The use of molecular biology methods for detection of phytopathogenic bacteria: the study case of Erwinia amylovora (Burrill) Winslow et al.
2001
Demsar, T. | Petrovic, N. | Stebih, D. | Dreo, T. | Blatnik, A. | Ravnikar, M. (National Inst. of Biology, Ljubljana (Slovenia))
Fireblight is a quarantine bacterial disease on fruit trees and ornamental plants recorded in most European countries. The fact that fireblight is present in all countries neighbouring Slovenia represents a very high possibility of it's introduction to Slovenia, which is a serious treat to Slovenian fruit growing, especially production of pears and apples. We have introduced a laboratory testing scheme for a reliable detection of Erwinia amylovora (Ea), the bacterial agent causing fireblight disease. A testing procedure reccomended by EPPO (European Plant Protection Organization) includes: isolation of Ea by plating on semiselective and non-selective media, serological methods (agglutination and immunofluorescence) and pathogenicity test on young pear fruits. As an addition to all these classical laboratory tests we have introduced a molecular biology based method, which both confirms the Ea in a bacterial culture, and enables to distinguish Ea from other bacteria such as Pseudomonas syringae pv. syringae, a causal agent of a disease similar to fireblight regarding the symptoms, and from other frequently present bacteria on fruit trees. The results of PCR testing with the use of different primers, which amplify bacterial DNA, and their use for a routine testing scheme of Ea will be discussed.
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