Enhancement of in situ detection and imaging of phytohormones in plant tissues by MALDIMSI using 2,4‐dihydroxy‐5‐nitrobenzoic acid as a novel matrix

Author:

Chen Lulu123ORCID,Zhang Yue123,Hao Qichen45,Fu Jinxiang45,Bao Zhibin45,Bu Yufen123,Sun Na123,Wu Xinyuan123,Lu Liang67,Kong Zhaosheng67ORCID,Qin Liang45,Zhou Yijun45,Jing Yanping123ORCID,Wang Xiaodong45ORCID

Affiliation:

1. State Key Laboratory of Tree Genetics and Breeding, College of Biological Sciences and Technology Beijing Forestry University Beijing 100083 China

2. National Engineering Research Center of Tree Breeding and Ecological Restoration Beijing Forestry University Beijing 100083 China

3. The Tree and Ornamental Plant Breeding and Biotechnology Laboratory of National Forestry and Grassland Administration Beijing Forestry University Beijing 100083 China

4. College of Life and Environmental Sciences, Centre for Imaging & Systems Biology Minzu University of China Beijing 100081 China

5. Key Laboratory of Mass Spectrometry Imaging and Metabolomics (Minzu University of China) State Ethnic Affairs Commission Beijing 100081 China

6. State Key Laboratory of Plant Genomics, Institute of Microbiology Chinese Academy of Sciences Beijing 100101 China

7. State Key Laboratory of Microbial Resources, Institute of Microbiology Chinese Academy of Sciences Beijing 100101 China

Abstract

Summary Phytohormones possess unique chemical structures, and their physiological effects are regulated through intricate interactions or crosstalk among multiple phytohormones. MALDI‐MSI enables the simultaneous detection and imaging of multiple hormones. However, its application for tracing phytohormones is currently restricted by low abundance of hormone in plant and suboptimal matrix selection. 2,4‐Dihydroxy‐5‐nitrobenzoic acid (DHNBA) was reported as a new MALDI matrix for the enhanced detection and imaging of multiple phytohormones in plant tissues. DHNBA demonstrates remarkable sensitivity improvement when compared to the commonly used matrix, 2,5‐dihydroxybenzoic acid (DHB), in the detection of isoprenoid cytokinins (trans‐zeatin (tZ), dihy‐drozeatin (DHZ), meta‐topolin (mT), and N6‐(Δ2‐isopentenyl) adenine (iP)), jasmonic acid (JA), abscisic acid (ABA), and 1‐aminocyclo‐propane‐1‐carboxylic acid (ACC) standards. The distinctive properties of DHNBA (i.e. robust UV absorption, uniform matrix deposition, negligible background interference, and high ionization efficiency of phytohormones) make it as an ideal matrix for enhanced detection and imaging of phytohormones, including tZ, DHZ, ABA, indole‐3‐acetic acid (IAA), and ACC, by MALDI‐MSI in various plant tissues, for example germinating seeds, primary/lateral roots, and nodules. Employing DHNBA significantly enhances our capability to concurrently track complex phytohormone biosynthesis pathways while providing precise differentiation of the specific roles played by individual phytohormones within the same category. This will propel forward the comprehensive exploration of phytohormonal functions in plant science.

Funder

National Natural Science Foundation of China

Publisher

Wiley

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