ASSESSING NITRIC OXIDE IN PLANT DEFENSE USING NO SPIN TRAPPING

不使用自旋捕获评估植物防御中的一氧化氮

• 批准号: 8364044
• 负责人: BORIS G DZIKOVSKI
• 金额: $ 0.05万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-01 至 2012-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8364044
• 关键词: Animals; Arabidopsis; Biology; Cells; Complex; Detection; Development; Disease Resistance; Eukaryota; Flagellin; Flowers; Funding; Grant; Growth and Development function; Host Defense; Infiltration; Measurement; Methods; Microbe; Mouse-ear Cress; National Center for Research Resources; Nitric Oxide; Oomycetes; Peptides; Physiological Processes; Plant Diseases; Plant Model; Plants; Play; Principal Investigator; Production; Prokaryotic Cells; Proteins; Pseudomonas syringae; Research; Research Infrastructure; Resources; Role; Signaling Molecule; Source; Spin Trapping; Stimulus; Stress; Suspension substance; Suspensions; Technology; Tobacco; United States National Institutes of Health; base; cost; defense response; ferro-N-methyl-d-glucamide dithiocarbamate; interest; pathogen; response; tool

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. NO acts as a signaling molecule in a wide range of physiological processes in both eukaryotes and prokaryotes. In plants, NO participates in growth, development, flowering, and response to biotic and abiotic stresses. It has been shown that nitric oxide plays an important role in plant disease resistance. However, unlike our appreciation of NO functioning in animals, the origin and the quantification of plant NO responses to stimuli remain unclear. A comprehensive understanding of NO biology is dependent on the development of reliable NO detection methods, and EPR represents a reliable tool in this respect. Stabilization of NO radical with spin trap complexes and its detection by ESR offer the advantage of being both selective and sensitive. Initial ESR study in ACERT using NO spin trap Fe(DETC)2 showed that the model plant Arabidopsis thaliana responds to the stress caused by infiltration with Pseudomonas syringae by more rapid and robust production of NO. The Dan Klessig group is interested in using lipophilic Fe(DETC)2 complex or hydrophilic Fe(MGD)2 spin traps to evaluate NO formation in the context of plant-microbe interactions in 1) plant cell suspensions (tobacco cells) elicited for NO production with cryptogein, a 10 kDa protein secreted by the oomycete Phytophtora cryptogea that induces host defense responses, and 2) in Arabidopsis plants infected with either a bacterial pathogen (Pseudomonas syringae) or a peptide elicitor corresponding to the conserved domain of bacterial flagellin. We currently use 15N substitution to exploit the ability of the ESR measurement to distinguish between 14NO and 15NO, and we are confident that an ESR-based approach will help to elucidate the origin(s) of this radical in planta.

该子项目是利用资源的众多研究子项目之一 由 NIH/NCRR 资助的中心拨款提供。子项目的主要支持 并且子项目的主要研究者可能是由其他来源提供的， 包括其他 NIH 来源。 子项目可能列出的总成本 代表子项目使用的中心基础设施的估计数量， NCRR 赠款不直接向子项目或子项目工作人员提供资金。 NO 在真核生物和原核生物的多种生理过程中充当信号分子。在植物中，NO 参与生长、发育、开花以及对生物和非生物胁迫的反应。 研究表明，一氧化氮在植物抗病性中发挥着重要作用。然而，与我们对动物中 NO 功能的认识不同，植物 NO 对刺激的反应的起源和定量仍不清楚。 对 NO 生物学的全面了解取决于可靠的 NO 检测方法的开发，而 EPR 是这方面的可靠工具。 使用自旋陷阱复合物稳定 NO 自由基及其通过 ESR 检测具有选择性和灵敏性的优点。 ACERT 中使用 NO 自旋捕获 Fe(DETC)2 进行的初步 ESR 研究表明，模型植物拟南芥通过更快速、更强劲地产生 NO 来响应丁香假单胞菌渗透引起的胁迫。 Dan Klessig 小组有兴趣使用亲脂性 Fe(DETC)2 复合物或亲水性 Fe(MGD)2 自旋陷阱来评估植物-微生物相互作用中 NO 的形成：1) 植物细胞悬浮液（烟草细胞）用隐菌素诱导产生 NO，隐菌素是一种由卵菌Phytophtora cryptogea 分泌的 10 kDa 蛋白质，可诱导宿主防御反应， 2) 在感染细菌病原体（丁香假单胞菌）或对应于细菌鞭毛蛋白保守结构域的肽引发子的拟南芥植物中。我们目前使用 15N 取代来利用 ESR 测量的能力来区分 14NO 和 15NO，我们相信基于 ESR 的方法将有助于阐明植物中该自由基的起源。