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.

随着人口达到近80亿，需要可持续的解决办法来确保粮食安全。造成作物产量和品质损失的一个主要因素是由病原菌引起的疾病。农民每年在杀虫剂上花费数十亿美元，尽管其中许多对环境不友好，对哺乳动物有毒，并可能导致杀虫剂抗药性病原体。另一种方法是微调植物用来对抗病原体的自然防御系统。这样的战略更可持续，对环境更安全。为了实现这一目标，世界各地的科学家，包括申请者实验室的科学家，正在联手研究病原体扰乱宿主免疫的毒力策略，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。准确地定量SA水平对于分析植物对病原体侵染的免疫反应至关重要。这相当于常规的白细胞计数来评估人类的免疫状况。因为我们在植物免疫和病原体感染策略的研究中经常测量SA水平，所以我们定制了一套高效液相系统来测量SA。它不仅被李实验室大量使用，而且还被邻近的克朗斯塔德和张小组使用。目前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，并识别SA调节的途径。使用这个高效液相系统产生的大量数据对最近在《自然》、《科学》、《细胞》和《植物细胞》等顶级期刊上发表的几篇高影响力的论文是有帮助的。SA测定仍将是剖析植物免疫信号途径和揭示病原菌毒力策略的重要手段。不幸的是，由于在过去21年中的大量使用，高效液相系统在2021年初崩溃了。不再提供旧的损坏部件，也不再为系统提供维修支持。没有高效液相系统的其他测量SA水平的方法，如生物发光生物分析，是不准确的，不适合高质量的出版物。高效液相系统的丢失严重影响了我们实验室的研究产出，我们迫切需要更换一个系统来处理我们现在积累的积压样本。这项申请的目的是获得资金，以升级更换系统，以促进准确的SA测量。高效液相色谱将在生物科学和UBC的迈克尔·史密斯大楼的多个实验室的研究人员之间共享。它的推出将继续帮助我们吸引优秀的多样化学员，并培训他们的分析化学技能。在使用中不允许任何形式的歧视。申请者承诺特别确保来自服务不足社区的用户公平和及时地获得设备和培训。