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研究了抗生素种类、继代和菌液浓度对农杆菌介导籼稻转化的影响,并将反义蜡质基因导入4个高产、直链淀粉含量高的籼稻品种中。结果显示,羧苄青霉素的抑菌效果优于头胞霉素,其适宜浓度为250￣350mg/L;4个品种继代愈伤比初代愈伤能获得更高的抗性愈伤得率;各个品种转化有其最佳的农杆菌菌液浓度(OD600值),茉莉新占和绿黄占为1.0,粤香占和矮秀占为0.6。应用此优化的农杆菌转化体系,我们得到了转化植株;PCR扩增和Southern杂交结果证明外源基因已经整合到转基因水稻的基因组中。
The effects of antibiotic species, subculture and bacterial concentration on Agrobacterium-mediated transformation of indica rice were studied. Antisense waxy genes were introduced into four high-yielding indica rice varieties with high amylose content. The results showed that the antibacterial effect of carbenicillin was better than that of cephalosporin, and its optimum concentration was 250-350 mg / L. The callus yield of four varieties was higher than that of primary callus. The best Agrobacterium tumefaciens concentration (OD600) was obtained for each variety transformation, accounting for 1.0 for jasmine, 15 for jasmine and 0.6 for jasminoides. Using this optimized Agrobacterium transformation system, we obtained transformed plants; PCR amplification and Southern hybridization results showed that foreign genes have been integrated into the genome of transgenic rice.