Simple fluorescence-based high throughput cell viability assay for filamentous fungi

Area of Science:

• Mycology
• Biotechnology
• Antimicrobial Research

Background:

• Filamentous fungi are crucial eukaryotic models and industrial resources.
• They also cause significant plant and human diseases, necessitating effective management.
• Current antifungal susceptibility testing methods are often slow and unsuitable for large-scale analysis.

Purpose of the Study:

• To develop a rapid, high-throughput microplate-based fluorescence method for assessing antifungal and stress agent toxicity on filamentous fungi.
• To compare the performance of this new assay with conventional methods like agar dilution.

Main Methods:

• Optimization of a resazurin reduction assay to measure fungal viability via resorufin fluorescence.
• Application of the assay to the filamentous fungus Magnaporthe oryzae.
• Comparison of results (inhibitory rates, IC50 values) with the agar dilution method.

Main Results:

• The resazurin fluorescence bioassay demonstrated comparable inhibitory rates and IC50 values to the agar dilution method.
• Results were consistent with previously reported data for Magnaporthe oryzae and other fungi.
• The assay proved effective in evaluating the toxicity of various antifungal and stress agents.

Conclusions:

• A simple, fluorescence-based, high-throughput method was successfully developed for evaluating antifungal and stress agent effects on filamentous fungi.
• This resazurin fluorescence assay offers advantages in simplicity, speed, and cost-effectiveness over traditional growth inhibition assays.
• The assay has significant potential for large-scale screening of antifungal agents and studying fungal population dynamics.