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

作　　者：Jie Wu Wentao Zhu Xiaotong Shan Jinyue Liu Lingling Zhao Qiao Zhao 

机构地区：[1]Shenzhen Key Laboratory of Synthetic Genomics,Guangdong Provincial Key Laboratory of Synthetic Genomics,CAS Key Laboratory of Quantitative Engineering Biology,Shenzhen Institute of Synthetic Biology,Shenzhen Institutes of Advanced Technology,Chinese Academy of Sciences,Shenzhen 518055,China [2]Innovation Center of Pesticide Research,Department of Applied Chemistry,College of Science,China Agricultural University,Beijing 100193,China [3]Center for Plant Biology,School of Life Sciences,Tsinghua University,Beijing 100084,China

出　　处：《Molecular Plant》2022年第10期1517-1532,共16页分子植物（英文版）

基　　金：the National Key R&D Program of China(2019YFA0903900);National Natural Science Foundation of China(grant number 31870273);Guangdong Provincial Key Laboratory of Synthetic Genomics(2019B030301006);Shenzhen Key Laboratory of Synthetic Genomics(ZDSYS201802061806209);Shenzhen Instituteof Synthetic Biology Scientific Research Program(ZTXM20190007).

摘　　要：Glycosylation by uridine diphosphate-dependent glycosyltransferases(UGTs)in plants contributes to the complexity and diversity of secondary metabolites.UGTs are generally promiscuous in their use of acceptors,making it challenging to reveal the function of UGTs in vivo.Here,we described an approach that combined glycoside-specific metabolomics and precursor isotopic labeling analysis to characterize UGTs in Arabidopsis.We revisited the UGT72E cluster,which has been reported to catalyze the glycosylation of monolignols.Glycoside-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 analysis.In 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 precursor.Our 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 conditions.Using the newly developed method,the function of UGT78D2 was also evaluated.These 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 

分 类 号：Q94[生物学—植物学]