ISHS Acta Horticulturae 657: XIX International Symposium on Virus and Virus-like Diseases of Temperate Fruit Crops - Fruit Tree Diseases
NUCLEOTIDE SEQUENCE ANALYSIS AND DETECTION OF CHERRY RASP LEAF VIRUS |
| Author: | D. James |
| Keywords: | CRLV, RT-PCR, nucleotide sequence analysis |
| Abstract:
Cherry rasp leaf virus (CRLV) is transmitted by the nematode Xiphinema americanum and has been classified as a species of the genus Nepovirus, within the family Comoviridae. Nucleotide sequence analysis of the RNA-1 and RNA-2 components of the genome indicate that this bipartite virus is more closely related to Apple latent spherical virus (ALSV), an unclassified virus with no known vector, than to any definitive species of the genus Nepovirus. Analysis of the nucleotide sequence and deduced amino acid residues of CRLV (flat apple isolate) RNA-1 and RNA-2 support reclassification of CRLV and ALSV into a new genus within the family Comoviridae. The RNA-1 and RNA-2 fragments of the CRLV genome are approximately 7.0 kb and 3.3 kb in size, respectively. Oligonucleotide primers based on sequences from both fragments were designed to amplify fragments of similar sizes in RT-PCR analysis, 447 bp and 429 bp for RNA-1 and RNA-2, respectively. RT-PCR targeting the RNA-1 fragment appears to be slightly more sensitive than RT-PCR targeting the RNA-2 fragment, suggesting that target size does not affect the sensitivity of CRLV detection. | |
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