We have identified a tobamovirus, isolated from pepper (Capsicum annuum cv. Tosajishi), as a Japanese isolate of Tobacco mild green mosaic virus (TMGMV-J). A cDNA clone of TMGMV-J from which infectious RNA transcripts can be synthesized in vitro was constructed. The RNA transcript was infectious to several plant species tested here including C. annuum cv. Tosajishi and Nicotiana benthamiana plants. No viral RNA accumulation was observed in either tomato plants or protoplasts inoculated with transcript or virion RNA, indicating that tomato plants were immune to TMGMV. This is the first report of a biologically active TMGMV cDNA clone and direct evidence of the presence of TMGMV in Japan. Key words Tobacco mild green mosaic virus · Tobamovirus · cDNA clone · Host range · TomatoTobacco mild green mosaic virus (TMGMV) is a member of the genus Tobamovirus, which includes Tobacco mosaic virus (TMV) as its type member. It has a genome consisting of a monopartite positive-sense single-stranded RNA (Wetter 1986). TMGMV is known as a strain of TMV and includes U2-TMV (TMGMV-U2), which causes mild symptoms on tobacco (Wetter 1986). Although the complete nucleotide sequence of the genomic RNA of TMGMV-U2 has been determined (Soils and García-Arenal 1990), no biologically active cDNA clones have been reported for TMGMV.We have isolated a tobamovirus from field grown pepper (Capsicum annuum cv. Tosajishi) in Kochi Prefecture in Japan. The virus was identified as TMGMV by sequencing a cDNA clone of the virus (this paper and accession no. AB078435). We named this virus TMGMV-J, a Japanese strain of TMGMV. To obtain a gene manipulation system for TMGMV, we constructed a cDNA clone of TMGMV-J from which infectious transcripts can be synthesized in vitro. The transcripts were infectious to C. annuum cv. Tosajishi, Nicotiana benthamiana, and Chenopodium quinoa but not to tomato plants. Our results indicate that early infection processes, including translation and RNA replication, comprise a primary step that limits TMGMV infection in tomato.First we determined nucleotide sequences at the 5Ј and 3Ј proximal regions of TMGMV-J genomic RNA by 5Ј-and 3Ј-RACE analysis (Frohman et al. 1988) as previously described (Mizumoto et al. 2002). For 3Ј-RACE, a poly(T)-adapter (5Ј-CGACTCGAGTCGACATCGAT 16 -3Ј) was used for the first-strand cDNA synthesis from polyadenylated viral RNA. The cDNA was amplified by polymerase chain reaction (PCR) using a set of primers of a modified adapter MA (5Ј-CGACTCGAGTCGACATCGA-3Ј) and a primer 5Ј-GGAATTCCAGCTGATCAATCTGTGT ACG-3Ј (nt 5732-5752) harboring an EcoRI site (shown in italics). The PCR products digested with EcoRI and SacI were cloned into pUC118 and sequenced with an automated DNA sequencer (model 373; Applied Biosystems, Foster City, CA, USA). For 5Ј-RACE, first-strand cDNAs synthesized with a primer TG758 5Ј-AAGCGTAACCTC CGTCTG-3Ј (nt 785-802) were polyadenylated and then amplified with the poly(T)-adapter and TG785. PCR
Ocular complications of chronic active Epstein-Barr virus infection (CAEBV) have rarely been reported. We describe a 7-year-old girl with CAEBV with associated uveitis. The patient was first observed to have recurrent fever and hepatos-plenomegaly in August 1991. She presented with left facial nerve palsy in June 1993. Ocular examination showed right iridocyclitis. Both optic disks were swollen, and the retinal vessels were dilated. Antibody titers to EBV were markedly elevated. Treatment with topical steroids, systemic interleukin-2 and splenectomy dramatically relieved all her symptoms, including the ocular ones.
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