Study On Culture And Propagation In Vitro Of Chinese Traditional Medicine Millettia Specisoa Champ

The seven aspects of culture and propagation in vitro of Chinese traditional medicine Millettia specisoa Champ were studied in this paper:(1) The test of explant sterilization;(2) The test of callus induction;(3) The test of callus differentiation;(4) The test of aseptic buds proliferation;(5) The test of aseptic seedlings rooting inside the bottle;(6) The test of aseptic seedlings rooting outside the bottle;(7) The test of aseptic rooting seedlings transplanting. The main results were as follows:1. The test effects of explant sterilizationThe best treatment of Millettia specisoa Champ seeds sterilization was0.1%HgCl220min+sterile water soak5h,the pollution rate was40%and seeds rooting rate was83.3%.The best treatment of Millettia specisoa Champ stem section explant sterilization was0.1%HgCl220min, the pollution rate was40%and bud induction rate was37.5%.2. The test effects of callus inductionThe best hormone combination of Millettia specisoa Champ callus induction was MS+2,4-D2.0mg/L+IBA0.2mg/L+TDZ0.03mg/L, the best effect of callus induction was tender petiole, the induction rate was100%and browning rate was0。3. The test effects of callus differentiationIn callus differentiation experiments, which was firstly induced by tender stems without bud in the induction medium group:MS+6-BA2.0mg/L+IBA0.2mg/L+TDZ0.03mg/L, and then transferd into the differentiation medium combination:MS+6-BA0.5mg/L+KT1.0mg/L+IAA0.3mg/L for90days, the effect of callus differentiation come to the best and the bud differentiation rate was10%.4. The test effects of aseptic buds proliferation(1) MS medium was more suitable than Miller medium and Improved White medium on aseptic buds proliferation of Millettia specisoa Champ, the proliferation times was3.60(30d)and5.33(60d) respectively. (2) Comparison of the different cytokinins of6-B A and KT and their concentration displayed that6-BA showed the best effect on buds proliferation. The best combination of Millettia specisoa Champ buds proliferation was MS+6-BA3.0mg/L+NAA0.2mg/L, the proliferation times was3.70(30d) and5.43(60d) respectively.(3) Comparison of the different auxins of NAA, IB A and IAA and their concentration displayed that in the combination of MS+IAA0.2mg/L+6-BA3.0mg/L, the Millettia specisoa Champ buds proliferation was as high as4.38(30d) and6.55(60d) respectively.(4) Comparison of the different cytokinins of CPPU+6-BA and TDZ+6-BA and their concentration displayed that the combination of MS+TDZ0.03mg/L+6-BA3.0mg/L+IAA0.2mg/L, the Millettia specisoa Champ buds proliferation was as high as6.00(30d) and9.65(60d) respectively.(5) Comparison of the different inoculation ways of aseptic buds displayed that the Single bud inoculation way was more suitable on aseptic buds proliferation of Millettia specisoa Champ, the proliferation times was5.34(30d)and10.63(60d) respectively.(6) Comparison of the different concentration of sucrose displayed that when the sucrose concentration was30g/L, the Millettia specisoa Champ buds proliferation times was as high as4.30(30d) and6.40(60d) respectively.(7) The adding of organic coconut water was benefits to the Millettia specisoa Champ buds proliferation, when the coconut water concentration was15%, the proliferation times as high as5.82(30d) and8.12(60d) respectively. The adding of potato juice and banana juice was not benefits to the Millettia specisoa Champ buds proliferation.5. The test effects of aseptic seedlings rootage inside the bottle(1) In single factor hormone comparison test,1/2MS+ABT2.0mg/L rooting rate was as high as27%.(2) In hormone combination double factor comparison test, the combination of1/2MS+IBA3.0mg/L+NAA1.0mg/L had the highest rooting rate was67.5%.6. The test of aseptic seedlings rooting outside the bottleThe best hormone combination of aseptic seedlings rooting outside the bottle was ABT500mg/L12h+transplanting to imported flowers matrix:pearlite matrix (2:1), the survival rate and rooting rate of30days, respectively was100%and81.3%. 7. The test of aseptic rooting seedlings transplantingThe best matrix combination of aseptic rooting seedlings transplanting was imported flowers matrix:pearlite matrix (2:1), transplanting of30days, the survival rate of septic rooting seedlings was94%.