Journal: Biotechnology for Biofuels

Article Title: Enhanced cellulase production by decreasing intercellular pH through H + -ATPase gene deletion in Trichoderma reesei RUT-C30

doi: 10.1186/s13068-019-1536-2

Figure Lengend Snippet: Effects of Ca 2+ channel inhibitor LaCl 3 on growth, cytosolic Ca 2+ concentration, and cellulase production. a Biomass dry weight of T. reesei RUT-C30 and Δ76238 was measured in 100 mL MA medium treated with different concentrations of LaCl 3 for 120 h, using glucose as the sole carbon source. b PNPCase activity of Δ76238 was examined after culture in MA medium for 120 h containing 0, 0.25, 0.5, or 1 mM LaCl 3 . c Fluorescence analysis of LaCl 3 influence on cytosolic Ca 2+ burst in Δ76238. Δ76238 was cultured in MA medium for 120 h and treated with 0 or 0.5 mM LaCl 3 , using glucose as the sole carbon source. For detection, 50 μM Fluo-3/AM was used, and its intensity was monitored using the Nikon A1R confocal microscope. Green fluorescence represents the free cytosolic Ca 2+ . “ a ” in the figure indicates that conidia of Δ76238 cannot germinate with 1 mM LaCl 3 . Values are the mean ± SD of the results from three independent experiments. Asterisks indicate significant differences (** p < 0.01, *** p < 0.001, Student’s t test)

Article Snippet: Images of Ca 2+ green fluorescence were obtained using an S Plan Fluor ELWD 40x, 0.45 numerical aperture (NA) objective and a digital sight camera on an Eclipse Ti inverted Nikon or Nikon A1R confocal microscope, comprising an FITC filter (420–490 nm band-pass excitation filter, and emission filter of 535 nm).

Techniques: Concentration Assay, Activity Assay, Fluorescence, Cell Culture, Microscopy