou city, Jiangsu Province, east China. Five leaf samples were collected from symptomatic tomato plants. All samples were assayed by western blotting using polyclonal antiserum to ChiVMV. The results suggest that two of five samples were positive for ChiVMV. Total RNA was extracted from leaf samples of five symptomatic tomato plants using Trizol reagent (Invitrogen, Carlsbad, CA), according to the manufacturer’s protocol. The first-strand cDNA was prepared using M-MLV reverse transcription (Invitrogen). Polymerase chain reaction (PCR) was performed using primers identical to nucleotides 8,969 to 8,985 (ChiVMV-F: 5′-GGATTGATGGTTTGGTG-3′) and complementary to nucleotides 9,202 to 9,219 (ChiVMV-R: 5′-CGCGCTAATGACATATCG-3′), which were designed to amplify the partial coat protein region of the ChiVMV (accession no. GQ981316). The expected 251-bp amplicon was amplified from the two symptomatic leaf samples that were positive for ChiVMV in western blotting assay using 35 cycles. The obtained products were purified with a TIANgel Midi purification kit (Tiangen, Beijing, China), cloned into the pMD19-T vector (TaKaRa, Dalian, China), and sequenced (Genewiz, Suzhou, China). The sequence alignment was performed using DNAMAN software. The results showed that the two isolate sequences were identical. Therefore, only one of the sequences was submitted to GenBank (accession no. MF537352) and thus represents a consensus sequence. Nucleotide BLAST analysis revealed that the sequence showed 99% nucleotide identity with a ChiVMV isolate available in GenBank (accession no. GQ981316). The virus was mechanically transmitted by inoculating sap from symptomatic tomato leaves to Nicotiana tabacum, which showed mottle, spots, and vein banding on leaves. This virus was also mechanically inoculated onto tomato by inoculating sap from symptomatic N. tabacum leaves. The same mottle and leaf distortion symptoms in tomato systemic leaves were observed. ChiVMV was also confirmed in tobacco and tomato by reverse-transcription PCR from total RNA extracted from tobacco or tomato leaves with the aforementioned primers. To our knowledge, this is the first report of ChiVMV in tomato in Jiangsu Province in China.