Exogenous DNA uptake via cellular permeabilization and expression of foreign gene in wheat zygotic embryos

Abstract

Exogenous DNA uptake and transient expression of the GUS reporter gene was studied in wheat (Triticum aestivum L.) zygotic embryos employing a simple procedure of cellular permeabilization by membrane interactive agents like saponin and toluene. Uptake of expression vectors with different promoters was detected in mature and immature embryos. The frequency of GUS expression was detected to be higher in mature embryos than immature embryos probably due to a certain degree of disorganization of the plasmalemma during the dehydrated state of the embryos. Of the various permeabilizing agents employed, saponin and toluene were detected as potential membrane permeabilizers without adversely affecting embryo viability. Variations in GUS gene expression were not significant among different genotypes of T. aestivum CPAN1676, PBW343 and HD2329, thus indicative of the relative genotype independence of this process. The results support the proposed idea of direct DNA uptake as an alternate and simple method for inserting foreign DNA into plant cells and its use for transformation of higher plants.