Development of an Effective In Vitro Regeneration System for Ukrainian Breeding Winter Rape Brassica napus L.

Abstract

An optimized regeneration method for commercial winter rape of the Bn1 line was proposed. Hypocotyl fragments of 6-day-old seedlings were used as explants. Regeneration occurred via organogenesis on an MS medium supplemented with 3 mg/L of 6-benzylaminopurine and 2 mg/L of 2-isopentyladenine. All obtained regenerated plants successfully formed roots on hormone-free media and were adapted to the soil conditions. The vernalization conditions were specified to promote budding and flowering at the level of 83.93 ± 5.33%. The PCR analysis using the ISSR 2 and ISSR 15 markers has shown that no somaclonal variations in Bn1 winter rape occur under the protocols based on the proposed methodology. Thus, the developed methodology is efficient and economically profitable for obtaining biotechnology-based winter rape plants.