Figure 5. The role of gamma-glutamyl cycle during Magnaporthe conidiation. (A) Oxp1-RFP and Snx41-GFP colocalize in aerial hyphae and conidiophore during conidiogenesis. Confocal microscopy was performed with conidiating culture at 6–16 h post photo-induction. Arrowheads denote colocalization of Oxp1-GFP and Snx41-GFP vesicles. Scale bar: 5 μm. (B) Snx41 physically interacts with Oxp1. Affinity purification with RFP-Trap beads using total protein lysates from Magnaporthe strains expressing Oxp1-RFP and Snx41-GFP, or Snx41-GFP alone, Immuno-blotting was performed with anti-GFP. (C) Oxp1-RFP is predominantly vacuolar in the snx41Δ mutant. In vegetative mycelia of snx41Δ, Oxp1-RFP was frequently seen inside the enlarged spherical vacuoles. In the aerial hyphae of snx41Δ, Oxp1-RFP was barely detectable, except for occasional vacuolar localization. (D) Bar chart depicting quantification of conidiation in the wild type (WT), oxp1Δ and snx41Δ grown on PA medium supplemented with 5-oxoproline. Mean values (± S.E.) presented as number of conidia were derived from three independent experiments (n = 30 colonies for each sample). Total conidia counts were performed five days post photo-induction.