![]() Plants that blocked the conversion of PPIX to Mg-PPIX displayed chlorophyll-deficient or albino- lethal phenotypes. The Chls biosynthetic pathway is a branch of tetrapyrrole biosynthesis, it begins with the insertion of Mg 2+ into the protoporphyrin IX (PPIX) to form magnesium protoporphyrin IX (Mg-PPIX) that catalyze by magnesium chelatase complex (Scheme 1). This procedure will facilitate studying chlorophyll metabolism and natural chlorophyll production.Ĭhlorophylls (Chls) underpin photosynthesis, which generates oxygen and fix carbon dioxide as carbohydrates that support life on Earth. Our study establishes a universal and reliable method for determining PPIX and Mg-PPIX in two plants using UPLC-MS/MS. kg − 1) and they were only detected in the leaf.kg − 1) in tea were significantly higher than in Arabidopsis (PPIX: 0.05 ± 0.02 mg.kg − 1 and the limit of quantification was 0.05 mg.Validation was performed in spiked samples and mean recoveries ranged from 70.5 to 916%, and the intra-day and inter-day variations were less than 7.5 and 10.9%, respectively. Interestingly, it could be easier to monitor these two compounds in dehydrated samples rather than in fresh samples. Since the Mg-PPIX could be substantially de-metalized into PPIX in acidic conditions, analysis was carried out by UPLC-MS/MS with 0.1% ammonia (v/v) and 0.1% ammonium acetonitrile (v/v) as mobile phases using negative ion multiple reaction monitoring modes. Two metabolites could be extracted by 80% acetone (v/v) and 20% 0.1 M NH 4OH (v/v) without hexane washing. ResultsĪn advanced and sensitive UPLC-MS/MS strategy for determining PPIX and Mg-PPIX was established in two metabolic different plants, Arabidopsis thaliana (Columbia-0) and Camellia sinensis var. However, the lack of systematic study of the detection method and the metabolic difference between species have caused the research on chloroplast retrograde signaling controversial for a long time. ![]() Plants that blocked the conversion of PPIX to Mg-PPIX displayed yellowish or albino- lethal phenotypes. Insertion of Mg 2+ into protoporphyrin IX (PPIX) to produce magnesium-protoporphyrin IX (Mg-PPIX) was the first step toward chlorophyll biosynthesis, which not only imparts plants green pigmentation but underpins photosynthesis. ![]()
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