水稻水杨酸代谢突变体高通量筛选方法的建立与应用

doi:10.11983/CBB24148

摘要/Abstract

摘要： 水杨酸(SA)是植物免疫的关键防御信号分子。植物SA的定量分析对于SA代谢途径及其生物学功能研究至关重要。高效液相色谱仪(HPLC)和液相-质谱联用仪(LC/MS)是测定SA含量的常用方法, 但难以实现高通量测定。水稻(Oryza sativa)合成代谢途径尚未完全解析, 高效筛选SA相关突变体对于解析其代谢途径具有重要意义。该文针对已有的基于SA生物传感菌株Acinetobacter sp. ADPWH_lux估算SA的分析方法进行改良, 建立了水稻SA高通量估算方法, 对样品采集以及提取过程进行了简化, 省去了样品称重、组织研磨和离心等耗时步骤, 整个操作流程便捷高效。我们利用已报道的水稻SA代谢相关遗传材料证明了该方法的可行性, 同时用该方法筛选钴-60诱变的水稻突变体库, 获得了一批水稻SA含量发生显著变化的突变体, 并利用HPLC法对突变体内源SA进行验证。该方法可用于SA代谢突变体的遗传筛选和SA代谢相关酶的鉴定, 对于水稻等作物的SA代谢及生物学功能研究具有重要应用价值。

关键词: 水杨酸测定, 生物传感器, 高通量方法, 水杨酸代谢突变体, 水稻

Abstract: Salicylic acid (SA) plays essential roles in plant immunity. Quantitative analysis of SA in plants is essential for studying SA metabolism and biological function. Although high-performance liquid chromatography (HPLC) and liquid chromatography-mass spectrometry (LC/MS) are widely used for SA determination, but it is difficult to achieve high throughput determination. However, SA synthesis pathway in rice has not yet to be fully elucidated, efficient screening method of SA-related mutants is of great significance for the analysis of the metabolic pathway. In this paper, we improved the analytical method for quantifying SA based on biosensor Acinetobacter sp.ADPWH_lux, and established a rapid high-throughput method for SA quantification in rice with simplifying the sample collection and extraction process. It avoids the time-consuming steps such as sample weighing, tissue grinding and centrifugation, and the entire process is convenient and efficient. We demonstrated the method's feasibility using the published rice SA metabolism mutants. Then, we screened some rice mutants from Co-60 mutant library with high or low SA metabolism using this method and confirmed the source SA level of the mutants by HPLC. Our results showed that this method could be used to screen SA related metabolic mutants, and has important application potential for studying the role of SA metabolism and biological could function in rice and other crops.

叶灿, 姚林波, 金莹, 高蓉, 谭琪, 李旭映, 张艳军, 陈析丰, 马伯军, 章薇, 张可伟. 水稻水杨酸代谢突变体高通量筛选方法的建立与应用. 植物学报, 2025, 60(4): 586-596.

Can Ye, Linbo Yao, Ying Jin, Rong Gao, Qi Tan, Xuying Li, Yanjun Zhang, Xifeng Chen, Bojun Ma, Wei Zhang, Kewei Zhang. Establishment and Application of a High-throughput Screening Method for Salicylic Acid Metabolic Mutants in Rice. Chinese Bulletin of Botany, 2025, 60(4): 586-596.

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链接本文: https://www.chinbullbotany.com/CN/10.11983/CBB24148

https://www.chinbullbotany.com/CN/Y2025/V60/I4/586

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