大豆论文改良的大豆子叶节转基因方法.docVIP

大豆论文：改良的大豆子叶节转基因方法 【中文摘要】随着基因工程技术的发展,采用遗传转化导入外源基因,改善大豆品质及抗病抗逆性,已成为大豆改良的重要方法之一。本文从多个层面对转基因方法的原理和进展作了详细综述,最终选取相对高效的农杆菌介导法作为大豆转基因的基础研究方法,并从种子消毒,外植体选择,农杆菌侵染和共培养,以及各培养时期植物激素种类和浓度等方面作了分析研究,以探讨大豆转基因优化研究步骤,以期在一定程度上达到提高转基因效率的。文章以苏云金芽孢杆菌(Bacillus thuringiensis)毒性蛋白基因Bt为基因,先研究未进行转染时期11种不同硼酸浓度处理对大豆子叶节丛生芽再生的影响,再从中选取代表性的硼酸浓度处理诱导期转染的大豆,观察三种硼酸浓度对大豆再生丛生芽转基因的影响,结果表明不加入硼酸时表现为严重缺硼症状,不仅生长点没有长出,而且根的生长也受到严重抑制。结果表明高浓度硼酸有较多的愈伤组织形成,外植体短期内容易死亡,不利于转基因丛生芽的再生。适宜浓度硼酸培养基诱导的大豆丛生芽长势最好,而且转基因效率也高达3.3%。对大豆抗性幼苗的根和叶片分别采用PCR、逆转录PCR和Southern blotting等方法进行筛选与鉴定,结果均获得阳性幼苗,证明外源基因已经成功转入大豆基因组中。 【英文摘要】Plant genetic engineering techniques have unprecedented potential for crop improvement. The technique of genetic transformation has been applied in the improvement of seed quality and disease resistance in soybean. Based on the principle and advatages of transgenetic technology the agrobacterium-mediated technique was eventually chosed as the basic transformation method. Sterile seedling, explant choice, agrobacterium infection and coincubation, and concentration of plant hormone during the training period were tried in the investigation of plant regeneration, in order to optimize the medium for regeneration and greatly improve the transgenic efficiency.The agrobacterium-mediated transgenic technique combined with the treatment of boric acid was studied for the transformation of foreign gene Bt to explant, soybean cotyledon nodes. 11 different boric acid concentration processing multiple shoot of soybean cotyledons section were chosen, then transfomed soybeans during the induction period were treated seperately by the concentration of typical boric acid. The results showed that the transformation seedlings without boric acid had no growing point, and the roots growth were also severely restrained.At the same time,the high boric acid concentration growed more callus, the explants short-term easily died, which were not conducive to the regeneration of transformed multiple shoots. However,the shoots regenerated from