[A simple and highly efficient Agrobacterium-mediated rice transformation system].
[A simple and highly efficient Agrobacterium-mediated rice transformation system].
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发表时间：
2003-08
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通讯作者：
Mei-Ru Li;Hong Li
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作者：
Mei-Ru Li;Hong Li
通过对影响农杆菌介导水稻遗传转化的因素的研究，建立了一套简单、高效的水稻遗传转化体系。将成熟胚的胚性愈伤组织与A.根癌农杆菌EHA 101，含有双元载体：pHQ 9、pHQ 10、pHQT 3。转化频率最高可达100个潮霉素抗性愈伤组织/克愈伤组织，其中85%以上的愈伤组织可再生为植株。通过GUS检测和Southern印迹分析，证实了转基因植株的存在。潮霉素抗性试验表明，转基因在T1代中的分离符合孟德尔比例3：1。该系统的建立将为利用T-DNA插入突变和基因打靶技术进行水稻功能基因组学研究奠定基础。
A simple and highly efficient rice transformation system was established based on the studying of factors influencing the Agrobacterium-mediated rice transformation. Embryogenic calli derived from mature embryos were infected and cocultivated with A. tumefaciens EHA101 harboring binary vectors: pHQ9, pHQ10, pHQT3. The highest transformation frequency was about 100 hygromycin resistance calli per gram of calli explants, and above 85% of these calli could be regenerated into plants. The putative transgenic plants were confirmed by GUS assay and southern-blot analysis. Hygromycin resistance tests indicated that the segregation of transgene in T1 progeny corresponded to the Mendelian ratio, 3:1. This system will benefit the functional genomic study of rice by using T-DNA insertion mutagenesis and gene targeting.