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Datum
2021-12-31Schlagwort
630 Landwirtschaft, Veterinärmedizin InfektionErkennungEchtzeitbildverarbeitungQuantifizierungErbseWeizenWurzelfäuleMetadata
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Aufsatz
A Novel Real Time PCR Method for the Detection and Quantification of Didymella pinodella in Symptomatic and Asymptomatic Plant Hosts
Zusammenfassung
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 environment.
Zitierform
In: Journal of Fungi Volume 8 / Issue 1 (2021-12-31) eissn:2309-608XFörderhinweis
Gefördert durch den Publikationsfonds der Universität KasselZitieren
@article{doi:10.17170/kobra-202202225796,
author={Šišić, Adnan and Oberhänsli, Thomas and Baćanović-Šišić, Jelena and Hohmann, Pierre and Finckh, Maria Renate},
title={A Novel Real Time PCR Method for the Detection and Quantification of Didymella pinodella in Symptomatic and Asymptomatic Plant Hosts},
journal={Journal of Fungi},
year={2021}
}
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2022-02-22T13:05:34Z 2022-02-22T13:05:34Z 2021-12-31 doi:10.17170/kobra-202202225796 http://hdl.handle.net/123456789/13649 Gefördert durch den Publikationsfonds der Universität Kassel eng Namensnennung 4.0 International http://creativecommons.org/licenses/by/4.0/ pea foot and root rot Didymella pinodella Phoma medicaginis var. pinodella qPCR wheat asymptomatic infections 630 A Novel Real Time PCR Method for the Detection and Quantification of Didymella pinodella in Symptomatic and Asymptomatic Plant Hosts Aufsatz 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 environment. open access Šišić, Adnan Oberhänsli, Thomas Baćanović-Šišić, Jelena Hohmann, Pierre Finckh, Maria Renate doi:10.3390/jof8010041 Infektion Erkennung Echtzeitbildverarbeitung Quantifizierung Erbse Weizen Wurzelfäule publishedVersion eissn:2309-608X Issue 1 Journal of Fungi Volume 8 false 41
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