【作者】 汤敏；

【导师】 刘忠松；

【作者基本信息】 湖南农业大学 ， 作物遗传育种， 2012， 硕士

【摘要】 研究表明,芥菜型油菜黄籽形成涉及类黄酮合成基因。初步研究表明芥菜型油菜种皮颜色黄黑籽差异的形成受到MYB类转录因子、bHLH类转录因子、WD40蛋白等协同调控,并且不断发现有新的调控基因影响原花色素的合成,如KAN4等转录因子。本研究以本实验室克隆的芥菜型油菜A9染色体上黄籽候选基因BjuA.YSC为研究对象,通过互补试验研究BjuA.YSC基因的功能,从而揭示油菜黄籽性状形成的分子机制,为甘蓝型黄籽油菜育种提供科学依据。本研究构建了含黄籽候选基因的表达载体,以芥菜型油菜四川黄籽无菌苗的下胚轴为外植体,利用农杆菌介导法进行转化,抗性筛选和PCR分析鉴定目的基因已整合至受体植株中。试验结果如下：1、通过酶切、连接等分子克隆技术,首次将BjuA.YSC整合入pBI121载体中,构建成表达载体pBI-BjuA.YSC。2、建立了以下胚轴为外植体、农杆菌介导法的芥菜型油菜遗传转化体系。3、从6400个芥菜型油菜四川黄籽下胚轴外植体中,共获得抗卡那霉素的再生苗24株,经PCR检测证实有外源基因整合的阳性植株为3株,转化效率为0.046%。3株转基因植株发育正常,能开花、结实,但T0代植株所结种子仍为黄色。更多还原

【Abstract】 The yellow seed is showed to be associated with the expression of the genes for flavonoid synthesis in Brassicajuncea. It is suggested that the transcription factors MYB、 bHLH and WD40are the key factors responsible for difference in seed coat color. In this study, the gene BjuA.YSC our lab has cloned from Brassica juncea was used to study its function by complementation, which will provide the foundations for molecular mechanism of the yellow-seeded trait.Expression vector of the gene BjuA. YSC was constructed and introduced into the Agrobacterium tumefaciens strain EHA105by the freeze-thaw method in this study. The hypocotyls from aseptic seedlings of B. juncea cv. Sichuan Yellow were used as explants for transformation. The main results are as follows:1. The expression vector pBl-BjuA. YSC was constructed and transferred to competent E. coli cells for amplification.2. The transformation system based on hypocotyl explants and Agrobacterium tumefaciens strain EHA105was established for the species Brassicajuncea.3. The24kanamycin-resistant plantlets were regenerated from6400explants and three transgenic plants confirmed by PCR analysis. The transformation rate was estimated to be0.046%. The transgenic plants developed normally, flowered and set fruits and seeds. However, their seeds remained yellow like the untransformed parent.更多还原