A Novel Real Time PCR Method for the Detection and Quantification of Didymella pinodella in Symptomatic and Asymptomatic Plant Hosts
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Publication date
2021-12-31ISSN
2309-608X
Abstract
Didymella pinodella is the major pathogen of the pea root rot complex in Europe. This wide host range pathogen often asymptomatically colonizes its hosts, making the control strategies challenging. We developed a real-time PCR assay for the detection and quantification of D. pinodella based on the TEF-1 alpha gene sequence alignments. The assay was tested for specificity on a 54- isolate panel representing 35 fungal species and further validated in symptomatic and asymptomatic pea and wheat roots from greenhouse tests. The assay was highly consistent across separate qPCR reactions and had a quantification/detection limit of 3.1 pg of target DNA per reaction in plant tissue. Cross-reactions were observed with DNA extracts of five Didymella species. The risk of cross contamination, however, is low as the non-targets have not been associated with pea previously and they were amplified with at least 1000-fold lower sensitivity. Greenhouse inoculation tests revealed a high correlation between the pathogen DNA quantities in pea roots and pea root rot severity and biomass reduction. The assay also detected D. pinodella in asymptomatic wheat roots, which, despite the absence of visible root rot symptoms, caused wheat biomass reduction. This study provides new insights into the complex life style of D. pinodella and can assist in better understanding the pathogen survival and spread in the environament.
Document Type
Article
Document version
Published version
Language
English
Subject (CDU)
632 - Plant damage, injuries. Plant diseases. Pests, organisms injurious to plants. Plant protection
Pages
18
Publisher
MDPI
Is part of
Journal of Fungi (JoF)
Citation
Šišić, Adnan, Thomas Oberhänsli, Jelena Baćanović-Šišić, Pierre Hohmann, and Maria Renate Finckh. 2021. "A Novel Real Time PCR Method For The Detection And Quantification Of Didymella Pinodella In Symptomatic And Asymptomatic Plant Hosts". Journal Of Fungi 8 (1): 41. doi:10.3390/jof8010041.
Program
Protecció Vegetal Sostenible
This item appears in the following Collection(s)
- ARTICLES CIENTÍFICS [2685]
Except where otherwise noted, this item's license is described as http://creativecommons.org/licenses/by/4.0/