Use of isotope mass probes for metabolic analysis of the jasmonate biosynthetic pathway

摘要: In order to quantitatively study the jasmonate biosynthetic pathway, we chemically synthesized a pair of isotope mass probes and established labeling protocol. The used in our work were ω-bromoacetonylpyridinium bromide (BPB) d5-ω-bromoacetonylpyridinium (d5-BPB), which contain carboxylic acid reactive groups, isotopically labeled groups permanent positive charges. High performance liquid chromatography (HPLC) electrospray ionization quadrupole-time flight spectrometry (ESI-QTOF-MS) for detection standard mixtures plant samples. comparison negative mode unlabeled analytes, ESI signal reverse charge compounds was shown improve by 20- 80-fold. Accurate relative quantification achieved as no isotopic effects different phytohormones during RP/SCX mixed-mode chromatographic separation observed. A data analysis method analyzing metabolic pathways using strategy. We then applied examined pathway rice under salt stress premature senescence mutant. Here found that conditions, showed up-regulation (13S)-hydroperoxyoctadecatrienoic (HOPT), cis-(+)-12-oxophytodienoic (OPDA), 3-oxo-2-(2′-pentenyl)-cyclopentane-1-octanoic (OPC-8) jasmonoyl-valine (JA-Val) levels, while α-linolenic (LA) jasmonic (JA) down-regulation, three components (HPOT, OPC-8 JA-Val) accumulated. mutant all major with exception LA, an accumulation HPOT, OPC-6 JA-Val. This demonstrates chemical stable strategy can be powerful tool plants.