【作者】 曲波；

【导师】 李庆章； 郭德栋；

【作者基本信息】 东北农业大学 ， 生物化学与分子生物学， 2003， 硕士

【摘要】 甜菜单体附加系M14品系是我国特有的无融合生殖材料。筛选和鉴定从M14后代中分离出来的带有白花甜菜染色体片段的易位系，将会加快从遗传性状到基因克隆和功能鉴定的速度，对克隆无融合生殖基因有重大意义。 本文采用γ射线照射的方法加速产生易位系。首先利用γ射线处理栽培甜菜单体附加系M14种子，然后在温室中播种、调查不同放射剂量种子的成活率，并进行细胞学观察，最后对发生染色体变异的株系利用白花甜菜特异探针进行DNA分子杂交。共得到11株易位系，易位系发生率4.54％，明显高于自发易位0.26％的发生率，诱导作用明显。研究结果表明γ射线的剂量对甜菜的成活率及易位系产生的几率有很大影响。辐射强度越大，甜菜成活率越低，而染色体变异率和易位系发生率越高。综合两方面因素，强度为60Kr和80Kr的辐射效果较好。本文还对16个株系共264株易位系的后代进行斑点杂交，计算其总传递率为50.4％，这与理论值相符合。其中株系11的传递率为100％，株系14也达到83.3％，基本属于高频传递。最后本文对荧光原位杂交的染色体制片、变性、杂交、洗涤、信号检测等过程的实验条件进行了各种对比实验，摸索出一套较成功的实验方案，建立了易位系鉴定的基本程序。更多还原

【Abstract】 Monosomic addition lines M14 of Beta corolliflora Zoss in sugar beet is a specific apomictic material of our country. It will facilitate the speed from genetic characters to gene cloning and function identifying that selection and characterization of the beet translocation lines with the fragment of chromosome from Beta corolliflora Zoss, which is very important to clone apomictic gene.The research accelerated the generation of translocation line with y -ray. Firstly, seeds of monosomic addition lines M14 were induced by y -ray, and then were planted in greenhouse.Secondly, The paper investigated the survival probability of seeds induced with different radiant intensity and had a cytological analysis by microscopical examination on surviving ones.Finally, this experiment made those plants have a dot hybridization by using the specific sequence 1054 of the Beta corolliflora Zoss as probe, of which chromosomes were aberrant. In that investigation, 11 translocation lines were found, and translocation line occurring frequency reached 4.54%, which obviously was more than that of spontaneous translocation and had a great effect on radiant inducement. The research results indicated that the intensity of -ray affect the survival probability of beet and the frequency of translocation line occurring deeply. The more strong radiant intensity was, the lower the survival probability of beet was, and the higher the frequency of chromosome differentiating and translocation line occurring was. According to that two factors, radiation with 60Kr and 80Kr had a better effect The experiment also had a research with dot hybridization to the 264 translocation plants of the 16 series, the research results indicated that the offsprings’general transfer-rate reached 50.4%, of which the transfer-rate of plant 11 and plant14 was up to 100% and 83.3%, belonging to high frequency transmission. Finally, a set of successful experiment project about fluorescence in situ hybridization was constructed through kinds of contrastive experiments which excluded spreading of plant chromosomes, denaturalization, hybridization,washing and detection of signals and so on.更多还原