A high-throughput chemical screen for resistance to Pseudomonas syringae in Arabidopsis
A high-throughput chemical screen for resistance to Pseudomonas syringae in Arabidopsis
复制标题
DOI：
10.1111/j.1365-313x.2008.03425.x
复制
发表时间：
2008-05-01
期刊：
影响因子：
7.2
通讯作者：
Desveaux, Darrell
中科院分区：
文献类型：
--
作者：
Schreiber, Karl;Ckurshumova, Wenzislava;Desveaux, Darrell
通过高通量的病原系统来评估病原体毒力抑制剂的小分子文库，可以极大地促进植物发病机理的研究和保护植物免受疾病的有效治疗的发展。革兰氏阴性细菌假单胞菌和拟南芥之间的相互作用是植物致病的模型。然而，一个强大的高通量测定，以评分这种相互作用的结果是目前缺乏。我们证明，拟南芥幼苗培养与P. P.在液体培养显示一个宏观上可见的“漂白”的症状在5天内感染。漂白与子叶组织叶绿素的损失有关，并且与细菌的毒性有关。在液体环境中不存在基因对基因抗性，这可能是因为在这些条件下抑制了过敏反应。重要的是，可以通过在接种前用已知的植物防御诱导剂如水杨酸（SA）或细菌鞭毛蛋白的基础防御诱导肽（fIg 22）处理幼苗来防止漂白。基于这些观察结果，我们设计了一种高通量的液体检测方法，使用标准的96孔板来研究拟南芥-拟南芥的相互作用。对拟南芥中活跃的小分子进行的初步筛选揭示了一个磺胺化合物家族，该家族提供了对漂白症状的保护。最活跃的化合物磺胺甲恶唑，也减少了植物中的细菌生长时，适用于成熟的土壤生长的植物。全生物体液体测定提供了一种以高通量方式探测化学文库以获得降低植物中细菌毒力的化合物的新方法。
The study of plant pathogenesis and the development of effective treatments to protect plants from diseases could be greatly facilitated by a high-throughput pathosystem to evaluate small-molecule libraries for inhibitors of pathogen virulence. The interaction between the Gram-negative bacterium Pseudomonas syringae and Arabidopsis thaliana is a model for plant pathogenesis. However, a robust high-throughput assay to score the outcome of this interaction is currently lacking. We demonstrate that Arabidopsis seedlings incubated with P. syringae in liquid culture display a macroscopically visible 'bleaching' symptom within 5 days of infection. Bleaching is associated with a loss of chlorophyll from cotyledonary tissues, and is correlated with bacterial virulence. Gene-for-gene resistance is absent in the liquid environment, possibly because of the suppression of the hypersensitive response under these conditions. Importantly, bleaching can be prevented by treating seedlings with known inducers of plant defence, such as salicylic acid (SA) or a basal defence-inducing peptide of bacterial flagellin (fIg22) prior to inoculation. Based on these observations, we have devised a high-throughput liquid assay using standard 96-well plates to investigate the A syringae-Arabidopsis interaction. An initial screen of small molecules active on Arabidopsis revealed a family of sulfanilamide compounds that afford protection against the bleaching symptom. The most active compound, sulfamethoxazole, also reduced in planta bacterial growth when applied to mature soil-grown plants. The whole-organism liquid assay provides a novel approach to probe chemical libraries in a high-throughput manner for compounds that reduce bacterial virulence in plants.