Genetic engineering of wheat for enhancement to resistant to fungal disease

Production of transgenic wheat (Triticum aestivum) was studied by Agrobacteriummediated transformation and gen gun method and also pBI121 plasmid containing neomycin phospho transferase selectable marker gene under control of Nos promoter and chitinase and glucanase as target genes. In addition, chitinase and glucanase genes were individually expressed under control of CaMV35S promoter. In gen gun method, Immature embryo explants from ( arta, mogan, line A, sisun, gascogen ) cultivars , excised from seeds and were placed on callus induction medium and they produced embryonic calluses. These were bombarded with gold nanoparticles coated by plasmids containing the transgenes. Thus they were transferred to selective callus induction medium containing 50 mg/l Kanamycin, and following to regeneration medium containing 25 mg/l Kanamycin to produce shoot and root. Maximum percent of transformation by gen gun method was 4.8% for line A and then 0.3% for mogan. There was not any transgenic plant in other cultivars and transformation percent was 0 Agrobacterium-mediated transformation was studied by strains (AGL1, EHA101, C58, LBA4404) containing recombinant pBI121 plasmid. Immature embryo explants, excised from seeds, then were co-cultivated with bacterial suspension containing the recombinant plasmid and they were placed to callus induction medium supplemented with 50 mg/l kanamycin. Embryonic Calli were selected and they were transferred to regeneration medium with 25 mg/l kanamycin in order to producing shoots and roots. Maximum percent of transformation by Agrobacterium-mediated transformation was 0.31% for transformed arta with C58 strain and then 0.14% for transformed arta with LBA4404. There was not any transgenic plant in other cultivars or strains and transformation percent was 0% the number of produced transgenic plants in this study were 18, include 3 plants from Agrobacterium-mediated transformation and 15 plants from gen gun method. PCR and Dot blot analysis showed that the putative transgenic plants consist at least one copy of either, chitinase, glucanase and neomycin phospho transferase genes in their genome in comparison with control plant. Integration of intact gene into the genome of putative transgenics was further confirmed by southern blot analysis.The results of PCR in the T1 transgenic plants confirmed integration of transgen. In study the effect of transgenic wheat plants and untransgenic on the fungus Fusarium graminearum it was revealed that transgenic and untransgenic cotton extracts had the most and least effects on fungus growth Fusarium graminearum.