蜡梅CpBEAT的克隆、表达分析及功能验证  

Cloning,Expression and Functional Identification of CpBEAT Gene in Chimonanthus praecox

作　　者：王飞龙 梁前艳 尚均忠 陈龙清 向林[1] WANG Feilong;LIANG Qianyan;SHANG Junzhong;CHEN Longqing;XIANG Lin(National Key Laboratory for Germplasm Innovation&Utilization of Horticultural Crops,Huazhong Agricultural University,Wuhan 430070,China;Yangtze River Biodiversity Research Center of China Three Gorges Corporation Limited,Wuhan 430014,China;National Engineering Research Center of Eco-environment Protection for Yangtze River Economic Belt,Beijing 100038,China;College of Landscape Architecture and Horticulture Sciences,Southwest Forestry University,Kunming 650224,China)

机构地区：[1]华中农业大学果蔬园艺作物种质创新与利用全国重点实验室,武汉430070 [2]中国长江三峡集团有限公司长江生物多样性研究中心,武汉430014 [3]长江经济带生态环境国家工程研究中心,北京100038 [4]西南林业大学园林园艺学院,昆明650224

出　　处：《园艺学报》2025年第1期160-170,共11页Acta Horticulturae Sinica

基　　金：国家自然科学基金项目(32372755)。

摘　　要：以蜡梅(Chimonanthus praecox)为材料,采用顶空固相微萃取-气相色谱质谱技术对其5个花期的花器官挥发物含量进行了测定。结果表明,随着花朵的发育,苯甲醇和乙酸苄酯的含量逐渐升高,在盛开期最高。从花中克隆出结构基因CpBEAT,Gen Bank登录号为OQ503489,开放阅读框为1 392 bp,编码463个氨基酸。CpBEAT的表达水平随着花发育逐渐升高,表达趋势与乙酸苄酯的挥发趋势基本一致。将CpBEAT导入烟草异源表达,在外施底物苯甲醇的条件下能够合成乙酸苄酯。进一步克隆CpBEAT的启动子,通过双荧光素酶试验发现CpMYC2正向调控CpBEAT启动子转录活性。In this study,the contents of benzyl alcohol and benzyl acetate in the tepals were determined from flower of Chimonanthus praecox at five developmental stages by headspace solid-phase microextraction coupled to gas chromatography-mass spectrometry(HS-SPME-GC-MS).These results showed that the content of benzyl alcohol and benzyl acetate continued to increase along with the development of flowers,and the highest content was in full flowering stage flower.The sequence of CpBEAT was cloned from C.praecox and the GenBank accession number was OQ503489.CpBEAT contained an open reading frame of 1392 bp encoding a putative protein of 463 amino acids.The expression level of CpBEAT showed a trend of rising during the flower development.The expression pattern was consistent with the volatilization of benzyl acetate in the five stages of flower development.CpBEAT could synthesize benzyl acetate under the condition of external substrate benzyl alcohol in transgenic plants.The promoter sequence of CpBEAT was also cloned.A dual luciferase assay showed that CpMYC2 positively regulated transcription activity on the CpBEAT promoter.

关 键 词：蜡梅 花香 苯甲醇乙酰基转移酶(BEAT) 乙酸苄酯 MYC2 

分 类 号：S685.99[农业科学—观赏园艺]