农杆菌介导鼠源轮状病毒vp4基因转化马铃薯条件优化研究

Optimal conditions for transforming murine vp4 gene into potatoes induced by Agrobacterium tumernfaciens

目的 获得马铃薯再生和农杆菌介导的遗传转化体系。方法 以马铃薯品种“台湾红”为受体材料 ,对其再生和农杆菌介导的遗传转化体系进行了研究。结果 诱愈培养基为MS + 0 1mg L 2 ,4-D + 1mg LBA ,出芽培养基为MS + 1mg LZT +5 0mg LGA3 ,卡那霉素浓度为 5 0mg L ,农杆菌液浓度为A6 0 0 =0 5。通过该体系将一鼠源轮状病毒外壳蛋白基因vp4导入马铃薯中 ,获得了具卡那霉素抗性的转化植株。经PCR验证 ,外源基因已导入马铃薯基因组中。结论 获得相对高效的马铃薯遗传转化体系 ,并成功地将外源基因转入马铃薯中。

Objective To obtain a system of transforming foreign gene into potatoes with regenerating ability Methods A potato cultivar 'taiwanhong' sensitive to Agrobacterium tumerfaciens, was used as the transformation receptor A regeneration system and a transformation system via Agrobacterium tumerfaciens were studied Results The optimal conditions for regeneration and transformation for foreign gene into potato were listed as following: the callus inducement medium: MS+0 1mg/L 2,4-D +1 mg/L BA; the budding medium: MS+1 mg/L ZT+5 0 mg/L GA3; kanamycin concentration: 50 mg/L; the bacterial concentration: A 600 =0 5 A murine rotavirus outer capsid protein gene vp4 was introduced into the potato under the condition as above mentioned PCR analysis of the kanamycin resistant transformants manifested that the foreign gene was integrated into the potato genome Conclusion A high effective genetic transformation system is obtained and successfully used in transforming the foreign gene into potatoes