Regeneration of Plants from Somatic Cell Cultures: Applications for in vitro Genetic Manipulation

Abstract

Plant regeneration from in vitro cultures of maize was first reported in 1975 by Green and Phillips. Steady improvements have been made since then in the culture systems and in applying them in genetic studies. Protoplasts have been regenerated into fertile plants by several groups (Prioli and Sondahl 1989; Shillito et al. 1989; Morocz et al. 1990). Selection experiments have yielded disease- and herbicide-resistant plants (Gengenbach et al. 1977; Shaner and Anderson 1985). Fertile transgenic plants are now reproducibly obtained in some laboratories through either direct DNA delivery into protoplasts or particle bombardment of intact, cultured cells. Successful applications of corn tissue culture technology have been largely limited to large research groups. The goal of this chapter is to present a simple how-to guide for successful establishment of maize somatic cell tissue cultures suitable for in vitro genetic manipulation. Attention is focused on model genotypes and also on minor technical details that are important, yet difficult to find elsewhere.