Single-cell RNA sequencing facilitates the elucidation of the complete biosynthesis of the antidepressant hyperforin in St. John's wort  

作　　者：Song Wu Ana Luisa Malaco Morotti Jun Yang Ertao Wang Evangelos C.Tatsis 

机构地区：[1]National Key Laboratory of Plant Molecular Genetics,CASs Center for Excellence in Molecular Plant Sciences,Shanghai lnstitute of Plant Physiology and Ecology,ChineseAcademy of Sciences,Shanghai 200032,China [2]University of Chinese Academy of Sciences,Shanghai 200032,China [3]UEPAMS-CAS-JICs Centre of Excellencefor Plant anamlcroblalSclence,Snangnal 200032,Cnina

出　　处：《Molecular Plant》2024年第9期1439-1457,共19页分子植物（英文版）

基　　金：supported by the National Natural Science Foundation of China,Research Fund for International Excellent Young Scientists(RFIS-Il)(grant 32150610477,awarded to E.C.T.);Strategic Priority Research Program of the Chinese Academy of Sciences(grant XDB27020204,awarded to E.C.T.);Chinese Academy of Sciences,International Partnership Program of CAS(grant 153D31 KYSB20160074,awarded to E.C.T.);National Key Laboratory of Plant Molecular Genetics Special Fund.

摘　　要：Hyperforin is the compound responsible for the effectiveness of St.John's wort(Hypericum perforatum)as an antidepressant,but its complete biosynthetic pathway remains unknown.Gene discovery based on co-expression analysis of bulk RNA-sequencing data or genome mining failed to discover the missing steps in hyperforin biosynthesis.In this study,we sequenced the 1.54-Gb tetraploid H.perforatum genome assem-bled into 32 chromosomes with the scaffold N50 value of 42.44 Mb.By single-cell RNA sequencing,we iden-tified a type of cell,“Hyper cells”,wherein hyperforin biosynthesis de novo takes place in both the leaves and flowers.Through pathway reconstitution in yeast and tobacco,we identified and characterized four transmembrane prenyltransferases(HpPT1-4)that are localized at the plastid envelope and complete the hyperforin biosynthetic pathway.The hyperforin polycyclic scaffold is created by a reaction cascade involving an irregular isoprenoid coupling and a tandem cyclization.Our findings reveal how and where hy-perforin is biosynthesized,enabling synthetic-biology reconstitution of the complete pathway.Thus,this study not only deepens our comprehension of specialized metabolism at the cellularlevel but also provides strategic guidance for elucidation of the biosynthetic pathways of other specializied metabolites in plants.

关 键 词：HYPERFORIN Hypericum perforatum de novo genome sequencing single-cell RNA sequencing biosyn-thetic pathway MEROTERPENOIDS 

分 类 号：Q946[生物学—植物学]