药百合LsSPX1基因的克隆及表达分析  

Cloning and expression analysis of LsSPX 1 gene in Lilium speciosum var.gloriosoides

作　　者：孙婷婷 刘筱琳 杨捷[1] 赵一燃 朱云涛 何恒斌[1] SUN Tingting;LIU Xiaolin;YANG Jie;ZHAO Yiran;ZHU Yuntao;HE Hengbin(College of Landscape Architecture,Beijing Forestry University,Beijing 100083,China)

机构地区：[1]北京林业大学园林学院,北京100083

出　　处：《河南农业大学学报》2024年第6期946-956,共11页Journal of Henan Agricultural University

基　　金：国家重点研发计划(2019YFD1000400);国家自然科学基金项目(31672191)。

摘　　要：【目的】对药百合的SPX基因进行克隆和表达模式的分析,探究其对药百合花色形成可能存在的调控功能。【方法】以药百合(Lilium speciosum var.gloriosoides)为材料,通过实时荧光定量PCR(RT-PCR)扩增的方法克隆LsSPX1基因。对该基因进行生物信息学、亚细胞定位和qRT-PCR分析,以探究LsSPX1基因在百合花色形成中的功能特征。【结果】药百合LsSPX1基因编码区序列长度822 bp,编码273个氨基酸;在LsSPX1蛋白的N端含有1个SPX结构域。系统进化分析显示,LsSPX1蛋白属于含1个SPX结构域的SPX亚家族蛋白,与铁皮石斛(Dendrobium officinale)和蝴蝶兰(Phalaenopsis equestris)的SPX1亲缘关系最近。亚细胞定位显示,LsSPX1蛋白主要定位于细胞核。qRT-PCR分析显示,LsSPX1基因在药百合花被片无色区的发育过程中上调表达,在同期有色区域表达相对较低。利用基于树的集成方法的基因网络推断(gene network inference with ensemble of trees,GENIE3)构建的潜在基因调控网络表明,LsSPX1与多个花青素合成负调控转录因子共表达。【结论】药百合LsSPX1基因可能对花青素生物合成具有负调控功能,在药百合花被片无色区域的形成过程中起到重要作用。【Objective】To clone and analyze the expression pattern of SPX gene of Lilium speciosum var.gloriosoides,and to explore its potential regulatory function on the formation of color of Lilium speciosum var.gloriosoides.【Methods】In this study,LsSPX 1 gene was cloned from Lilium speciosum var.gloriosoides using RT-PCR.Bioinformatics,subcellular localization and quantitative real-time PCR(qRT-PCR)analyses were performed to explore the functional characteristics of LsSPX 1 gene in lily flower color formation.【Results】The coding region of LsSPX 1 gene was 822 bp in length,encoding 273 amino acids.LsSPX1 protein contains an N-terminal SPX domain.Phylogenetic analysis showed that LsSPX1 belonged to the SPX subfamily with one SPX domain,and was most closely related to the SPX1 of Dendrobium officinale and Phalaenopsis equestris.Subcellular localization showed that LsSPX1 was mainly located in the nucleus.qRT-PCR fluorescence quantitative analysis showed that LsSPX 1 was up-regulated during the development of the colorless regions of the tepals,and remained at relatively low levels in the colored regions at the same stage.A potential gene regulatory network constructed using GENIE3 indicated that LsSPX 1 is co-expressed with multiple transcription factors that negatively regulate anthocyanin synthesis.【Conclusion】LsSPX 1 may have a negative regulatory function on anthocyanin biosynthesis and play an important role in the formation of the colorless regions in the tepals of Lilium speciosum var.gloriosoides.

关 键 词：药百合 SPX基因 花青素 克隆 基因表达 

分 类 号：S68[农业科学—观赏园艺]