共培养条件对农杆菌转化大豆子叶节的影响  

Conditions of Co-Culture Affecting on the Efficiency of AgrobacteriumMediated Transformation of Cotyledonary Node of Soybean

作　　者：李桂兰[1] 刘晨光[1] 乔潇[2] 杨晓倩[1] 王迪[1] 乔亚科[1] 

机构地区：[1]河北科技师范学院生命科技学院,河北昌黎066600 [2]河北科技师范学院物理系,河北秦皇岛066004

出　　处：《核农学报》2014年第9期1567-1575,共9页Journal of Nuclear Agricultural Sciences

基　　金：转基因生物新品种培育科技重大专项(20142X08004-004B);河北省科技支撑计划(11220107D);河北省自然基金(C2014407051)

摘　　要：为提高大豆的遗传转化效率,利用根癌农杆菌对大豆成熟种子的子叶节进行遗传转化,从影响农杆菌和大豆子叶节外植体双方的因素出发,重点研究共培养阶段培养基的pH值、光照条件和共培养时间三个因素对转化的影响,以寻找适宜农杆菌和外植体的平衡点。分析单因素试验和正交试验的研究结果表明,在共培养阶段,培养基pH值适宜范围为6.0~6.6,pH值以6.6为佳;提供10~16h·d-1光照均提高了转化效率,与完全黑暗差异显著;延长共培养时间显著提高转化效率,以7~10d为宜。本研究范围内以冀豆12、豫豆22和秦黄小粒12三个基因型大豆获得了较高转化效率,供试的6个大豆品种均获得了转基因植株,PCR和Southern分析表明植酸酶基因phyA基因已经整合到大豆基因组中。该结果对提高大豆的遗传转化效率有重要参考价值。The factors affecting transformation efficiency in the co-culture stage were studied to improve Agrobacterium- mediated transformation of cotyledonary node of soybean. The results showed that the medium pH, illumination and culture duration significantly affected the transformation efficiency in the stage of co-culture. The optimal pH value of coculture medium was 6.6, the appropriate illumination duration ranged from 10 to16 h.d^-1, and the optimal co - culture time duration ranged from 7 - 10 d. PCR and southern analyses confirmed the stable integration of the phyA gene in the primary transgenic plants. Soybean varieties of ‘Jidou 12＇ , ‘ Yudou 22 ＇ and ‘ Qinhangxiaolil2＇ acquired higher transformation efficiency than others in this study.

关 键 词：农杆菌 大豆子叶节 共培养基pH值 共培养时间 共培养光照 

分 类 号：S565.1[农业科学—作物学]