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农杆菌介导的玉米转化

Agrobacterium-mediated transformation of maize.

作者信息

Ishida Yuji, Hiei Yukoh, Komari Toshihiko

机构信息

Plant Innovation Center, Japan Tobacco Inc., 700 Higashibara, Iwata, Shizuoka, Japan.

出版信息

Nat Protoc. 2007;2(7):1614-21. doi: 10.1038/nprot.2007.241.

Abstract

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.

摘要

利用根癌农杆菌介导的方法可以非常高效地转化玉米。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根癌农杆菌共同培养;胚应仅从生长在条件良好的温室中的健壮植株上采集。采用此处所述的方案,自交系A188中约50%的未成熟胚以及自交系A634、H99和W117中15%的未成熟胚将产生转化体。预计约一半的转化植株携带一到两个转基因拷贝,这些转基因以孟德尔方式由后代遗传。预计超过90%的转化体在形态上是正常的。该方案从共同培养开始到将转化体种植到花盆中大约需要3个月。

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