油菜转录因子BnWRI1提高种子含油量的研究

文档编号:SW097  文档字数:11263,页数:24

摘  要：油菜是我国和国际上最主要的油料作物之一，食用和能源两大战略需求都对提高油菜含油量提出了新的要求。转录因子也称为反式作用因子，是能够与真核基因启动子区域中顺式作用元件发生特异性相互作用的DNA结合蛋白，以序列特异的方式识别并结合靶基因启动子的位点，调节(激活或抑制)启动子活性进而调控基因转录。油脂合成代谢涉及众多酶系，对单一结构基因的表达调控，难以充分发挥油脂高效积累的潜能。利用转录因子提高脂肪酸合成整体代谢水平，提供了高油育种的新途径。WRI1是拟南芥脂肪酸合成关键调控基因，该基因突变导致种子含油量下降80%。我们已从油菜中克隆了WRI1的同源基因BnWRI1，将BnWRI基因籽粒特异表达载体NAPIN Pr::BnWRI1转化拟南芥，并对转化植株进行PCR检测，通过对其中35个T3株系的检测结果表明BnWRI1可提高种子含油量和重量，（提高幅度最高可达27%）表现种子油脂积累水平提高。综合以上结果，我们认为BnWRI1基因与拟南芥WRI1基因功能相似，在种子油脂积累过程中发挥转录因子的作用；在种子中特异超量表达BnWRI1是基因工程提高含油量的有效途径。上述结果为后继基因工程高油育种提供关键的分子依据。
关键词：BnWRI1；油菜；转录因子；种子含油量；拟南芥

Increasing Seed Oil Content by Seed-specific Over-expression of
 Transcription Factor BnWRI1
Abstract: Rape is one of the most important oil-bearing crops in my country and the international community，Edible and energy are the two major strategies to improve the oil content of rape made new demands . Transcription factor, also known as trans-acting factor , with eukaryotic gene promoter regions of cis-acting element specificity of the interaction of DNA-binding protein, A sequence-specific manner to identify and target genes with the promoter sites, regulation (activation or inhibition) promoter activity thereby regulating gene transcription .Oil biosynthesis involves many enzymes Structure of a single gene expression regulation Oil is difficult to give full play to the potential of efficient accumulation The use of transcription factors increase the overall synthesis of fatty acid metabolism Provide high oil breeding new avenues .WRI1 is the key regulatory genes of Arabidopsis fatty acid synthesis, The mutation resulted in seed oil content decreased 80%. We cloned from the rape of a WRI1 homologous gene BnWRI1, Grain will BnWRI specific gene expression vector rapeseed Pr : : BnWRI1 into Arabidopsis , Transformed plants with PCR , Through which 35 T3 lines to the test results showed BnWRI1 can raise the seed oil content and weight ,( Increases of up to 27% ) Seed oil accumulation performance level . These results , We believe BnWRI1 genes similar whit WRI1 Arabidopsis genes, In the seed oil accumulation process play a role of transcription factors; in Seeds, specific overexpressed BnWRI1 is an effective way of genetic engineering to increase oil content. The results of subsequent genetic engineering breeding high oil provide key molecular basis.
Keywords: BnWRI1; Rape;Transcription factor ;Seed oil content ;Arabidopsis

目录
中文摘要……………………………………………………………………………………… I
英文摘要……………………………………………………………………………………  Ⅱ
目录…………………………………………………………………………………………  Ⅲ
1．  绪论………………………………………………………………………………………1
 1.1  油菜研究的背景与意义……………………………………………………………1
 1.2  我国油菜研究的现状………………………………………………………………1
 1.3  基因工程技术在提高油菜含油量方面的应用……………………………………1
 1.4  利用转基因技术进一步提高油菜含油量的研究…………………………………3
2.  实验部分………………………………………………………………………………… 4
 2.1  实验材料……………………………………………………………………… ……4
  2.2  构建BnWRI1种子特异表达载体…………………………………………………5
 2.3  拟南芥的转化过程及其筛选……………………………………………………… 7
 2.4  BnWRI1种子特异表达结构拟南芥转化植株的PCR鉴定…………………………8
 2.5  拟南芥种子重量和种子含油量的测定…………………………………………… 9
3.  结果与讨论……………………………………………………………………………… 12
 3.1  BnWRI1超表达载体构建结果分析…………………………………………………12
 3.2  拟南芥抗性植株筛选结果………………………………………………………… 13
 3.3  琼脂糖凝胶电泳结果分析………………………………………………………… 13
 3.4  拟南芥种子重量和种子含油量的测定结果分析………………………………… 14
 3.5  RT-PCR结果分析……………………………………………………………………17
4.  前景与展望……………………………………………………………………………… 18
致谢…………………………………………………………………………………………… 19
参考文献……………………………………………………………………………………… 20

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