Luu Van T, Stiebner Melissa, Maldonado Paula Emmerich, Valdés Sandra, Marín Didier, Delgado Gerardo, Laluz Virginia, Wu Lin-Bo, Chavarriaga Paul, Tohme Joe, Slamet-Loedin Inez H, Frommer Wolf B
Institute for Molecular Physiology, Heinrich Heine University of Düsseldorf (HHU), Düsseldorf, Germany.
Max-Planck-Institute for Plant Breeding Research (MPI), Köln, Germany.
Bio Protoc. 2020 Sep 5;10(17):e3739. doi: 10.21769/BioProtoc.3739.
Genetic transformation is crucial for both investigating gene functions and for engineering of crops to introduce new traits. Rice ( L.) is an important model in plant research, since it is the staple food for more than half of the world's population. As a result, numerous transformation methods have been developed for both and rice. Since breeders continuously develop new rice varieties, transformation protocols have to be adapted for each new variety. Here we provide an optimized transformation protocol with detailed tips and tricks for a new African variety Komboka using immature embryos. In Komboka, we obtained an apparent transformation rate of up to 48% for GUS/GFP reporter gene constructs using this optimized protocol. This protocol is also applicable for use with other elite rice varieties.
遗传转化对于研究基因功能以及通过工程手段培育具有新性状的作物都至关重要。水稻(Oryza sativa L.)是植物研究中的重要模式植物,因为它是世界上一半以上人口的主食。因此,已经为粳稻和籼稻开发了多种转化方法。由于育种者不断培育新的水稻品种,转化方案必须针对每个新品种进行调整。在此,我们提供了一种优化的转化方案,并针对一种新的非洲品种孔博卡(Komboka)使用未成熟胚给出了详细的提示和技巧。在孔博卡品种中,使用这种优化方案,我们获得了GUS/GFP报告基因构建体高达48%的明显转化率。该方案也适用于其他优良粳稻品种。