【作者】 曲晓言；

【导师】 林良斌；

【作者基本信息】 云南农业大学 ， 生物化学与分子生物学， 2012， 硕士

【摘要】 油菜作为我国及世界范围内的一种重要的油料经济作物，菜籽油已然是人类食用油的重要来源，其产量及质量对其经济价值有着非常重要的影响。油菜不仅是食用植物油的最主要来源，也是潜在的仅次于豆粕的大宗饲用蛋白源，而且还是目前全球用于解决能源危机方向之一的生物柴油的理想原料。近几年我国南部地区已出现百年不见得干旱灾害，严重影响了油菜的产量，同时也在一定程度上严重影响了油菜作为经济作物的经济效益。因此，随着油菜在人们生活中越来越重要的地位，其种植生产也越来越受到人们的关注，提高油菜的含油量，改善菜籽油的品质，已是目前油菜育种的主要目标。为此，本研究分别以油菜下胚轴、愈伤组织、花序等组织器官为受体进行农杆菌介导转化OSWRKY45、AtWRKY57、BrACCD基因的研究，对获得的卡那霉素抗性植株经PCR、RT-PCR、Northern杂交检测以及性状检测等，证明获得了转基因植株，为转基因改良油菜品种提供了技术支撑和育种材料。研究结果如下：（1）通过农杆菌介导法，分别以甘蓝型油菜湘油15号及花油7号的下胚轴为外植体进行遗传转化，获得了转OSWRKY45基因的油菜植株。转OSWRKY45基因植株较未转基因植株有一定的抗旱性增加。（2）通过农杆菌介导法，以27号云南小油菜的花序为外植体进行基因转化，获得了转AtWRKY57基因植株。转AtWRKY57基因油菜植株较未转基因植株有一定的抗旱性增加。并初步建立起花序浸泡转基因方法，为油菜大规模转基因并改良油菜品种提供了条件。（3）通过农杆菌介导法，分别以甘蓝型油菜湘油15号及花油7号的愈伤组织为外植体进行基因转化，获得了转BrACCD基因的油菜植株。转BrACCD基因的油菜植株较未转基因的油菜植株其含油量有不同程度的增加，增加幅度高达14.6%。更多还原

【Abstract】 Rapeseed is a kind of the important oil-bearing and economic crops all over theworld, and rapeseed oil is already become the important source for human cooking oil, theyield and quality impact its economic value importantly. Rapeseed is not only the mainsource of edible vegetable oil, but also the potential protein source for large foragefollowed with soybean, and it is the ideal raw materials of the biodiesel used to solve theenergy crisis.The severe drought disaster has appeared in our southwest in recent years, which wasnot occur even in a hundred yearsand which influenced the production of rapeseedseriously and, to a certain extent, the economic benefits of rapeseed. Therefore, with moreand more important position of rape in people’s live, the crop production become more andmore attentioned by people, improving the rape oil content and the quality of rapeseed oil,is already turn into the main target of rapeseed breeding.So, this study using the hypocotyls, callus and inflorescence of the rape to transformthe genes of AtWRKY57, OSWRKY45,BrACCD by agrobacterium-mediated, and detectionthe kanamycin-resistance plants through PCR, RT-PCR, Northern blotting and charactersto get transgenic plants, which can provide technical support and breeding materials forimproving rape varieties by transgenosis. Get the following results,(1) Transformation of OSWRKY45using hypocotyls from xiangyou15and huayou7into Brassica napus by Agrobacterium-mediated and get the transgenic plants.Transgenicplants have more drought-resistance than non-transgenic plants.(2)Transformation of AtWRKY57into yunnan small rape27through inflorescence byAgrobacterium-mediated, and get transgenic plants. Transgenic plants have more drought-resistance than non-transgenic plants. Moreover, build the genetically modified methodthrough inflorescence preliminary, for expending transgenic scale and improving rapevarieties provides conditions. (3) Transformation of BrACCD using callus from xiangyou15and huayou7intoBrassica napus by Agrobacterium-mediated and get the transgenic plants. Improve the oilcontent of the rape and its economic benefits as oil-bearing crops. Transgenic plants ofBrACCD gene has a different oil content increased than nontransgenic plants the, theincrease rate as high as14.6%.更多还原