The Biological Reactor Research Of Salt Tolerance Safflower Transformed With BFGF And H~+-ATPase

The plant bioreactor used plant cell suspension cultures or whole plants for the factory mass production of a protein with important functions, such as human or animal vaccines, antibodies t o essential amino acids, which had important medicinal value, or can be used as food additives, i ndustrial raw materials of plant secondary metabolites. It also referred as the whole plants of tra nsgenic plant mass production of a variety of high-value bio-products.Safflower (Carthamus tinctorius L.), a member of the family Compositae, is a branching, thistle-like herbaceous annual or winter annual plant. The tubular flower has analgesic efficacy, which is used to treat uterine hyperemia, cardiovascular, thrombosis. With the rapid development of the safflower industry, people have focued on safflower genetic transformation from the traditional production plant safflower. Gene transfer techniques are currently used for genetic modification of agronomically important characters engineering herbicide-resistant or pests and diseases-resistant cultivars.The purpose of this research was to improve the salt-tolerance of safflower by transfering bFG and V-H+-ATPase subunits B(ScVHA-B) from Suaeda corniculata.1. Safflower (Carthamus tinctorius L.) in "XINJIANG-TACHENG" was used in this study. The bFGF and ScVHA-B genes were transferred into safflower byAgrobacterium-mediated methods.2. By finding the optional hormonal conditions, using different concentrations of NAA and6-BA, GA3and TDZ,6-BA and TDZ effect the hormone safflower cotyledon callus, regeneration bud induction and expression, the experimental results showed that in the presence of sucrose (3%),6-BA (1.Omg-L-1) seed germination medium, germination rates of up to89%; containing NAA (1.0mgL-1),6-BA (1.0mg.L-1)adventitious bud differentiation rate of43%;in the presence of6-BA (2.0mg·L-1) and NAA(0.5mg-L-1), ABTC(1000mg-L-1), KNO3(3800mg·L-1) rooting culturebased on the rooting rate of15%.3. Safflower had tolerance to hygromycin and grass ammonium phosphate sensitivity.The experiments showed that the the hyg concentration of5mg.L-1, and concentrations of Basta are greater than3mg.L-1.4. Determine the most suitable concentration of buds induced differentiation, shoot elongation, and rooting culture bacteriostatic antibiotics.Bud induction of200mg.L-1carbencillin differentiation medium,100mg.L-1Cef and regeneration shoot elongation rooting medium using100mg.L-1carbencillin and250mg.L-1Cef.5. Detect genetically modified safflower with PCR, the PCR results showed that the target gene had been transferred to the safflower genome. Southern blot analysis showed that the ScVHA-B gene had been successfully integrated into the safflower genome.