Real-Time PCR for Diagnosing and Quantifying Co-infection by Two Globally Distributed Fungal Pathogens of Wheat

Araz S. Abdullah, Chala Turo, Caroline S. Moffat, Francisco J. Lopez-Ruiz, Mark R. Gibberd, John Hamblin, Ayalsew Zerihun

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Co-infections - invasions of a host-plant by multiple pathogen species or strains are common, and are thought to have consequences for pathogen ecology and evolution. Despite their apparent significance, co-infections have received limited attention; in part due to lack of suitable quantitative tools for monitoring of co-infecting pathogens. Here, we report on a duplex real-time PCR assay that simultaneously distinguishes and quantifies co-infections by two globally important fungal pathogens of wheat: Pyrenophora tritici-repentis and Parastagonospora nodorum. These fungi share common characteristics and host species, creating a challenge for conventional disease diagnosis and subsequent management strategies. The assay uses uniquely assigned fluorogenic probes to quantify fungal biomass as nucleic acid equivalents. The probes provide highly specific target quantification with accurate discrimination against non-target closely related fungal species and host genes. Quantification of the fungal targets is linear over a wide range (5000-0.5 pg DNA mu l(-1)) with high reproducibility (RSD

Original languageEnglish
Article number1086
Number of pages10
JournalFrontiers in Plant Science
Publication statusPublished - 9 Aug 2018

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