Micro-scale Experimental System Coupled with Fluorescence-based Estimation of Fungal Biomass to Study Utilisation of Plant Substrates

Abstract

The degradation capacity and utilisation of complex plant substrates are crucial for the functioning of saprobic fungi and different plant symbionts with fundamental functions in ecosystems. Measuring the growth capacity and biomass of fungi on such systems is a challenging task. We established a new micro-scale experimental setup using substrates made of different plant species and organs as media for fungal growth. We adopted and tested a reliable and simple titration-based method for the estimation of total fungal biomass within the substrates using fluorescence-labelled lectin. We found that the relationship between fluorescence intensity and fungal dry weight was strong and linear but differed among fungi. The effect of the plant organ (i.e. root vs. shoot) used as substrate on fungal growth differed among plant species and between root endophytic fungal species. The novel microscale experimental system is useful for screening the utilisation of different substrates, which can provide insight into the ecological roles and functions of fungi. Furthermore, our fungal biomass estimation method has applications in various fields. As the estimation is based on the fungal cell wall, it measures the total cumulative biomass produced in a certain environment.

Poaceascoma zborayi sp. nov. and Agrorhizomyces patris gen. et spec. nov. – Two novel dark septate endophytes colonizing wheat (Triticum aestivum) roots from a cropland in Hungary

“…To visualize the hyphae within the roots we used two methods. A wheat germ agglutinin (WGA) conjugate, WGA-Alexafluor®488 (Thermo Fisher Scientific, Lithuania) was used that stains chitin and is widely used as a fluorescent dye for fungal cell walls (Németh et al 2022). The other method applied aniline blue as a stain for in planta visualization of fungal endophytes (Andrade-Linares et al 2011; Knapp et al 2019).…”