Although the plant kingdom provides an enormous diversity of metabolites with potentially beneficial applications for humankind,a large fraction of these metabolites and their biosynthetic pathways remain unknown.Reso...Although the plant kingdom provides an enormous diversity of metabolites with potentially beneficial applications for humankind,a large fraction of these metabolites and their biosynthetic pathways remain unknown.Resolving metabolite structures and their biosynthetic pathways is key to gaining biological understanding andto allow metabolic engineering.In orderto retrieve novel biosynthetic genes involved in specialized metabolism,we developed a novel untargeted method designated as qualitative trait GWAs(QT-GWAS)that subjects qualitative metabolic traits to a genome-wide association study,while the conventional metabolite GWAS(mGWAS)mainly considers the quantitative variation of metabolites.As a proof of the validity of QT-GWAS,23 and 15of the retrieved associations identified in Arabidopsis thaliana by QTGWAS and mGWAS,respectively,were supported by previous research.Furthermore,seven genemetabolite associations retrieved by QT-GWAS were confirmed in this study through reverse genetics combined with metabolomics and/or in vitro enzyme assays.As such,we established that CYTOCHROME P450706A5(CYP706A5)is involved in the biosynthesis of chroman derivatives,UDP-GLYCOSYLTRANSFERASE 76C3(UGT76C3)is able tohexosylate guanine in vitro and in planta,and SULFOTRANSFERASE 202B1(SULT202B1)catalyzes the sulfation of neolignans in vitro.Collectively,our study demonstrates that the untargeted QT-GWAS method can retrieve valid gene-metabolite associations at the level of enzyme-encoding genes,even new associations that cannot be found by the conventional mGwAs,providing a new approach for dissecting qualitative metabolic traits.展开更多
基金The Research Foundation-Flanders(FWO,personal PhD fellowship grant 1S38920N,awarded to M.B.)Marie Sktodowska-Curie Actions(MSCA,Individual Fellowship CHORPATH-897918,awarded to M.P.)+4 种基金R.H.was funded by Marie Sktodowska-Curie Actions COFUND(OMICS@VIB)I.E.H.was funded by FWO personal PhD fellowship grant(1S04020N)and iBOF(Next-BIOREF,011B4220)C.D.and W.B.were funded by Stanford University's Global Climate and Energy Project"Towards New Degradable Lignin Types"and W.B.by the ERC-Advanced Grant POPMET.We also thank the Bijzonder Onderzoeksfonds-Zware Apparatuur of Ghent University for the Fourier transform ion cyclotron resonance mass spectrometer(174PZA05)the Hercules program of Ghent University for the Synapt QTOF High Definition MS(grant AUGE/014)V.I.T.and J.R.were funded by the DOE Great Lakes Bioenergy Research Center(DOE BER Office of Science DESC0018409).
文摘Although the plant kingdom provides an enormous diversity of metabolites with potentially beneficial applications for humankind,a large fraction of these metabolites and their biosynthetic pathways remain unknown.Resolving metabolite structures and their biosynthetic pathways is key to gaining biological understanding andto allow metabolic engineering.In orderto retrieve novel biosynthetic genes involved in specialized metabolism,we developed a novel untargeted method designated as qualitative trait GWAs(QT-GWAS)that subjects qualitative metabolic traits to a genome-wide association study,while the conventional metabolite GWAS(mGWAS)mainly considers the quantitative variation of metabolites.As a proof of the validity of QT-GWAS,23 and 15of the retrieved associations identified in Arabidopsis thaliana by QTGWAS and mGWAS,respectively,were supported by previous research.Furthermore,seven genemetabolite associations retrieved by QT-GWAS were confirmed in this study through reverse genetics combined with metabolomics and/or in vitro enzyme assays.As such,we established that CYTOCHROME P450706A5(CYP706A5)is involved in the biosynthesis of chroman derivatives,UDP-GLYCOSYLTRANSFERASE 76C3(UGT76C3)is able tohexosylate guanine in vitro and in planta,and SULFOTRANSFERASE 202B1(SULT202B1)catalyzes the sulfation of neolignans in vitro.Collectively,our study demonstrates that the untargeted QT-GWAS method can retrieve valid gene-metabolite associations at the level of enzyme-encoding genes,even new associations that cannot be found by the conventional mGwAs,providing a new approach for dissecting qualitative metabolic traits.
