Agrobacterium-Mediated Transformation of Oilseed Rape (Brassica napus)

Q1 Agricultural and Biological Sciences

Current protocols in plant biology Pub Date : 2018-02-13 DOI:10.1002/cppb.20060

Ruth Bates, Melanie Craze, Emma J. Wallington

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引用次数: 15

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.

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农杆菌介导的油菜转化研究

油菜(Brassica napus)是芸苔科植物中具有重要商业价值的成员。种植它是为了食用和工业用油，以及作为动物饲料。利用遗传转化技术研究油菜基因功能，生产出具有改良农艺性状的油菜。本文描述了一种由农杆菌介导的油菜子叶叶柄转化的方法。该方法重复性好，已用于春、冬品种的转化。对生根期进行了改造，减少了枝条的玻璃化。这不仅增加了表型正常的芽的数量，而且还增加了转化效率，同时大大减少了再生的脱芽数量。转化频率通常在5%到10%之间，使用双单倍体模型品种的平均转化频率为12%，但最高效率为20%。©2017 by John Wiley &儿子,Inc。

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来源期刊

Current protocols in plant biology Agricultural and Biological Sciences-Plant Science

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期刊介绍： Sound and reproducible laboratory methods are the foundation of scientific discovery. Yet nuances that are critical for an experiment''s success are not captured in the primary literature but exist only as part of a lab''s oral tradition. Current Protocols in Plant Biology provides reproducible step-by-step instructions for protocols relevant to plant research. Furthermore, Current Protocols content is thoughtfully organized by topic for optimal usage and to maximize contextual knowledge. Quarterly issues allow Current Protocols in Plant Biology to constantly evolve to keep pace with the newest discoveries and developments. Current Protocols in Plant Biology is the comprehensive source for protocols in the multidisciplinary field of plant biology, providing an extensive range of protocols from basic to cutting edge. Coverage includes: Extraction and analysis of DNA, RNA, proteins Chromosome analysis Transcriptional analysis Protein expression Metabolites Plant enzymology Epigenetics Plant genetic transformation Mutagenesis Arabidopsis, Maize, Poplar, Rice, and Soybean, and more.