Fig. 7

Influence of TPMR1 on Mn²⁺-induced cytosolic Ca²⁺ burst and cellulase production. A Fluorescence analysis of the influence of TPMR1 on the cytosolic Ca²⁺ burst induced by Mn²⁺. The T. reesei Rut-C30 and Δtpmr1 strains were cultured in liquid minimal medium for 48–60 h with 0 or 10 mM MnCl₂ (0 or 10 Mn, respectively). For detection, 50 μM Fluo-3/AM was used, and the intensity was monitored using Automatic Inverted Fluorescence Microscopy. Green fluorescence represents the free cytosolic Ca²⁺. DIC, differential interference contrast. B Comparative fluorescence ratio analysis of TPMR1 influence on cytosolic Ca²⁺ burst induced by Mn²⁺. The y-axis represents the Ca²⁺ fluorescence ratio measured by CLSM and the x-axis the different treatments. The CMCase activity (C) and pNPCase activity (D) of T. reesei Rut-C30 and Δtpmr1 strains were examined after culture in medium containing 0 or 10 mM MnCl₂. The expression levels of cbh1 (E) and egl1 (F) in T. reesei Rut-C30 and Δtpmr1 strains were analyzed after culture in medium containing 0 or 10 mM MnCl₂. Values are the mean ± SD of the results from three independent experiments. Asterisks indicate significant differences from untreated strains (*p < 0.05, **p < 0.01, Student’s t test). Different letters indicate significant differences between the columns (p < 0.05, according to Duncan’s multiple-range test)