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Establishment of a quadruplex real-time PCR assay to distinguish the fungal pathogens Diaporthe longicolla, D. caulivora, D. eres, and D. novem on soybean

Hosseini, Behnoush; Voegele, Ralf T. and Link, Tobias I (2021) Establishment of a quadruplex real-time PCR assay to distinguish the fungal pathogens Diaporthe longicolla, D. caulivora, D. eres, and D. novem on soybean. PLOS ONE, 16 (9), e0257225.

[thumbnail of journal.pone.0257225.pdf] PDF - Published Version - English
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Document available online at: https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0257225


Diaporthe species are fungal plant pathogens of many important crops. Seed decay is one of the most important diseases on soybean. It is caused by various species of the genus Diaporthe and responsible for significant economic damage. In central Europe the four species D. longicolla, D. caulivora, D. eres, and D. novem are considered the principal species of Diaporthe on soybean. Fast and accurate detection of these pathogens is of utmost importance. In this study four species-specific TaqMan primer-probe sets that can be combined into a quadruplex assay were designed based on TEF sequences. The specificity and efficiency of the primer-probe sets were tested using PCR products and genomic DNA from pure cultures of the four Diaporthe species and other soybean fungal pathogens. Our results indicate that the primer-probe sets DPCL, DPCC, DPCE, and DPCN allow discrimination of D. longicolla, D. caulivora, D. eres, and D. novem, respectively, and can be used to detect and quantify these four Diaporthe species in parallel using quadruplex real-time PCR. In addition, the quadruplex real-time PCR assay was evaluated on different plant materials including healthy and infected soybean seeds or seed lots, soybean stems, and soybean leaves. This assay is a rapid and effective method to detect and quantify Diaporthe species from samples relevant for disease control.

EPrint Type:Journal paper
Keywords:BOEL, BÖL, BOELN, BÖLN, EPS, FKZ 15EPS082, SoySound, Diaporthe, DPC, qPCR, Pflanzenschutz, Soja
Agrovoc keywords:
Glycine max
diagnostic techniques
Subjects: Crop husbandry > Production systems > Cereals, pulses and oilseeds
Crop husbandry > Crop health, quality, protection
Research affiliation: Germany > Federal Agency for Agriculture and Food - BLE - Protein Strategy
Germany > University of Hohenheim > Institute for Phytomedicine
DOI:https://doi. org/10.1371/journal.pone.0257225
Related Links:https://www.bundesprogramm.de, https://orgprints.org/cgi/search/advanced?addtitle%2Ftitle=&keywords=15EPS082&projects=BOEL&_order=bypublication&_action_search=Suchen
Deposited By: Link, Dr. Tobias
ID Code:42375
Deposited On:17 Sep 2021 09:51
Last Modified:17 Sep 2021 09:51
Document Language:English
Refereed:Peer-reviewed and accepted

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