白桦BpMADS基因在烟草中的遗传转化及功能分析

发布时间：2018-03-07 23:04

本文选题：白桦　切入点：烟草　出处：《东北林业大学》2016年硕士论文　论文类型：学位论文

【摘要】：一止匕MADS-box基因参与了植物的开花过程,对于植物的生殖发育起到了非常重要的作用。在本研究中,我们从白桦中克隆了一个MADS-box基因,命名为BpMADS。序列比对显示该基因与拟南芥AP1基因同源。我们进行了该基因的分析及烟草的遗传转化研究,主要结果如下：亚细胞定位研究表明,BpFLC-GFP蛋白在细胞核与细胞质中表达,推测该基因在细胞质和细胞核内均有表达。荧光实定时定量PCR结果表明,该基因在花序中表达量显著高于叶片,说明此基因可能参与了开花调控。将BpMADS基因转入烟草使其过量表达,结果显示BpMADS基因的过量表达能显著促进转基因烟草开花,并导致植株矮小。转基因烟草的叶片在组培条件下,不定芽迅速形成花序,此外由叶片也可直接形成花器官。暗处理结果显示,转基因烟草与野生型烟草表现相近,没有产生花器官。由此说明BpMADS基因是白桦开花发育过程的一个重要基因,并且需要在光照的辅助下才能完成其开花促进功能。实验结果表明,BpMADS基因的过量表达能显著促进转基因烟草叶绿体的形成,并能促进淀粉的合成。利用实时定量PCR检测了转基因烟草中多个与开花相关基因的表达,结果说明BpMADS的过量表达,显著提高了以上基因的表达,说明BpMADS可通过提高烟草中开花基因的表达来促进转基因烟草的开花。将BpMADS基因导入拟南芥apl突变体,发现BpMADS能够恢复突变体的表型,说明BpMADS可能参与了开花过程及花器官的调控。
[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.
【学位授予单位】：东北林业大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：Q943.2;S792.153

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