Studies On The Transformation Of Wheat With Genes Of AGD2 And CTB

Fusarium head blight (FHB) causes serious losses to wheat production. Due to the shortage of natural FHB-resistant germplasms, progress in breeding FHB-resistant wheat varieties is slow. Improving wheat resistance to FHB by transgenic technology has become a hot topic. Type?diabetes is a human organ-specific autoimmune disease, mainly treated with insulin injection to control high level of blood glucose. P277 can be used as vaccines to type?diabetes since it is an autoantigen in autoimmune diabetes. Co-expressiion of cholera toxin B subunit with other partners can enhance their immunogenicity. Avian influenza caused by avian influenza virus is a type of poultry avian disease. M1, M2, NA and HA are the major surface antigens of avian influenza virus, and thus these antigens can be used to produce vaccines. However, medical vaccines in current use are expensie and have side effects. Therefore, transgenic plants as a green bioreactor can produce diabete's vaccines and bird flu vaccines using seeds as carriers. This strategy provides a new way for the treatment of Type?diabetes and avian influenza. Transgenic wheat expressing a FHB resistance-related gene AGD2 can provide materials for FHB resistance breeding.In this study, an AGD2 gene, and a gene containing p277 fused with CTB were transferred into wheat through biolistic bombardment. We had obtained trangenic wheat plants comprising those genes. Besides, we identified a number of wheat transgenic lines that carried surface antigen genes of avian influenza virus. The main results were as follows:(1) The AGD2 was succesfully transferred into wheat variety Z9023. PCR identification results showed that there were 6 plants of transgenic wheat plants in To generation, and the target gene was inherited stably to the T1 generation.(2) According to the plant codon usage, we optimized CTB and p277 sequences, including addition of the plant signal peptide, endoplasmic reticulum resident signal KDEL, His-tag labeling and restriction enzyme sites. We synthetized a 576 bp sequence and inserted the fusion genes into the monocot seed expression vectors. (3) The P277 peptide fused with CTB sequence was successfully transferred to wheat varieties Y158 and Y15. PCR identification results indicated that there were 10 plants of transgenic wheat in To generation. Among them 6 plants contained CTB-3x-p277 gene targeting to the protein body and 3 plants contained CTB-p277 gene targeting to endoplasmic reticulum in Y158, while 1 plants contained CTB-p277 gene targeting to protein body in Y15.(4) According to PCR identification, in the transgenic wheat plants with the avian influenza surface antigen genes, the target genes in T2 and T3 generation displayed a non-Mendelian segregation, suggesting a linkage inheritance of these genes,. By selection, we obtained 6 lines of genetically stable transgenic wheat in Z9023 that contained 4 target genes of M1, M2, NA and HA, as well as 5 lines in Y12 that contianed 3 target genes of M2, NA and HA.