A high-performance liquid chromatography (HPLC) system for analyzing plant defense hormone salicylic acid

用于分析植物防御激素水杨酸的高效液相色谱 (HPLC) 系统

• 批准号: RTI-2023-00119
• 负责人: Li, Xin
• 金额: $ 6.48万
• 依托单位: University of British Columbia
• 依托单位国家: 加拿大
• 项目类别: Research Tools and Instruments
• 财政年份: 2022
• 资助国家: 加拿大
• 起止时间: 2022-01-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=760916
• 关键词: performance; liquid; chromatography; HPLC; system

With the human population reaching almost eight billion, sustainable solutions are needed to ensure food security. One major factor contributing to crop yield and quality loss is diseases caused by pathogens. Every year farmers spend billions of dollars on pesticides, even though many of them are environmentally unfriendly, toxic to mammals, and can lead to pesticide-resistant pathogens. An alternative method is to fine-tune the natural defense system that plants employ to fight against pathogens. Such strategy is more sustainable and environmentally safe. To achieve this, scientists around the globe, including the ones in the applicants' labs, are teaming up to study the virulence strategies of pathogens in disturbing host immunity, and the underlying general defense mechanisms of plants. Salicylic acid (SA) is an essential plant defense hormone. Quantifying SA levels accurately is crucial for analyzing plant immune responses upon pathogen infections. It is equivalent of a routine white blood cell count to evaluate human immunity status. Because we routinely measure SA levels in studies of plant immunity and pathogen infection strategies, an HPLC system was customized for SA measurements. It had been heavily used not only by the Li lab, but also other neighboring Kronstad and Zhang groups. The current system served us for over 20 years. Its usage allowed us to solve the SA biosynthesis pathway, uncover the SA receptors, and identify the pathways SA is regulating. A large amount of data generated using this HPLC system was instrumental for several high-impact papers recently published in top journals such as Nature, Science, Cell, and Plant Cell. SA measurement will remain an essential assay to dissect plant immune signaling pathways and uncover virulence strategies of pathogens. Unfortunately, due to its heavy usage over the past 21 years, the HPLC system broke down early 2021. Old broken parts are not available and servicing support for the system is no longer provided. Alternative methods for measuring SA levels without the HPLC system, such as a bioluminescence bioassay, are not accurate and unsuitable for high quality publications. The loss of the HPLC system significantly impacts the research output from our labs and there is an immediate need for a replacement to process the backlog of samples we have now accumulated. The purpose of this application is to obtain funds for an upgraded replacement system to facilitate accurate SA measurement. The HPLC will be shared among researchers of multiple labs in the Biological Sciences and Michael Smith buildings at UBC. Its availability will continue to help us attract excellent diverse trainees and train them with analytical chemistry skills. No discrimination of any kind will be allowed in its usage. The applicants pledge to especially ensure that users from underserved communities receive fair and timely access to the equipment and training.

随着人口接近80亿，需要可持续的解决方案来确保粮食安全。造成作物产量和品质损失的一个主要因素是由病原体引起的疾病。每年，农民在农药上花费数十亿美元，尽管其中许多农药对环境不友好，对哺乳动物有毒，并可能导致抗药性病原体。另一种方法是微调植物用来对抗病原体的自然防御系统。这种战略更具可持续性和环境安全性。为了实现这一目标，世界各地的科学家，包括申请人实验室的科学家，正在合作研究病原体在干扰宿主免疫方面的毒力策略，以及植物潜在的一般防御机制。水杨酸（SA）是一种重要的植物防御激素。准确定量SA水平对于分析植物对病原体感染的免疫反应至关重要。它相当于常规白细胞计数来评估人体免疫状态。由于我们在植物免疫和病原体感染策略研究中经常测量SA水平，因此我们为SA测量定制了高效液相色谱系统。它不仅被李实验室大量使用，而且被邻近的Kronstad和Zhang团队大量使用。现行的制度为我们服务了20多年。它的使用使我们能够解决SA的生物合成途径，揭示SA受体，并确定SA调节的途径。使用该高效液相色谱系统生成的大量数据对最近在Nature， Science， Cell和Plant Cell等顶级期刊上发表的几篇高影响力论文至关重要。SA测量将仍然是解剖植物免疫信号通路和揭示病原体毒力策略的必要分析。不幸的是，由于过去21年的大量使用，HPLC系统在2021年初出现了故障。旧的损坏部件不可用，不再提供系统的维修支持。没有高效液相色谱系统测量SA水平的替代方法，如生物发光生物测定，不准确，不适合高质量的出版物。HPLC系统的丢失严重影响了我们实验室的研究成果，我们迫切需要一个替代系统来处理我们目前积累的积压样品。本申请的目的是为升级的替代系统获得资金，以促进准确的SA测量。HPLC将在UBC生物科学和Michael Smith大楼的多个实验室的研究人员之间共享。它的可用性将继续帮助我们吸引优秀的不同的学员，并培养他们的分析化学技能。在使用上不允许有任何形式的歧视。申请人承诺特别确保来自服务不足社区的用户公平及时地获得设备和培训。