Molecular characterization of Erwinia Amylovora strains originated from pome fruits and indigenous plant in montenegro

Abstract

In the period from 2012-2015 plant samples with fireblight symptoms were collected from pome fruits and indigenous plant, in main fruitgrowing regions of Montenegro. After succesfull isolation, pathogenicity of the obtained strains was tested by artificial inoculation of immature pear fruits, variety Viljamovka. Hypersensitive reaction was tested on tobacco leaves, variety White Burley. Identification and genetic diversity studies were performed using several molecular techniques on 18 Erwinia amylovora strains originating from quince, pear, apple and hawthorn. Bacterial identity was confirmed by nested PCR in which all studied strains produced the expected amplification fragment of plasmid pEA29. To detect potential genetic variations in E. amylovora population, rep-PCR was conducted. Using REP, ERIC and BOX primers, in all three PCR reactions, differences between studied strains were detected, i.e. pear strains had different genetic profiles from all other studied strains, including reference strain. Genetic variability of selected E. amylovora strains was studied by RAPD-PCR as well. Both of the used random primers, CUGEA-3 and CUGEA-5, showed discriminatory potential by separating genetic profiles of pear strains from all other studied strains, including reference strain. This is the first study of genetic variability of E. amylovora in Montenegro.