High-frequency embryogenesis in Brassica campestris microspore culture

Microspore culture is a very important and useful tool in plant breeding for haploid production and has been developed for many years. Brassica campestris (Brassica rapa L. ssp. oleifera) is an important oilseed crop, but it is relatively recalcitrant in tissue cullum including microspore culture. The microspore culture in our laboratory is based on the Canadian protocol. Thirty genotypes of B. campestris were included in this study; twenty produced embryos. The highest yield was 5930 embryos per 100 buds from Canadian genotype Cv-2, this result was one of the best that had been reported in microspore culture in B. campestris. The buds measuring 2.0 mm to 3.9 mm in length responded best to produce embryos, the optimum timing for microspore culture was confirmed to be during the mid-late to very-late uninucleate stage. The buds could be removed from either the main raceme or lateral racemes. Activated charcoal (150 mg l) was added to the liquid NLN medium, it promoted embryogenesis significantly; embryo development was faster and the embryo yield was significantly higher than those cultures without activated charcoal. Tire donor plant condition was considered an important factor influencing embryogenesis; older donor plants (older than five weeks) and a cold treatment are recommended.