‘海黄’牡丹香叶醇合酶基因PsGES的克隆及功能验证  

Cloning and Functional Verification of Geraniol Synthase Gene PsGES in Tree Peony‘High Noon’

作　　者：王佩云 李子昂 白杨 杨萍 尹承芃 宋秀华[1] WANG Pei-yun;LI Zi-ang;BAI Yang;YANG Ping;YIN Cheng-peng;SONG Xiu-hua(College of Horticulture Science and Engineering,Shandong Agricultural University,Taian,Shandong,271018)

机构地区：[1]山东农业大学园艺科学与工程学院,山东泰安271018

出　　处：《林业科学研究》2024年第4期52-61,共10页Forest Research

基　　金：教育部产学合作协同育人项目(202101280004)。

摘　　要：[目的]探究植物释放的萜类化合物合成代谢的机理,为进一步解析牡丹萜类合酶基因家族的生物学功能提供理论依据。[方法]以芳香型牡丹品种‘海黄’(Paeonia×lemoinei‘High Noon’)为材料,根据转录组测序所得片段,克隆得到PsGES基因。同时利用瞬时表达方法将PsGES基因过表达载体注射进烟草叶片和‘凤丹’花瓣中验证基因功能。[结果]该结构基因的完整CDS序列长897 bp,编码298个氨基酸,氨基酸序列分析发现该蛋白具有较高的保守性,有1个保守结构域Terpene_cyclase_plant_C1。PsGES基因在盛开期花瓣中表达量最高且明显高于其他器官。亚细胞定位分析表明PsGES主要定位于叶绿体中。通过GC-MS方法可检测到烟草叶片和‘凤丹’花瓣中香叶醇的释放,同时烟草叶片和‘凤丹’花瓣中PsGES基因呈现高表达。[结论]本研究表明PsGES基因在植物体内调控香叶醇的合成。[Objective]To explore the mechanism of synthesis and metabolism of terpenoids released by plants,and to provide theoretical basis for further elucidating the biological functions of the terpenoid synthase gene family in tree peony.[Method]Based on the aromatic tree peony variety‘High Noon’,the PsGES gene was cloned based on transcriptome sequencing of the obtained fragments.Simultaneously,the PsGES gene overexpression vector was injected into tobacco leaves and‘Fengdan’petals using transient expression methods to verify gene function.[Result]The complete CDS sequence of this structural gene was 897 bp long,encoding 298 amino acids.Amino acid sequence analysis showed that the protein had high conservation,with a conserved domain Terpene_Cyclase_Plant_C1.The expression level of PsGES gene was highest in the blooming petals and significantly higher than other organs.Subcellular localization analysis showed that PsGES was mainly localized in chloroplasts.The release of geraniol in tobacco leaves and‘Fengdan’petals could be detected by GC-MS method,while the PsGES gene was highly expressed in tobacco leaves and‘Fengdan’petals.[Conclusion]This study indicates that the PsGES gene regulates the synthesis of geraniol in plants.

关 键 词：牡丹 香叶醇 香叶醇合酶基因 基因克隆 功能验证 

分 类 号：S685.11[农业科学—观赏园艺]