|
Pinellia ternata (Pinellia ternata (Thunb.) Breit.) The Araceae Banxia species, medication history more than 2,000 years, is one of the most commonly used one of the traditional Chinese medicine. The paper through the artificial division growth period and tissue culture process optimization, standardization and standardization of operational processes. And the shortcomings of a lot of manpower and resources for traditional organizational culture, design and assembly intermittently submerged culture reactor, and clover Banxia for initial culture experiments, it is desirable to provide a new way for of Pinellia ternata large-scale automation production. 1 Standard System Tissue Culture: the Pinellia ternata tissue culture operation standard processes as follows: petiole processing 20 s by 75% alcohol, mercuric chloride treatment 8 min, inoculated into MS, 6-BA1.0 mg / L NAA0.05 mg / L 3% sucrose agar 5.0 g / L of culture medium to induce callus and multiple shoots, let it grow to the petiole elongation cutting a single bud calls to MS, 6-BA1.0 mg / of L NAA0.02 mg / L 3% sucrose, 5.0 g of agar / L of culture medium for proliferation. During the planting seasons, take the blade mature buds inoculated to MS IBA0.03 mg / L NAA 0.01 mg / L sucrose, 3% agar 5.0 g / L AC 0.3 g / L of culture medium to induce rooting 30 d hardening transplanting. Should not be planted, blade proliferation of multiple shoots inoculated to, MS IBA0.03 mg / L NAA 0.01 mg / L sucrose 5% agar 5.0 g / L AC 0.3 g / L medium, rooted in a bottle within inverted seedlings in vitro tubers. In vitro tubers long-term storage, planted wash, can be planted in the bottle. Intermittent immersion training reactor design and assembly and develop Banxia the research: Design a batch the submerged culture reactor and assembly, commissioning and stability. Subsequent use of the reactor Pinellia propagation. On the one hand, biomass, proliferation coefficient, the plantlets state, guard cells and chloroplasts number contrast, research the different explants growth in the reactor. The experimental multiple shoots and petioles as explants each reactor number of plantlets were 1957 and 1382, the multiplication coefficient of 39.15 and 27.65. And petiole explant tissue culture is more robust, the explant types suitable reactor. On the other hand petiole explants, the reactor intermittently submerged culture and traditional solid culture and liquid shake flask culture plantlets were compared. Intermittently submerged reactor multiplication coefficient of 24.73, 14.75 solid culture, liquid shake flask cultures 12.26; the plantlets form solid culture seedlings is the most robust; the chloroplast number solid culture and culture undifferentiated reactor, better than liquid shake flask cultures. This shows that relative to traditional solid culture, the reactor culture Pinellia has the advantages of high multiplication coefficient, a single vaccination, and the medium was replaced with convenient, high degree of automation. Intermittently submerged culture the reactor fully for Pinellia ternata tissue culture production.
|