‘海黄’牡丹芳樟醇合酶基因PsTPS14的克隆及功能验证

Cloning and Functional Verification of Linalool Synthase Gene PsTPS14 in Tree Peony‘High Noon’

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PsTPS14基因,该结构基因的完整CDS序列长903 bp,共编码300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。PsTPS14在‘海黄’牡丹半开期花瓣中表达量最高,与其花香挥发物释放规律一致。亚细胞定位分析表明PsTPS14主要定位于叶绿体中。利用瞬时表达方法将PsTPS14过表达载体注射到烟草叶片和淡香型牡丹‘凤丹’切花花瓣中,通过GC–MS方法可检测到烟草叶片和‘凤丹’花瓣中芳樟醇的释放,同时PsTPS14高表达,且芳樟醇合酶含量及其活性明显升高。研究结果表明PsTPS14在植物体内调控芳樟醇的合成。

The linalool synthase gene PsTPS14 was cloned from the aromatic tree peony‘High Noon’based on the transcriptome sequencing.The complete CDS sequence of this gene was 903 bp in length encoding 300 amino acids.The analysis of the amino acids sequence showed that the protein has one conservative domain(Terpene_cyclase_plant_C1).The expression level of PsTPS14 was the highest in half opening stage of petals in tree peony‘High Noon’,consistent with the release pattern of floral volatiles.Subcellular localization analysis showed that PsTPS14 was mainly localized in the chloroplast.The overexpression vector of PsTPS14 was injected into tobacco leaves and the petals of cut flowers of‘Fengdan’,a light scent tree peony cultivar,by using the transient expression method.The release of linalool in tobacco leaves and‘Fengdan’petals could be detected by GC-MS method.At the same time,the expression level of PsTPS14 was higher in tobacco leaves and‘Fengdan’petals,and the content and activity of linalool synthase were significantly increased.This study showed that PsTPS14 regulated the synthesis of linalool in plants.