东方百合LoABI1基因的克隆及表达分析  

Cloning and Expression Analysis of LoABI1 in Lilium Oriental Hybrid

作　　者：赵一燃 胡钧舒 朱云涛 孙婷婷 何恒斌[1] ZHAO Yiran;HU Junshu;ZHU Yuntao;SUN Tingting;HE Hengbin(Beijing Key Laboratory of Ornamental Plants Germplasm Innovation and Molecular Breeding,National Engineering Research Center for Floriculture and Beijing Laboratory of Urban and Rural Ecological Environment,College of Landscape Architecture,Beijing Forestry University,Beijing 100083,China)

机构地区：[1]花卉种质创新与分子育种北京市重点实验室,国家花卉工程技术研究中心,城乡生态环境北京实验室,教育部林木花卉育种实验室,北京林业大学园林学院,北京100083

出　　处：《西北植物学报》2023年第2期190-201,共12页Acta Botanica Boreali-Occidentalia Sinica

基　　金：国家自然科学基金(31672191);中央高校基本科研业务费专项(YX2015-16)。

摘　　要：该研究以东方百合‘索邦’为材料,采用RT-PCR扩增方法克隆ABI1基因,对其进行了生物信息学分析,并采用实时荧光定量PCR检测其组织表达特性和低温处理过程及定植后的表达特征,以明确ABI1基因的功能特性,为解析百合ABA信号转导途径及其调控低温解除休眠过程的机理奠定基础。结果表明:(1)成功克隆得到东方百合LoABI1基因,其编码序列长度为1 341 bp,共编码446个氨基酸;LoABI1氨基酸序列中含有1个蛋白磷酸酶2C(PP2C)保守结构域。(2)系统进化分析显示,LoABI1蛋白与水稻PP2C家族成员OsPP2C06的同源进化关系最近,且与拟南芥AtABI1聚为一支,同属于PP2C基因家族中的A亚群。(3)亚细胞定位发现,LoABI1蛋白定位于烟草表皮细胞的细胞核和细胞质。(4)qRT-PCR荧光定量分析显示,LoABI1基因在百合茎生根、嫩茎、叶片及各花部组织中均有表达,且在幼嫩组织中表达量较高;LoAB1I基因在冷藏期间的表达量呈先升高后降低的趋势,并于冷藏期第5周达到峰值,但在定植期间持续下降且保持较低水平。(5)经4℃低温处理60 d的百合鳞茎在定植后14~28 d能发芽、28~48 d可见花蕾,而不进行低温处理的鳞茎既不萌芽也不开花。研究推测,百合LoABI1基因可能在低温解除种球休眠过程中具有重要作用,LoABI1蛋白可能在百合ABA信号转导过程中发挥作用。In this study, ABI1 gene was cloned from Lilium oriental hybrid ‘Sorbonne’ by RT-PCR, and the bioinformatics analysis were performed. Meanwhile, the qRT-PCR was used to detect the expression patterns of LoABI1 in lily at different tissues and in the process of low temperature treatment and after transplanting, for clarifying the functional characteristics of LoABI1 gene and lay a foundation for the analysis of the ABA signal transduction pathway and the mechanism of breaking dormancy at low temperature in lily. The results showed that:(1) the LoABI1 gene of lily was cloned successfully. The coding sequence of LoABI1 was 1 341 bp long and LoABI1 encoded 446 amino acids containing a conserved phosphatase 2C(PP2C) domain.(2) Phylogenetic analysis showed that LoABI1 was closest to Oryza sativa and clustered with AtABI1, which is a member of Arabidopsis thaliana PP2C gene family, and both of them belong to PP2C A group.(3) The subcellular localization results showed that LoABI1 is localized in the nucleus and cytoplasm of tobacco epidermal cells.(4) Real time RT-PCR expression analysis showed that LoABI1 expressed in stem root, tender stem, leaf, and floral organ, and the expression level is higher in tender tissues;The expression of LoABI1 increased at first reaching the peak at the fifth week of cold storage, and then decreased during cold storage. Finally, it remained at a low-level during colonization.(5) The lily bulbs treated at 4 ℃ for 60 days could germinate at 14-28 days, and the buds could be seen at 28-48 days after transplanting, while the bulbs without low temperature treatment neither germinated nor flowered. It is speculated that LoABI1 gene may play an important role in the process of breaking bulbs dormancy at low temperature, and LoABI1 protein may play an important role in lily ABA signal transduction.

关 键 词：东方百合‘索邦’ ABI1基因 克隆 基因表达 

分 类 号：Q785[生物学—分子生物学] Q786