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Journal of Virology, April 2000, p. 3149-3155, Vol. 74, No. 7
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Complete Nucleotide Sequence and Genome Organization of Hibiscus Chlorotic Ringspot Virus, a New Member of the Genus Carmovirus: Evidence for the Presence and Expression of Two Novel Open Reading Frames

Mei Huang,1 Dora Chin-Yen Koh,2 Li-Juan Weng,2 Min-Li Chang,2 Yun-Kiam Yap,2 Lee Zhang,1 and Sek-Man Wong2,*

Institute of Molecular Agrobiology, Singapore 117604,1 and Department of Biological Sciences, The National University of Singapore, Singapore 117543,2 Republic of Singapore

Received 30 August 1999/Accepted 3 January 2000

The complete nucleotide sequence of hibiscus chlorotic ringspot virus (HCRSV) was determined. The genomic RNA (gRNA) is 3,911 nucleotides long and has the potential to encode seven viral proteins in the order of 28 (p28), 23 (p23), 81 (p81), 8 (p8), 9 (p9), 38 (p38), and 25 (p25) kDa. Excluding two unique open reading frames (ORFs) encoding p23 and p25, the ORFs encode proteins with high amino acid similarity to those of carmoviruses. In addition to gRNA, two 3'-coterminated subgenomic RNA (sgRNA) species were identified. Full-length cDNA clones derived from gRNA and sgRNA were constructed under the control of a T7 promoter. Both capped and uncapped transcripts derived from the full-length genomic cDNA clone were infectious. In vitro translation and mutagenesis assays confirmed that all the predicted ORFs except the ORF encoding p8 are translatable, and the two novel ORFs (those encoding p23 and p25) may be functionally indispensable for the viral infection cycle. Based on virion morphology and genome organization, we propose that HCRSV be classified as a new member of the genus Carmovirus in family Tombusviridae.

* Corresponding author. Mailing address: Department of Biological Sciences, The National University of Singapore, 10 Science Dr. 4, Singapore 117543, Republic of Singapore. Phone: 65-874-2976. Fax: 65-779-5671. E-mail: dbswsm{at}nus.edu.sg.

Journal of Virology, April 2000, p. 3149-3155, Vol. 74, No. 7
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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