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| 国槐遗传转化体系的优化 | |
| 引用本文： | 张晓英,甘敬,尹伟伦,朱祯,王华芳.国槐遗传转化体系的优化[J].林业科学,2009,45(5). |
| 作者姓名： | 张晓英 甘敬 尹伟伦 朱祯 王华芳 |
| 作者单位： | 1. 北京林业大学生物科学与技术学院,北京,100083;北京市琅山苗圃,北京,100039 2. 北京市园林绿化局,北京,100029 3. 北京林业大学生物科学与技术学院,北京,100083 4. 中国科学院遗传与发育生物学研究所,北京,100101 |
| 基金项目： | 国家高技术研究发展计划(863计划) |
| 摘 要： | 采用根癌农杆菌介导的国槐叶盘转化法,在建立修饰的豇豆胰蛋白酶抑制剂基因(sck)转化体系过程中,对影响农杆菌转化频率的各种因素进行研究.结果表明:国槐叶片需在黑暗条件下在MS培养基上预培养5天,农杆菌菌株选用GV3101,农杆菌共培养3天较合适;农杆菌菌液浓度OD600约为0.7,感染时间10 min;侵染前的菌液和共培养基中添加200/μmol·L-1乙酰丁香酮(AS)对国槐遗传转化有明显的促进作用;抑制农杆菌生长的抗生素浓度以头孢霉素(Cef)500mg·L-1最好.PCR和Southern blotting检测证实sck基因已整合到国槐基因组中. |
| 关 键 词： | 国槐 根癌土壤农杆菌 乙酰丁香酮 遗传转化 槐尺蠖 抗虫基因 |
An effective Transformation System of Sophora japonica |
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| Zhang Xiaoying,Gan Jin,Yin Weilun,Zhu Zhen,Wang Huafang.An effective Transformation System of Sophora japonica[J].Scientia Silvae Sinicae,2009,45(5). | |
| Authors: | Zhang Xiaoying Gan Jin Yin Weilun Zhu Zhen Wang Huafang |
| Institution: | 1.College of Biological Science and Biotechnology;Beijing Forestry University Beijing 100083;2.Beijing Langshan Nursery Beijing 100039;3. Beijing Gardening and Greening Bureau Beijing 100029;4. Institute of Genetics and Developmental Biology;Chinese Academy of Sciences Beijing 100101 |
| Abstract: | This paper investigated several factors affecting transformation frequency of Sophora japonica in establishing the modified sck gene transfer system process with Agrobacterium tumefaciens-mediated leaf disc transformation method. The results showed that S. japonica leaves were required to pre-culture on MS medium for 5 d in the dark. Agrobacterium strains,GV3101,were selected,and the Agrobacterium was needed to co-cultivation for 3 d. Agrobacterium bacterium concentration was about 0.7 in OD 600 ,and the in... |
| Keywords: | Sophora japonica Agrobacterium tumefaciens acetosyringone genetic transformation Semiothisa cinerearia insect-resistant genes |
| 本文献已被 CNKI 维普 万方数据 等数据库收录！ | |