Optimization of genetic transformation protocol mediated by biolistic method in some elite genotypes of wheat (Triticum aestivum L.)

Abstract

We report here an efficient genotype-independent genetic transformation system in wheat. Highly regenerable embryogenic calli obtained from mature seeds were employed as the target tissue for the genetic transformation of three bread wheat varieties viz C306, HDR77 and PBW343 representing diverse genetic background. The plasmid pDM803 containing GUS and BAR genes driven by rice Act1 and maize Ubiqutin promoter, respectively was transferred into a month-old calli employing particle delivery system. The bombarded calli were transferred to medium supplemented with phosphinothricin at 4 mg L^-1 for the selection of the transformed calli. The transgenic calli were confirmed for the expression of GUS by histochemical analysis of β-glucuronidase. Transformation efficiency of the genotypes was calculated based on the number of calli bombarded and the number of plants that were resistant to Basta. Among the three genotypes studied, C306 had a higher efficiency of 0.56% followed by HDR77 with 0.5% and PBW343 with 0.22%. The transformation system developed in this study may facilitate studies on functional genomics and crop improvement via transgenic development.

Keywords: Wheat, biolistic, transgenics, bar, phosphinothricin, transformation efficiency

African Journal of Biotechnology Vol. 12(6), pp. 531-538