国家自然科学基金项目(面上项目,重点项目,重大项目)
以3个水稻品种的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。
Abstract:Callus induced from mature embryos of 3 rice varieties were transformed by Agrobactorium tumefaciens. Transformation efficiency was affected by factors of pre-culture time, density of bacteria, duration for inoculation and co-cultivation. The transformation conditions were optimized with the combination of pre-culture of 4 days, infection in bacterium with OD600 0.7-1.0 for 5-10 minutes., followed by co-culture of 2 days. Plantlets with resistance to hygromycin were regenerated. PCR and PCR-Southern analysis on them confirmed the integration of the LycB be into the rice genome.
抗艳红.农杆菌介导类胡萝卜素合成酶基因LycB转化水稻的研究[J].植物遗传资源学报,2011,12(4):605-611.
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