摘要
以新黑田五寸胡萝卜下胚轴为外植体,研究对胡萝卜抗性植株的影响因素及目的基因的整合情况,建立了较为完善的转化体系。实验结果表明:以胡萝卜下胚轴为材料,用苗龄7d,共培养2d,浸染15min的体系,Kan抗性芽的分化频率可达52%。获得抗性愈伤培养基:B5+0.1mg/L2,4-D+0.2mg/LKT+75mg/LKan+500mg/LCef,抗性芽诱导培养基:B5+100mg/LKan+500mg/LCef;抗性植株生根培养基:B5+0.1mg/LIBA+75mg/LKan+300mg/L:Cef;对转基因植株进行了PCR分子学检测,初步证明外源基因已整合到胡萝卜基因组中。
The hypocotyls of carrot were used as explants in the experiment. Factors that affected carrot resistant plants and integration performance of target gene were studied, and a stable transformation system was established. Result showed that differentiation frequency of Kan resistant shoots reached to 52 % in the system of 7 d seedling, 2 d of co cultivation and 15 minutes of infection. Media for resistant callus： B5＋ 0.1 mg/L 2, 4-D ＋ 0.2 mg/L KT ＋ 75 mg/L Kan ＋500 mg/L Cef. Media for resistant shoots： B5 ＋ 100 mg/L Kan ＋ 500 mg/L Cef. Media for resistant roots： B5 ＋ 0.1 mg/L IBA ＋ 75 mg/L Kan ＋ 300 mg/L Cef. PCR detection of transgenic plant DNA confirmed that the target gene had been integrated into the carrot genome.
出处
《甘肃农业大学学报》
CAS
CSCD
2005年第4期535-539,共5页
Journal of Gansu Agricultural University
基金
国家高技术研究发展计划(863)资助项目(2001 AA212171.A)
