Application of tetra primer ARMS-PCR approach for detection of Fusarium graminearum genotypes with resistance to carbendazim

Abstract

Tetra-primer ARMS-PCR (amplification refractory mutation system-PCR) was developed to detect isolates of Fusarium graminearum with resistance to carbendazim, a methyl benzimidazole carbamate-group fungicide. Resistance to carbendazim in F. graminearum is caused by point mutations at codon 167, 198, and 200 of the β 2-tubulin gene (FGSG_06611.3). According to single nucleotide polymorphisms on codon 167, 198, 200 of the resistance gene β 2-tubulin gene, two pairs of primer designed on each codon respectively, were used for amplification. SNP genotyping was made successfully based on the different patterns in agarose gels between point mutations and wild type strain. Tetra-primer ARMS-PCR protocol developed in this study is a simple and useful technique to identify genotypes and has a potential application in management of carbendazim resistance in F. graminearum. It will help researchers and growers select the best antifungal strategy to use in the field.