Salvia species native to the Americas are rich in valuable bioactive furanoclerodanes like the psychoactive salvinorin A found in Salvia divinorum,which is used in the treatment of opioid addiction.However,rela-tively...Salvia species native to the Americas are rich in valuable bioactive furanoclerodanes like the psychoactive salvinorin A found in Salvia divinorum,which is used in the treatment of opioid addiction.However,rela-tively little is known about their biosynthesis.To address this,we investigated the biosynthesis of salviarin,the most abundant furanoclerodane structure in the ornamental sage Salvia splendens.Using a self-organizing map and mutual rank analysis of RNA-seq co-expression data,we identified three cytochrome P450 enzymes responsible for the consecutive conversion of kolavenol into the salviarin precursors:anno-nene,hardwickiic acid,and hautriwaic acid.Annonene and hardwickiic acid have been proposed as inter-mediates in the biosynthesis of salvinorin A,and we therefore tested for a common evolutionary origin of the furanoclerodane pathway in these Salvia species by searching for homologous genes in available data for S.divinorum.The enzymes encoded by orthologous genes from S.divinorum displayed kolavenol synthase,annonene synthase,and hardwickiic acid synthase activity,respectively,supporting the view that these are intermediate steps in the biosynthesis of salvinorin A.We further investigated the origin of annonene synthase and the role of gene duplication in the evolution of this specific activity.Our work shows how S.splendens can serve as a model species for the study of furanoclerodane biosynthesis in Salvia species,contributes to understanding the evolution of specialized metabolism in plants,and provides new tools for the production of salvinorin A in biotechnological chassis.展开更多
基金supported by the Strategic Priority Research Program of the Chinese Academy of Sciences(XDB27020204 awarded to E.C.T.)the International Partnership Program of the Chinese Academy of Sciences(153D31KYSB20160074 awarded to E.C.T.)+3 种基金the National Natural Science Foundation of China,Research Fund for International Excellent Young Scientists(RFIS-II)(grant 32150610477,awarded to E.C.T.)H.L.acknowledges support from the Youth Fund of the National Natural Science Foundation of China(32200313)R.L.and C.M.gratefully acknowledge support from the BBSRC ISP Grant"Harnessing Biosynthesis for Sustainable Food and Health(HBio)"(BB/X01097X/1)C.M.and E.C.T.gratefully acknowledge the Royal Society for the Newton Advanced Fellowship awarded to E.C.T.(NAF\R2\192001).
文摘Salvia species native to the Americas are rich in valuable bioactive furanoclerodanes like the psychoactive salvinorin A found in Salvia divinorum,which is used in the treatment of opioid addiction.However,rela-tively little is known about their biosynthesis.To address this,we investigated the biosynthesis of salviarin,the most abundant furanoclerodane structure in the ornamental sage Salvia splendens.Using a self-organizing map and mutual rank analysis of RNA-seq co-expression data,we identified three cytochrome P450 enzymes responsible for the consecutive conversion of kolavenol into the salviarin precursors:anno-nene,hardwickiic acid,and hautriwaic acid.Annonene and hardwickiic acid have been proposed as inter-mediates in the biosynthesis of salvinorin A,and we therefore tested for a common evolutionary origin of the furanoclerodane pathway in these Salvia species by searching for homologous genes in available data for S.divinorum.The enzymes encoded by orthologous genes from S.divinorum displayed kolavenol synthase,annonene synthase,and hardwickiic acid synthase activity,respectively,supporting the view that these are intermediate steps in the biosynthesis of salvinorin A.We further investigated the origin of annonene synthase and the role of gene duplication in the evolution of this specific activity.Our work shows how S.splendens can serve as a model species for the study of furanoclerodane biosynthesis in Salvia species,contributes to understanding the evolution of specialized metabolism in plants,and provides new tools for the production of salvinorin A in biotechnological chassis.
