Isolated microspore culture of oat ( Avena sativa L.) for the production of doubled haploids: effect of pre-culture and post-culture conditions

摘要: The production of doubled haploid (DH) plants from microspores is an important technique used in plant breeding programs and basic research. Although haploidy efficiencies wheat barley are sufficient for purposes, oat (Avena sativa L.) considered recalcitrant. objective this project was to develop a protocol the microspore-derived embryos further these into fertile DH plants. A number experiments were conducted evaluating factors influencing microspore embryogenesis, i.e. donor conditions, pretreatments, media composition, culture conditions. initial studies yielded little response, it not until high densities (106 microspores/mL greater) that embryogenesis achieved. Depending on treatment, yields over 5,000 embryos/106 obtained line 2000QiON43. includes: 0.3 M mannitol pretreatment tillers 7 days, W14 basal medium with pH 6.5–7.5, density 106 microspores/mL, continuous incubation at 28 °C incubation. resulting observed after 28 days plated onto solidified 0.8 or 1.0 g/L activated charcoal. colchicine treatment 0.2 % 4 h resulted conversion 80 % DH. This successful green minimal albinism. seed produced planted evaluation field nursery.