Studies On Anther Culture Of Non-heading Chinese Cabbage (Brassica Campestris Ssp.chinensis Makino)

Anthers from ten genotypes of non-heading Chinese cabbage were cultured in vitro. The effects of genotype and types of medium on callus formation and differentiation in anther culture were analyzed. Factors that plant regeneration concerned.such as different growth season, developmental stage of pollen, culture conditions, pretreatment and AgNO3 concentration during the culturing process, were investigated using hybrids of YHY-2 and YHY-4. Methods for transplanting regeneration plant from medium to soil were simultaneously studied. Meanwhile, the cytology of regerenation plant via anther culture in non-heading Chinese cabbage was observed.The results showed that:(1)The ability of different genotypes of non-heading Chinese cabbage to anther culture was discrepancy. All ten genotypes could formate callus on three induction medium, but the further differentiation was difficult. Four of ten genotypes could differenate green plant on the employed shoot regeneration medium, and the frequency differentiation was comparatively low, even the highest percentage of average differentiation was only 1.515%. The other six were failed to differentiate.(2)No differences were noted between the members of each pairs of reciprocal crosses.(3)The growth environmental conditions of donor plant affected on callus formation and differentiation in anther culture. Anthers produced in spring showed a higher response frequency than those in autumn.The development stage of microspore appeared to play an important role in anther culture. For non-heading Chinese cabbage, stage of uninuclear-binucleate is optimum to get higher rate of callus formation and differentiation. At thiscase, the morphological characteristics of bud were as follows: bud size was 1~2mm, petal/anther length ratio was 1/2, the color of anther was thin yellow-greenand translucent. The darkness was good for callus formation, which was in the interest of callus formation, but powerful light was benefical to the differentiation of green plant. Pollen pretreated with 0 - 4'C low temperature could abort the inducing stage that androgenesis needed,but most pretreated pollen still kept vigor when abortion occurrred. On the contrary, most unpretreated pollen degenerated and died. The due frequency of symmetry nuclear fission appeared in the first mitosis of pollen after pretreated by 33C for 24 hours. And higher temperature pretreatment (33 C,24hr) could significantly increase the frequency of plant regeneration. Adding 6~10mg/L AgNO3 to medium could promote the frequency of callus formation and increase ratio of regenerated green plant and control the root differentiation of callus.(4)The regenerated plants showed polyploid in karyotype analysis,in which included diploid, tetraploid and aneuploid. It was also notable that survival rate of transplant could be increased when thizome of regenerated plant was immersed by OOmg/L NAA.