Department of Molecular & Systems Biology, Geisel School of Medicine at Dartmouth, Hanover, NH, USA.
Department of Biochemistry & Cell Biology, Geisel School of Medicine at Dartmouth, Hanover, NH, USA.
Fungal Biol. 2020 May;124(5):398-406. doi: 10.1016/j.funbio.2019.11.004. Epub 2019 Nov 20.
Work on the filamentous fungus Neurospora crassa has contributed to or pioneered many aspects of research on circadian clock mechanism, a process that is functionally conserved across eukaryotes. Biochemical assays of the fungal circadian clock typically involve growth in liquid medium where Neurospora forms a spherical ball of submerged mycelium. Here, we revive a method for dispersed growth of Neurospora in batch culture using polyacrylic acid as an additive to the medium. We demonstrate that dispersed growth cultures utilize more carbon than mycelial balls, but nonetheless retain a functional circadian clock. This culturing method is suited for use in circadian experiments where uniform exposure to nutrients and/or increased biomass is required.
丝状真菌粗糙脉孢菌(Neurospora crassa)的研究工作为昼夜节律机制的许多方面做出了贡献或开创了先河,这一过程在真核生物中具有功能保守性。真菌昼夜节律的生化分析通常涉及在液体培养基中进行生长,其中粗糙脉孢菌形成一个浸没菌丝体的球形球。在这里,我们恢复了一种使用聚丙烯酸作为培养基添加剂在分批培养中分散生长粗糙脉孢菌的方法。我们证明,分散生长培养物比菌丝球消耗更多的碳,但仍然保留了功能正常的昼夜节律。这种培养方法适用于需要均匀暴露于营养物质和/或增加生物量的昼夜节律实验。