茶梅ACO基因的克隆及表达分析  

Cloning and Expression Analysis of ACO Gene from Camellia sasanqua

作　　者：高园 宫树森 解元 李佳莹 杨自云[1] 陈龙清 吴田[1] GAO Yuan;GONG Shusen;XIE Yuan;LI Jiaying;YANG Ziyun;CHEN Longqing;WU Tian(School of Landscape Architecture and Horticulture Sciences,Southwest Forestry University,Yunnan Province Engineering Research Center for Functional Flower Resources and Industralization,Kunming,650224)

机构地区：[1]西南林业大学园林园艺学院,云南省功能性花卉资源及产业化技术工程研究中心,昆明650224

出　　处：《基因组学与应用生物学》2023年第3期308-317,共10页Genomics and Applied Biology

基　　金：国家重点研发计划课题(2019YFD1001005);云南省教育厅研究生项目(2022Y615)共同资助。

摘　　要：为研究1-氨基环丙烷-1-羧酸氧化酶(1-aminocyclopropane-1-carboxylic acid oxidase,ACO)基因在茶梅(Camellia sasanqua)花朵衰老过程中的功能,本研究在考察不同品种茶梅单花花期和ACO总酶活、ACO基因表达相关性的基础上,通过简并引物结合RACE法从茶梅‘小玫瑰’中克隆ACO基因,并进行生物信息学分析、实时定量PCR表达分析和不同花期ACO总酶活测定分析。结果表明,不同品种茶梅单花花期不同,且与ACO总酶活、ACO基因表达量显著相关,即茶梅单花花期越长,ACO酶活就越低,ACO基因表达量也越低。茶梅ACO基因cDNA全长1316 bp,其开放阅读框为963 bp,编码320个氨基酸。系统进化树表明,茶梅ACO与茶树(Camellia sinensis)ACO聚为一支。实时定量PCR分析表明,随着‘小玫瑰’花朵的开放,ACO基因的表达量逐渐升高,在花朵衰败期达最高值。不同花期‘小玫瑰’的ACO总酶活变化、乙烯释放量与茶梅ACO基因表达量变化趋势基本一致。因此,茶梅ACO基因可能是控制茶梅花期的相关基因之一。本研究为深入解析ACO基因参与茶梅花期调控机制奠定了基础。In order to study the function of 1-aminocyclopropane-1-carboxylic acid oxidase(ACO)gene in the senescence process of Camellia sasanqua flowers,this study cloned ACO gene from C.sasanqua′Xiaomeigui′by degenerate primers combined with RACE method on the basis of investigating the correlation between single flowering time,total ACO enzyme activity and ACO gene expression of different C.sasanqua varieties.Bioinformatics analysis,real time quantification PCR expression analysis and total ACO enzyme activity analysis were performed.The results showed that the single flowering period of different C.sasanqua varieties was different,and it was significantly correlated with total ACO enzyme activity and ACO gene expression.In other words,the longer the single flowering period of C.sasanqua,the lower the activity of ACO enzyme and the lower the expression of ACO gene.The full length of C.sasanqua ACO gene cDNA is 1316 bp,with the open reading frame of 963 bp,encoding 320 amino acids.The phylogenetic tree showed that the ACO from Camellia sasanqua and the ACO from Camellia sinensis were clustered in the same clade.The real time quantification PCR analysis showed that the expression of ACO gene increased gradually with the flowering of′Xiaomeigui′,and reached the highest level in the declining stage of flowers.The change of ACO activity and ethylene release of′Xiaomeigui′at different flowering stages was consistent with the change of ACO gene expression.Therefore,the ACO gene may be one of C.sasanqua the related genes controlling the plum blossom stage.This study laid a foundation for further analysis of the regulation mechanism of ACO gene involved in C.sasanqua blossom stage.

关 键 词：茶梅 单花花期 ACO酶活 基因克隆 表达分析 

分 类 号：S685.14[农业科学—观赏园艺]