High-Resolution melting assay for identification of Colletotrichum species from strawberry and endpoint genotyping for detection of the G143A mutation
Bruna Forcelini: Gulf Coast Research and Education Center, University of Florida
<div><em>Colletotrichum</em> species cause major diseases of strawberry and disease management depends on the species present. However, species identification based on symptoms and spore morphology is difficult. Therefore, development of molecular techniques for trustworthy and high-throughput identification of <em>Colletotrichum</em> species are desirable. A High-Resolution Melting (HRM) assay was developed for simultaneous identification and differentiation of <em>Colletotrichum </em>species from fungal colonies or from symptomatic strawberry tissues. HRM markers were designed based on the ITS region of <em>C. acutatum</em> and <em>C. gloeosporioides</em> from strawberry and accurately identified and differentiated <em>C. acutatum</em> from <em>C. gloeosporioides</em>. In addition, an endpoint genotyping analysis was developed for the rapid detection of a single nucleotide polymorphism at codon 143 of the cytochrome <em>b</em> (<em>cytb</em>) gene of <em>C. acutatum</em> and <em>C. gloeosporioides</em> associated with resistance to quinone-outside inhibitor fungicides. The results of this study suggest that the HRM and endpoint genotyping analyses are useful tools that can be implemented in plant diagnostic clinics for the rapid and accurate identification of <em>Colletotrichum</em> spp. and detection of the G143A mutation in the <em>cytb</em> gene of <em>C. acutatum</em> and <em>C. gloeosporioides</em>.</div>
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