Itani Ayaka, Masuo Shunsuke, Yamamoto Riho, Serizawa Tomoko, Fukasawa Yu, Takaya Naoki, Toyota Masatsugu, Betsuyaku Shigeyuki, Takeshita Norio
Microbiology Research Center for Sustainability (MiCS), Faculty of Life and Environmental Sciences, University of Tsukuba, Tennodai 1-1-1, Tsukuba, 305-8572, Japan.
Graduate School of Agricultural Science, Tohoku University, 232-3 Yomogida, Naruko, Osaki, Miyagi, 989-6711, Japan.
PNAS Nexus. 2023 Mar 7;2(3):pgad012. doi: 10.1093/pnasnexus/pgad012. eCollection 2023 Mar.
Many fungi live as mycelia, which are networks of hyphae. Mycelial networks are suited for the widespread distribution of nutrients and water. The logistical capabilities are critical for the extension of fungal survival areas, nutrient cycling in ecosystems, mycorrhizal symbioses, and virulence. In addition, signal transduction in mycelial networks is predicted to be vital for mycelial function and robustness. A lot of cell biological studies have elucidated protein and membrane trafficking and signal transduction in fungal hyphae; however, there are no reports visualizing signal transduction in mycelia. This paper, by using the fluorescent Ca biosensor, visualized for the first time how calcium signaling is conducted inside the mycelial network in response to localized stimuli in the model fungus . The wavy propagation of the calcium signal inside the mycelium or the signal blinking in the hyphae varies depending on the type of stress and proximity to the stress. The signals, however, only extended around 1,500 μm, suggesting that the mycelium has a localized response. The mycelium showed growth delay only in the stressed areas. Local stress caused arrest and resumption of mycelial growth through reorganization of the actin cytoskeleton and membrane trafficking. To elucidate the downstream of calcium signaling, calmodulin, and calmodulin-dependent protein kinases, the principal intracellular Ca receptors were immunoprecipitated and their downstream targets were identified by mass spectrometry analyses. Our data provide evidence that the mycelial network, which lacks a brain or nervous system, exhibits decentralized response through locally activated calcium signaling in response to local stress.
许多真菌以菌丝体的形式存在,菌丝体是由菌丝构成的网络。菌丝体网络适合于营养物质和水分的广泛分布。这种后勤能力对于真菌生存区域的扩展、生态系统中的养分循环、菌根共生以及致病性至关重要。此外,预计菌丝体网络中的信号转导对于菌丝体的功能和稳健性至关重要。许多细胞生物学研究已经阐明了真菌菌丝中的蛋白质和膜运输以及信号转导;然而,尚无关于可视化菌丝体中信号转导的报道。本文通过使用荧光钙生物传感器,首次可视化了在模式真菌中,菌丝体网络内部如何响应局部刺激进行钙信号传导。菌丝体内钙信号的波浪式传播或菌丝中的信号闪烁因应激类型和与应激的接近程度而异。然而,这些信号仅延伸约1500μm,这表明菌丝体具有局部反应。菌丝体仅在受应激的区域表现出生长延迟。局部应激通过肌动蛋白细胞骨架的重组和膜运输导致菌丝体生长的停滞和恢复。为了阐明钙信号传导、钙调蛋白和钙调蛋白依赖性蛋白激酶的下游机制,对主要的细胞内钙受体进行了免疫沉淀,并通过质谱分析鉴定了它们的下游靶点。我们的数据提供了证据,表明缺乏大脑或神经系统的菌丝体网络通过响应局部应激的局部激活钙信号传导表现出分散反应。