Articles
AMERICAN PLUM LINE PATTERN VIRUS IS A DISTINCT ILARVIRUS
Article number
550_31
Pages
221 – 228
Language
English
Abstract
The complete nucleotide sequence for the genome of an isolate of American plum line pattern virus (APLPV) was determined.
The genomic segments resemble those of other ilarviruses in both size and organization.
Like the majority of other ilarviruses that infect fruit trees (Prunus necrotic ringspot virus [PNRSV], Prune dwarf virus [PDV], Apple mosaic virus [ApMV]), the virus does not possess a 2b ORF. Previous serological analyses had shown no relationship between APLPV and these other ilarviruses.
This lack of relationship was supported when putative translations of the coat protein genes of the respective viruses were compared.
Sequence data for the RNA 3 of APLPV were used to design pairs of primers for detection of the virus using RT-PCR. The product from a reaction using purified viral RNA was cloned into pCR®II-TOPO® (Invitrogen) and used to generate an antisense cRNA probe.
The probe was labelled with digoxigenin and used to detect the virus using a chemiluminescent substrate (CDP-Star™). Both PCR and tissue blots are being evaluated for the detection of APLPV in plant tissues in order to provide an alternative to the systems currently used for the detection of the virus in quarantine programs and certification schemes.
The sequences of the genomic molecules RNA 1, RNA 2, and RNA 3 have been deposited in GenBank as accessions AF235033, AF235165, and AF235166, respectively
The genomic segments resemble those of other ilarviruses in both size and organization.
Like the majority of other ilarviruses that infect fruit trees (Prunus necrotic ringspot virus [PNRSV], Prune dwarf virus [PDV], Apple mosaic virus [ApMV]), the virus does not possess a 2b ORF. Previous serological analyses had shown no relationship between APLPV and these other ilarviruses.
This lack of relationship was supported when putative translations of the coat protein genes of the respective viruses were compared.
Sequence data for the RNA 3 of APLPV were used to design pairs of primers for detection of the virus using RT-PCR. The product from a reaction using purified viral RNA was cloned into pCR®II-TOPO® (Invitrogen) and used to generate an antisense cRNA probe.
The probe was labelled with digoxigenin and used to detect the virus using a chemiluminescent substrate (CDP-Star™). Both PCR and tissue blots are being evaluated for the detection of APLPV in plant tissues in order to provide an alternative to the systems currently used for the detection of the virus in quarantine programs and certification schemes.
The sequences of the genomic molecules RNA 1, RNA 2, and RNA 3 have been deposited in GenBank as accessions AF235033, AF235165, and AF235166, respectively
Publication
Authors
S.W. Scott, M.T. Zimmerman
Keywords
American plum line pattern, Ilarvirus, Sequence, PCR
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