ASSESSING NITRIC OXIDE IN PLANT DEFENSE USING NO SPIN TRAPPING

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。次级项目的主要支助 子项目的主要研究者可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 NO在真核生物和原核生物中作为信号分子参与了广泛的生理过程。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响应。 研究表明，一氧化氮在植物抗病性中起着重要作用。然而，与我们对动物NO功能的理解不同，植物NO对刺激的反应的起源和量化仍然不清楚。 全面了解NO生物学依赖于可靠的NO检测方法的发展，EPR在这方面是一个可靠的工具。 用自旋陷阱络合物稳定NO自由基并用ESR检测它提供了选择性和灵敏度的优点。在ACERT中使用NO自旋陷阱Fe（DETC）2的初始ESR研究表明，模式植物拟南芥通过更快速和稳健地产生NO来响应由假单胞菌（Pseudomonasarabidingae）渗透引起的胁迫。 Dan Klessig小组对使用亲脂性Fe（DETC）2络合物或亲水性Fe（MGD）2自旋捕集器来评估1）植物细胞悬浮液中植物-微生物相互作用背景下的NO形成感兴趣（烟草细胞）用隐地蛋白（一种由卵菌隐地疫霉（Phytophtora cryptogea）分泌的诱导宿主防御反应的10 kDa蛋白）引发NO产生，和2）在用细菌病原体（假单胞菌属（Pseudomonaslingae））或对应于细菌鞭毛蛋白保守结构域的肽激发子感染的拟南芥属植物中。我们目前使用15 N取代来利用ESR测量区分14 NO和15 NO的能力，并且我们相信基于ESR的方法将有助于阐明这种自由基在植物中的起源。

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.