Functional Analyses Of The Terpenoid Biosynthetic Genes From Two Salvia Species And Glycine Max

Terpenoid is considered form the largest group of natural products and a class of secondary metabolites found in a lot of plants and many other organisms with more than 40,000 different structures.The plant produces multiple terpenoid compounds with highly diverse structures.These compounds play critical role and function in the interactions with environmental factors and fundamental biological processes.Terpenoid,which is a characteristic feature of plants,is of particular importance and can protect plants against insect and a wide variety of microorganisms(i.e.,viruses,bacteria,fungi).Numerous studies have been carried out to understand in-depth the mechanisms of terpene functions in the transgenic plant.Garden sage(Salvia officinalis L.)and Blue Anise Sage(Salvia guaranitica L),perennial herb plants of Lamiaceae family,and an important genus for their medicinal,aromatic and ethnobotanical properties,which have various terpenes components in different tissues.However,the genetic basis of terpenes biosynthesis pathways in these two species is virtually unknown.In this study,a transcriptome database established for S.officinalis,and S.guaranitica leaves using next-generation sequencing(NGS)technology to identify and characterize related genes in the terpenoid biosynthesis pathway.After completed sequencing,we characterized the functions of terpene synthase genes(SoNEOD,SoCINS,SoSABS,SoLINS,SoFPPS,SoTPS6,SoFARD,SgFPPS,SgGPS,and SgLINS)from S.officinalis and S.guaranitica using transgenic Nicotiana tabacum,Arabidopsis thaliana,and in vitro Glycine max hairy root to determine their biological significance for terpenoid production in various tissues.And we found most of the transgenic Tobacco,Arabidopsis plants and Glycine max hairy root expressing these terpene synthetic genes produced increased amounts of terpenoids compared with wild-type and GUS as a control.In addition,we cloned and functional characterized two terpene biosynthesis genes,GmFDPS and GmGGPP,from Glycine max.Furthermore,twelve terpene genes(GUS as a control SoNEOD,SoCINS,SoGPS,SoSABS,SoLINS,SoFARD,SoTPS6,SgFPPS,SgGPS,SgLINS,GmFDPS and GmGGPP)from S.officinalis L,S.guaranitica L,and Glycine max were studied for their functions in nodulation biosynthesis pathway by using in vivo transgenic soybean hairy roots.And we found most of the transgenic soybeans hairy root lines expressing these terpene genes and increased the number of nodules and weight compared with GUS as a control.One the other hand,Dual-choice feeding-preference and force-feeding tests were conducted to assess the insect resistance of transgenic N.tabacum plants.And we found most of the insect leafworms preferred to the wild-type tobacco leaves rather than the SoNEOD,SoCINS,SoSABS,SoLINS or SoTPS6-expressing N.tabacum leaves.Based on our results,it is believed that the resistance came from the accumulation of various types of terpenoids.This study provide new insights into our understanding of the complex metabolic genes in terpenoid-rich sage,and our study paves the way for the future metabolic engineering of the biosynthesis of useful terpene compounds in S.officinalis,S.guaranitica,Nicotiana tabacum,Arabidopsis thaliana,and Glycine max hairy root.Moreover,our study provides new insight into our understanding the epistatic relationship between terpene genes and the nodulation signaling gene pathway.Furthermore,this study paves the way for the future metabolic engineering for increasing insect-resistant soybean cultivars.