Baseline sensitivity and resistance risk of Sclerotinia sclerotiorum to glabridin and the possible anti-fungal mechanism.

Sclerotinia stem rot caused by Sclerotinia sclerotiorum is one of the most serious diseases of oilseed rape. Chemical control is an important method to control this disease, however, development of fungal resistance to commonly used fungicides has led to severe yield losses in recent years. Therefore, development of novel fungicides against S. sclerotiorum is urgently needed. Glabridin is one of the major flavonoids in Glycyrrhiza L. plants, and we previously found that it is very effective against S. sclerotiorum. Nevertheless, the baseline sensitivity and resistance risk of S. sclerotiorum to glabridin as well as the possible anti-fungal mechanism need further elucidation. In this study, we revealed that the EC50 (median effective concentration) values of glabridin against 109 S. sclerotiorum isolates collected from Jiangsu Province of China ranged from 0.51 to 8.03 μg/mL with a mean EC50 value of 3.05 ± 1.27 μg/mL. No cross-resistance was observed between glabridin and carbendazim, and no glabridin-resistant mutants were obtained by chemical induction. RNA profiling result showed that tyrosine metabolism of S. sclerotiorum were evidently affected by glabridin. qRT-PCR, enzyme activity assay, and molecular docking proved that glabridin greatly reduced both the expression level and enzyme activity of tyrosinase in S. sclerotiorum. Furthermore, S. sclerotiorum incurred certain impairment in its membrane integrity after glabridin treatment at 10 μg/mL. This study is the first report on baseline sensitivity and resistance risk of S. sclerotiorum to glabridin, and it is revealed that glabridin may interfere tyrosine metabolism and membrane integrity of S. sclerotiorum.