Study On The Anther Culture Technological System Of Brassica Rapa Ssp. Pekinensis

Anthers from 9 genotypes of Chinese cabbage were cultured. The effects of genotype were analyzed in anther induction and differentiation culture. 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 F1D-1和F1D-3. Methods for transplanting regeneration plant from medium to soil were simultaneously studied. Meanwhile, the cytology of regeneration plant via anther culture in non-heading Chinese cabbage was observed.The results showed that:(1) The ability of different genotypes of Chinese cabbage to anther culture was discrepancy. In all nine genotypes, two genotypes couldn't form callus; two genotypes formed embryoid; five genotypes formed callus.(2) The growth environmental conditions of donor plant affected on embryoid formation 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 Chinese cabbage, stage of uninuclear-binucleate is optimum to get higher rate of embryoid formation and differentiation. At this case, the morphological characteristics of bud were as follows: bud size was 2~3mm, petal/anther length ratio was 1/2, the color of anther was thin yellow-green and 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. The treatment that buds were pretreated with 4℃low temperature during 3 days before at initial stage, could obtain better inducing result. The induction rate of 8% sucrose treatmeng was the best, high concentration sucrose could stimulate germinating microspores and increase induction rate. Adding 10mg/L AgNO3 to medium could promote the frequency of embryoid formation.(3) Embryoid which plated in differentiation medium became green after 3 days, and formed regenerated plantlets; Callus which plated in differentiation medium formed roots, and formed a little regenerated plantlets.(4) The regenerated plantlets from anther culture were easy to grow roots. The effect of inducing roots was the best in the medium inducing roots which was 1/2MS medium supplemented with NAA0.1mg/L, Agar0.8% and 3% Sucrose. Regeneration plants could be easy to adapt to natural environmental conditions by the method that regeneration plant first grew in condition of hydroponics, and then grew in condition of aggregate culture, which survival rate was 100%.(5) The regenerated plant karyotype analysis showed the existence of diploid, aneuploid and polyploid.