ISHS Acta Horticulturae 657: XIX International Symposium on Virus and Virus-like Diseases of Temperate Fruit Crops - Fruit Tree Diseases
PEAR DECLINE PHYTOPLASMA DETECTION IN PEAR TREES AND INSECT VECTORS BY DOT-BLOT HYBRIDIZATION AND NESTED-PCR |
| Authors: | M. García-Chapa, A. Batlle, A. Laviņa, L. Galipienso, G. Firrao |
| Keywords: | Pear decline phytoplasma, nested-PCR, dot-blot hybridization, Cacopsylla pyri, non-ribosomal probes |
| Abstract:
A non-isotopic hybridisation procedure was used to detect pear decline (PD) phytoplasma. Two new digoxigenin labelled DNA probes obtained from putative thymidilate kinase and peptide release genes of the PD phytoplasma were used. Hybridisations of these non-ribosomal probes with dot-blotted total nucleic acid extracts on nylon membranes allowed the detection of PD phytoplasma in infected plants and individual Cacopsylla pyri vectors. Assays to compare dot-blot hybridisation and nested-PCR procedures showed that the dot-blot hybridisation is a more reliable assay than nested-PCR. We propose the use of dot-blot hybridisation as a suitable technique for specific PD detection in epidemiological studies where there are a large number of samples to be tested. | |
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