Cochliobolus sativus (S. Ito & Kurib.) Drechsler ex Dastur, the causal agent of spot blotch disease, is a necrotrophic pathogen affecting barley production worldwide. Traditional methods to control this disease are commonly used fungicide applications including triadimefon, a triazole compound. C. sativus have a great interest since it might expand resistance against triadimefon due to its high genetic variability and abundant spore production. Therefore, understanding of resistance strategies in this fungus is imperative to developing more effective defenses against the spread of resistance. Major Facilitator Superfamily (MFS) transporters play an important role in resistance in fungi. In this study, expression patterns of MFS transporter gene in two C. sativus virulent and avirulent pathotypes were monitored at early time periods of triadimefon treatments using quantitative PCR (qPCR) method. Results demonstrated 50% mycelial growth inhibition (EC₅₀) for both C. sativus pathotypes was recorded after 96h at 0.25 µg mL^-1 triadimefon treatments. qRT-PCR revealed that MFS expression increased in both pathotypes at 24 h post triadimefon treatments as compared with the corresponding controls. The most outstanding variations in this gene expression were 6.11 and 3.55-fold in the virulent and avirulent pathotypes, respectively, 96h of 0.25 µg mL^-1 triadimefon treatments. This is the first report on the changes of MFS expression in the fungus C. sativus. According to these findings, it is likely to suggest that MFS gene might has a function in signaling events during treatment C. sativus with triazole fungicide.