莲类胡萝卜素裂解双加氧酶4(NnCCD4)基因家族鉴定与功能分析  

Identification and Functional Analysis of NnCCD4 Gene Family From Nelumbo nucifera

作　　者：王志文 杨宁宁 张晨 秦莉莉 曹泷云 李海锋 董臣[1] WANG Zhiwen;YANG Ningning;ZHANG Chen;QIN Lili;CAO Longyun;LI Haifeng;DONG Chen(College of Biological Engineering,Henan University of Technology,Zhengzhou,Henan 450001)

机构地区：[1]河南工业大学生物工程学院,河南郑州450001

出　　处：《核农学报》2023年第10期1938-1946,共9页Journal of Nuclear Agricultural Sciences

基　　金：国家自然科学基金资助项目(31601366);河南省科技厅自然科学基金面上项目(202300410114)。

摘　　要：莲以其独特香气深受人们喜爱。为了研究莲的香气物质形成机制,本研究通过生物信息学对莲香气物质形成的关键酶NnCCD4基因家族进行分析,分析NnCCD4的催化产物,进一步探索莲属植物香气物质形成的分子机制,并通过同源克隆从莲基因组中获得3个NnCCD4基因家族成员(NnCCD4a、NnCCD4b和NnCCD4c)。结果表明,植物CCDs家族结构相似且都含有保守结构域,与NnCCD4c相比,NnCCD4a与NnCCD4b之间具有较高的同源性;实时荧光定量PCR(qRT-PCR)结果发现,NnCCD4家族成员在莲子、叶片和花中表达量存在显著差异;构建NnCCD4原核表达载体,细菌颜色互补试验结果证明NnCCD4b可以催化β-胡萝卜素氧化裂解,产生挥发性物质β-紫罗兰酮。本研究为NnCCD4的生物学功能研究提供了理论基础。Nelumbo nucifera is widely popular with people for its specific aroma.In order to unveil the mechanism of forming specific aroma,we analyzed the NnCCD4 gene family of lotus and investigated its catalytic reaction,which is helpful to further explore the molecular mechanism of forming aroma substance in lotus.The seeds,young leaves and mature leaves of TaiKong lotus 36 and flowers from TaiKong lotus 36,Jinsenianhua and Jinlinghuodu were used.The CCDs family of high plants contained the conserved structural domains,NnCCD4a and NnCCD4b exhibited higher homology compared to NnCCD4c.The expression of NnCCD4 was significantly different among lotus seeds,leaves and flowers by real-time PCR.The pMAL-c5x-NnCCD4 vectors were constructed,and bacterial color complementation experiments demonstrated that NnCCD4b could catalyze the oxidative cleavage ofβ-carotene to produceβ-ionone.This study provided a theoretical basis for the industrial application of NnCCD4.

关 键 词：莲 类胡萝卜素 类胡萝卜素裂解双加氧酶 Β-紫罗兰酮 

分 类 号：S645.1[农业科学—蔬菜学]