Agrobacterium-mediated transformation of maize
Agrobacterium-mediated transformation of maize
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DOI：
10.1038/nprot.2007.241
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发表时间：
2007-01-01
期刊：
影响因子：
14.8
通讯作者：
Komari, Toshihiko
中科院分区：
文献类型：
--
作者：
Ishida, Yuji;Hiei, Yukoh;Komari, Toshihiko
使用根癌农杆菌介导的方法可以非常有效地转化玉米。转化方案中最关键的因素是将处于正确发育阶段的健康未成熟胚与A.根癌农杆菌;胚应仅从生长在条件良好的温室中的旺盛植物中收集。采用本文所述的方案，来自近交系A188的约50%的未成熟胚和来自近交系A634、H99和W117的15%的未成熟胚将产生转化体。预计约有一半的转化植物携带一个或两个拷贝的转基因，这些转基因以孟德尔方式遗传给后代。预期超过90%的转化体在形态上是正常的。该方案从共培养开始到将转化体种植到盆中花费约3个月。
Maize may be transformed very efficiently using Agrobacterium tumefaciens-mediated methods. The most critical factor in the transformation protocol is the co-cultivation of healthy immature embryos of the correct developmental stage with A. tumefaciens; the embryos should be collected only from vigorous plants grown in well-conditioned glasshouses. With the protocol described here, approximately 50% of immature embryos from the inbred line A188 and 15% from inbred lines A634, H99 and W117 will produce transformants. About half of the transformed plants are expected to carry one or two copies of the transgenes, which are inherited by the progeny in a mendelian fashion. More than 90% of transformants are expected to be normal in morphology. The protocol takes about 3 months from the start of co-cultivation to the planting of transformants into pots.