文献类型：期刊文献

中文题名：油楠SgTPS7的克隆及其在萜类生物合成和非生物胁迫中的功能

英文题名：Cloning of SgTPS7 in Sindora glabra and Its Function in Terpene Synthesis and Abiotic Stress

作者：余纽[1] 柳帆[1] 杨锦昌[1]

第一作者：余纽

机构：[1]中国林业科学研究院热带林业研究所,广州510520

年份：2024

卷号：40

期号：8

起止页码：164-173

中文期刊名：生物技术通报

外文期刊名：Biotechnology Bulletin

收录：CSTPCD;;北大核心:【北大核心2023】；CSCD:【CSCD2023_2024】；

基金：广东省自然科学基金青年提升项目(2024A1515030223)。

语种：中文

中文关键词：萜类合成酶;酶活;非生物胁迫;油楠

外文关键词：terpene synthase;enzymatic activity;abiotic stress;Sindora glabra

分类号：Q943.2;S794.9

摘要：【目的】萜类化合物在植物生理和防御中发挥着重要作用。萜类合成酶基因(TPS)是萜类生物合成途径中的关键酶,热带泌油树种油楠(Sindora glabra)茎部含有大量萜类树脂油,探究油楠SgTPS在萜类物质合成和非生物胁迫响应中的功能,为通过合成生物学生产萜类提供重要基因资源,有助于拓展植物萜类合成途径与调控机制的认识。【方法】利用油楠基因组和转录组数据从油楠茎部组织克隆SgTPS7,并对其进行生物信息学分析;表达纯化SgTPS7蛋白利用GC-MS鉴定其催化功能;采用RT-qPCR技术分析SgTPS7的表达模式。【结果】油楠SgTPS7编码区序列全长1650 bp。经同源比对分析发现,SgTPS7与苏木亚科古巴香胶树(Copaifera officinalis)CoTPS2的相似性最高,达92.3%,与其他科属植物的亲缘关系较远,属于苏木亚科特异的TPS基因。体外酶活试验表明,SgTPS7主要催化法尼基焦磷酸产生大根香叶烯D,同时催化香叶基焦磷酸产生芳樟醇。在非生物胁迫处理条件下,SgTPS7的表达差异显著。高温胁迫处理后,SgTPS7在叶片和茎中的表达均在24 h达到峰值,而在根部的表达则呈下降趋势;长期干旱处理显著影响SgTPS7的表达,在处理5 d后SgTPS7在叶和茎中的表达均达到最高,而根部SgTPS7的表达在10 d后达到最高。【结论】SgTPS7为多功能萜类合成酶基因,其在响应高温和干旱两种非生物胁迫中可能具有重要的作用。
【Objective】Terpenes play important roles in plant physiology and defense.The terpene synthase gene(TPS)is a key enzyme in the terpene biosynthetic pathway.The stems of the tropical oil-secreting tree species Sindora glabra contain a large amount of terpenoid resin oil.This study explores the role of S.glabra SgTPS in terpene synthesis and abiotic stress response.These will provide important genetic resources for the production of terpenes through synthetic biology,and help expand the understanding of plant terpene biosynthesis pathways and regulatory mechanisms.【Method】The SgTPS7 gene was cloned from the stem tissue of S.glabra using genome and transcriptome data,and bioinformatics analysis was performed on it.The SgTPS7 protein was expressed and purified and its catalytic function was identified using GC-MS,and RT-qPCR technology was used to analyze the expression pattern of SgTPS7.【Result】The full length sequence of the coding region of SgTPS7 of S.glabra was 1650 bp.Homology comparison analysis found that SgTPS7 had the highest similarity with CoTPS2 of Copaifera officinalis,at 92.3%.It was distantly related to plants of other families and genera,and it belonged to the unique TPS genes of Caesalpinioideae.In vitro enzyme activity experiments showed that SgTPS7 mainly catalyzed farnesyl pyrophosphate to produce germacrene D,and catalyzed geranyl pyrophosphate to produce linalool.The expression of SgTPS7 gene was significantly different under different stress treatment conditions.After treatment with high-temperature stress,the expression of SgTPS7 in the leaves and stems reached the peak at 24 h,while the expression in the roots showed a downward trend.Long-term drought treatment significantly affected the expression of SgTPS7,and the expression of SgTPS7 in the leaves and stems reached the highest after 5 days of treatment,while the expression of SgTPS7 in the roots reached the highest after 10 d.【Conclusion】SgTPS7 is a multifunctional terpene synthase gene,which plays an important role in responding to abiotic stresses such as high temperature and drought.