Fig. 1. Production of PpoA oxylipins and complementation of ΔppoA sporulation phenotype by 5,8-diHODE.

a Developmental steps in Aspergillus. Asexual spores swell, germinate, form branched hyphae, and undergo conidiophore development. Different oxylipin signals are proposed to regulate germination, degree of branching, and development of spore-bearing conidiophores. b Quantification of PpoA oxylipins 5,8-diHODE and 8R-HODE in Af293 WT and ΔppoA fungal biomass, when cultures were grown at 25 °C for 120 h under constant shaking at 250 rpm (n = 4). Fungal biomass was separated from supernatant and extracted independently, followed with UHPLC-MS/MS analysis. c Quantification of PpoA oxylipins 5,8-diHODE and 8R-HODE in Af293 WT and ΔppoA culture supernatant obtained in the same experiment described in (b). d Sporulation quantification of Af293 WT and ∆ppoA cultures (n = 3) treated in EtOH, 0.5 µg/mL and 5 µg/mL of 5,8-diHODE. Three independent cultures of Af293 WT and ΔppoA were grown in liquid shaking GMM at 25 °C for 65 h before the addition of 5,8-diHODE or EtOH and cultured up to 120 h before homogenization and spore quantification. A two-way ANOVA test followed by multiple comparison tests was performed to detect the difference between WT and ΔppoA in the three different treatment conditions. Multiple two-sided t tests were performed to compare the production of each oxylipin in WT and ΔppoA in (a, b), and two-way ANOVA was performed to test the effect of strain and treatment, followed by Holm–Šídák multiple comparison tests to compare two strains with the same treatment or two treatments in the same strain in (c). All values represent mean ± SEM. ns not significant (P > 0.05).