Retrospective survey of Dickeya fangzhongdai using a novel validated real-time PCR assay

Front Microbiol. 2024 Feb 13:14:1249955. doi: 10.3389/fmicb.2023.1249955. eCollection 2023.

Abstract

Dickeya fangzhongdai, an aggressive plant pathogen, causes symptoms on a variety of crops and ornamental plants including bleeding canker of Asian pear trees. Historical findings stress the need for a specific detection tool for D. fangzhongdai to prevent overlooking the pathogen or assigning it to general Dickeya spp. Therefore, a qualitative real-time PCR for specific detection of D. fangzhongdai has been developed and validated. The developed assay shows selectivity of 100%, diagnostic sensitivity of 76% and limit of detection with 95% confidence interval in plant matrices ranging from 311 to 2,275 cells/mL of plant extracts. The assay was successfully used in a retrospective survey of selected host plants of relevance to Europe and environmental niches relevant to D. fangzhongdai. Samples of potato tubers and plants, plants from the Malinae subtribe (apple, pear, quince, and Asian pear tree) and fresh surface water from Slovenia were analyzed. D. fangzhongdai was not detected in any plant samples, however, 12% of surface water samples were found to be positive.

Keywords: Dickeya; diagnostics; molecular testing; plant pathogen; real-time PCR; survey; water.

Grants and funding

This study was financed by the Slovenian Research and Innovation Agency (contract numbers: P4-0165 and Z1-3190) and by the Euphresco Phytosanitary ERA-NET project on taxonomy and epidemiology of Pectobacterium and Dickeya spp. in Europe, North America, and South Africa (2020-F-336). The plant material used in this study was collected by the Administration of the Republic of Slovenia for Food Safety, Veterinary and Plant Protection, of the Ministry of Agriculture and Environment and Phytosanitary Inspectorate.