Division of Microbial Genetics and Pathogen Interaction, Department of Applied Genetics and Cell Biology, BOKU University of Natural Resources and Life Science, Campus Tulln, Tulln/Donau, Austria.
Departamento de Genética, Universidad de Sevilla, Seville, Spain.
Sci Rep. 2017 Jun 27;7(1):4289. doi: 10.1038/s41598-017-03598-9.
Filamentous fungi naturally grow on solid surfaces, yet most genetic and biochemical analyses are still performed in liquid cultures. Here, we report a multiplexing platform using high-throughput photometric continuous reading that allows parallel quantification of hyphal growth and reporter gene expression directly on solid medium, thereby mimicking natural environmental conditions. Using this system, we have quantified fungal growth and expression of secondary metabolite GFP-based reporter genes in saprophytic Aspergillus and phytopathogenic Fusarium species in response to different nutrients, stress conditions and epigenetic modifiers. With this method, we provide not only novel insights into the characteristic of fungal growth but also into the metabolic and time-dependent regulation of secondary metabolite gene expression.
丝状真菌自然生长在固体表面上，但大多数遗传和生化分析仍在液体培养基中进行。在这里，我们报告了一种使用高通量光度连续读数的多路复用平台，该平台允许直接在固体培养基上平行定量菌丝生长和报告基因表达，从而模拟自然环境条件。使用该系统，我们已经定量了腐生曲霉属和植物病原菌镰刀菌属在不同营养物质、胁迫条件和表观遗传修饰剂作用下的生长和基于 GFP 的次生代谢物报告基因的表达。通过这种方法，我们不仅提供了对真菌生长特性的新见解，还提供了对次生代谢物基因表达的代谢和时变调节的新见解。
