ISHS Acta Horticulturae 386: XVI International Symposium on Fruit Tree Virus diseases
DETECTION OF PRUNE DWARF ILARVIRUS FROM INFECTED STONE FRUITS USING REVERSE TRANSCRIPTION-POLYMERASE CHAIN REACTION |
| Authors: | D.R. Parakh, A.M. Shamloul, A. Hadidi, H.E. Waterworth, S.W. Scott, H.E. Howell, G.I. Mink |
| Abstract:
Prune dwarf ilarvirus (PDV) was detected in imported and domestic stone fruit germplasm by RT-PCR assay using DNA primers for the viral coat protein region. The expected size of the amplified products was 172 bp. Samples (10–20 mg/sample) from bark, young green bud and/or base of leaf petiole were used. Samples were extracted in TE buffer (10 mM Tris, 1 mM EDTA, pH 8.0) and then treated with GeneReleaser. Similar samples were treated with phenol/chloroform and the isolated nucleic acids were either directly treated with GeneReleaser or concentrated before treating with GeneReleaser. Treatment of sap extract in TE buffer with GeneReleaser was suitable for PDV detection from leaf tissue by RT-PCR. However, nucleic acid extraction was required for sensitive detection of the virus from bark or bud tissue. The detection of PDV by RT-PCR was correlated with biological and/or ELISA assays. RT-PCR assays were successful in the detection of several isolates of PDV and appear useful for testing stone fruit germplasm in quarantine or budwood certification programs. | |
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