【摘要】：CYC同源基因作為控制花瓣對(duì)稱性形成的關(guān)鍵基因,在菊科頭狀花序中主要調(diào)控著舌狀花(兩側(cè)對(duì)稱)的生長(zhǎng)發(fā)育。菊科植物中舌狀花的有或無及其分子調(diào)控機(jī)制和演化過程一直備受關(guān)注。本研究從花型不同的川甘亞菊、戈壁短舌菊和神農(nóng)香菊中分別同源克隆了轉(zhuǎn)錄因子基因CYC2d。3個(gè)序列與菊花CmCYC2d基因氨基酸序列的比對(duì)結(jié)果顯示其同源性均超過90%,且均含有保守的TCP和R結(jié)構(gòu)域。半定量RT-q PCR結(jié)果顯示,CYC2d在地被菊品種‘毛香玉’幼嫩花序中的表達(dá)量最高,而在川甘亞菊和戈壁短舌菊中的表達(dá)量非常微弱。因此,進(jìn)一步通過實(shí)時(shí)熒光定量檢測(cè)了CmCYC2d在‘毛香玉’6個(gè)發(fā)育時(shí)期舌狀花和管狀花中的表達(dá)情況,結(jié)果表明,CmCYC2d各個(gè)時(shí)期管狀花中的表達(dá)量均很低,而在相應(yīng)時(shí)期舌狀花中的表達(dá)均很豐富。在不同花型的雜交F1代優(yōu)株中,CmCYC2d也主要在不同位置的舌狀花中高表達(dá)。通過農(nóng)桿菌轉(zhuǎn)化重組質(zhì)粒p SUPER1300-CmCYC2d-GFP在煙草表皮細(xì)胞瞬時(shí)表達(dá),亞細(xì)胞定位結(jié)果顯示其定位于表皮細(xì)胞核。分別在野生型擬南芥和tcp1突變體(SALK-022364)中過表達(dá)CmCYC2d基因的結(jié)果表明,轉(zhuǎn)基因陽性株系的營(yíng)養(yǎng)生長(zhǎng)受到抑制,花期延遲,花瓣大小和排列均發(fā)生了變化,使原本輻射對(duì)稱的花瓣呈現(xiàn)兩側(cè)對(duì)稱的趨勢(shì)。從研究結(jié)果可知,轉(zhuǎn)錄因子基因CmCYC2d對(duì)菊花舌狀花的發(fā)育有重要調(diào)控作用。本研究為菊科舌狀花演化的分子調(diào)控機(jī)制研究奠定了基礎(chǔ)。
[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.
【作者單位】： 花卉種質(zhì)創(chuàng)新與分子育種北京市重點(diǎn)實(shí)驗(yàn)室國(guó)家花卉工程技術(shù)研究中心城鄉(xiāng)生態(tài)環(huán)境北京實(shí)驗(yàn)室教育部林木花卉育種實(shí)驗(yàn)室北京林業(yè)大學(xué)園林學(xué)院;
【基金】：“十二五”國(guó)家科技支撐計(jì)劃項(xiàng)目(2013BAD01B07) 北京市共建項(xiàng)目(2016)
【分類號(hào)】：S682.11
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本文編號(hào)：2291233