Development of a Matrix-Assisted Laser Desorption Ionization High Resolution Mass Spectrometry Method for the Quantification of Camalexin and Scopoletin in Arabidopsis thaliana.

Abstract

Rationale Understanding plant defense mechanisms against pathogens is essential for enhancing agricultural productivity and crop protection. This study focuses on the quantification of camalexin and scopoletin, two critical phytoalexins in Arabidopsis thaliana, using mass spectrometry techniques. Precise measurement of these compounds provides insights into plant resistance and supports agricultural research. Methods Camalexin and scopoletin were quantified using matrix-assisted laser desorption ionization high-resolution mass spectrometry (MALDI-HRMS). The matrix and solvent conditions were optimized to maximize sensitivity and accuracy. MS/MS experiments confirmed compound identification with high mass accuracy (mass error Results The method exhibited high linearity for scopoletin (R 2 = 0.9992) and camalexin (R 2 = 0.9987) across concentration ranges of 0.16-5 and 0.31-5 μM, respectively. Limits of detection (LOD) were 0.16 μM for camalexin and 0.04 μM for scopoletin, with limits of quantification (LOQ) at 0.2 μM and 0.08 μM, respectively. Samples analysis demonstrated reliable quantification in WT and mutant lines, with significant reductions in camalexin and scopoletin levels observed in the atwrky33-2 and atmyb15-1 mutants, respectively. Additionally, the method detected sub-physiological concentrations, confirming its sensitivity and robustness for low-level detection. Conclusions This study presents a validated, precise, and accurate MALDI-HRMS method for the quantification of camalexin and scopoletin in Arabidopsis thaliana. The approach not only enhances understanding of plant defense mechanisms but also offers potential applications for biotechnological and agricultural research, especially for investigating genetic variations and stress-induced phytoalexin production.

Plant phenotyping relevance

植物の防御状態・ストレス応答に関連するファイトアレキシン量を対象としたMALDI-HRMS定量法を開発・最適化し、直線性、検出限界、定量限界、感度、頑健性を検証しているため、化学測定が中心的なフェノタイピング手法研究に該当する。

Code and data availability

The paper reports MALDI-HRMS quantification of camalexin and scopoletin in Arabidopsis, with supporting information (Table S1) containing the analytical replicate and ion intensity measurements. However, the Data Availability Statement says data are available only from the corresponding author upon reasonable request;,

No evidence-backed public reproduction asset is currently recorded.