In Vitro Fertilization System Using Wheat Gametes by Electric Fusion.

In vitro fertilization (IVF) systems using isolated gametes have been used to dissect post-fertilization events in angiosperms, as female plant gametophytes are deeply embedded within the ovaries. In addition, hybrid and polyploid zygotes can be produced by using IVF systems. Complete IVF systems of maize and rice, two out of three major energy-providing crops, have been established in order to acquire detailed knowledge of mechanisms of fertilization and early embryogenesis. Following in the footsteps of previous success, a wheat IVF system was developed to introduce the advantages of this technology to wheat research. Fusion of gametes was performed via a modified electrofusion method, and the zygote formed a cell wall and two nucleoli. The zygotes divided into symmetric two-celled embryos, globular-like embryos and multicellular club-shaped embryos which are mostly consistent with those in the embryos in planta. IVF-produced club-shaped embryos developed into compact embryonic calli and subsequently regenerated into fertile plants. In this chapter, we provide a detailed description of wheat IVF system that might become an important technique for generating new genotypes of wheat and/or new hybrids as well as for investigating fertilization-induced events in wheat.