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Detection and Visualization of an Exopolysaccharide Produced by Xylella fastidiosa In Vitro and In Planta

机译:Xylella fastidiosa体外和植物体内产生的胞外多糖的检测和可视化

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摘要

Many phytopathogenic bacteria, such as Ralstonia solanacearum, Pantoea stewartii, and Xanthomonas campestris, produce exopolysaccharides (EPSs) that aid in virulence, colonization, and survival. EPS can also contribute to host xylem vessel blockage. The genome of Xylella fastidiosa, the causal agent of Pierce's disease (PD) of grapevine, contains an operon that is strikingly similar to the X. campestris gum operon, which is responsible for the production of xanthan gum. Based on this information, it has been hypothesized that X. fastidiosa is capable of producing an EPS similar in structure and composition to xanthan gum but lacking the terminal mannose residue. In this study, we raised polyclonal antibodies against a modified xanthan gum polymer similar to the predicted X. fastidiosa EPS polymer. We used enzyme-linked immunosorbent assay to quantify production of EPS from X. fastidiosa cells grown in vitro and immunolocalization microscopy to examine the distribution of X. fastidiosa EPS in biofilms formed in vitro and in planta and assessed the contribution of X. fastidiosa EPS to the vascular occlusions seen in PD-infected grapevines.
机译:许多植物致病细菌,例如青枯菌(Ralstonia solanacearum),潘托亚氏菌(Pantoea stewartii)和油菜黄单胞菌(Xanthomonas campestris),都会产生胞外多糖(EPS),这些多糖有助于毒力,定植和生存。 EPS也可能导致宿主木质部血管阻塞。葡萄皮尔斯氏病(PD)的病原体Xylella fastidiosa的基因组包含一个操纵子,该操纵子与X. campestris胶操纵子非常相似,后者负责产生黄原胶。基于该信息,已经假设,X.fastidiosa能够产生与黄原胶相似的结构和组成但没有末端甘露糖残基的EPS。在这项研究中,我们提出了一种抗黄原胶聚合物的多克隆抗体,该聚合物类似于预测的X. fastidiosa EPS聚合物。我们使用酶联免疫吸附测定法定量了体外生长的法氏疟原虫细胞的EPS产量,并通过免疫定位显微镜检查了法氏疟原虫EPS在体外和植物体内形成的生物膜中的分布,并评估了法氏疟原虫EPS对生物膜的贡献。被PD感染的葡萄中发现的血管阻塞。

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