Factors influencing Agrobacterium-mediated transformation of CPT antisense gene in soybean

Cholinephosphotransferase (CPT) is an important enzyme reversibly catalyzing a reaction, in which diacylglycerol (DAG) is converted to phosphatidycholine (PtdCho). Monounsaturated-PtdCho is in turn the substrate for polyunsaturated-PtdCho. Reversal of the CPT reaction with polyunsaturated-PtdCho results in increase of polyunsaturated fatty acids in soybean seeds. DAG is mainly the substrate for triacylglycerol (TAG) synthesis. Therefore, the down regulation of CPT catalysis is expected to increase monounsaturated fatty acids content and improve the quality of soybean oil. This purpose may be achieved by the insertion of a CPT antisense gene.; To transfer the CPT antisense gene into soybean, plasmid pCB-GF1 was constructed previously. Plasmid pCB-GF1, 6293bp, harboring a CPT antisense gene and a marker gene, bialaphos resistance gene (Bar), located between the T-DNA borders. This plasmid was designed for Agrobacterium-mediated transformation, which is a biological regulation process.; After the plasmids were transferred into Agrobacterium tumefaciens EHA105, six experiments were constructed, and the factors including bacterium inoculum density, AS concentration, light intensity, photoperiod, transferred genes, auxins, and antibiotics were investigated using a standard protocol for Agrobacterium-mediated soybean transformation of cotyledonary nodes. (Abstract shortened by UMI.)...