期刊
PLANT JOURNAL
卷 106, 期 6, 页码 1791-1806出版社
WILEY
DOI: 10.1111/tpj.15261
关键词
aluminum toxicity; Arabidopsis thaliana; high resolution mass spectrometry; Marchantia polymorpha; mass spectrometry; mass spectrometry imaging; organic acids; phosphorus deficiency; root exudates; technical advance
资金
- Consejo Nacional de Ciencia y Tecnologia (CONACyT), Mexico [454143, 455156, 724164]
Low-molecular-weight organic acid (OA) extrusion by plant roots plays a critical role in plant nutrition, tolerance to cations toxicity, and plant-microbe interactions. This study developed two complementary methodologies for OA determination, which were used to evaluate the effect of inorganic ortho-phosphate (Pi) deficiency and aluminum toxicity on OA excretion by Arabidopsis roots. The quantification of six OAs in root exudates and the spatial location of citric and malic acid in roots and exudates were successfully achieved using targeted ultra-performance liquid chromatography coupled with high-resolution tandem mass spectrometry (UPLC-HRMS/MS) and targeted matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging (MSI).
Low-molecular-weight organic acid (OA) extrusion by plant roots is critical for plant nutrition, tolerance to cations toxicity, and plant-microbe interactions. Therefore, methodologies for the rapid and precise quantification of OAs are necessary to be incorporated in the analysis of roots and their exudates. The spatial location of root exudates is also important to understand the molecular mechanisms directing OA production and release into the rhizosphere. Here, we report the development of two complementary methodologies for OA determination, which were employed to evaluate the effect of inorganic ortho-phosphate (Pi) deficiency and aluminum toxicity on OA excretion by Arabidopsis roots. OA exudation by roots is considered a core response to different types of abiotic stress and for the interaction of roots with soil microbes, and for decades has been a target trait to produce plant varieties with increased capacities of Pi uptake and Al tolerance. Using targeted ultra-performance liquid chromatography coupled with high-resolution tandem mass spectrometry (UPLC-HRMS/MS), we achieved the quantification of six OAs in root exudates at sub-micromolar detection limits with an analysis time of less than 5 min per sample. We also employed targeted (MS/MS) matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging (MSI) to detect the spatial location of citric and malic acid with high specificity in roots and exudates. Using these methods, we studied OA exudation in response to Al toxicity and Pi deficiency in Arabidopsis seedlings overexpressing genes involved in OA excretion. Finally, we show the transferability of the MALDI-MSI method by analyzing OA excretion in Marchantia polymorpha gemmalings subjected to Pi deficiency.
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