【作者】 聂利红；

【导师】 崔党群；

【作者基本信息】 河南农业大学 ， 作物遗传育种， 2004， 硕士

【摘要】 本试验研究了离子束介导大豆DNA转化普通小麦当代转化效果和转化二代遗传特点；采用正交设计，综合考察了影响转化效果的4个因素；并将SDS—PAGE和A—PAGE电泳用于检测所获转化后代的高蛋白质植株。结果表明： 1．单纯的离子注入对不同小麦品种影响不同，对同一品种的各品质性状影响也不同。离子注入对小麦品质性状有所影响，但幅度不大。 2．转化当代（T₁代）小麦各品质性状的平均值与对照无显著差异，变幅增大，并出现了一定比例的高蛋白植株；T₂代开始分离，其蛋白变幅进一步增大，大量单株退回到正常对照水平，但仍保留有相当比例的高蛋白植株。 3．在受体、供体、DNA浓度、浸种时间这4个因素中，影响转化效果的主次顺序为：受体小麦品种、供体大豆品种、浸种时间和DNA浓度。4个受体小麦品种中，株系豫农92349后代转化率最高，是优良的受体；4个供体大豆品种中，郑14青豆是较好的供体；在浸种时间的这4个梯度中，浸种时间为48h时，转化效果最好；4个DNA浓度的水平分析，浓度为200μg／ml时转化效果最好。 4．同一受体对不同供体和同一供体不同受体间转化率差异显著。在供试的16个组合中，筛选了4个优良组合，排序为豫农92349／豫豆19、豫农92349／郑多枝、豫农92349／郑14青豆、豫麦52／郑14青豆。 5．SDS-PAGE电泳分析，获得了15株高蛋白变异株，变异株与对照间的差异表现在高分子量谷蛋白亚基（HMW-GS）和谱带的着色上。A-PAGE电泳分析，获得了9株变异株，变异主要在迁移率低的慢带区即ω区，而且谱带的染色较对照加深。在谷蛋白上有变异的植株在醇溶蛋白上与对照并没有差异，而在醇溶蛋白上的变异株在谷蛋白上与对照也没有差异。更多还原

【Abstract】 Using the induction method of ion beam, the exogenous DNA from soybean was introduced into wheat mature seeds and the first and the second transformation generations were studied in this paper. Orthogonal design was used to investigate the transformation effect of receptor, servant, density of DNA, the time of seeds soaked by exogenous DNA, and SDS-PAGE and A-PAGE were used for testing storage protein of the transformation offspring with high protein content. The results were as follows:1. Ion beam induction influenced wheat quality in some degree and its effects were different with different varieties and quality traits.2. The quality of the first transformation generation (T1 generation) showed no significant difference with CK, but its variation increased and some plants with high protein content could be found. Separation of protein content appeared in T2 generation, and the variation increased again. Most plants returned to the CK level, but there were many plants left with high protein content.3. In the four factors which affected transformation rate, the most influencing was receptor, then servant, time of seeds soaked by exogenous DNA and density of DNA. Yunong92349 had the highest transformation rate in four receptors, so it was the best receptor. Zhengl4qingdou was the best servant in four soybean cultivars, 48h was the best time of soaking seeds and 200μg/ml was the best density of DNA.4. There were significant differences among the receptors and servants. Four good combinations had been found, and the rank was Yunong92349/Yudoul9,Yunong92349 /zhengduozhi, Yunong92349/Zheng 14qingdou, Yumai52/Zheng 14qingdou.5. By means of SDS-PAGE, fifteen mutants with high protein content were found and they were different with CK in high molecular weight glutenin subunits and the band color. Nine mutants were found by A-PAGE, and their differences existed in the region of low migration speed or " area and their band color was darker than CK’s. Compared with CK, the mutants in glutenin showed no difference in gliadin, and that in gliadin composition showed no difference in glutenin.更多还原