Occurrence of Chilli veinal mottle virus in Nicotiana tabacum in Yunnan, China

Abstract

Flue-cured tobacco (Nicotiana tabacum) is an important crop in Yunnan Province, China. During a survey in July 2010, tobacco plants (N. tabacum cv. Yunyan 85) in the fields near Dali County in the northwest Yunnan Province of China had symptoms of chlorosis along leaf veins and later showed symptoms of white or brown necrosis along the veins. In 10 surveyed fields in Baizhishu Village in the city of Weishan, a commercial tobacco field (10 ha) developed virus-like disease symptoms; the incidence of affected plants ranged from 0.5 to 3%, which caused obvious economic losses. An isolate (YN75) was collected at random from five symptomatic leaves sampled from five plants. Negative staining of crude extracts of the infected leaves and subsequent electron microscopy revealed flexuous rods of 12 to 13 × 750 nm. Pinwheel-like inclusion bodies were abundant in thin sections of infected leaves. The particle size suggested a species of Potyviridae. Thus, the isolate was assayed in double antibody sandwich-ELISA (Agdia, Elkhart, IN) for the presence of Potato virus Y, Tobacco etch virus, and Tobacco vein mottling virus. All antigens gave negative results. Total RNA was extracted from leaves and tested by reverse transcription (RT)-PCR. The primer M4-T (5'-GTT TTC CCA GTC ACG ACT TTT TTT TTT TTT TT-3') was employed for cDNA synthesis described by Chen et al (1). The primer set ChiVMV-F (5'-TAG TTG YGC ATA C (C/G) C AGG AGA GAG-3')/M4 (5'-GTT TTC CCA GTC ACG AC-3') is complimentary to the region of coat protein and 3'-untranslated region of Chilli veinal mottle virus (ChiVMV), respectively. The expected 1,189-bp fragments were amplified from RNA templates and the amplicon was cloned and sequenced (GenBank Accession No. HQ218936). Comparisons of amplicons with the amino acid sequence available in the NCBI database using BLAST showed 91.4% identity with ChiVMV from India (GenBank Accession No. EF213688) and 90.7% with ChiVMV from Taiwan (Accession No. DQ854950). The virus particle size, RT-PCR results, and sequence data revealed that these tobacco plants were infected by ChiVMV. To our knowledge, this is the first report of ChiVMV infecting N. tabacum in China. Reference: (1) J. Chen et al. Arch. Virol. 146:757, 2001.