In vitro pollen culture and the regeneration of Brassica campestris L. plants

Authors

• Yang-Dong Guo Agricultural Research Centre of Finland, Institute of Crop and Soil Science, FIN-31600 Jokioinen, Finland
• Seppo Pulli Agricultural Research Centre of Finland, Institute of Crop and Soil Science, FIN-31600 Jokioinen, Finland

DOI: https://doi.org/10.23986/afsci.72627

Abstract

Brassica campestris (Brassica rapa L. ssp. oleifera) is an important oilseed crop, particularly in Finland. Pollen culture techniques for haploid production have been developed, but B. campestris is relatively recalcitrant in pollen culture. Twenty eight genotypes of B. campestris were included in this study. The donor plants were grown in the greenhouse and transferred to the growth cabinet before bolting. Buds (2-4 mm long) were selected, macerated in B 5 medium, then NLN liquid culture medium was added. The microspores were incubated in the dark at 32°C for 72 h, then at 25°C for a further three weeks. Nineteen genotypes produced microspore-derived embryos. The highest yield was more than 300 embryos per 100 buds. Activated charcoal (150 mg/L) promoted embryogenesis, pollen development was faster and the embryo yield was higher. Plants were regenerated after transferring embryos to a solid B 5 medium. Colchicine solution was used to double the chromosome complements. About 100 regenerate plants have been obtained in our laboratory, and these haploids will be useful for the oilcrop breeding.