海巴戟ACO基因启动子的克隆及功能初步验证  

Cloning and Functional Verification of Promoter of ACO Gene in Morinda citrifolia

作　　者：张正雪 王青芬 胡丹焱 蓝增全[2] 吴田[3] Zhang Zhengxue;Wang Qingfen;Hu Danyan;Lan Zengquan;Wu Tian(College of Ecology and Environment,Southwest Forestry University,Kunming,650224;Southwest Institute of Ecology Development,Southwest Forestry University,Kunming,650224;Southwest Landscape Architecture Engineering Research Center of State Forestry Administration,Southwest Forestry University,Kunming,650224)

机构地区：[1]西南林业大学生态与环境学院,昆明650224 [2]西南林业大学西南绿色发展研究院,昆明650224 [3]西南林业大学,国家林业与草原局西南风景园林工程技术研究中心,昆明650224

出　　处：《基因组学与应用生物学》2021年第1期342-348,共7页Genomics and Applied Biology

基　　金：国家林业与草原局《诺丽果高产优质生产及加工技术推广示范》([2019]27号);国家留学基金([2019]75号)共同资助。

摘　　要：为了进一步明确海巴戟(Morinda citrifolia Linn.) ACO基因(GenBank登录号:ARB05681.1)功能,本研究通过染色体步移法克隆得到2 066 bp启动子。该区域除了含有大量TATA-box、CAAT-box等启动子核心元件外,还含有乙烯等植物激素响应元件、胁迫和防御相关元件、光相关元件等,转录起始位点预测在-96 bp处。为了验证启动子序列中的植物激素响应元件,分别用乙烯利、茉莉酸甲酯、赤霉素、生长素、脱落酸分别处理海巴戟叶片,荧光定量PCR结果表明,5种激素均能上调ACO基因表达,与启动子中存在相应的元件一致,暗示着这些元件的存在能够调控海巴戟ACO基因的表达;此外,ACO受乙烯诱导表达最强,表明启动子中确实存在乙烯应答元件。In order to explore the function of ACO gene(GenBank Accession number ARB05681.1) in Morinda citrifolia, the 2 066 bp promoter of the ACO gene was cloned by adopting genome walking methodology. The promoter not only contains a large number of promoter core elements such as TATA-box and CAAT-box, but also contains plant hormone response components such as ethylene, stress and defense related components and optical related components. Besides there also has a new finding that the transcription initiation site was predicted at-96 bp. Then, the leaves of Morinda citrifolia were treated with ethylene, methyl jasmonate, gibberellin, auxin, and abscisic acid respectively, and then qPCR was conducted to verify the plant hormone response elements in the promoter sequence. The results show that all the five hormones could up-regulate the expression of ACO gene expression which is the same as the presence of corresponding components in the promoter, suggesting that the presence of these components could regulate the expression of ACO gene in Morinda citrifolia. Furthermore, the ethylene induced expression of ACO was strongest, indicating the presence of ethylene response elements in the promoter.

关 键 词：海巴戟 乙烯 启动子元件 染色体步移 

分 类 号：Q943.2[生物学—植物学]