The BioActives Lab, Center for Desert Agriculture, Division of Biological and Environmental Sciences and Engineering, King Abdullah University of Science and Technology, Thuwal, Kingdom of Saudi Arabia.
Curr Protoc. 2022 Feb;2(2):e375. doi: 10.1002/cpz1.375.
Carotenoid oxidative cleavage products, apocarotenoids (APOs), are a class of important plant secondary metabolites, which include phytohormones abscisic acid (ABA) and strigolactones (SLs), and growth regulators and signaling molecules such as β-cyclocitral, zaxinone, anchorene, β-apo-11-carotenoids, and retinal. Qualitative and quantitative analysis of these bioactive compounds is crucial for understanding their metabolism and may also enable discovering further regulatory APOs. The state-of-the-art mass spectrometry (MS) technology has advanced the detection of plant APOs; however, it is still challenging to perform an accurate analysis of the low-level phytohormones ABA and SL and the structurally diverse APOs from complex plant matrices. Here, we describe ultrahigh-performance liquid chromatography-MS (UHPLC-MS) methods to determine carotenoid-derived hormones and APOs from plants by integrating ultrasound-assisted extraction and solid-phase extraction. These assays enable an accurate quantification of carotenoid-derived hormones and APOs from plant tissues by using an UHPLC hybrid quadrupole-Orbitrap mass spectrometer. © 2022 Wiley Periodicals LLC. Basic Protocol 1: UHPLC-MS analysis of APOs from rice roots Support Protocol: Preparation of dried plant root powder Basic Protocol 2: UHPLC-MS analysis of SLs from rice roots Basic Protocol 3: UHPLC-MS analysis of ABA from rice roots.
类胡萝卜素氧化裂解产物，即脱辅基类胡萝卜素 (APO)，是一类重要的植物次生代谢产物，包括植物激素脱落酸 (ABA) 和独脚金内酯 (SL)，以及生长调节剂和信号分子，如β-环柠檬醛、玉米黄质、栓皮酮、β-apo-11-类胡萝卜素和视黄醛。这些生物活性化合物的定性和定量分析对于了解它们的代谢至关重要，也可能有助于发现更多的调节 APO。最先进的质谱 (MS) 技术已经推进了植物 APO 的检测；然而，从复杂的植物基质中对低水平的植物激素 ABA 和 SL 以及结构多样的 APO 进行准确分析仍然具有挑战性。在这里，我们描述了通过整合超声辅助提取和固相萃取，使用超高效液相色谱-MS (UHPLC-MS) 方法从植物中测定类胡萝卜素衍生的激素和 APO。这些检测方法可通过使用 UHPLC 混合四极杆-Orbitrap 质谱仪对植物组织中的类胡萝卜素衍生激素和 APO 进行准确的定量。© 2022 威利父子公司。基本方案 1：水稻根中 APO 的 UHPLC-MS 分析 支持方案：干植物根粉的制备 基本方案 2：水稻根中 SL 的 UHPLC-MS 分析 基本方案 3：水稻根中 ABA 的 UHPLC-MS 分析。
