The responses and detoxification mechanisms of dark septate endophytes (DSE), Exophiala salmonis, to CuO nanoparticles

To understand the tolerance mechanisms of dark septate endophytes (DSE), Exophiala salmonis, to CuO nanoparticles (CuO-NPs) with different sizes (40 and 150 nm), we investigated the morphology, antioxidant response, Cu subcellular distribution, and the melanin gene expression in the mycelia of E. salmonis. E. salmonis was cultured in liquid and solid media under the stress of increasing CuO-NP concentrations (0, 50, 100, 150, and 250 mg/L). Results showed that (1) E. salmonis showed good CuO-NP tolerance, and the tolerance to CuO-NPs at 150 nm was stronger than that at 40 nm. A large number of agglomeration structures were observed on the mycelia surface with the exception of 50 mg/L CuO-NPs with a diameter of 150 nm. (2) CuO-NP stress significantly stimulated the production of antioxidant enzymes, particularly the CuO-NPs with small particle size ( 40 nm). ( 3) Cu uptaken by E. salmonis increased proportionally with the increase of CuO-NP concentration in the medium. More than 80% Cu was absorbed in cell wall of mycelia treated with a small particle size (40 nm). (4) FTIR analysis revealed that hydroxyl, amine, carboxyl, and phosphate groups were associated with CuONP binding regardless of particle size. (5) Fungal melanin content increased with the addition of CuO-NPs; the increase of melanin induced by CuO-NPs with small particle size (40 nm) was more significant. (6) The expression of 1,3,6,8-tetrahydroxynaphthalene reductase (Arp2) in the melanin synthesis pathway increased under the stress of CuO-NPs, and CuO-NPs with a small particle size ( 40 nm) caused a significant change in the expression level of Arp2 gene than those with a large particle size (150 nm). In conclusion, E. salmonis had a strong tolerance to CuO-NPs and mitigated the toxic effects of CuO-NPs through the antioxidant system, the expression of genes related to melanin synthesis, and the synthesis of melanin.