A high throughput Brassica napus microspore culture system: influence of percoll gradient separation and bud selection on embryogenesis

摘要: Microspore culture for the purpose of developing doubled haploid plants is routine numerous plant species; however, embryo yield still very low compared with total available microspore population. The ability to select and isolate highly embryogenic microspores would be desirable high in culture. To maximize efficiency canola culture, a combination bud size selection fractionation using Percoll gradient was followed. This approach has consistently given yields uniform development. Microspores isolated from buds 1.5 4.4 mm length Brassica napus genotypes Topas 4079, DH12075, Westar 0025 formed embryos at different frequencies. most range varied each cultivar: 4079 3.5–3.9 mm, DH12075 2.0–2.4 2.5–2.9 mm. When 2.0 2.4 were carefully layered on top discontinuous 10, 20 40%, subsequently spun through layers by centrifugation, bands containing populations developmental stage. middle layer contained late uninucleate early binucleate that embryogenic. In addition, relationship between size, stage microspores, concentration frequency cultivar studied. Optimization these factors required genotype evaluated.