Supplemental Data

Files in this Data Supplement:

• Supplemental Figure 1 - Growth of endophyte infected perennial ryegrass. Plants were infected with E. festucae Fl1 (WT), noxA mutant (FR2) or complemented strains (C5, C8). Up to 10 tillers were harvested from 3 individual plants for each strain 9 weeks after inoculation and the length of the sheath and blade measured. Bars show the standard error for each sample, N=30.
• Supplemental Figure 2 - Growth of epiphyllous hyphae on the surface of leaf-blades. Plants of perennial ryegrass were infected with E. festucae Fl1 (A) or FR2 (B). Plants were grown on Murashige and Skoog medium containing 0.6% Phyto agar (Duchefa Biochemie, Haarlem, The Netherlands) in axenic culture. Photographs were taken 8 weeks after the inoculation.
• Supplemental Figure 3 - Symbiotic Phenotype of E. festucae FR2. (A) Phenotype of tall fescue infected with E. festucae wild-type Fl1 and symbiotic mutant FR2. The photograph was taken 9 weeks after inoculation. (B) Phenotype of meadow fescue infected with E. festucae wild-type Fl1 and symbiotic mutant FR2. The photograph was taken 9 weeks after inoculation.
• Supplemental Figure 4 - Relative biomass of wild type and noxA mutant in perennial ryegrass. Pseudostems of perennial ryegrass infected with wild type (WT) or noxA mutant (A44) were harvested 9 weeks after inoculation. Total pseudostem DNA was isolated from 3 individual plants for each strain and the endophyte DNA biomass determined by real time PCR. The y axis value represents endophyte biomass relative to the average of the wild type for three independent reactions. Each bar represents the standard error of the mean, N=3
• Supplemental Figure 5 - Lactophenol trypan blue stained perennial ryegrass tissue. Tissue was infected with E. festucae Fl1 (A and C) or noxA mutant A44 (B and D). Plants were grown in axenic culture on MS media for 8 weeks after endophyte inoculation. Pseudostem (A and B) or leaf blade tissues (C and D) were stained with lactophenol trypan blue stain. (D) Some leaf blade infected with noxA mutant showed a few clusters of dead mesophyll cells (arrow heads), which are correlated with precocious senescence of perennial ryegrass infected with noxA mutant. Bar = 60 μm.
• Supplemental Table 1
• Supplemental Table 2