Construction of an infectious cDNA clone of the apple stem grooving capillovirus (isolate Li-23) genome containing a cauliflower mosaic virus 35S RNA promoter
1997
Terauchi, H. (Iwate Univ., Morioka (Japan). Faculty of Agriculture) | Magome, H. | Yoshikawa, N. | Takahashi, T. | Inouye, N.
The complete nucleotide sequence of the genome of apple stem grooving capillovirus (ASGV) isolate Li-23 from lily was determined. The isolate Li-23 genome consists of 6496 nucleotides excluding poly(A) tail and contains two overlapping open reading frames (ORFs 1 and 2). ORF1 encodes a protein with a Mr 241,768 (242K) and ORF2, in a different frame within ORF1, encodes a Mr 36,179 (36K) protein. The number of nucleotides and the arrangement of ORFs of the isolate Li-23 genome are in perfect agreement with those of isolate P-209 from apple and isolate L from lily. Identities of the nucleotide sequence were 98.4% (Li-23/L) and 83.0% (Li-23/P-209) in whole genomes excluding poly(A) tail. A full-length cDNA clone (pCT35SF) of the isolate Li-23 genome was cloned into a plasmid vector containing cauliflower mosaic virus 35S RNA promoter which was designed to transcribe the isolate Li-23 genome from its first nucleotide. Mechanical inoculation of pCT35SF onto Chenopodium quinoa induced symptoms typical of isolate Li-23 infection. Virus particles, viral RNA and coat protein could be detected in pCT35SF-inoculated plants, demonstrating that pCT35SF is infectious
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