Synthetic and analytical routes to the L-amino acid conjugates of cis-OPDA and their identification and quantification in plants.

Václav Mik, Tomáš Pospíšil, Federica Brunoni, Jiří Grúz, Vladimíra Nožková, Claus Wasternack, Otto Miersch, Miroslav Strnad, Kristýna Floková, Ondřej Novák, Jitka Široká

Journal: Phytochemistry 2023;215():113855

PMID: 37690699

Full Text on Elsevier Science View on PubMed

Abstract

Cis-(+)-12-oxophytodienoic acid (cis-(+)-OPDA) is a bioactive jasmonate, a precursor of jasmonic acid, which also displays signaling activity on its own. Modulation of cis-(+)-OPDA actions may be carried out via biotransformation leading to metabolites of various functions. This work introduces a methodology for the synthesis of racemic cis-OPDA conjugates with amino acids (OPDA-aa) and their deuterium-labeled analogs, which enables the unambiguous identification and accurate quantification of these compounds in plants. We have developed a highly sensitive liquid chromatography-tandem mass spectrometry-based method for the reliable determination of seven OPDA-aa (OPDA-Alanine, OPDA-Aspartate, OPDA-Glutamate, OPDA-Glycine, OPDA-Isoleucine, OPDA-Phenylalanine, and OPDA-Valine) from minute amount of plant material. The extraction from 10 mg of fresh plant tissue by 10% aqueous methanol followed by single-step sample clean-up on hydrophilic-lipophilic balanced columns prior to final analysis was optimized. The method was validated in terms of accuracy and precision, and the method parameters such as process efficiency, recovery and matrix effects were evaluated. In mechanically wounded 30-day-old Arabidopsis thaliana leaves, five endogenous (+)-OPDA-aa were identified and their endogenous levels were estimated. The time-course accumulation revealed a peak 60 min after the wounding, roughly corresponding to the accumulation of cis-(+)-OPDA. Our synthetic and analytical methodologies will support studies on cis-(+)-OPDA conjugation with amino acids and research into the biological significance of these metabolites in plants.

Address: Department of Experimental Biology, Faculty of Science, Palacký University in Olomouc, Šlechtitelů 27, Olomouc, 783 71, Czech Republic. Electronic address: [email protected].; Department of Chemical Biology, Faculty of Science, Palacký University in Olomouc, Šlechtitelů 27, Olomouc, 783 71, Czech Republic. Electronic address: [email protected].; Laboratory of Growth Regulators, Palacký University in Olomouc & Institute of Experimental Botany AS CR, Šlechtitelů 27, Olomouc, 783 71, Czech Republic. Electronic address: [email protected].; Department of Experimental Biology, Faculty of Science, Palacký University in Olomouc, Šlechtitelů 27, Olomouc, 783 71, Czech Republic. Electronic address: [email protected].; Department of Chemical Biology, Faculty of Science, Palacký University in Olomouc, Šlechtitelů 27, Olomouc, 783 71, Czech Republic. Electronic address: [email protected].; Laboratory of Growth Regulators, Palacký University in Olomouc & Institute of Experimental Botany AS CR, Šlechtitelů 27, Olomouc, 783 71, Czech Republic. Electronic address: [email protected].; Laboratory of Growth Regulators, Palacký University in Olomouc & Institute of Experimental Botany AS CR, Šlechtitelů 27, Olomouc, 783 71, Czech Republic. Electronic address: [email protected].; Laboratory of Growth Regulators, Palacký University in Olomouc & Institute of Experimental Botany AS CR, Šlechtitelů 27, Olomouc, 783 71, Czech Republic. Electronic address: [email protected].; Laboratory of Growth Regulators, Palacký University in Olomouc & Institute of Experimental Botany AS CR, Šlechtitelů 27, Olomouc, 783 71, Czech Republic. Electronic address: [email protected].; Laboratory of Growth Regulators, Palacký University in Olomouc & Institute of Experimental Botany AS CR, Šlechtitelů 27, Olomouc, 783 71, Czech Republic. Electronic address: [email protected].; Laboratory of Growth Regulators, Palacký University in Olomouc & Institute of Experimental Botany AS CR, Šlechtitelů 27, Olomouc, 783 71, Czech Republic. Electronic address: [email protected].

DOI: 10.1016/j.phytochem.2023.113855

Link outs

Subscription / membership required

Full Text Sources:

https://linkinghub.elsevier.com/retrieve/pii/S0031...

MeSH Terms: Amino Acids, Oxylipins, Diazonium Compounds, Cyclopentanes