ISHS Acta Horticulturae 568: X International Symposium on Virus Diseases of Ornamental Plants
CLONING OF COMPLETE CARNATION MOTTLE VIRUS cDNA AND ITS INFECTIVITY |
| Authors: | A-P. Zhang, H-Q. Zhu, S-Q. Yu, L. Yang, Y. Lou |
| Keywords: | Shanghai isolate; full-length sequence; CaMV 35S promoter; plasmid pCass2; symptoms on Chenopodium |
| Abstract:
A full-length cDNA clone of the RNA genome of carnation mottle carmovirus Shanghai isolate (CarMV-sh) was obtained by ligating four fragments of cDNA together, and constructed downstream from partially doubled cauliflower mosaic virus 35S RNA promoter, generating a transient expression plasmid pCaCarMV derived from a pUC-based pCass2. By routine mechanical inoculation, local chlorotic lesions, similar to those induced by wild-type carnation mottle virus, or its native RNA, were observed on inoculated leaves of Chenopodium amaranticolor and C. murale. The appearance of symptoms in leaves inoculated with pCaCarMV was slightly delayed for 1-2 days compared to leaves inoculated with wild-type virus or its native RNA. The progeny virions recovered from infected leaves had the same morphological properties as the parental virus. The progeny virions also reacted positively with CarMV antiserum in double diffusion on agarose gel and indirect enzyme-linked immunosorbent assay (ELISA). This may be the first case of an infectious full-length cDNA clone of carmovirus members. Successful construction of infectious cDNA clone of carnation mottle virus makes it possible to develop a molecular tool used in better understanding of gene strategy of this virus, and a virus-based vector for gene transfer into and expression in cytoplasmic system . | |
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