Fungicides are extensively used in agriculture to control fungal pathogens which are responsible for significant economic impact on plant yield and quality. Discovery and development of new fungicides face great challenges including a) screening of large libraries of antifungal candidates to evaluate their efficacy, phytotoxicity, and other possible effects, b) high cost of product development driven by the extensive studies, and c) complex in vitro and in vivo experiments. The conventional antifungal screening techniques, such as water ager and 96-well plates, are based on laborious protocols and bulk analysis, are time consuming and have closed system designs (no flow through reagents or culture medium). The classical approaches are also limiting in terms of single spore analysis and particular difficulties, such as handling and reproducibility can lead to considerable variability between observations within experimental runs. Therefore, a droplet-based microfluidic platform tackling these challenges was developed to progressively transition to single spore analysis for fungicide screening for routine assays.
The developed technology can be used not only in the context of any large-scale fungicide screening studies, but is also well-suited to analyze the mode of action of antifungal agents and resistance-development in fungi. The platform can be further extended for analysis of the molecular mechanisms of fungicides which play a crucial role in cellular mechanisms. A deeper understanding of the mode of action of fungicide will help to optimize their application, which in turn contributes to their successful use in food and feed production. The optimized fungicide application can reduce the cost of growers’ budget, lowers the risk of resistance development and pesticide residue in harvest and can positively impact the environmental and beneficial organisms.
The main objectives of this MSCA project were to establish a simple microfluidic device capable of (a) encapsulating single spores of fungi, (b) performing antifungal screening, and (c) quantifying gradient-based antifungal-dose response.
