| Penetration Assays, Fungal Recovery and Pathogenicity Assays for Verticillium dahliae
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Author:
Date:
2017-02-20
[Abstract] Verticillium dahliae is a soil-borne phytopathogenic fungus that infects host roots and proliferates in vascular tissues. The great loss of economically important crop caused by V. dahliae has raised worldwide concern, however, little is known about the mechanism of its pathogenicity (Klosterman et al., 2011; Yadeta and Thomma, 2013). Our recent work has shown that V. dahliae develops hyphopodium as an infection structure to breach plant root cell wall (Zhao et al., 2016). Here, we provide a detailed protocol to analyze the penetration ability and the pathogenicity of V. dahliae as well as recover fungal hyphae from infected cotton stems developed from our previous studies (Zhang et al., 2016a and 2016b; Zhao et al., ...
[摘要] 大丽轮枝菌属是土壤传播的植物病原真菌，其感染宿主根并在血管组织中增殖。经济上重要的作物造成的巨大损失。大丽花已经引起了全世界的关注，然而，其致病性机制知之甚少（Klosterman等人，2011; Yadeta和Thomma，2013）。我们最近的工作表明，大丽花开发hyphopodium作为感染结构以破坏植物根细胞壁（Zhao等人，2016）。在这里，我们提供了一个详细的方案来分析渗透能力和致病性。大丽花，以及从以前的研究（Zhang等人，2016a和2016b; Zhao等人，2016年）开发的感染棉花茎中恢复真菌菌丝）。玻璃纸膜已被用于诱导叶面病原体的发育，而不是根系病原体（Bourett和Howard，1990）。我们采用玻璃纸膜的方法诱导和评估hyphopodium的发展。希望能大力促进调控传染性发育的宿主的分子生物学研究。该协议还有助于识别控制V的致病性的关键成分。大丽花，并扩大了我们对植物与微生物相互作用机理的理解。
背景 玻璃纸膜被广泛用于研究叶病原体感染结构的发展（Bourett和Howard，1990; Kleemann等人，2012; Gu等人， ...
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| In vitro Cell Wall Stress Assay for Fusarium oxysporum
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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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| Establishing a Biofilm Co-culture of Pseudomonas and Aspergillus for Metabolite Extraction
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Author:
Date:
2015-12-05
[Abstract] Filamentous fungi and bacteria form mixed-species biofilms in nature and diverse clinical contexts (Frey-Klett et al., 2011; Peleg et al., 2010). The interactions between fungi and bacteria, often mediated by secreted metabolites, have important ramifications for the biology of the interacting partners (Frey-Klett et al., 2011). This is particularly true for the bacterium Pseudomonas aeruginosa (P. aeruginosa) and the fungus Aspergillus fumigatus (A. fumigatus) which often reside in the same niche such as lungs of cystic fibrosis (CF) patients. Some studies have reported that co-infection with P. aeruginosa and A. fumigatus could lead to a decrease in lung function relative to their respective single species ...
[摘要] 丝状真菌和细菌在自然界和不同的临床背景中形成混合物种生物膜（Frey-Klett等人，2011; Peleg等人，2010）。真菌和细菌之间的相互作用，通常由分泌的代谢物介导，对相互作用的伴侣的生物学具有重要的影响（Frey-Klett等人，2011）。对于绿脓假单胞菌（铜绿假单胞菌）和真菌烟曲霉（ A。fumigatus >），其常常位于相同的位置，例如囊性纤维化（CF）患者的肺。一些研究报道了与p的共感染。铜绿和 烟曲霉可能导致相对于它们各自的单一物种感染的肺功能降低（Amin等人，2010; Peleg等人，2010）。代谢物提取和分析允许表征多微生物生物膜中的特定微生物代谢物。该方案描述了如何在固体培养基上制备假单胞菌属 - 曲霉菌共培养生物膜以准备代谢物提取。
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