Novel Detection Protocol for Erwinia amylovora in Orchard Soil after Removal of Infected Trees.

ABSTRACT

Fire blight is a bacterial disease caused by Erwinia amylovora. In Korea, fire blight was first reported in 2015 in an orchard. If the infection is confirmed, all trees in the orchard must be removed and the orchard must remain closed for 3 years. Since 2020, if the number of trees infected with fire blight is less than 5% of the total trees in the orchard, only the infected tree and adjacent trees are removed in Korea. Three years after removal, the trees can be replanted after confirming that the orchard soil is free from E. amylovora. In this study, a protocol was established for detecting E. amylovora in soil via selective enrichment, using tryptic soy broth with 0.05% bile salts and 50 µg/ml cycloheximide, and real-time polymerase chain reaction. This protocol resulted in a 1,000-times improved detection limit for E. amylovora in soil samples compared to that in unenriched samples. Soil monitoring was performed for orchards where fire blight-infected trees had been removed 3-27 months prior; the selected orchards were monitored every 3 months. Monitoring confirmed that E. amylovora was not present in the soil at any site in any of the orchards. A new detection protocol facilitates the monitoring of E. amylovora in soil and could help permit the replanting of trees in orchards. Also monitoring results provide evidence that trees can be planted earlier.