Construction Of Bt Cry3Aa Gene Expression Vector And Agrobacterium Tumefaciens-mediated Genetic Transformation Of Sweet Potato Stems

Sweet potato(Ipomoea batatas Lam.) is an important crop and industrial crops in the world, and both function of anticancer, health care.It is widely cultivated throughout the world. Sweet potato pest species diversity, endanger the exact same way every year due to the loss of production of sweet potato pests cause to bring the country can not be estimated. The lack of conventional sweet potato varieties in the field of natural resistance to pests, it is difficult to rely on ordinary breeding methods to solve these problems. At present, more than a means of controlling pests of sweet potato rely on chemical pesticides, increased production costs and environmental pollution, and because of the consumption of residual chemicals causing a safety hazard. Hence, the urgent need for the emergence of a new means of transgenic insect-resistant sweet potato. Bt gene is considered a specific insecticidal activity against pests, and for people, animals and other non-target organisms harmless insect-resistant genes, because of their excellent properties, now widely used in the world. Bt cry3 Aa gene is a kind of Coleoptera pests gene-specific insecticidal activity, the use of the gene into sweet potato, sweet potato cultivation and transgenic potato can coleopteran pests have some weevil resistant to reach insect the purpose of increasing the yield of sweet potato.The main results of this study are:1. In the upstream end of the Bt cry3 Aa 5’ gene sequence introduced KpnI, BamHI restriction sites and PTD joints, 3’ end add SalI restriction endonuclease, and the use of codon preference for sweet potato transformation after cry3 Aa the company synthesis.2. synthesized gene was ligated, transformed and other steps to eventually obtain binary vector pCAMBIA1301-cry3 Aa and pCAMBIA1301-PTD-cry3 Aa.3. Plant tissue culture without the exploratory phase of Agrobacterium-mediated genetic transformation system of sweet potato, the initial establishment of Guangshu87 wide transformation strategy sweet potato varieties in bacteria OD600=0.4, infection time10 min, chances of AS concentration of between 25-50 mg/L, Carb concentration of 200mg/L, to obtain the maximum positive strains of sweet potato.4. After the conversion to the new leaves of plants grown for PCR testing. The results showed that the plants turn cry3 Aa strains detected 7, a total of 2 positive strains;the plants in the test strains turn PTD-cry3 Aa 11, a total of 3 positive strains.