One of the major limitations of maize transformation is the isolation of a large number of immature embryos using the time-consuming manual extraction method. In this article, we describe a novel bulk embryo extraction method for fast isolation of a large number of embryos suitable for both biolistic- and Agrobacterium-mediated transformation. Optimal gene delivery and tissue culture conditions are also described for achieving high efficiency in Agrobacterium-mediated maize transformation using phosphomannose isomerase (PMI) as a selectable marker.
Keywords: Agrobacterium-mediated transformation; Bulk embryo isolation; Elite inbred line; Immature embryos; Maize transformation; Mannose selection; Phosphomannose isomerase (PMI) gene; Transgenic event.