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High-Resolution Melting (HRM) Curve Assay for the Identification of Eight Fusarium Species Causing Ear Rot in Maize

dc.creatorSchiwek, Simon
dc.creatorBeule, Lukas
dc.creatorViñas Meneses, María
dc.creatorPfordt, Annette
dc.creatorvon Tiedemann, Andreas
dc.creatorKarlovsky, Petr
dc.date.accessioned2021-02-01T13:49:48Z
dc.date.available2021-02-01T13:49:48Z
dc.date.issued2020
dc.description.abstractMaize plants are often infected with fungal pathogens of the genus Fusarium. Taxonomic characterization of these species bymicroscopic examination of pure cultures or assignment to mating populations is time-consuming and requires specific expertise. Reliable taxonomic assignment may be strengthened by the analysis of DNA sequences. Species-specific PCR assays are available for most Fusarium pathogens, but the number of species that infect maize increases the labor and costs required for analysis. In this work, a diagnostic assay for major Fusarium pathogens of maize based on the analysis of melting curves of PCR amplicons was established. Short segments of genes RPB2 and TEF-1 , which have been widely used in molecular taxonomy of Fusarium, were amplified with universal primers in a real-time thermocycler and high-resolution melting (HRM) curves of the products were recorded. Among major Fusarium pathogens of maize ears, F. cerealis, F. culmorum, F. graminearum, F. equiseti, F. poae, F. temperatum, F. tricinctum, and F. verticillioides, all species except for the pair F. culmorum/F. graminearum could be distinguished by HRM analysis of a 304 bp segment of the RPB2 gene. The latter two species could be differentiated by HRM analysis of a 247 bp segment of the TEF-1 gene. The assay was validated with DNA extracted from pure cultures of fungal strains, successfully applied to total DNA extracted from infected maize ears and also to fungal mycelium that was added directly to the PCR master mix (“colony PCR”). HRM analysis thus offers a cost-efficient method suitable for the diagnosis of multiple fungal pathogens.es_ES
dc.description.procedenceUCR::Vicerrectoría de Investigaciónes_ES
dc.description.procedenceUCR::Vicerrectoría de Investigación::Unidades de Investigación::Ciencias Agroalimentarias::Centro para Investigaciones en Granos y Semillas (CIGRAS)es_ES
dc.description.sponsorshipGerman Federal O ce for Agriculture and Food/[2818208315]/BLE/Alemaniaes_ES
dc.description.sponsorshipGerman Federal Ministry of Education and Research/[031A562]/BMBF/Alemaniaes_ES
dc.description.sponsorshipGerman Federal Ministry of Education and Research/[031B0510A]/BMBF/Alemaniaes_ES
dc.identifier.citationhttps://www.mdpi.com/2076-0817/9/4/270
dc.identifier.doi10.3390/pathogens9040270
dc.identifier.issn2076-0817
dc.identifier.urihttps://hdl.handle.net/10669/82679
dc.language.isoen_USes_ES
dc.rightsacceso abiertoes_ES
dc.sourcePathogens, vol.9(4), pp.270es_ES
dc.subjectFusariumes_ES
dc.subjectHRM analysises_ES
dc.subjectRPB2es_ES
dc.subjectTEF-1es_ES
dc.subjectHigh-resolution melting (HRM) curveses_ES
dc.subjectMaize ear rotes_ES
dc.subjectFungal colony PCRes_ES
dc.titleHigh-Resolution Melting (HRM) Curve Assay for the Identification of Eight Fusarium Species Causing Ear Rot in Maizees_ES
dc.typeartículo originales_ES

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