Selection of redox dye and inoculum conditions for the optimisation of respirometric indices in Verticillium and Trichoderma
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Institute of Agrophysics, Polish Academy of Sciences, Doświadczalna 4, 20-290 Lublin, Poland
Institute for Biological Systems (IBS), Council of National Research of Italy (CNR), Area della Ricerca di Roma 1, via Salaria Km 29, 300, 00015 Monterotondo (RM), Italy
Final revision date: 2023-08-07
Acceptance date: 2023-08-09
Publication date: 2023-08-28
Corresponding author
Karolina Oszust   

Institute of Agrophysics, Polish Academy of Sciences, Doświadczalna 4, 20-290, Lublin, Poland
Int. Agrophys. 2023, 37(3): 279-292
  • Fungal competition for nutrients is measurable and exploitable in biocontrol.
  • Toxic and nutritional effects of dyes affect the results of metabolic profiling.
  • 1% F and 0.5% D BiologTM dyes are reliable with Verticillium and Trichoderma spp.
Understanding which carbon or nitrogen sources the competing fungal species prefer is pivotal for specific biotechnological applications. However, this is not straightforward, as each strain sometimes behaves differently under the experimental conditions adopted. To analyse the trophic overlap of two species, it is necessary to refine diagnostic techniques and exclude variables that may interfere with the measurements. A protocol for establishing the suitability of chromogenic dyes in the analysis of filamentous fungi with phenotype microarrays is described here. The research goal was to determine the most suitable redox dye indicator and its optimal concentration that reacts quantitatively to the respiratory activity of both Verticillium spp. and Trichoderma spp. isolates in the presence of a nitrogen source. The commercial BiologTM Redox Dye Mixes D, E, and F and also TTC (2,3,5-Triphenyltetrazolium chloride), INT (2-(4-iodophenyl)-3-(4-nitrophenyl)-5-phenyl-2H-tetrazolium chloride), and MTT solutions were evaluated in selected concentrations. Both their nutritive and toxic effects were quantified. Results showed that 1% “F” and 0.5% “D” BiologTM dyes were appropriate for the functional evaluation of isolates belonging to the Verticillium and Trichoderma genera. They can be used in BiologTM PM-nitrogen plate respiratory assays for a comprehensive functional characterisation of these organisms.
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