药百合LsSPX1基因的克隆及表达分析OA北大核心CSTPCD
Cloning and expression analysis of LsSPX1 gene in Lilium speciosum var.gloriosoides
[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 spe-ciosum var.gloriosoides.[Methods]In this study,LsSPX1 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 LsSPX1 gene in lily flower color formation.[Results]The coding region of LsSPX1 gene was 822 bp in length,enco-ding 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 relat-ed 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 LsSPX1 was up-regulated during the development of the colorless regions of the tepals,and re-mained at relatively low levels in the colored regions at the same stage.A potential gene regulatory net-work constructed using GENIE3 indicated that LsSPX1 is co-expressed with multiple transcription fac-tors that negatively regulate anthocyanin synthesis.[Conclusion]LsSPX1 may have a negative regula-tory function on anthocyanin biosynthesis and play an important role in the formation of the colorless re-gions in the tepals of Lilium speciosum var.gloriosoides.
孙婷婷;刘筱琳;杨捷;赵一燃;朱云涛;何恒斌
北京林业大学园林学院,北京 100083
园艺学与植物营养学
药百合SPX基因花青素克隆基因表达
Lilium speciosum var.gloriosoidesSPX geneanthocyaninscloninggene expression
《河南农业大学学报》 2024 (006)
946-956 / 11
国家重点研发计划(2019YFD1000400);国家自然科学基金项目(31672191)
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