Immature Embryo-Derived of Two Bread Wheat (Triticum aestivum L.) Varieties Transformation Using Particle Bombardment Method
D. C. Tinak Ekom
Biotechnology Research Unit, National Institute for Agronomic Research (INRA), BP 415, Avenue de la Victoire, Rabat, Morocco and Laboratory of Virology, Microbiology and Quality / Eco Toxicology and Biodiversity, Faculty of Sciences and Techniques Mohammedia, Casablanca Hassan II University Mohammedia, Mohammedia PO 146 20650, Morocco.
S. M. Udupa
ICARDA-INRA Cooperative Research Project, International Center for Agricultural Research in the Dry Areas (ICARDA), B.P. 6299, Rabat, Morocco.
M. N. Benchekroun
Laboratory of Virology, Microbiology and Quality / Eco Toxicology and Biodiversity, Faculty of Sciences and Techniques Mohammedia, Casablanca Hassan II University Mohammedia, Mohammedia PO 146 20650, Morocco.
M. M. Ennaji
Laboratory of Virology, Microbiology and Quality / Eco Toxicology and Biodiversity, Faculty of Sciences and Techniques Mohammedia, Casablanca Hassan II University Mohammedia, Mohammedia PO 146 20650, Morocco.
R. Abdelwahd
Biotechnology Research Unit, National Institute for Agronomic Research (INRA), BP 415, Avenue de la Victoire, Rabat, Morocco.
D. Iraqi *
Biotechnology Research Unit, National Institute for Agronomic Research (INRA), BP 415, Avenue de la Victoire, Rabat, Morocco.
*Author to whom correspondence should be addressed.
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’.
Keywords: Bar gene, genetic transformation, bread wheat, plasmid pBY520