A versatile microfluidic platform measures hyphal interactions between Fusarium graminearum and Clonostachys rosea in real-time

Abstract

Routinely, fungal–fungal interactions (FFI) are studied on agar surfaces. However, this format restricts high-resolution dynamic imaging. To gain experimental access to FFI at the hyphal level in real-time, we developed a microfluidic platform, a FFI device. This device utilises microchannel geometry to enhance the visibility of hyphal growth and provides control channels to allow comparisons between localised and systemic effects. We demonstrate its function by investigating the FFI between the biological control agent (BCA) Clonostachys rosea and the plant pathogen Fusarium graminearum. Microscope image analyses confirm the inhibitory effect of the necrotrophic BCA and we show that a loss of fluorescence in parasitised hyphae of GFP-tagged F. graminearum coincides with the detection of GFP in mycelium of C. rosea. The versatility of our device to operate under both water-saturated and nutrient-rich as well as dry and nutrient-deficient conditions, coupled with its spatio-temporal output, opens new opportunities to study relationships between fungi.

Spatial scales of competition and a growth-motility tradeoff interact to determine bacterial coexistence

“…This also limits our ability to investigate deeper into the interactions between network type and different ways of inoculation, for example, by performing a two-factor ANOVA test. Therefore, a valuable next step would be to tease apart the large variations of our current results under the abiotic and biotic network settings by manipulating their structures, for example, by using 3D printed abiotic hyphae models with fixed structure and surface physicochemical properties, or microfluidic platforms that constrain the growth of fungal hyphae networks into pre-defined topology [76,77].…”

Microfabrication and its use in investigating fungal biology

“…These systems can incorporate mechanisms to generate concentration gradients through complex mixing or parallel flow diffusion channels within the chip in order to fine-tune exposure to drugs and other reagents (Lee et al, 2020;Saka et al, 2017;Witkowski et al, 2018). Bespoke designs have also been used to study multispecies interactions, including biocontrol approaches and metabolite exchange in fungus-bacteria symbiosis (Gimeno et al, 2021;Uehling et al, 2019). However, in this review, we focus on how microfabrication has been used to enhance our understanding of yeast and hyphal growth.…”

Methods for Studying Bacterial–Fungal Interactions in the Microenvironments of Soil

“…Microfluidic platforms allow for the precisely organization and monitoring of small heterogeneous microbial populations [114,115] in a three-dimensional geometry [116]. They can be constructed in different dimensions (in volumes as small as~100 fl [117]), materials (e.g., transparent polydimethylsiloxane (PDMS) [116,118,119], hydrogels, proteins crosslinked by multiphoton lithography [120], lipid-silica containers [121,122]), or shapes (e.g., soil micromodels [123,124], arenas [125], channels [126], mazes [127], or single droplets [128]). Some of the microfabricated biomaterials used for constructing a microfluidic system can be responsive to external stimuli, hence acting as both physical barriers, as well as an additional function for active control, manipulation, and observation of the microbes in real time.…”