本文選題：白樺　切入點(diǎn)：煙草　出處：《東北林業(yè)大學(xué)》2016年碩士論文　論文類型：學(xué)位論文

【摘要】：一止匕MADS-box基因參與了植物的開花過(guò)程,對(duì)于植物的生殖發(fā)育起到了非常重要的作用。在本研究中,我們從白樺中克隆了一個(gè)MADS-box基因,命名為BpMADS。序列比對(duì)顯示該基因與擬南芥AP1基因同源。我們進(jìn)行了該基因的分析及煙草的遺傳轉(zhuǎn)化研究,主要結(jié)果如下：亞細(xì)胞定位研究表明,BpFLC-GFP蛋白在細(xì)胞核與細(xì)胞質(zhì)中表達(dá),推測(cè)該基因在細(xì)胞質(zhì)和細(xì)胞核內(nèi)均有表達(dá)。熒光實(shí)定時(shí)定量PCR結(jié)果表明,該基因在花序中表達(dá)量顯著高于葉片,說(shuō)明此基因可能參與了開花調(diào)控。將BpMADS基因轉(zhuǎn)入煙草使其過(guò)量表達(dá),結(jié)果顯示BpMADS基因的過(guò)量表達(dá)能顯著促進(jìn)轉(zhuǎn)基因煙草開花,并導(dǎo)致植株矮小。轉(zhuǎn)基因煙草的葉片在組培條件下,不定芽迅速形成花序,此外由葉片也可直接形成花器官。暗處理結(jié)果顯示,轉(zhuǎn)基因煙草與野生型煙草表現(xiàn)相近,沒(méi)有產(chǎn)生花器官。由此說(shuō)明BpMADS基因是白樺開花發(fā)育過(guò)程的一個(gè)重要基因,并且需要在光照的輔助下才能完成其開花促進(jìn)功能。實(shí)驗(yàn)結(jié)果表明,BpMADS基因的過(guò)量表達(dá)能顯著促進(jìn)轉(zhuǎn)基因煙草葉綠體的形成,并能促進(jìn)淀粉的合成。利用實(shí)時(shí)定量PCR檢測(cè)了轉(zhuǎn)基因煙草中多個(gè)與開花相關(guān)基因的表達(dá),結(jié)果說(shuō)明BpMADS的過(guò)量表達(dá),顯著提高了以上基因的表達(dá),說(shuō)明BpMADS可通過(guò)提高煙草中開花基因的表達(dá)來(lái)促進(jìn)轉(zhuǎn)基因煙草的開花。將BpMADS基因?qū)霐M南芥apl突變體,發(fā)現(xiàn)BpMADS能夠恢復(fù)突變體的表型,說(shuō)明BpMADS可能參與了開花過(guò)程及花器官的調(diào)控。
[Abstract]:The MADS-box gene is involved in the flowering process of plants and plays a very important role in the reproductive development of plants. In this study, we cloned a MADS-box gene from Bai Hua. Sequence alignment showed that the gene was homologous to the AP1 gene of Arabidopsis thaliana. We analyzed the gene and studied the genetic transformation of tobacco. The main results were as follows: subcellular localization showed that BpFLC-GFP protein was expressed in the nucleus and cytoplasm of Arabidopsis thaliana, and BpFLC-GFP protein was expressed in the cytoplasm of Arabidopsis thaliana. It was speculated that the gene was expressed in cytoplasm and nucleus. The results of PCR showed that the expression of the gene in inflorescence was significantly higher than that in leaf. The results showed that the overexpression of BpMADS gene could significantly promote the flowering of transgenic tobacco. Under the condition of tissue culture, the adventitious buds formed inflorescence rapidly, and the floral organs were formed directly from the leaves. The results of dark treatment showed that transgenic tobacco was similar to wild-type tobacco. BpMADS gene is an important gene in the flowering and development of Bai Hua. The results showed that the overexpression of BpMADS gene could significantly promote chloroplast formation in transgenic tobacco. The expression of several genes related to flowering in transgenic tobacco was detected by real-time quantitative PCR. The results showed that the overexpression of BpMADS significantly increased the expression of the above genes. The results showed that BpMADS could promote the flowering of transgenic tobacco by increasing the expression of flowering genes in tobacco, and introduced BpMADS gene into Arabidopsis thaliana apl mutants, and found that BpMADS could restore the phenotype of the mutant. It is suggested that BpMADS may be involved in the process of flowering and the regulation of floral organs.
【學(xué)位授予單位】：東北林業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：Q943.2;S792.153

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本文編號(hào)：1581337