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

Routinely, fungal-fungal interactions (FFIs) are studied on agar surfaces. However, this format restricts high-resolution dynamic imaging. To gain experimental access to FFIs 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) <em>Clonostachys rosea </em>and the plant pathogen <em>Fusarium graminearum. </em>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 <em>F. graminearum </em>coincides with the detection of GFP in mycelium of <em>C. rosea</em>. 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.真菌-真菌互作（fungal-fungal interactions, FFIs）的常规研究多在琼脂平板表面开展，但该实验范式无法支持高分辨率动态成像。为实现菌丝水平真菌-真菌互作的实时观测，本研究开发了一款微流控平台（microfluidic platform）——FFI装置。该装置通过优化微通道结构提升菌丝生长的可视性，并配备对照通道，可实现局部效应与系统效应的对比分析。本研究通过探究生物防治因子（biological control agent, BCA）玫烟色棒束孢（*Clonostachys rosea*）与植物病原菌禾谷镰孢菌（*Fusarium graminearum*）之间的真菌-真菌互作，验证了该装置的功能。显微镜成像分析证实了该死营养型生物防治因子的抑菌效果；研究同时发现，带有绿色荧光蛋白（green fluorescent protein, GFP）标记的禾谷镰孢菌被寄生菌丝的荧光信号消失之际，玫烟色棒束孢菌丝中可检测到GFP信号。该装置可在水饱和富营养、干燥贫营养等多种环境下稳定运行，结合其时空维度的观测输出能力，为真菌间互作关系的研究开辟了全新路径。