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Author:
Date:
2016-09-05
[Abstract] In this protocol we describe a cell wall stress assay for the fungal pathogen F. oxysporum, based on exposure to the two anionic dyes Calcofluor White (CFW) and Congo Red (CR). Both compounds have been used to exert stress upon the fungal cell wall in vitro (Perez-Nadales and Di Pietro, 2015; Perez-Nadales and Di Pietro, 2011; Leach et al., 2012; Heilmann et al., 2013; Garcia et al., 2015). CFW perturbs chitin assembly, whereas CR interferes with β-glucan synthesis, resulting in cell wall-weakening and activation of the cell wall stress response (Ram and Klis, 2006; Kopecka and Gabriel, 1992; Roncero and Duran, 1985). Presumably, the signaling pathways and cell wall changes associated with this response reflect cell wall homeostasis during ...
[摘要] 在该协议中，我们描述了真菌病原体F的细胞壁应激测定。 ，基于暴露于两种阴离子染料Calcofluor White（CFW）和刚果红（CR）。两种化合物都已经用于在体外对真菌细胞壁施加应激（Perez-Nadales和Di Pietro，2015; Perez-Nadales和Di Pietro，2011; Leach等人， 2012; Heilmann等人，2013; Garcia等人，2015）。 CFW干扰几丁质装配，而CR干扰β-葡聚糖合成，导致细胞壁弱化和细胞壁应激反应的激活（Ram和Klis，2006; Kopecka和Gabriel，1992; Roncero和Duran，1985）。据推测，与该反应相关的信号通路和细胞壁变化反映了正常生长期间的细胞壁稳态以及响应于真菌 - 宿主相互作用期间遇到的应力的细胞壁重塑事件。本方案中规定的CFW和CR培养基的制备条件是基于Ram和Klis的论文"Identification of fungal cell wall mutants using susceptibility assays based on Calcofluor white and Congo red"，published in Nature protocols（Ram and ...
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