Microspore Culture And Ploidy Identification In Brassica Napus

In recent years, with the continuous development of biotechnology, isolated microspore culture technology has increasingly been in the widespread application of breeding, which greatly accelerated the process of rapeseed breeding. In this study, the microspores were systematically studied with big seed line and small seed line in Brassica napus from the ordinary greenhouses, meanwhile microspore embryogenesis condition, plant ploidy methods detection were investigated, the results are as follows:1. Sampling time has more effect on microspore embryogenesis. The study is discovered that the best time to take bud is the primary flowering period, flourishing florescence is the better.2. Inflorescence length markedly affect rapeseed microspore embryogenesis, the cytological observation showed that when the bud length is 2.5-3.5mm,75% of microspore development was in the late uninucleate stage, and embryoids were more obtained.3. About isolated microspore culture, the investigation is that NLN liquid medium isolated microspore culture is better, the application of cytokinin 0.1mg/l6-BA can promote microspore embryogenesis, while the activated carbon had no significant effect on the embryos, only improve environment for the survival of small spores.4. Low temperature pretreatment affects production of microspore embryo rate. When the bud was pretreated at 4℃low temperature 6h, it would promote microspore embryogenesis capacity; But with the inflorescence low temperature pretreatment time processing, the rate of microspore embryo production had a downward trend.5. Microspore culture ploidy was rapidly identified with the method of morphological and anatomical characteristics, and the results is that the chromosome of Haploid plantlet n=19, and the normal diploid plantlet 2n=38; It is the first time identified the microspore plants and doubled plant ploidy using flow cytometry, the results show that the consequence of flow cytometry are consistent of morphological observation, chromosome count, it is a fast, simple and accurate method of detecting plant ploidy, providing the reference of the early identification of rapeseed microspore ploidy for genetic breeding.