Research Of Embryogenic Cells Suspension Culture On Picea Mongolica

In this paper,the materials was the mature zygotic embryos of Picea mongolica, induced embryogenic callus and subculture, which established a good dispersion and a high dynamic suspension cell line. That provided an experimental basis for the protoplast culture, gene transfer and cell fusion study and so on of Picea mongolica.1. There were two types callus of Picea mongolica, embryogenic callus and non-embryogenic callus. Soft texture, glossy surface of the callus was embryogenic callus, and the remaining was non-embryogenic callus. The experiments callus induction rate was relatively high, most induction rate in more than 60%, in addition to no added hormones based on the cultivation of callus did not been produced. The highest rates of induction was 93.75% in the concentration ratio of 2mg/L6-BA and 1mg/L2,4-D. Embryogenic callus of all organizations in the maintenance and proliferation of the phenomenon when the browning, but not the same as the degree. Without any hormone, the callus browning rate was 100%; the minimum rate of callus browning was 16.67% in hormones of 1mg/L6-BA and 0.2mg/L2,4-D; overall the case of 2,4-D concentration in the 0.1mg / L and 0.2mg / L at the rate of callus browning small,6-BA and 2,4-D with the additional time than a single additional rate of callus browning small. The embryogenic calli to maintain a good when the same concentration of 2,4-D of 0.1mg / L and 0.2mg / L , and embryogenic calli to maintain the good with 6-BA and 2,4-D than a singlethe of additional surcharge at the time.2. The growth cycle of Picea mongolica embryogenic cell suspension culture was 12d, and the best culture conditions was that: DCR+0.1mg/L2,4-D+1.0mg/L6-BA+30g/L sucrose, pH6.0, inoculum 3.0%, shaker speed 110r/min, 25±1℃, dark under the conditions of culture shock.3. In the kinetics research process, the cell fresh weight and dry weight was the same trend, the delay period on the 0³d; logarithmic phase on 4⁸d; the first phase was beginning at the 9d. Cell viability of the OD value reaches the maximum when the first 4d, 0.74; then begin to decline, to minimize the first time 10d, 0.34; followed by pick-up. Taking into account the cell viability and cell number of the total, following the generation period was 6⁸d. In the growth cycle the pH has been reduced, the first range of 1⁹d decreased than between 10¹2d larger. NH⁴⁺consumption in the development of the course the fastest in the first 10d has been exhausted; total sugar consumption decreased in the 3⁸d fastest, NO₃^-and PO₄^3- trends and sucrose consumption of basically were same, but the total sugar consumption preand post than in NO₃^-and PO₄^3-consumption faster. NO₃^-reduction rate of 63.79%, NH⁴⁺ was 100%, sugar and PO₄^3-were 80% and 84.62%. The first phase beginning 9d due to the lack of cell growth of nutrition4. Cell viability and other parameters in the 0.05 and 0.01 tested levels were not related; and the rest of the parameters in the tested level of 0.01 were significantly related. The cell dry weight all assumes the inverse correlation with other various parameters between. In addition to the OD values of cell viability, the other parameters with cell dry weight in the correlation coefficient above -0.95. Not only the total sugar and other nutrients directly affected cell proliferation, the pH of the culture medium have a direct impacted on cell proliferation.