Exploring the Potential Mechanism of Prothioconazole Resistance in <i>Fusarium graminearum</i> in China

The Fusarium head blight (FHB) caused by <i>Fusarium graminearum</i> is one of the most important diseases threatening wheat production in China. However, the triazole sterol 14α-demethylation inhibitor (DMI), prothioconazole, is known to exhibit high activity against <i>F. graminearum</i>. The current study indicated that three highly resistant laboratory mutants exhibited significantly (<i>p</i> < 0.05) altered growth and sporulation, although contrary to expectation, only one of the mutants exhibited reduced growth and sporulation, while the other two exhibited significant (<i>p</i> < 0.05) increases. Despite this, pathogenicity tests revealed that all of the mutants exhibited significantly (<i>p</i> < 0.05) reduced pathogenicity, indicating a substantial cost to fitness. Sequence analysis of the prothioconazole target protein, CYP51, of which <i>F. graminearum</i> has three homologues (FgCYP51A, FgCYP51B, and FgCYP51C), identified three mutations in the FgCYP51B sequence with a high likelihood of being associated with the observed resistance, as well as another three mutations in the FgCYP51B sequence, and two in the FgCYP51A sequence that are worthy of further investigation. Two of the prothioconazole-resistant mutants were also found to have several amino acid substitutions in their FgCYP51C sequences, and it was interesting to note that these two mutants exhibited significantly (<i>p</i> < 0.05) reduced pathogenicity compared to the other mutant. Expression analysis revealed that prothioconazole treatment (0.1 μg/mL) resulted in altered expression of all the <i>FgCYP51</i> target genes, and that expression was also altered in the prothioconazole-resistant mutants compared to their wild-type parental isolates. Meanwhile, no evidence was found of any cross-resistance between prothioconazole and other commonly used fungicides, including carbendazim, pyraclostrobin, and fluazinam, as well as the triazole tebuconazole and the imidazole DMI prochloraz. Taken together, these results not only provide new insight into potential resistance mechanism in <i>F. graminearum</i>, and the biological characteristics associated with them, but also convincing evidence that prothioconazole can offer effective control of FHB.