Enhancement of Agrobacterium-mediated transformation efficiency in immature embryo of Triticum aestivum, cv. Arya

An efficient Agrobacterium-mediate transformation method was developed by employing different duration of sonication, Agrobacterium strains, type of inoculation medium and concentrations of acetosyringone in Arya cultivar of wheat. Immature embryos were used as an explant for inoculation with Agrobacterium tumefaciens harboring the recombinant pBI121 plasmid. Among the durations of sonication the highest percentage of GUS positive immature embryos (54.58 ± 1.14%) and transformation (0.78 ± 0.07%) was observed in 10-seconds of sonication. Among the Agrobacterium strains, the highest GUS expression was 62.50 ± 1.55% and 0.39 ± 0.04% with LBA4404 strain. Between types of inoculation medium, the highest GUS positive immature embryos and transformation (1.86 ± 0.14% and 0.75 ± 0.04%, respectively) was observed in using the IM inoculation medium. Between concentrations of acetosyringone, the highest transformation was 0.75 ± 0.04% obtained at 200 µM acetosyringone. Also, the studying of simultaneous effects showed that the highest transformation efficiency (1.56 ± 0.06%) obtained from immature embryos inoculated with LBA4404 strain followed by 10-seconds sonication, immature embryos inoculated with Agrobacterium in IM inoculation medium after 10-seconds sonication and immature embryos inoculated with Agrobacterium in inoculation medium containing 200 µM acetosyringone after 10-seconds of sonication.