北京林业大学学报2026，Vol.48Issue(6)：49-58,10.DOI:10.12171/j.1000-1522.20250429
檀香烯合酶基因启动子克隆及其活性分析
Cloning and its activity analysis of santalene synthase gene(SaSSy)promoter from Santalum album
摘要
Abstract
[Objective]The primary active component of sandalwood essential oil is santalol,yet the transcriptional regulatory mechanism of key biosynthetic enzyme gene santalene synthase(SaSSy)remains unclear.This study aimed to clone and preliminarily characterize the promoter of SaSSy gene in Santalum album,and to explore its transcriptional regulatory features.[Method]The 1 437 bp promoter sequence upstream of ATG start codon of SaSSy gene was cloned from Santalum album using Tail-PCR.Cis-regulatory element analysis was performed to identify functional motifs.Transcription factor binding sites were predicted,and KEGG enrichment analysis was conducted to explore associated regulatory pathways.The transcriptional activity of the promoter was verified via transient expression assays in tobacco leaves.Sandalwood calli were treated with 100 µmol/L methyl jasmonate(MeJA),and SaSSy expression was analyzed by RT-qPCR.GUS activity driven by the promoter was measured to assess MeJA responsiveness.Truncated promoter versions were constructed to identify functional regions.[Result]A 1 437 bp promoter sequence of SaSSy was successfully cloned,containing light-,hormone-,and stress-responsive cis-elements.Transcription factor binding site prediction and KEGG analysis revealed enrichment in pathways related to plant hormone signaling,MAPK signaling,plant-pathogen interaction and circadian rhythm.The promoter demonstrated transcriptional activity in tobacco transient assays.MeJA treatment significantly upregulated SaSSy expression and enhanced promoter-driven GUS activity.A truncated 264 bp core promoter region(PSaSSy-D4)retained full transcriptional activity,comparable to full-length promoter.[Conclusion]The present study reveals fundamental characteristics of PSaSSy,confirming that its expression is positively regulated by MeJA,and identifies a core promoter region(-264 bp)containing key cis-acting elements.The MeJA-inducible characteristics of PSaSSy provide experimental evidence for enhancing sandalwood essential oil biosynthesis through exogenous signalling molecules,and offer a candidate target for further elucidating the transcriptional regulatory network of santalol synthesis.关键词
檀香烯合酶/SaSSy启动子/遗传转化烟草/茉莉酸甲酯诱导Key words
santalene synthase/SaSSy promoter/transgenic Nicotiana tabacum/MeJA induction分类
农业科技引用本文复制引用
周慧文,丘立杭,闫海锋..檀香烯合酶基因启动子克隆及其活性分析[J].北京林业大学学报,2026,48(6):49-58,10.基金项目
国家自然科学基金项目(32060358),广西自然科学基金项目(2019GXNSFAA185005). （32060358）
