Agrobacterium-mediated transformation of wheat (Triticum aestivum) using chitinase and glucanase genes
2010
Negin Mohammadi zadeh | Masoud tohidfar | Motahareh Mohsenpooor
Production of transgenic wheat (<em>Triticum Aestivum</em>) was studied by <em>chitinase</em> and <em>glucanase</em> genes by<em> Agrobacterium</em>-mediated transformation and also pBI121 plasmid containing <em>neomycin phospho transferasa</em> selectable marker gene under control of Nos promoter. In addition, <em>chitinase</em> and <em>glucanase</em> genes were individually expressed under control of CaMV35S promoter. 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 calluses, 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 <em>Agrobacterium</em>-mediated transformation was 0.31% for transformed ARTA with C58 strain and then 0.074% for transformed ARTA with LBA4404 . There was not any transgenic plant in other cultivars or strains and transformation percent was 0%. PCR and Dot blot analysis showed that the putative transgenic plants consist at least one copy of either, <em>chitinase</em>, <em>glucanase </em>and <em>neomycin phospho transferase</em> genes in their genome in comparison with control plants.
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