An improved Agrobacterium-mediated transformation protocol for recalcitrant elite indica rice cultivars
An improved Agrobacterium-mediated transformation protocol for recalcitrant elite indica rice cultivars
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DOI：
10.1007/bf02772648
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发表时间：
2005-03-01
影响因子：
2.1
通讯作者：
Balasubramanian, P
中科院分区：
文献类型：
--
作者：
Kumar, KK;Maruthasalam, S;Balasubramanian, P
本文报道了一种利用选择标记基因hph和gusA报告基因对籼稻品种IR 64和IR 72进行高效转化的方法。有利于高效转化的因素被发现是使用2个月大的成熟种子来源的胚性愈伤组织，麦芽糖作为碳源，较高浓度的2，4-二氯苯氧乙酸，和phytagel和琼脂作为胶凝剂。通过聚合酶链反应和Southern印迹分析来分析推定的转基因（T-0）植物的转基因整合。被认为是负责提高转化效率的各种因素进行了讨论。
We report here a high-efficiency transformation protocol for recalcitrant indica rice cultivars IR64 and IR72 with the selectable marker gene hph and the gusA reporter gene. Factors that favor high-efficiency transformation were found to be use of 2-month-old mature seed-derived embryogenic calli, maltose as a source of carbon, a higher concentration of 2,4-dichlorophenoxyacetic acid, and both phytagel and agar as gelling agents. The putative transgenic (T-0) plants were analyzed for integration of the transgene through polymerase chain reaction and Southern blotting analyses. Various factors thought to be responsible for increased transformation efficiency are discussed.