籼粳稻对农杆菌遗传转化反应的差异及提高转化率的研究

Differences in response of indica and japonica rice variety to Agrobacterium tumefaciens-mediated transformation in relation to its improvement

通过对粳稻品种日本晴及5个籼稻品种的研究,发现籼稻、粳稻在成熟胚愈伤组织培养、农杆菌遗传转化的最适条件方面存在差异.在MS,CC,NB培养基上,日本晴的愈伤组织诱导率达86.37%以上,远高于各籼稻品种;与农杆菌液的共培养时间,籼、粳稻均以3 d为宜;转化后的籼稻抗性愈伤组织用40 mg/L的潮霉素筛选效果较好,日本晴则需用50 mg/L的潮霉素筛选;幼苗生根阶段,籼稻的潮霉素适宜质量浓度为20～30 mg/L,日本晴的潮霉素适宜质量浓度为50 mg/L;日本晴的gus基因瞬时表达率强于各籼稻品种,hpt基因转化率也高于各籼稻品种.

Different response to A.tumefaciens-mediated transformation of Nipponbare（japonica rice） and five indica rice isolates were investigated. The results are asfollows ： the inducement rate of Nipponbare reached 86.37%, on MS, CC, NB medium significantly higher than that of the indica rice isolates examined. For both indica and japonica variety, co-culture with tumefaciens for 3 days was beneficial to the target gene＇s transformation; 40 mg/L hygromycin could be used for selection of indica rice transformants selection, but 50 mg/L was needed for Nipponbare; 20 - 30 mg/L hygromycin is beneficial for selecting positive transplants of incica rice, and 50 mg/L for Nipponbare; Gus gene transient expression of Nipponbare and hpt gene transformation efficiency were better than that ofindica isolates.