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以籼稻品种9311为材料,研究了不同诱导培养基对籼稻9311的诱导率,以及农杆菌浓度、浸染时间和干燥处理时间对抗性愈伤率的影响,进而采用农杆菌介导的共转化方法将含有反义Waxy基因的p13W8质粒和含有潮霉素抗性标记基因的p1300质粒同时转化受体材料.结果表明:籼稻9311成熟胚在含质量浓度为3.0mg/L的2,4-D的N6基本培养基上具有较高的愈伤组织诱导率,且愈伤的胚性性状良好;愈伤组织在OD600值为0.6的农杆菌菌液中浸染10min、干燥1.5～2.0h处理后能够获得较多的抗性愈伤组织;对转化水稻的再生植株进行PCR检测表明,反义Waxy基因已整合进T0代水稻基因组中.
Indica rice 9311 was used to study the effects of different induction medium on the induction rate of indica 9311, the effect of Agrobacterium tumefaciens concentration, the time of dipping and the time of drying treatment on the resistant callus. Then Agrobacterium-mediated cotransformation The p13W8 plasmid containing the antisense Waxy gene and the p1300 plasmid containing the hygromycin resistance marker gene were simultaneously transformed into the recipient material.The results showed that the mature embryos of indica rice 9311 were cultured in a medium containing 2,4-D N6 at a concentration of 3.0 mg / L The callus induction rate on the basic medium was good, and the embryogenic traits of the callus were good. The callus in the Agrobacterium solution with an OD600 value of 0.6 was immersed for 10 minutes and dried for 1.5-2.0 hours. More resistant callus; PCR analysis of the transformed rice transformed plants showed that the antisense Waxy gene has been integrated into the T0 generation of rice genome.