Agrobacterium-mediated transformation of wheat (Triticum aestivum) using chitinase and glucanase genes

Abstract

Production of transgenic wheat (Triticum Aestivum) was studied by chitinase and glucanase genes by Agrobacterium-mediated transformation and also pBI121 plasmid containing neomycin phospho transferasa selectable marker gene under control of Nos promoter. In addition, chitinase and glucanase 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 Agrobacterium-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, chitinase, glucanase and neomycin phospho transferase genes in their genome in comparison with control plants.

Keywords

• agrobacterium
• chitinase
• glucanase
• transgenic wheat