Ff microplate assay

Variations in growth based on diverse carbon sources were assessed using the BIOLOG FF Microplate assay (Biolog Inc., Hayward, CA), as described previously^{90 (link)}. Inoculated microplates were incubated at 28°C in constant darkness, spanning a timeframe of up to 144 h. Measurements of absorbance at 750 nm, indicative of biomass accumulation, were taken at 24-h intervals, starting at 72-h. To evaluate the statistical significance of growth differences, a T-test was employed (with a threshold p-value of ≤ 0.05) using Excel 2016 (Microsoft, Redmond, USA).

Growth on different carbon sources were analyzed using BIOLOG FF Microplate assay (Biolog Inc., Hayward, CA) as described previously [94 (link)]. Inoculated microplates were incubated at 28 °C in constant dark or constant light (1700 lx) for up to 144 h and absorbances measured at 750 nm reflecting biomass accumulation in 24 h intervals starting at 72 h. Analyses were repeated in triplicates for each strain. Statistical significance of growth differences was analyzed by the T-test (p-value threshold ≤ 0.05) as implemented in Excel 2016 (Microsoft, Redmond, USA).

The Biolog FF MicroPlate assay (Biolog Inc., Hayward, CA) which comprises 95 wells with different carbon-containing compounds and one well with water was used to investigate growth rates on pre-filled carbon sources. Conidia were collected from the Trichoderma strains (parental strain, Δtmk1, Δste12) and used as inoculums as described [33] (link). Inoculated microplates were incubated in darkness at 28°C, and OD750 readings determined after 18, 24, 42, 48, 66, 72, 96 and 168 hours using a microplate reader (Biolog), which measures the turbidity and reflects mycelia production on the tested substrate. All analyses were performed in triplicate. For comparative analysis, values from the 48 hours time point was used as this was within the linear growth phase of T. atroviride on the majority of carbon sources. Values were quantitatively illustrated using the Hierarchical Clustering Explorer 3 (HCE3) [34] . For all analyses the hierarchical clustering algorithm with average linkage and Euclidean distance measure was applied.