一种高效的农杆菌介导大豆子叶节转化体系的建立

Optimization of Agrobacterium-Mediated Soybean Transformation of Cotyledonary-Node

以子叶节为外植体的农杆菌介导的大豆遗传转化体系,其转化效率受到受体基因型、无菌苗状态、丛生芽诱导及伸长和生根过程涉及的多种因素的影响。作者筛选了10个大豆栽培品种,选择诱导率最高的东农50作为转化的受体品种,建立子叶节为外植体的遗传转化化体系。还对萌发培养基中6-BA添加量和生根方式进行了研究。结果表明:萌发培养基中不添加6-BA伸长率可达2.51%～20.59%;伸长芽切下后浸入1mg/L的IBA中处理1min后于1/2MS培养基中生根,生根时间缩短,生根率提高至85.25%。经PCR检测,本转化体系的转化率最高可达7.10%。

The efficiency of Agrobacterium-mediated soybean transformation of cotyledonary-node was affected by some factors including receptor genotype,germinating condition,shoot induction,elongation and rooting.10 soybean cultivars were chosen,Dongnong 50 in 10 tested cultivars was used as the transformed receptor species,and the system of cotyledonary-node transformation was established.We investigated the proper dosage of 6-BA in germination medium and rooting mode.The results showed that different concentration of 6-BA was added in germination medium and the rate of elongation was 2.51%~20.59% with 0 concentration of 6-BA in germination stage.Dipping the stem in 1 mg/L IBA for 1 minute and rooting in 1/2 MS medium shortened the rooting time and increased rooting rate by 85.25%.The transforming rate of the system could achieve 7.10% by PCR assay.