Fig. 3

Trrab7 knock-down influenced the fungal development and vacuolar maintenance of T. reesei. A Effect of Trrb7 knock-down on the vegetative growth, cell wall integrity, stress response and conidiation of T. reesei strains. Growth of QM9414 and Ptcu-rab7^KD strains was examined on plates with various carbon sources (1% w/v) as indicated. The conidiation of QM9414 and Ptcu-rab7^KD was performed on malt extract plates. Cell wall integrity was determined on minimal medium (MM) plates containing 250 mg/L Congo red (CR). Osmotic and oxidative stress response tests were performed on minimal medium (MM) plates containing 0.5 M NaCl and 30 mM H₂O₂, respectively. QM9414 and Ptcu-rab7^KD on different agar plates were kept at 30 °C for 3 days. B Number of spores in QM9414 and Ptcu-rab7^KD strains on the malt extract was counted by a hemocytometer. C Statistical quantitation of percentage in colony diameter of each strain under different agar plates relative to QM9414 cultured with glucose. Significant differences were determined by a two-tailed student's t test. *, P < 0.05; **P < 0.01; *** P < 0.001. D Biomass accumulation of the QM9414 and Ptcu-rab7^KD in MA medium containing 1% (wt/v) glucose as the sole carbon source. E Effect of Trrab7 knock-down on vacuole formation. QM9414 and Ptcu-rab7^KD were grown in MA medium with 1% (w/v) Avicel and mycelia were collected and stained with 10 μM FM4-64 for fluorescence microscopy analysis at 24 h. The fluorescence was examined with a Nikon Eclipse 80i fluorescence microscope. The structure of the normal vacuole in T. reesei QM9414 is indicated by arrows in the figure. Scale bar, 10 μm