| Objective:. Using seeds of B.R. as material,to establish a new tissue culture propagation system and seeds storage system of B.R.,then to compare the total PHenolic content and antioxidant capacity of them.Methods:(1) To determinating the seeds Viability by TTC staining method,study the optimum conditions of the TTC staining method.(2) The seeds of B.R.were drying different times,then moisture content of them were measured, seeds with different moisture content were storing at room temperature,4℃for different times or storing in liquid nitrogen, then to determinating the seeds Viability by TTC staining method.(3) The seeds with different viability were sowing to seeds germination medium, then study the relationship of seed viability and germination rate.(4) To study the different combinations and concentrations of hormones on seeds germination, plant growing, callus induction, propagation and differentiation,and of this basic to research the hardening process of B.R.(5) To compare the total PHenolic content and antioxidant capacity of B.R.which is propagation by tissuse culture method and traditional method.(6) Using Clear diPHenyl bitter acyl (DPPH) free radicals, iron reduction/antioxidant capacity (FRAP) assay to compare the antioxidant activity of B.R.with different propagation methods.Results:(1) The best testing conditions for TTC staining method is at the temperature of40℃with the concentration of TTC is0.5%, and staining for24hour.(2) The moisture content is the key factor of B.R. seeds storing,the seeds can storing for half a year at room temperature with the viability above90%and storing for one year at4 ℃with the viability more than89%while the moisture content is less than10%. The seeds with high moisture content storing in liquid nitrogen will lose viability while don’t lose viability with the moisture content less than10%.(3) The higher the viability of B.R. seeds,the higher seed germination rate,there are a certain linear relationship between them.(4) We studyed the tissue culture condition of B.R., fing that the best germination and proliferation medium are MS+1.0mg/L NAA+1.0mg/LKT, the seeds growth in this medium for three months can have a height of7.656±1.173cm, MS+0.1mg/L NAA+0.1mg/L TDZ+1.5mg/L2,4-D is the best medium for callus induction and propagation. The seedlings of B.R. in a suitable matrix control more than70%humidity and temperature is between22℃-27℃can has a survival rate of80%or more.(5)The total PHenolic content of the B.R. which were propagation by tissue culture method is0.401±0.002%while propagation by traditional method is0.394±0.002%,their Clear diPHenyl bitter acyl (DPPH) free radicals, iron reduction/antioxidant capacity (FRAP) ability is no statistical difference.Conclusion:We established a new tissue culture propagation system and seeds storing system successfully,The total PHenolic content and antioxidant activity of B.R. which were propagation bying this system and tradition method were no statistical difference..Using this system to propagate B.R.can ensure the quality of the medicine. The system we estabished is suitable for industrial development... |
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