Thole Vera, Vain Philippe
Department of Crop Genetics, John Innes Centre, Norwich, UK.
Methods Mol Biol. 2012;847:137-49. doi: 10.1007/978-1-61779-558-9_12.
Brachypodium distachyon is an attractive genomics and biological model system for grass research. Recently, the complete annotated genome sequence of the diploid line Bd21 has been released. Genetic transformation technologies are critical for the discovery and validation of gene function in Brachypodium. Here, we describe an efficient procedure enabling the Agrobacterium-mediated transformation of a range of diploid and polyploid genotypes of Brachypodium. The procedure relies on the transformation of compact embryogenic calli derived from immature embryos using either chemical selection alone or a combination of chemical and visual screening of transformed tissues and plants. Transformation efficiencies of around 20% can routinely be achieved using this protocol. In the context of the BrachyTAG programme (BrachyTAG.org), this procedure made possible the mass production of Bd21T-DNA mutant plant lines.
短柄草是禾本科研究中一个具有吸引力的基因组学和生物学模式系统。最近，二倍体系 Bd21 的完整注释基因组序列已发布。遗传转化技术对于短柄草基因功能的发现和验证至关重要。在此，我们描述了一种高效方法，可实现农杆菌介导的一系列短柄草二倍体和多倍体基因型的转化。该方法依赖于使用单独的化学筛选或转化组织和植物的化学与视觉筛选相结合的方式，对从未成熟胚衍生的紧密胚性愈伤组织进行转化。使用该方案通常可实现约 20%的转化效率。在 BrachyTAG 计划（BrachyTAG.org）的背景下，此方法使得 Bd21 T-DNA 突变体植株系的大规模生产成为可能。
