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中国农学通报

中国农学通报 ›› 2011, Vol. 27 ›› Issue (21): 87-92.

所属专题： 马铃薯

• 农学 农业基础科学 • 上一篇    下一篇

加工型马铃薯品种Shepody农杆菌介导的转化体系研究

辛翠花 黄三文 郭江波 刘杰 冷小云 屈冬玉   

  • 收稿日期:2011-02-28 修回日期:2011-05-12 出版日期:2011-08-25 发布日期:2011-08-25

Research of Agrobacterium-mediated Transformation System for Economically Important Potato Cultivar Shepody

  • Received:2011-02-28 Revised:2011-05-12 Online:2011-08-25 Published:2011-08-25

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摘要：

马铃薯Shepody是经济价值很高的炸条加工品种，但Shepody对逆境胁迫非常敏感，为确立Shepody的遗传转化体系并获得转基因马铃薯，以叶片、茎段和微型薯为外植体，分别比较了不同转化条件（菌液浓度、侵染时间及激素配比）对遗传转化效率的影响。结果表明：叶片为最理想的外植体，以OD600=0.5的菌液侵染10 min，在加有1.0 mg/L萘乙酸（NAA）和1.0 mg/L反式玉米素（ZT-t）的MS20选择培养基上的遗传转化率最高，其愈伤诱导率可达到95%，分化率可达到20%。通过该转化体系将抗晚疫病基因R1导入马铃薯，获得了具有卡那霉素抗性的转化植株，经PCR鉴定外源基因已经成功导入马铃薯基因组中。

关键词: 半夏, 半夏, 3414肥料试验, 产量, 有效成分

Abstract:

Potato Shepody is economically important potato cultivar for french fries in the world. However, this cultivar exhibits very susceptible to severe environmental stress. The aim of this study was to develop an efficient plant transformation system for potato cultivar Shepody and to obtain transgenic potatoes. Leaf disks, stem segments and minitubers from potato cultivar Shepody were used as receptor materials to compare transformation efficiency with different Agrobacterium tumefaciens content, duration of infection and the proportion of hormones. The results showed that leaf disks was most suitable for the induction of callus and shoots; Leaf disks explants infected by A.tumefaciens strain Agl-0 (OD600=0.5, 10 min) and selected on MS20 containing 1.0 mg/L NAA and 1.0 mg/L zeatin-trans(ZT-t), yielded the highest transformation efficiency with 95% of callus induction and 20% of differentiation. The gene resistant to potato late blight (R1) was introduced into potato through the above transformation system. PCR analysis to the kanamycin-resistant transformants manifested that the R1 gene was integrated into the potato genome.