Isolated microspore culture is the most efficient technique among those used to induce microspore embryogenesis. In the particular case of Brassica napus, it is also the most widely used and optimized. In this chapter, we describe a protocol for microspore culture in B. napus which includes the steps necessary to isolate and culture microspores, to induce microspore-derived embryos, to produce doubled haploid plants from them, as well as to check for the developmental stage of the microspores isolated, their viability, and the ploidy level of regenerated plantlets.
Keywords: Androgenesis; Doubled haploid; Embryogenesis; Haploid; Microspores; Pollen; Rapeseed; Tissue culture.