An Efficient Agrobacterium-Mediated Genetic Transformation Method for a Monocot Model Plant, Brachypodium distachyon

Abstract

Model plants are necessary and important for advances in various research areas of plant biology. Compared with <italic>Arabidopsis thaliana</italic>, a dicot model plant that is widely used thus far, monocot model plants are not popular in using plant biology research. In this regard, <italic>Brachypodium distachyon</italic> is suggested as a new and alternative monocot model plant for economically valuable temperate grasses and cereals such as wheat and barley, and also for bioenergy crops such as switchgrass and miscanthus. One of the major constraints in performing plant research would be the difficulty of genetic transformation and its low frequency. Here, we provide a detailed and improved method for <italic>Agrobacterium</italic>-mediated genetic transformation of <italic>B. distachyon</italic>, based on our experimental experience. With hygromycin resistance as a selectable marker, we could obtain approximately 40%−60% of the transformation efficiency using this method, in which the process takes approximately 26 weeks to obtain T₁ seeds starting from co-cultivation of immature embryos with <italic>Agrobacterium</italic> cells. Here, we also describe about important factors possibly contributing to the genetic transformation of <italic>B. distachyon</italic>.