Immature Embryo-Derived of Two Bread Wheat (Triticum aestivum L.) Varieties Transformation Using Particle Bombardment Method

Abstract

Aims: Wheat cultivation is still difficult to control because it faces several biotic and abiotic stresses. The transfer of resistance in wheat plants to these stresses through traditional approaches remains limited. However, genetic transformation allows the acquisition of this tolerance while overcoming the difficulties of classical improvement. Methodology: In this context, immature embryos of two Moroccan bread wheat varieties (‘Massira’ and ‘Arréhane’) and the pBY520 plasmid containing the HVA1 gene for drought tolerance as well as the gene bar selection marker for resistance to the herbicide phosphinothricin were used in this study for bombardment protocol. Results: The percentage of calli bombarded with regenerated plantlets was 23.92 % for the variety ‘Massira’ and 5.26% for the variety ‘Arréhane’. The selection was carried out on ½ MS rooting medium lacking hormones and supplemented with phosphinothricin (3 mg l-1); 4.36% and 6% of plantlets of varieties ‘Massira’ and ‘Arréhane’ respectively survived. The resistant plantlets were transferred to the greenhouse and the evaluation of the expression of the bar gene in leaves was positive about 60%. The confirmation by molecular analysis revealed only a transformation efficiency of 0.52 % for the variety ‘Arréhane’.