Li Mei Ru, Li Hong Qing
South China Institute of Botany, Chinese Academy of Sciences, Guangzhou 510650.
Shi Yan Sheng Wu Xue Bao. 2003 Aug;36(4):289-94.
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.
基于对影响农杆菌介导的水稻转化因素的研究,建立了一种简单高效的水稻转化系统。将来自成熟胚的胚性愈伤组织与携带二元载体pHQ9、pHQ10、pHQT3的根癌农杆菌EHA101进行感染和共培养。最高转化频率约为每克愈伤组织外植体产生100个潮霉素抗性愈伤组织,且这些愈伤组织中85%以上能够再生为植株。通过GUS检测和Southern杂交分析对推定的转基因植株进行了鉴定。潮霉素抗性试验表明,转基因在T1代子代中的分离符合孟德尔比例3:1。该系统将通过T-DNA插入诱变和基因打靶有利于水稻功能基因组学研究。