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以3个水稻品种成熟胚诱导的良好胚性愈伤组织为受体,以LycB为目的基因,应用根癌农杆菌介导法对水稻进行遗传转化,同时以抗性愈伤率为依据,对影响转化的几个因素进行优化研究。结果表明,预培养4d、侵染5~10min、农杆菌菌液浓度OD600值0.7~1.0、共培养2d有利于提高转化率。经潮霉素筛选获得的抗性植株经PCR和PCR-Southern分析鉴定,初步证明外源基因LycB已整合到水稻的基因组中。
The good embryogenic callus induced by three mature rice embryos was used as the receptor. LycB was used as the target gene and the Agrobacterium tumefaciens-mediated method was used to transform the rice. Based on the resistant callus rate, Several factors that influence the conversion are optimized. The results showed that the pre-culture 4d, infection 5 ~ 10min, the Agrobacterium OD600 value of 0.7 to 1.0, co-cultured for 2d is conducive to improving the conversion rate. The resistant plants obtained by hygromycin screening were identified by PCR and PCR-Southern analysis, which initially proved that the foreign gene LycB has been integrated into the genome of rice.