重瓣百合LiSEP3基因克隆与表达分析

发布时间：2018-07-21 14:28

【摘要】：【目的】克隆百合LiSEP3基因并确定其在百合不同地上器官及不同花瓣中的表达差异,探讨SEP3基因在重瓣花中的表达模式。【方法】利用RACE方法从重瓣百合品种‘比罗尼卡’中克隆得到LiSEP3基因核苷酸序列,利用SMART软件对其蛋白结构进行分析,以MEGA 5.0软件进行系统进化树分析,用基因枪轰击法进行亚细胞定位分析,采用荧光定量PCR进行不同地上器官及不同花瓣中LiSEP3基因表达差异分析。【结果】LiSEP3基因属于MADS-box家族E类基因,其开放阅读框为729 bp,共编码242个氨基酸;LiSEP3基因编码的蛋白具有典型的MADS结构域、K-box区及2个SEP基序;LiSEP3与同为单子叶植物的六出花的SEP3亲缘关系最近;LiSEP3蛋白定位于细胞核中;LiSEP3基因在花中的相对表达量最高,在茎、叶中几无表达;在1~7轮花瓣中均能检测到LiSEP3表达,且位于最内侧的第7轮花瓣中相对表达量最高。【结论】LiSEP3基因属于MADS-box家族E类基因,主要在花中表达,其中在最内侧花瓣中表达量最高,其表达模式与双子叶植物花中SEP3基因表达模式存在不同。
[Abstract]:[objective] to clone the LiSEP3 gene and determine the difference of its expression in different shoot organs and petals of lily. To investigate the expression pattern of SEP3 gene in double flower. [methods] the nucleotide sequence of LiSEP3 gene was cloned from double lily variety 'Bironica' by race method, and its protein structure was analyzed by smart software. The phylogenetic tree analysis was carried out by MEGA5.0 software, and subcellular localization was analyzed by gene gun bombardment. Fluorescence quantitative PCR was used to analyze the difference of LiSEP3 gene expression in different aboveground organs and petals. [results] LiSEP3 gene belongs to MADS-box family E gene. Its open reading frame is 729bp.The protein encoding 242 amino acids LiSEP3 gene has a typical MADS domain, K-box region and two SEP motifs, LiSEP3 is closely related to SEP3 of six flowers in monocotyledonous plants. The relative expression of LiSEP3 gene was the highest in the flower. LiSEP3 was not expressed in stem and leaf, LiSEP3 could be detected in 1 and 7 petal, and the relative expression of LiSEP3 was highest in the innermost seventh petal. [conclusion] LiSEP3 gene belongs to MADS-box family E gene and is mainly expressed in flower. The expression pattern of SEP3 gene was different from that in dicotyledonous flowers.
【作者单位】：阜阳师范学院生物与食品工程学院;中国农业大学观赏园艺与园林系花卉发育与品质调控北京市重点实验室;聊城大学农学院;
【基金】：国家自然科学基金项目(31471904) 安徽省教育厅自然科学研究重点项目(KJ2016A871) 安徽省高校质量工程项目(2015jyxm221)
【分类号】：S682.29

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