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农杆菌介导的油菜(甘蓝型油菜)转化

Agrobacterium-Mediated Transformation of Oilseed Rape (Brassica napus).

作者信息

Bates Ruth, Craze Melanie, Wallington Emma J

机构信息

The John Bingham Laboratory, National Institute of Agricultural Botany, Cambridge, United Kingdom.

出版信息

Curr Protoc Plant Biol. 2017 Dec;2(4):287-298. doi: 10.1002/cppb.20060.

Abstract

Oilseed rape (Brassica napus) is a commercially important member of the Brassicacea family. It is grown for its edible and industrial oils as well as for animal feed. Genetic transformation technology has been used to study gene function and produce oilseed rape with improved agronomic characteristics. This protocol describes a method for the Agrobacterium tumefaciens-mediated transformation of oilseed rape cotyledonary petioles. The method is reproducible and has been used to transform both spring and winter cultivars. Modifications have been made to the rooting stage, which have reduced the vitrification of shoots. This has not only increased the number of phenotypically normal shoots but has also resulted in an increase in transformation efficiency, concomitant with a dramatic reduction in the number of escapes regenerated. Transformation frequencies typically range from 5% to 10%, with an average of 12% using doubled haploid model varieties, but a maximum efficiency of 20% has been achieved. © 2017 by John Wiley & Sons, Inc.

摘要

油菜(甘蓝型油菜)是十字花科中具有重要商业价值的成员。它因其可食用和工业用油以及动物饲料而被种植。遗传转化技术已被用于研究基因功能,并培育出具有改良农艺性状的油菜。本方案描述了一种通过根癌农杆菌介导转化油菜子叶叶柄的方法。该方法具有可重复性,已用于转化春、冬品种。在生根阶段进行了改进,减少了芽的玻璃化现象。这不仅增加了表型正常芽的数量,还提高了转化效率,同时显著减少了再生的逃逸植株数量。转化频率通常在5%至10%之间,使用双单倍体模式品种时平均为12%,但已实现的最高效率为20%。© 2017约翰威立父子公司版权所有

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