蜡梅花发育相关基因CpUFO表达特性与功能分析

doi:10.11707/j.1001-7488.LYKX20230227

摘要/Abstract

摘要： 目的 UFO是被子植物中首个被发现的F-box蛋白，主要参与调控花分生组织特性和花器官发育。本文通过对蜡梅CpUFO基因表达特性分析及异源表达拟南芥表型分析，初步鉴定CpUFO的功能，为阐明蜡梅花发育分子调控机制提供参考。方法基于前期构建的蜡梅转录组数据，克隆蜡梅CpUFO基因并对其编码蛋白进行同源比对及进化树分析。根据qRT-PCR分析CpUFO基因在不同组织、器官及全年花发育各阶段中的相对表达量，比较35S::CpUFO转基因拟南芥株系及Col-0野生型表型差异，并通过qRT-PCR检测过表达拟南芥株系内源开花相关基因的表达特性，分析过表达CpUFO对拟南芥开花时间以及花器官发育的影响。结果获得的CpUFO基因cDNA序列为1 665 bp，编码403个氨基酸，含有保守的F-box结构域。CpUFO在外花被片中表达量相对最高，其次是内花被片，在果、茎、叶、腋芽及雌雄蕊中表达水平较低；而在全年花芽中的表达量分析结果显示，低温打破休眠后的露瓣和始花以及盛开期花芽中的表达量显著高于其他时期。与野生型拟南芥相比，35S::CpUFO转基因株系莲座叶数目减少，开花提前，且拟南芥内源基因TFL1表达量下调，成花关键基因FUL和AP1的表达量上调。结论 CpUFO在不同组织、器官及花发育各阶段均有表达，不同组织中在外花被片中表达量最高，全年花芽中打破休眠的花蕾开放阶段表达量最高。同时，过表达CpUFO会促进拟南芥早花并影响其花器官发育。

关键词: 蜡梅, UFO基因, F-box家族, 花发育

Abstract: Objective UFO is the first F-box protein discovered in angiosperms, and plays a role in determining floral meristem identity and floral organ growth. In this study, the expression profile and heterologous expression phenotypic traits in A. thaliana were analyzed to explore the gene function of CpUFO and provide a basis for molecular mechanism of floral development in wintersweet (Chimonanthus praecox). Method Based on the constructed transcriptome data of C. Praecox, the CpUFO gene was cloned and the sequence alignment of its encoded protein and phylogenetic analysis were conducted. Quantitative real-time RT-PCR (qRT-PCR) was performed to identify transcript level of CpUFO in different tissues, organs and annual flower buds in different flower development stages. The effects of CpUFO on flowering time and flower organ development of Arabidopsis thaliana were explored by analyzing the phenotypes and relative gene expression of endogenous flowering related gene between the 35S∷CpUFO transgenic Arabidopsis lines and the wild type. Result The obtained cDNA of CpUFO gene was 1 655 bp in full-length, encoding a protein of 403 amino acids residues with highly conserved F-box domain. The expression level of CpUFO was relatively highest in the outer tepal, followed by the inner tepal, and the expression level was lower in fruit, stem, leaf, axillary bud and stamen. Transcript level of CpUFO in annual flower buds showed that relative expression of CpUFO in flower buds of display tepals (DT), initial blooming (IB) and open flower (OF) stages after chilling-induced dormancy breaking was obviously higher than that in the other developmental periods. Compared with Arabidopsis Col-0, 35S::CpUFO transgenic Arabidopsis lines had less rosette leaves and early flowering. Moreover, the endogenous genes of AP1 and FUL in A. thaliana were significantly up-regulated, while TFL1 was significantly down-regulated. Conclusion CpUFO is expressed in different tissues, organs and annual flower buds, and the highest relative expression level is found in the outer tepals and flower buds of open period after dormancy breaking. Meanwhile, overexpression of CpUFO promotes early flowering in Arabidopsis and affects its floral organ development．

Key words: Chimonanthus praecox, UFO gene, F-box family, flower development

中图分类号:

S718.46

车冰雁, 余尚妍, 李志能, 李先源. 蜡梅花发育相关基因CpUFO表达特性与功能分析[J]. 林业科学, 2024, 60(2): 87-96.

Che Bingyan, Yu Shangyan, Li Zhineng, Li Xianyuan. Expression Characteristics and Functional Analysis of Gene CpUFO Related to Flower Development of Chimonanthus praecox[J]. Scientia Silvae Sinicae, 2024, 60(2): 87-96.

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