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

【摘要】：一止匕MADS-box基因參與了植物的開花過程,對于植物的生殖發(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MADS-box基因,命名為BpMADS。序列比對顯示該基因與擬南芥AP1基因同源。我們進行了該基因的分析及煙草的遺傳轉(zhuǎn)化研究,主要結果如下：亞細胞定位研究表明,BpFLC-GFP蛋白在細胞核與細胞質(zhì)中表達,推測該基因在細胞質(zhì)和細胞核內(nèi)均有表達。熒光實定時定量PCR結果表明,該基因在花序中表達量顯著高于葉片,說明此基因可能參與了開花調(diào)控。將BpMADS基因轉(zhuǎn)入煙草使其過量表達,結果顯示BpMADS基因的過量表達能顯著促進轉(zhuǎn)基因煙草開花,并導致植株矮小。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。暗處理結果顯示,轉(zhuǎn)基因煙草與野生型煙草表現(xiàn)相近,沒有產(chǎn)生花器官。由此說明BpMADS基因是白樺開花發(fā)育過程的一個重要基因,并且需要在光照的輔助下才能完成其開花促進功能。實驗結果表明,BpMADS基因的過量表達能顯著促進轉(zhuǎn)基因煙草葉綠體的形成,并能促進淀粉的合成。利用實時定量PCR檢測了轉(zhuǎn)基因煙草中多個與開花相關基因的表達,結果說明BpMADS的過量表達,顯著提高了以上基因的表達,說明BpMADS可通過提高煙草中開花基因的表達來促進轉(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能夠恢復突變體的表型,說明BpMADS可能參與了開花過程及花器官的調(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.
【學位授予單位】：東北林業(yè)大學
【學位級別】：碩士
【學位授予年份】：2016
【分類號】：Q943.2;S792.153

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本文編號：1581337