Uptake of Exogenous DNA by Brassica napus Dry Seed Embryos via Cellular Permeabilization and Transient Expression of NPTII Activity

Exogenous DNA Uptake via Cellular Permeabilization and Expression of Foreign Gene in Wheat Zygotic Embryos.

“…1). This indicates that this method is almost independent of the genotypic influence (see also Parihar et al, 1994) probably due to the underlying biochemical/biophysical principle involved during cellular permeabilization. It also offers a relative advantage over the other techniques, which have limited utility for recalcitrant genotypes (Tyagi et al, 1989;DeBlock and DeBrouwer, 1991;Maheshwari et al, 1995).…”

“…The effect on germination could be the principal reason for the ineffectiveness of both DMSO and Triton X-100 in the wheat system, which was not so in the case of Brassica spp. (Parihar et al, 1994). Interestingly, the embryos treated with toluene, a widely known inducer of transient `pore' formation of specific sizes (Lerner et al, 1978), exhibited significantly higher frequency of GUS expression (78%) at 1:20 v/v dilution with permeabilization buffer (Fig.…”

“…the molecular sieving-properties of the cell wall in conjunction with the impermeant lipid bilayer of plasmalemma offer resistance to the entry of exogenous macromolecules. Nevertheless, by challenging such barriers through effective membrane permeabilizers such as DMSO, toluene and saponin, the permeation of exogenous DNA molecules in the target plant tissue has been achieved (TOpfer et al, 1989;Parihar et al, 1994;Yoo and Jung 1995;Luan et al, 1996). membrane appear to play a vital role in DNA imbibition.…”