| The 2a seedlings of Fraxinus mandschurica were treated by cutting(treated or untreated),spraying hormones(treated or untreated)and covering with sawdust(treated or untreated).In order to explore the influence of different treatments on the seedlings and optimize the culture condition of tissue,the apical and lateral buds that germinated from the treated shoots were cultured in the same medium with different hormones combinations.The results showed that:(1)the branches number,length and diameter of coppice shoot that came from young seedlings that treated by cutting and hormones,were better than other treatments.Under this treatment,each of seedlings will produce six to seven coppice shoots and about twenty stem explants for tissue culturing during that year.(2)The average germinate rate of explants were higher than the seedlings without treated.Under the WPM medium with 1.0mg/L BA,the sprouting rate of the explants was onetime higher than control group.The sprouted buds amount and length was also significantly higher than control group.(3)Based on Membership Functions sorting,the tissue culture seedlings from four different medium were evaluated,the WPM medium with 2%sugar and 1.0mg/L ZT were best for bud to bud proliferation.The multiplication coefficient was 9.61,which is significant higher than any other treatments.(4)IAA/ABA indicate that the coppice shoot produced in July and Septmber are better rejuvenated than coppice shoot produced in June.(5)Results of Realtime PCR shows that,the relative expression level of ARR15 was down expressed-4.15 folds,and the relative expression fo ABP,AOS and WOX4 was up expressed 3.35,2.36 and 3.71folds.The results proved the fact that the sprouting shoot rejuvenation treatment obtained good effects. |
