Whiskers-mediated maize transformation

Abstract

Summary: There has been rapid progress in recent years in extending gene transfer capabilities to include plant species that fall outside the normal host range of Agrobacterium. Methods that allow direct DNA delivery into plant cells have contributed significantly to this expanded capability. Whiskers treatment is one means of delivering macromolecules, including DNA, to plant cells. Using relatively simple equipment and inexpensive materials, whiskers-mediated transformation of maize is possible. A critical prerequisite, however, is the establishment and maintenance of embryogenic tissue cultures as a source of totipotent, transformation-competent cells. Within hours of agitation in the presence of silicon carbide whiskers and DNA, embryogenic maize tissue cultures display transient gene expression, providing evidence for DNA uptake. Using appropriate selectable marker genes, following in vitro selection on inhibitory levels of a corresponding selection agent, stably transgenic tissue cultures can be generated from which fertile plants can be recovered. The timeline from whiskers treatment of embryogenic maize tissue cultures to fertile seed recovery is approximately 9 months, which is competitive with other methods of maize transformation. © 2009 Humana Press.