Metabolite Profiling and Transcriptome Analysis Explains Difference in Accumulation of Bioactive Constituents in Licorice (Glycyrrhiza uralensis) Under Salt Stress.

Chengcheng Wang,Mengxia Tan,Lihong Chen,Jiali Chen,Lifang Wei,Xunhong Liu,Yuqi Mei,Shengjin Liu,Lisi Zou,Juan Liang,Zhichen Cai,Cuihua Chen,Jine Chen,Zixiu Liu

doi:10.3389/fpls.2021.727882

Abstract

Salinity stress significantly affects the contents of bioactive constituents in licorice Glycyrrhiza uralensis. To elucidate the molecular mechanism underlying the difference in the accumulation of these constituents under sodium chloride (NaCl, salt) stress, licorice seedlings were treated with NaCl and then subjected to an integrated transcriptomic and metabolite profiling analysis. The transcriptomic analysis results identified 3,664 differentially expressed genes (DEGs) including transcription factor family MYB and basic helix-loop-helix (bHLH). Most DEGs were involved in flavonoid and terpenoid biosynthesis pathways. In addition, 121 compounds including a triterpenoid and five classes of flavonoids (isoflavone, flavone, flavanone, isoflavan, and chalcone) were identified, and their relative levels were compared between the stressed and control groups using data from the ultrafast liquid chromatography (UFLC)–triple quadrupole–time of flight–tandem mass spectrometry (TOF–MS/MS) analysis. Putative biosynthesis networks of the flavonoids and triterpenoids were created and combined with structural DEGs such as phenylalanine ammonia-lyase (PAL), 4-coumarate-CoA ligase [4CL], cinnamate 4-hydroxylase [C4H], chalcone synthase [CHS], chalcone-flavanone isomerase [CHI], and flavonoid-3′,5′ hydroxylase (F3′,5′H) for flavonoids, and CYP88D6 and CYP72A154 for glycyrrhizin biosynthesis. Notably, significant upregulation of UDP-glycosyltransferase genes (UGT) in salt-stressed licorice indicated that postmodification of glycosyltransferase may participate in downstream biosynthesis of flavonoid glycosides and triterpenoid saponins. Accordingly, the expression trend of the DEGs is positively correlated with the accumulation of glycosides. Our study findings indicate that key DEGs and crucial UGT genes co-regulate flavonoid and saponin biosynthesis in licorice under salt stress.

Highlights

Licorice (Glycyrrhiza uralensis) is considered to be one of the most economically important medicinal plants worldwide
We investigated the molecular mechanisms involved in the biosynthesis of the chemical components of licorice cultivated under salt-stress conditions with a focus on qualitative differences, using an integrative transcriptomic analysis and metabolomic approach
On the basis of the chemical features of flavonoid glycosides and triterpenoid saponins, we focused on the expression changes of UDP-glycosyltransferase genes (UGT), which have been demonstrated to regulate the synthesis of various secondary metabolites and resistance to environmental stress (Liang et al, 2015)

Summary

Introduction

Licorice (Glycyrrhiza uralensis) is considered to be one of the most economically important medicinal plants worldwide. Research on key genes regulating the biosynthesis of these constituents in licorice growing naturally under high salinity conditions still remains limited. The whole-genome sequence of G. uralensis has been published and potential genomic information about molecules involved in the biosynthesis of bioactive constituents has been identified (Mochida et al, 2017). These findings provide necessary and important genetic clues for developing strategies for effective molecular breeding and crop-quality control (Mochida et al, 2017). It is extremely important to characterize the key genes in the biosynthetic pathways of these constituents of licorice

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