根癌农杆菌介导的水稻转基因技术体系的优化

Condition optimization of Agrobacterium-mediated transgenic rice

对根癌农杆菌介导水稻遗传转化的各重要因素逐一分析,比较不同基因型、培养基成份、光照、继代次数、菌液浓度、侵染时间、干燥情况等因素对遗传转化的影响。通过培养条件的优化,建立了一个高效的水稻转化技术体系。研究结果表明,相比较于暗培养,光照培养进行愈伤诱导可以使诱导天数缩短4~5d;诱导培养基中加入适量的激素可以使籼稻愈伤诱导提高25%~30%;分化培养基中加入适量的氨基酸和甘露醇可以使植株再生频率提高15%~20%。

The important factors of A.tumefacienens mediated transformation of rice were analyzed one by one.The effects of different factors like genotype,culture component,light intensity,sub-cultures,inoculation and infection condition on the transformation frequency were investigated.By the optimization of the conditions,an efficient genetic transformation system of rice was developed.The main results showed that illumination culture for callus induction could reduce 4-5 days comparing to dark culture.Induction medium supplemented with appropriate hormone can significantly increase frequency of Indica callus induction of 25%-30%.Differentiation medium supplemented with appropriate amino acids and mannitol can enhance the regeneration frequency of 15%-20%.