中国野生葡萄芪合成酶基因STS19及其启动子的克隆与功能分析  

Cloning and Functional Analysis of the CDS and Promoter of Synthase Gene STS19 in Chinese Wild Grapevine

作　　者：刘慧 殷向静[1] 方景浩 高敏 李智[1] 王西平[1] LIU Hui;YIN Xiangjing;FANG Jinghao;GAO Min;LI Zhi;WANG Xiping(State Key Laboratory of Crop Stress Biology in Arid Areas,State Key Laboratory of Horticultural Plant Biology and Germplasm Innovation in Northwest China,Ministry of Agriculture and Rural Affairs,College of Horticulture,Northwest A&F University,Yangling,Shaanxi 712100,China)

机构地区：[1]西北农林科技大学园艺学院,旱区作物逆境生物学国家重点实验室,农业农村部西北园艺作物生物学与种质创新重点开放实验室,陕西杨凌712100

出　　处：《园艺学报》2023年第7期1389-1401,共13页Acta Horticulturae Sinica

基　　金：国家自然科学基金面上项目(31872071,32272693)。

摘　　要：以中国野生山葡萄(Vitis amurensis Rupr.)‘通化3号’和刺葡萄(V.davidii Foex.)‘塘尾’为材料,克隆得到芪合成酶基因VaSTS19和VdSTS19。序列分析发现VaSTS19和VdSTS19的编码区均为1179 bp,编码392个氨基酸,定位于16号染色体,二者核苷酸和氨基酸序列相似性分别为98.81%和98.21%。亚细胞定位结果显示VaSTS19和VdSTS19均定位于细胞膜、细胞质和细胞核。将35S强启动子连接VaSTS19和VdSTS19转入番茄能显著促进STS19的表达及番茄中白藜芦醇的合成和积累。分别从两种材料中克隆得到VaSTS19和VdSTS19的启动子。顺式作用元件分析表明,二者启动子中均包含多个胁迫应答元件、激素应答元件及光应答元件。序列分析发现二者启动子序列与欧洲葡萄VST2启动子序列同源性为60%。转基因烟草叶片中二者不同长度启动子片段对SA和MeJA诱导响应明显。结果表明STS19的表达模式和白藜芦醇的合成可能与其上游启动子类型和调控有关。VaSTS19 and VdSTS19 were cloned from Vitis amurensis Rupr.‘Tonghua 3’and V.davidii Foex.‘Tangwei’.Sequence analysis showed that the open reading frame of VaSTS19 and VdSTS19 were 1179 bp,encoding 392 amino acid,and they were located on chromosome 16.The identity between VaSTS19 and VdSTS19 nucleotide and amino acid was 98.81%and 98.21%,respectively.Both VaSTS19 and VdSTS19 are located on the cytomembrane,cytoplasm and nucleus.The expression of STS19 gene and the synthesis and accumulation of resveratrol were significantly promoted when VaSTS19 and VdSTS19 were transferred into tomato by Agrobacterium-mediated method.The promoters of VaSTS19 and Liu Hui,Yin Xiangjing,Fang Jinghao,Gao Min,Li Zhi,Wang Xiping.Cloning and functional analysis of the CDS and promoter of synthase gene STS19 in Chinese wild grapevine.1390 Acta Horticulturae Sinica,2023,50(7):1389–1401.VdSTS19 were cloned from‘Tonghua 3’and‘Tangwei’,respectively.cis-Acting elements analysis showed that both promoters contained stress-related elements,hormone-related elements and light-related elements.A sequence alignment showed that the VaSTS19 promoter was demonstrated 60% homology with the VST2 promoter cloned from V.vinifera.The promoters of VaSTS19 and VdSTS19 and their fragments were significantly responsive to SA and MeJA induction in transgenic tobacco leaves.The results indicated that the expression pattern of STS19 and the production of resveratrol may be related to the regulation of the upstream region.

关 键 词：葡萄 白藜芦醇 STS19 功能分析 启动子 

分 类 号：S663.1[农业科学—果树学]