Improved somatic embryogenesis in wheat by partial simulation of the in-ovulo oxygen, growth-regulator and desiccation environments

Abstract

The effects of O2, growth-regulators and desiccation on callus growth and somatic embryo (embryoid) development were investigated in cultures of immature embryos of two lines of Triticum aestivum L. Callus and embryoid formation were induced on media that contained N(6)-furfurylamin-opurine (kinetin) and either 2,4-dichlorophenoxyacetic acid or 3,6-dichloro-o-anisic acid, either with or without abscisic acid (ABA). Cultures containing differentiated embryoids were then exposed to high concentrations of both ABA and indole-3-acetic acid, after which samples were desiccated to approx. 10% tissue moisture. Incubating cultures in 3.2 mmol·l(-1) O2 (approx. 9%, low-O2) increased embryoid formation sixfold in one wheat line and nearly threefold in another. In the former line low-O2 caused the formation of mostly embryogenic callus. Low-O2 also decreased precocious germination of immature embryos, decreased callus growth, and improved development and viability of the resultant embryoids. Including 1.9 μmol·l(-1) ABA in the callus-induction medium reduced germination of immature embryos and reduced the incidence of embryoids with visible abnormalities. Despite the improved morphology, significantly fewer of the embryoids produced on ABA-containing medium germinated. Desiccation significantly enhanced germination of these embryoids as well as those produced on ABA-free medium.