Articles
MOLECULAR CHARACTERIZATION OF PLUM POX VIRUS ISOLATED FROM APRICOTS, PLUMS AND PEACHES IN CHILE
Article number
472_47
Pages
401 – 406
Language
Abstract
PPV is the causal agent of sharka disease of stone fruits.
This pathogen has been identified in Europe, the Middle East and recently in South America.
In Chile, it was first detected in 1991 in a localized apricot orchard, producing the typical symptoms of this disease on the fruit.
The presence of PPV was confirmed later by serological and molecular diagnosis.
In this investigation, we show the results of the molecular characterization of PPV, isolated from apricot, plum and peach trees.
We amplified two segments of the PPV genome using specific primers, a 220 bp fragment from the 3′-non coding region (NCR) and a 243 bp fragment from the 5′-terminal of the coat protein gene.
These fragments were cloned and sequenced.
Nucleotide sequences showed the presence of a conserved RsaI site, characteristic of the PPV-D isolates, as well as an AluI site, present in almost all PPV isolates.
Comparing all the sequenced isolates, homology ranged from 92.6% to 99.2% for the CP-fragment, having the Chilean isolates 99% homology to the PPV-D strain.
The homology is reduced to 96–97% when the 3′-NCR region is included in the comparison with the D strain.
This evidence strongly supports that the Chilean isolate examined correspond to the PPV-D strain.
Additionally, we have found samples without symptoms of sharka disease that tested positive by ELISA and RT-PCR analyses.
So far, these symptomless samples have not yet been characterized.
This pathogen has been identified in Europe, the Middle East and recently in South America.
In Chile, it was first detected in 1991 in a localized apricot orchard, producing the typical symptoms of this disease on the fruit.
The presence of PPV was confirmed later by serological and molecular diagnosis.
In this investigation, we show the results of the molecular characterization of PPV, isolated from apricot, plum and peach trees.
We amplified two segments of the PPV genome using specific primers, a 220 bp fragment from the 3′-non coding region (NCR) and a 243 bp fragment from the 5′-terminal of the coat protein gene.
These fragments were cloned and sequenced.
Nucleotide sequences showed the presence of a conserved RsaI site, characteristic of the PPV-D isolates, as well as an AluI site, present in almost all PPV isolates.
Comparing all the sequenced isolates, homology ranged from 92.6% to 99.2% for the CP-fragment, having the Chilean isolates 99% homology to the PPV-D strain.
The homology is reduced to 96–97% when the 3′-NCR region is included in the comparison with the D strain.
This evidence strongly supports that the Chilean isolate examined correspond to the PPV-D strain.
Additionally, we have found samples without symptoms of sharka disease that tested positive by ELISA and RT-PCR analyses.
So far, these symptomless samples have not yet been characterized.
Authors
M. Rosales, P. Hinrichsen, G. Herrera
Keywords
potyvirus, detection, sequencing, PPV, RT-PCR, Prunus spp.
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