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Določanje in identifikacija bakterije Erwinia amylovora (Burrill) Winslow et al. v okuženi hruški = Detection and identification of Erwinia amylovora (Burrill) Winslow et al. in infected pear tree
Štebih, Dejan ...
In the frame of systematic survey, conducted by official plant protection service in Slovenia, National Institute of Biology performs laboratory diagnosis of bacteria Erwinia amylovora (Burrill) ...Winslow et al., the causal agent of fire blight. Frequently, samples with characteristic symptoms arrive to the laboratory. However, similar symptoms can be caused by other plant pathogens such as Pseudomonas syringae pv. syringae which has already been demonstrated in the past. In 2001, Erwinia amylovora was detected in a pear sample with symptoms of fire blight for the first time in Slovenia. Different methods that can be used for detection of Erwinia amylovora were used on infected material with and without symptoms. Isolation on media (KB, NSA), pathogenicity test on pear slices, indirect immunofluorescence (IIF), agglutination test and PCR with different sets of primers were performed. Isolated bacteria were sent for confirmation with analysis of fatty acids profile to Plant Protection Service in Wageningen, the Netherlands. As expected, Erwinia amylovora was detected with IIF test in all samples with symptoms, where concentration of bacteria is high but also in asymptomatic samples with lower concentration of bacteria that have been collected from thesame pear tree. Other tests were less reliable particularly in case of latent infection. We established, that the DNA extraction using DNeasyTM Tissue Kit reduces inhibition in PCR reaction compared to extraction by boiling of the sample. Avoiding inhibition is essential, particularly when theconcentration of bacteria in sample is low. For reliable and accurate laboratory determination of Erwinia amylovora, the use of several properly combined detection methods is recommended.
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