Itraconazole

A. fumigatus strains Af293 and IFM 63432 were used to generate the srbA deletion mutants. Clinical strain IFM 63432 contains mutations TR46/Y121F/T289A within the cyp51A promoter region, and was isolated in Japan in 201313 (link). All strains used in the present study were cultivated in PDA and GMM containing 1% glucose at 37 °C. Strains were grown on PDA to collect conidia. Commercially available antifungal chemicals itraconazole, miconazole, bromuconazole, difenoconazole, propiconazole and tebuconazole were obtained from Wako Pure Chemical Industries (Osaka, Japan). Iprodione, fludioxonil and kresoxim-methyl were generously provided by Dr. Keietsu Abe, Tohoku University, Sendai, Japan.

Aspergillus fumigatus strain Af293 was used in this study. The strain was cultured in PDB at 37 °C on a rotary shaker at 120 rpm throughout the study. In some experiments, glucose minimal medium (GMM) (1%(w/v) glucose, 0.052% KCl, 0.052% MgSO₄·7H₂O, 0.152% KH₂PO₄, 0.0022% ZnSO₄·7H₂O, 0.0011% H₃BO₃, 0.0005% MnCl₂·4H₂O, 0.0005% FeSO₄·7H₂O, 0.00016% CoCl₂·5H₂O, 0.00016% CuSO₄·5H₂O, 0.00011% (NH₄)6Mo₇O₂₄·4H₂O, 0.005% Na₄EDTA, pH 6.5) containing variable nitrogen sources, such as sodium nitrate, ammonium sulphate, and proline (70 mM) were used, which were designated GMM(NO₃), GMM(NH₄), and GMM(Pro), respectively. The antifungal chemicals used were commercially obtained, as follows: thymol (NACALAI TESQUE, Inc., Kyoto, Japan), farnesol (Sigma-Aldrich Co., St. Louis, MO, USA), citral (NACALAI TESQUE, Inc.), nerol (Tokyo Chemical Industry Co., Ltd, Tokyo, Japan), salicylic acid (NACALAI TESQUE, Inc.), PCA (Wako Pure Chemicals Industries, Osaka, Japan), pyocyanin (Cayman Chemical Company, Ann Arbor, MI, USA), itraconazole (Wako Pure Chemicals Industries), voriconazole (Tokyo Chemical Industry Co., Ltd), and amphotericin B (Sigma-Aldrich Co.). Micafungin was a generous gift from Dr. Keietsu Abe, Tohoku University.