TY - JOUR
T1 - Mass spectrometry-based quantification and spatial localization of small organic acid exudates in plant roots under phosphorus deficiency and aluminum toxicity
AU - Gomez-Zepeda, David
AU - Frausto, Moises
AU - Nájera-González, Héctor Rogelio
AU - Herrera-Estrella, Luis
AU - Ordaz-Ortiz, José Juan
N1 - Funding Information:
We are very grateful to Ms. Sofia Avila‐Guzmán (Waters Corporation, Mexico) and Dr. Hernando J. Olivos (Waters Corporation, USA) for lending us HDI software for MSI experiments. The authors acknowledge Félix Rico‐Reséndiz for providing gemmalings for the MALDI‐MSI experiments. This work was partly funded by Consejo Nacional de Ciencia y Tecnología (CONACyT), Mexico, with master’s scholarships for MF (454143) and HRNG (455156) and a postdoctoral fellowship for DGZ (724164). M. polymorpha
Funding Information:
We are very grateful to Ms. Sofia Avila-Guzm?n (Waters Corporation, Mexico) and Dr. Hernando J. Olivos (Waters Corporation, USA) for lending us HDI software for MSI experiments. The authors acknowledge F?lix Rico-Res?ndiz for providing M. polymorpha gemmalings for the MALDI-MSI experiments. This work was partly funded by Consejo Nacional de Ciencia y Tecnolog?a (CONACyT), Mexico, with master?s scholarships for MF (454143) and HRNG (455156) and a postdoctoral fellowship for DGZ (724164).
Publisher Copyright:
© 2021 Society for Experimental Biology and John Wiley & Sons Ltd
PY - 2021/6
Y1 - 2021/6
N2 - 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.
AB - 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.
KW - Arabidopsis thaliana
KW - Marchantia polymorpha
KW - aluminum toxicity
KW - high resolution mass spectrometry
KW - mass spectrometry
KW - mass spectrometry imaging
KW - organic acids
KW - phosphorus deficiency
KW - root exudates
KW - technical advance
UR - http://www.scopus.com/inward/record.url?scp=85105924371&partnerID=8YFLogxK
U2 - 10.1111/tpj.15261
DO - 10.1111/tpj.15261
M3 - Article
C2 - 33797826
AN - SCOPUS:85105924371
VL - 106
SP - 1791
EP - 1806
JO - Plant Journal
JF - Plant Journal
SN - 0960-7412
IS - 6
ER -