Novel Multiplex and Loop-Mediated Isothermal Amplification Assays for Rapid Species and Mating-Type Identification of <i>Oculimacula acuformis</i> and <i>O. yallundae</i> (Causal Agents of Cereal Eyespot), and Application for Detection of Ascospore Dispersal and In Planta Use

Eyespot, caused by the related fungal pathogens Oculimacula acuformis and O. yallundae, is an important cereal stem-base disease in temperate parts of world. Both species are dispersed mainly splash-dispersed conidia but also known to undergo sexual reproduction, yielding apothecia containing ascospores. Field diagnosis eyespot can be challenging, with other causing similar symptoms, which complicates management strategies. Differences between yallundae (e.g., host pathogenicity fungicide sensitivity) require that both targeted for effective management. Here, we develop apply two molecular methods species-specific mating-type (MAT1-1 or MAT1-2) discrimination isolates. First, a multiplex PCR-based diagnostic assay targeting MAT idiomorph region was developed, allowing simultaneous determination mating type. This PCR successfully applied type global collection Second, development loop-mediated isothermal amplification (LAMP) assays β-tubulin sequences, allow fast (&lt;9 min) isolates, described. The LAMP detect very small amounts target DNA (1 pg) planta. In addition, mating-type-specific were developed rapid (&lt;12 genotyping Finally, applied, conjunction spore trapping field experiments, provide evidence wind dispersal ascospores from diseased crop. results indicate role cycle eyespot.