春黄菊族部分植物CYC2d同源基因的分离与功能初步分析

发布时间：2018-10-24 11:15

【摘要】：CYC同源基因作为控制花瓣对称性形成的关键基因,在菊科头状花序中主要调控着舌状花(两侧对称)的生长发育。菊科植物中舌状花的有或无及其分子调控机制和演化过程一直备受关注。本研究从花型不同的川甘亚菊、戈壁短舌菊和神农香菊中分别同源克隆了转录因子基因CYC2d。3个序列与菊花CmCYC2d基因氨基酸序列的比对结果显示其同源性均超过90%,且均含有保守的TCP和R结构域。半定量RT-q PCR结果显示,CYC2d在地被菊品种‘毛香玉’幼嫩花序中的表达量最高,而在川甘亚菊和戈壁短舌菊中的表达量非常微弱。因此,进一步通过实时荧光定量检测了CmCYC2d在‘毛香玉’6个发育时期舌状花和管状花中的表达情况,结果表明,CmCYC2d各个时期管状花中的表达量均很低,而在相应时期舌状花中的表达均很丰富。在不同花型的杂交F1代优株中,CmCYC2d也主要在不同位置的舌状花中高表达。通过农杆菌转化重组质粒p SUPER1300-CmCYC2d-GFP在烟草表皮细胞瞬时表达,亚细胞定位结果显示其定位于表皮细胞核。分别在野生型拟南芥和tcp1突变体(SALK-022364)中过表达CmCYC2d基因的结果表明,转基因阳性株系的营养生长受到抑制,花期延迟,花瓣大小和排列均发生了变化,使原本辐射对称的花瓣呈现两侧对称的趋势。从研究结果可知,转录因子基因CmCYC2d对菊花舌状花的发育有重要调控作用。本研究为菊科舌状花演化的分子调控机制研究奠定了基础。
[Abstract]:CYC homologous gene, as a key gene controlling the symmetry of petals, mainly regulates the growth and development of glossy flowers (bilateral symmetry) in the heads of Compositae. In Compositae, the existence or absence of glossy flowers and their molecular regulatory mechanisms and evolutionary processes have attracted much attention. In this study, different flower types of Chamomile were studied. CYC2d.3 sequence of transcription factor gene and amino acid sequence of CmCYC2d gene of Chrysanthemum chrysanthemi were homologous cloned respectively. The results showed that the sequence homology was more than 90% and both contained conserved TCP and R domains. The results of semi-quantitative RT-q PCR showed that the expression of CYC2d was the highest in young inflorescence of ground cover chrysanthemum 'Maoxiangyu', but very weak in Chamomile and Gobi chrysanthemum. Therefore, the expression of CmCYC2d in the tube flower and tongue flower of 'Maoxiangyu' was further detected by real-time fluorescence quantitative analysis. The results showed that the expression of CmCYC2d in the tube flower was very low in each stage of CmCYC2d. In the corresponding period, the expression of tongue flower is very rich. In F _ 1 hybrids with different flower types, CmCYC2d was also highly expressed in different positions of glossy flowers. The recombinant plasmid p SUPER1300-CmCYC2d-GFP transformed by Agrobacterium tumefaciens was transiently expressed in tobacco epidermal cells, and the subcellular localization showed that it was located in the epidermal nucleus. The results of overexpression of CmCYC2d gene in wild type Arabidopsis thaliana and tcp1 mutants (SALK-022364) showed that the vegetative growth of transgenic positive lines was inhibited, flowering was delayed, and the size and arrangement of petals changed. Causes the original radiation symmetry petal to present the bilateral symmetry tendency. The results indicate that transcription factor gene CmCYC2d plays an important role in the development of chrysanthemum tongue flower. This study laid a foundation for the study of molecular regulation mechanism of tongue flower evolution in Compositae.
【作者单位】： 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;
【基金】：“十二五”国家科技支撑计划项目(2013BAD01B07) 北京市共建项目(2016)
【分类号】：S682.11
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本文编号：2291233