浙贝母ACO基因克隆、表达及生物信息学分析  

Cloning,Expression and Bioinformatics Analysis of ACO Gene of Fritillaria thunbergii

作　　者：丁楚蔚 李梓铭 陈志[2] 范小平 金泽兰 江建铭[3] 王志安[3] 王忠华[1] Ding Chuwei;Li Ziming;Chen Zhi;Fan Xiaoping;Jin Zelan;Jiang Jianming;Wang Zhian;Wang Zhonghua(Institute of Biotechnology,ZhejiangWanli University,Ningbo,315100;Institute of Virology and Biotechnology,Zhejiang Academy of Agricultural Sciences,Hangzhou,310021;Zhejiang Institute of Traditional Chinese Medicine,Hangzhou,310023)

机构地区：[1]浙江万里学院,生物技术研究所,宁波315100 [2]浙江省农业科学院病毒学与生物技术研究所,杭州310021 [3]浙江省中药研究所,杭州310023

出　　处：《分子植物育种》2023年第10期3229-3236,共8页Molecular Plant Breeding

基　　金：国家重点研发计划项目(2017YFC1700700);现代农业产业技术体系建设专项(CARS-21);浙江省中药材新品种选育重大科技专项(2016C02058);浙江省重点实验室建设项目(2011E10015);浙江省重点研发计划项目(2017C02011)共同资助。

摘　　要：本研究利用RT-PCR和RACE技术克隆出浙贝母1-氨基环丙烷-1-羧酸氧化酶(1-aminocyclo-propane-1-carboxylateoxidase,ACO)基因全长,对该基因进行生物信息学分析,并以‘浙贝3号’浙贝母花败期根、茎、叶及幼苗期至成熟期鳞茎为材料,采用RT-qPCR测定ACO基因在浙贝母中的时空表达。成功克隆得到的序列全长1185 bp,开放阅读框为951 bp,编码317个氨基酸;浙贝母ACO氨基酸序列与同属百合科的麝香百合相似性最高,为95.21%。浙贝母不同部位ACO基因表达量依次为茎>根>叶>鳞茎,在鳞茎发育过程中ACO基因表达量逐渐增高,并在成熟期达到最高,比幼苗期提高了193.2%。This study about the full length of ACO gene in Fritillaria thunbergii was cloned by RT-PCR and RACE,and the gene was analyzed in bioinformatics.The roots,stems,leaves of deflorate stage and bulbs from seedling to mature stage of Fritillaria thunbergii'Zhebei 3'were used to explore the expression of ACO in Fritillaria thunbergii by RT-qPCR.The full-length of ACO gene of Fritillaria thunbergii was successfully cloned,which included 1185 bp,and the open reading frame was 951 bp,encoding 317 amino acids.The ACO amino acid sequence of Fritillaria thunbergii had the highest identity with Lilium longiflorum belongs to the same family Liliaceae(95.21%).The results showed that the expression abundance of ACO gene in different parts of Fritillaria thunbergii was stem>root>leaf>bulb.The expression abundance of ACO gene increased with bulb development and the peak expression was achieved in the mature stage which was 193.2%higher than in seedling stage.

关 键 词：浙贝母 ACO基因 克隆 荧光表达 鳞茎 

分 类 号：S567.231[农业科学—中草药栽培]