牡丹膜联蛋白基因PsANN1的克隆及表达分析  

Cloning and expression analysis of PsANN1 gene from tree peony

作　　者：徐娟娟 刘鑫[1] 贺丹[1] 逯久幸[1] 栗燕[1] XU Juanjuan;LIU Xin;HE Dan;LU Jiuxing;LI Yan(College of Landscape Architecture and Art, Henan Agricultural University, Zhengzhou 450002, China)

机构地区：[1]河南农业大学风景园林与艺术学院,河南郑州450002

出　　处：《河南农业大学学报》2021年第2期266-272,313,共8页Journal of Henan Agricultural University

基　　金：国家自然科学基金项目(31600568);河南省科技攻关项目(192102110062);河南省高等学校重点科研项目(19A220003);河南农业大学科技创新基金(KJCX2019A05);河南农业大学科技创新基金(KJCX2018A05)。

摘　　要：以盆栽‘凤丹’砧‘洛阳红’牡丹为试验材料,通过T-A克隆技术得到牡丹膜联蛋白基因(Paeonia suffruticosa Annexin1,PsANN1)并对其进行生物信息学分析,利用实时荧光定量PCR(qRT-PCR)技术对该基因在不同部位及不同花期花瓣的表达情况进行分析。结果表明,PsANN1基因编码序列(CDS)全长为939 bp,编码312个氨基酸;该蛋白相对分子质量为35.54 kD,二级结构以α-螺旋为主;功能结构域分析表明,该蛋白有4个重复的保守结构域,属Annexin蛋白家族;系统进化分析表明,牡丹ANN与月季花(Rosa chinensis)和野草莓(Fragaria vesca subsp.vesca)亲缘关系较近。qRT-PCR结果分析显示,PsANN1基因在‘洛阳红’牡丹根、茎、叶、花中均有表达,在花瓣中表达量最高,在叶片中表达量最低;PsANN1基因在不同花期的花瓣中均有表达,在盛花期的表达量最高,小风铃期的表达量最低。Taking‘Luoyanghong’tree peony with‘Fengdan’stock as material the peony annexin 1gene(PsANN1)and analyzed its bioinformatics characteristics.Real-time fluorescent quantitative PCR(qRT-PCR)was then used to analyze the expression pattern of PsANN1 in different organs and flowering stages.The results showed that PsANN1 had 939 bp coding sequence(CDS)that encoded 312 amino acids.The relative molecular weight of PsANN1 was 35.54 kD,and its secondary structure was mainlyα-helix.Functional domain analysis showed that PsANN1 had 4 conserved domains,which was consistent with the characteristics of annexin protein family.According to the phylogenetic analysis,PsANN1 is closely related to rose(Rosa chinensis)and wild strawberry(Fragaria vesca subsp.vesca).Analysis of qRT-PCR results showed that PsANN1 was expressed in roots,stems,leaves and petals of‘Luoyanghong’tree peony,with the highest expression in petals,and the lowest expression in leaves;PsANN1 gene was expressed in petals of different flowering stages,among which the expression level was the highest in the full flowering stage,and the lowest in the wind bell stage.

关 键 词：‘洛阳红’牡丹 PsANN1基因 克隆 生物信息学分析 表达分析 

分 类 号：S685[农业科学—观赏园艺] Q785[农业科学—园艺学]