摘要
为提高农杆菌介导的苜蓿遗传转化效率,获得转K88ac-STⅡ-LTA2/LTB基因苜蓿植株,以苜蓿下胚轴为外植体,对影响遗传转化的预培养时间、农杆菌菌液浓度、筛选方式等进行研究。结果表明,苜蓿下胚轴预培养3 d,菌液浓度为OD6000.5,筛选剂Kan浓度第一阶段和第二阶段分别为30和50 mg/L,延迟筛选7 d时转化效率最高。对转基因苜蓿进行PCR和PT-PCR检测,初步证实目的基因已经整合到苜蓿基因组中,转化率达28.4%。
The objective of this study was to optimize the condition of the Agrobacterium-mediated alfalfa transformation so that the transformation plants can be obtained. With alfalfa hypocotyls as explants, the factors affecting the transformation frequency （preculture time, concentration of Agrobacterium and antibiotics） were studied. High transformation rate was gained when alfalfa hypocotyls precultured for 5d were immersed into the OD60o 0.5 Agrobacterium suspension and finally were subcultured on selection medium containing Kan 30 and 50 mg/L during the first and second stages after reinforcing cultivation for 20 d. Results of PCR and RT-PCR proved that the aim gene had integrated into alfalfa genome; the transformation rate was 28.4%.
出处
《西北农业学报》
CAS
CSCD
北大核心
2009年第3期147-151,共5页
Acta Agriculturae Boreali-occidentalis Sinica
基金
新疆建设兵团博士基金(兵博02)
关键词
苜蓿
转化
农杆菌
融合基因
Alfalfa
Transformation
Agrobacterium tumefaciens
Fusion gene
