الفهرس 
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Abstract
We report new transfection methods for intact plant cells that uses laser beam to introduce foreign gene-coated gold nanoparticles into wheat mature embryos cells (Triticum aestivum) through the cell wall and cell membrane. This homemade setup secures the transformation of as many as 30 mature embryo derived calli in less than one hour using secondharmonic of Nd:YAG laser at 532nm(150mW) with two dimensional translation stages, a suitable computer program and a proper optical system. Seven days old calli were irradiated by a focused laser microbeam to puncture momentarily made self healing holes (~0.5 μm) in the cell wall and membrane to allow uptake of foreign genes coated gold nanoparticles. The plant expression vector PBI-121 harboring the green fluorescent protein gene (gfp) under the control of 35-S promoter and nos terminator and the neomycin phosphotransferase-II gene (npt-II) as a selectable marker for the herbicide kanamycin sulfate resistance. Treated wheat tissue samples were then cultured for several weeks on the selection medium. The resulted putative transgenic plantlets grew on kanamycin containing medium and transformation was assessed by confocal laser scanning microscopy (CLSM) and PCR. TEM examination indicated that Nd:YAG laser beam is less damaging for gene transfer than combined treatment ”gold nano-lasic method” .In conclusion: 1)Both novel approaches Nd:YAG laser beam micro puncture and gold nano – lasic succeeded in transforming wheat mature embryos. 2) Transformation efficiency (TE %) of Nd:YAG laser beam micro puncture was 5.5% and 4.49% for the cultivars sakha 93 and miser 1respectively. 3) TE% of gold nano lasic method was 1.46% and 2.33% for the cultivars sakha 93 and miser 1 respectively. 4) Nd:YAG laser beam micro puncture (532nm) was more safe in generating transgenic wheat than gold nano-lasic.