Nishimura Asuka, Aichi Ikuko, Matsuoka Makoto
Honda Research Institute Japan Co. Ltd, 2-1-4 Kazusa-Kamatari, Kisarazu, Chiba 292-0818, Japan.
Nat Protoc. 2006;1(6):2796-802. doi: 10.1038/nprot.2006.469.
Agrobacterium-mediated transformation of rice is an important method that has been widely adopted by many laboratories. However, because current approaches rely on culture systems, routine protocols have been established only in japonica rice, especially those varieties with higher regeneration potential. Some very efficient methods have been developed for japonica varieties that enable high-throughput functional analysis in rice; however, many elite japonica, and most indica, varieties are difficult to regenerate, leading to low transformation efficiencies. Much effort has been devoted to improving transformation efficiency for all rice genotypes. Here, we describe an Agrobacterium-mediated rice transformation method that is applicable to easily cultured varieties in addition to elite japonica varieties that are more difficult to culture. Using this method, transgenic rice plants can be obtained in about 2-3 months with a transformation frequency of 30-50%, both in easily cultured varieties and recalcitrant elite japonica rice.
农杆菌介导的水稻转化是一种重要方法，已被许多实验室广泛采用。然而，由于目前的方法依赖于培养系统，常规方案仅在粳稻中建立，特别是那些具有较高再生潜力的品种。已经为粳稻品种开发了一些非常有效的方法，能够在水稻中进行高通量功能分析；然而，许多优良粳稻品种以及大多数籼稻品种难以再生，导致转化效率较低。人们已付出很多努力来提高所有水稻基因型的转化效率。在此，我们描述一种农杆菌介导的水稻转化方法，该方法除适用于更难培养的优良粳稻品种外，还适用于易于培养的品种。使用这种方法，无论是易于培养的品种还是难培养的优良粳稻品种，都可以在大约2至3个月内获得转基因水稻植株，转化频率为30%至50%。
