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 CuO-NP 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.

Transcriptome analysis reveals the molecular mechanisms of Phragmites australis tolerance to CuO-nanoparticles and/or flood stress induced by arbuscular mycorrhizal fungi.

The molecular mechanism of arbuscular mycorrhizal fungi (AMF) in vertical flow constructed wetlands (VFCWs) for the purification of copper oxide nanoparticles (CuO-NPs) contaminated wastewater remains unclear. In this study, transcriptome analysis was used to explore the effect of AMF inoculation on the gene expression profile of Phragmites australis roots under different concentrations of CuO-NPs and/or flood stress. 551, 429 and 2281 differentially expressed genes (DEGs) were specially regulated by AMF under combined stresses of CuO-NPs and flood, single CuO-NPs stress and single flood stress, respectively. Based on the results of DEG function annotation and enrichment analyses, AMF inoculation under CuO-NPs and/or flood stress up-regulated the expression of a number of genes involved in antioxidant defense systems, cell wall biosynthesis and transporter protein, which may contribute to plant tolerance. The expression of 30 transcription factors (TFs) was up-regulated by AMF inoculation under combined stresses of CuO-NPs and flood, and 44 and 44 TFs were up-regulated under single CuO-NPs or flood condition, respectively, which may contribute to the alleviating effect of symbiosis on CuO-NPs and/or flood stress. These results provided a theoretical basis for enhancing the ecological restoration function of wetland plants for metallic nanoparticles (MNPs) by mycorrhizal technology in the future.