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Glycoside-specific metabolomics combined with precursor isotopic labeling for characterizing plant glycosyltransferases

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Molecular Plant 2022年第10期15卷 1517-1532页

作者： Jie Wu Wentao Zhu Xiaotong Shan Jinyue Liu Lingling Zhao Qiao Zhao Shenzhen Key Laboratory of Synthetic Genomics Guangdong Provincial Key Laboratory of Synthetic GenomicsCAS Key Laboratory of Quantitative Engineering BiologyShenzhen Institute of Synthetic BiologyShenzhen Institutes of Advanced TechnologyChinese Academy of SciencesShenzhen 518055China Innovation Center of Pesticide Research Department of Applied ChemistryCollege of ScienceChina Agricultural UniversityBeijing 100193China Center for Plant Biology School of Life SciencesTsinghua UniversityBeijing 100084China

Glycosylation by uridine diphosphate-dependent glycosyltransferases(UGTs)in plants contributes to the complexity and diversity of secondary *** are generally promiscuous in their use of acceptors,making it challenging to reveal the function of UGTs in ***,we described an approach that combined glycoside-specific metabolomics and precursor isotopic labeling analysis to characterize UGTs in *** revisited the UGT72E cluster,which has been reported to catalyze the glycosylation of ***-specific metabolomics analysis reduced the number of differentially accumulated metabolites in the ugt72e1e2e3 mutant by at least 90%compared with that from traditional untargeted metabolomics *** addition to the two previously reported monolignol glycosides,a total of 62 glycosides showed reduced accumulation in the ugt72e1e2e3 mutant,22 of which were phenylalanine-derived glycosides,including 5-OH coniferyl alcohol-derived and lignan-derived glycosides,as confirmed by isotopic tracing of[^(13)C_(6)]-phenylalanine *** method revealed that UGT72Es could use coumarins as substrates,and genetic evidence showed that UGT72Es endowed plants with enhanced tolerance to low iron availability under alkaline *** the newly developed method,the function of UGT78D2 was also *** case studies suggest that this method can substantially contribute to the characterization of UGTs and efficiently investigate glycosylation processes,the complexity of which have been highly underestimated.

关键词： glycosyltransferases glycosides-specific metabolomics precursor isotopic labeling glucoside Arabidopsis thaliana phenylpropanoid coumarins

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