|
|||||
|
|
| 农杆菌浸种法介导中华结缕草遗传转化体系的建立 | |
| 引用本文： | 雷江丽,王丹,吴燕民,刘水,吕慧.农杆菌浸种法介导中华结缕草遗传转化体系的建立[J].农业生物技术学报,2009,17(5):865-871. |
| 作者姓名： | 雷江丽 王丹 吴燕民 刘水 吕慧 |
| 作者单位： | 1. 深圳市园林科学研究所,深圳,518003 2. 中国农业科学院生物技术研究所,北京,100081 3. 深圳市铁汉园林绿化有限公司,深圳,518034 |
| 基金项目： | 深圳市科技和信息局资助项日 |
| 摘 要： | 以中华结缕草萌动种子为转化受体，利用NPT-Ⅱ筛选基因对影响农杆菌介导的遗传转化外部因素进行探索，通过优化转化条件，建立了农杆菌浸种途径介导的中华结缕草遗传转化体系。结果表明：利用催芽6-7d的种子，加入OD600值为0.9的农杆菌浸染菌液（在浸染菌液中添加100μmol.L-1乙酰丁香酮），附加5min+30s真空处理，振荡浸染48h，共培养3d后，用75mg.L-1卡那霉素筛选能够获得较好的转化效果。通过对卡那霉素抗性苗进行PCR检测和GUS组织化学染色，初步验证NPT-Ⅱ和GUS基因已经整合进植物基因组，阳性转化率为55.21%。 |
| 关 键 词： | 中华结缕草 农杆菌浸种法 遗传转化 |
| 收稿时间： | 2008-10-31 |
| 修稿时间： | 2009-5-27 |
Establishment of Genetic Transformation System of Chinese Zoysia(Zoysia sinica)Mediated by Agrobacterium tumefaciens Soaked Seeds |
|
| Abstract: | In this study, a high-frequency transformation system has been established by Agrobacterium tumefaciens soaked seeds transformation technology, Chinese zoysia germinating seed were used as transformation receptor. External factors that affect the efficiency of genetic transformation were studied and optimized, on the base of NPTⅡ. The major results as followed: seed was germinated for 7 days, then was infected with Agrobacterium which OD600 was 0.9 with 100μmol.L-1 AS for 48 hours after pumping 5min+30s vacuum treatment. Then co-cultivation for 3 days, selected the transferred plant by 75mg.L-1 kanamycin. It was originally proved by the PCR and GUS examination that NPT-Ⅱand GUS gene had been transferred into the genome. The positive transformation rate was 55.21% by the analysis of PCR. |
| Keywords: | |
| 本文献已被 维普 万方数据 等数据库收录！ | |
| 点击此处可从《农业生物技术学报》浏览原始摘要信息 | |