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.
Highlights
The degradation capacity and utilisation of different complex plant substrates play crucial roles in the functioning of saprobic fungi as well as various plant symbionts with fundamental roles in ecosystems [1,2,3]
A wide range of methods have been developed to estimate the quantity of different microorganisms: counting and culturing microbes [10, 11], light microscopy using chemically cleared plant tissues combined with specific staining [12], measuring microorganism-derived contents or structures [13], colorimetric methods [14], microarray analyses [15], rRNA targeted fluorescence in situ hybridization (FISH) [16], FISH in combination with microautoradiography [17], isotope tracking [18], proteomic analyses [19], enzyme activity assays and numerous assays based on respiration or utilisation of different substrates [20, 21], and measuring the concentration of microbe-derived bioindicator molecules, such as phospholipid phosphate (PL-P) [22], ATP, and dehydrogenases [23]
We evaluated differences in the growth capacity of different dark septate endophytes (DSEs) fungi with respect to plant and tissue type
Summary
The degradation capacity and utilisation of different complex plant substrates play crucial roles in the functioning of saprobic fungi as well as various plant symbionts with fundamental roles in ecosystems [1,2,3]. These features might contribute to in planta colonisation [4, 5] and to nutrient mobilisation from the soil in the case of mutualistic symbionts [6, 7]. PCR-based techniques [24, 25] such as RNA-or DNA-based qPCR, have been used for microbial quantification [26,27,28,29]
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