摘要
以兰州百合试管苗鳞片叶为受体,通过基因枪转化法将脱氢抗坏血酸还原酶基因(DHAR)转入兰州百合,并用GUS染色、PCR扩增和Southern杂交等技术进行转基因株系的检测.结果表明:以兰州百合鳞片叶为受体材料,在轰击压力1 100 psi、轰击距离6 cm时,抗性率高达14%,转化率为1.5%;10 mg/L的潮霉素对鳞片叶具有很好筛选作用.GUS分析发现,转化株系叶片为蓝色;PCR检测和Southern杂交进一步证实抗性株系为转基因植株,且为单拷贝.
For the establishment of a simple multi-channel transgenic technology of Lilium davidii var.unicolor Salisb.,a biolistic protocol for the stable genetic transformation of L.davidii var.unicolor Salisb.was developed.Scale leaves of Lanzhou lily were used as explants and transformed the dehydroascorbate reductase（DHAR） gene into the Lanzhou lily.The resistant lines were analyzed by GUS staining,PCR and Southern blot.The results showed that the resistance rate was 14% when scale leaves of Lanzhou lily were used as explants and transformed by gene gun under the conditions of 6 cm shoot distance,1 100 psi helium pressure.The better selective efficiency was gotten when the concentration of hygromycin was 10 mg/L.The results of PCR and Southern blot proved the resistant lines were transgenic plants,and the transgene numbers was single copy.
出处
《西北植物学报》
CAS
CSCD
北大核心
2010年第4期677-682,共6页
Acta Botanica Boreali-Occidentalia Sinica
基金
国家自然科学基金(30871700)
西北农林科技大学拔尖人才支持计划
