The Preliminary Study On Tissue Culture And Cell Suspension Culture Condition Of Androsace Longifolia

Androsace longifolia is a kind of perennial herbs, flowering in May. Though the plant is short and small and single flower is small, the color is pure and fresh. It is a rare early spring wild flowers in the northeast, which owns very high ornamental value. But at present, the research about it also very few, in view of it in the garden on the application prospect, it is necessary to explore the germplasm resource conservation and development of new varieties through the rapid propagation.The seeds with different disinfection treatment were taken as explants and cultured into plants. The plant regeneration system of Androsace longifolia can be established by the ways of the callus, leaf inducing adventtious buds, clustered bud multiplication.The primary results are as follows:1.The best seed disinfection method is using75%alcohol quickly to rinse30s, then flushing3times with sterile water, immersing with10%NaClO for12min. Pollution rate was5.0%, the average germination rate was66.7%.2. Inducing callus with leaves and hypocotyl of Androsace longifolia, and the results show that:the best medium for the hypocotyl callus induced was MS+0.3mg/L6-BA+0.1mg/L NAA, the induction rate was85%; The best medium for leaf callus induced was MS+1.0mg/L2,4-D+0.2mg/L6-BA, The best carbon source was source, and agar concentration is8.0g/L, induction rate was83.3%.3. The best successive transfer condition for callus was MS+0.5mg/L6-BA, which was in darkness and with3%sucrose, pH was5.4,and the browning rate was only2.8%.4. The study of inducing adventitious buds via leaf callus and hypocotyl callus show that:the best medium for inducing adventitious buds via leaf callus was MS+0.07mg/L TDZ+0.1mg/L KT, each callus average differentiation bud number was6.3. The suitable hormone combina-tion for hypocotyl callus to induce adventitious buds was0.5mg/L6-BA+0.2mg/L KT. The best rooting medium for adventitious buds was MS+Activated carbon1g/L+0.8mg/L NAA, the survival rate reached92.5%.5. Leaf directly induced adventitious bud process found:the upper, middle and underpart three parts of leaf, the underpart had the strongest differentiation capacity; the best medium for adventitious bud differentiation was MS+2mg/L6-BA+2mg/L TDZ. The upper and the middle parts did not show differentiation ability. The best medium for cluster bud multiplica-tion was MS (added1g/L Activated carbon)+0.12mg/L TDZ+0.01mg/L NAA, the highest multiplica-tion coefficient was11.9. The best rooting medium for cluster bud was MS+Activated carbon1g/L+1.0mg/L IBA, the survival rate reached70.6%. 6. The optimum condition for cell of Androsace longifolia suspension culture:MS+0.3mg/L6-B A with2%sugar, rotate speed for105rp/min, on the basis of which, the best initial inocula-tion quantity of cell is2.0g in100ml triangle bottle with50ml nutrient solution.