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 pBI۱۲۱ plasmid. Among the durations of sonication the highest percentage of GUS positive immature embryos (۵۴.۵۸ ± ۱.۱۴%) and transformation (۰.۷۸ ± ۰.۰۷%) was observed in ۱۰-seconds of sonication. Among the Agrobacterium strains, the highest GUS expression was ۶۲.۵۰ ± ۱.۵۵% and ۰.۳۹ ± ۰.۰۴% with LBA۴۴۰۴ strain. Between types of inoculation medium, the highest GUS positive immature embryos and transformation (۱.۸۶ ± ۰.۱۴% and ۰.۷۵ ± ۰.۰۴%, respectively) was observed in using the IM inoculation medium. Between concentrations of acetosyringone, the highest transformation was ۰.۷۵ ± ۰.۰۴% obtained at ۲۰۰ µM acetosyringone. Also, the studying of simultaneous effects showed that the highest transformation efficiency (۱.۵۶ ± ۰.۰۶%) obtained from immature embryos inoculated with LBA۴۴۰۴ strain followed by ۱۰-seconds sonication, immature embryos inoculated with Agrobacterium in IM inoculation medium after ۱۰-seconds sonication and immature embryos inoculated with Agrobacterium in inoculation medium containing ۲۰۰ µM acetosyringone after ۱۰-seconds of sonication.

Keywords:

Acetosyringone , Agrobacterium , Inoculation medium , Sonication , Transformation