Survival of Erwinia amylovora in mature apple fruit calyces through the viable but nonculturable (VBNC) state
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Survival of Erwinia amylovora, causal agent of fire blight in pome fruits and other rosaceous plants, was monitored inside mature apples calyces under some storage conditions utilized in fruit. Apple fruit calyces inoculated with two E. amylovora strains and their respective GFP-marked strains were maintained at 26 degrees and 5 degrees C, and the effect of copper treatment was assayed at 0.01 and 0.1 mmol l(-1) CuSO(4). In nontreated apples at 26 degrees C, part of the population of E. amylovora survived in the 'viable but nonculturable' (VBNC) state, whereas at 5 degrees C the majority of the population retained culturability. In copper-treated apples, the whole population adopted the VBNC state irrespective of temperature. Regardless of temperature, copper and inoculum dose, VBNC cells recovered culturability and pathogenicity in King's B broth or by host plant passage. Erwinia amylovora survived for at least 35 days in mature apple calyces. Besides, the ability of the pathogen in the VBNC state to regain culturability and pathogenicity suggests that the apple fruit could be a potential carrier of E. amylovora contributing to the spreading of fire blight disease. The risk of E. amylovora dissemination through mature fruit transport, although low, has been demonstrated, and should be considered in pest risk assessments.