一种安全高效大豆转化筛选体系的建立

以灭草烟作为筛选剂,利用基因枪法建立一种安全高效的大豆遗传转化体系.比较不同筛选剂对大豆胚尖外植体丛生芽诱导数目的影响.与卡那霉素、潮霉素和草胺膦等传统筛选剂相比,以灭草烟作为筛选剂可使丛生芽的数目增加1倍以上.克隆了拟南芥突变体csrl-2中突变的乙酰羟基酸合成酶基因(ahas),以其作为筛选标记基因,构建可利用灭草烟作为筛选剂的植物表达载体.利用基因枪法将该载体转化大豆,获得6棵灭草烟抗性植株,分子检测证明外源ahas基因整合到5棵转基因大豆植株的基因组中.

A High-efficient and Safe Selection System of Soybean Transformation

We used imazapyr as selection agent to establish a high-efficient and safe system of soybean transformation by biolistics.The effects of different selection agents including imazapyr,hygromycin,kanamycin and glufosinate in soybean shoot induction were investigated.It found that the number of multiple shoot increased twice using imazapyr compared with the other selection agents.The mutated acetohydroxyacid synthase gene(ahas)from Arabidopsis mutant csr1-2 was cloned and utilized in constructing a binary vector with imidazolinone herbicide resistance.We have successfully used this binary vector to transform soybean embryo tips via biolistics.Six resistant transformants were screened with imazapyr after transformation,and five of them were proved to harbor foreign ahas gene by PCR and sequencing analysis.