Biolistic transformation of Moroccan durum wheat varieties by using mature embryo-derived calli
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Environmental stresses are estimated to have reduced global crop yields of wheat by 5.5%. However, traditional approaches for the transfer of resistance to these stresses in wheat plants have yielded limited results. In this regard, genetic transformation has undoubtedly opened up new avenues to overcome crop losses due to various abiotic stresses. Particle bombardment has been successfully em- ployed for obtaining transgenic wheat. However, most of these procedures employ immature embryos, which are not available throughout the year. Therefore, the present investi- gation utilized mature seeds as the starting material and used the calli raised from three Moroccan durum wheat varieties as the target tissue for genetic transformation by the biolistic approach. The pANIC-5E plasmid containing the SINA gene for drought and salinity tolerance was used for genetic trans- formation. To enhance the regeneration capacity and trans- formation efficiency of the tested genotypes, the study compared the effect of copper supplementation in the induc- tion medium (up to 5 μM) with the standard MS medium. The results show that the genotypes displayed different sensitivities to CuSO4, indicating that the transformation efficiency was highly genotype-dependent. The integration of transgenes in the T0 transformants was demonstrated by polymerase chain reaction (PCR) analysis of the obtained resistant plantlets with primers specific to the SINA gene. Among the three genotypes studied, ‘Isly’ showed the highest efficiency of 9.75%, followed by ‘Amria’ with 1.25% and ‘Chaoui’ with 1%.
Abstract
Environmental stresses are estimated to have reduced global crop yields of wheat by 5.5%. However, traditional approaches for the transfer of resistance to these stresses in wheat plants have yielded limited results. In this regard, genetic transformation has undoubtedly opened up new avenues to overcome crop losses due to various abiotic stresses. Particle bombardment has been successfully em- ployed for obtaining transgenic wheat. However, most of these procedures employ immature embryos, which are not available throughout the year. Therefore, the present investi- gation utilized mature seeds as the starting material and used the calli raised from three Moroccan durum wheat varieties as the target tissue for genetic transformation by the biolistic approach. The pANIC-5E plasmid containing the SINA gene for drought and salinity tolerance was used for genetic trans- formation. To enhance the regeneration capacity and trans- formation efficiency of the tested genotypes, the study compared the effect of copper supplementation in the induc- tion medium (up to 5 μM) with the standard MS medium. The results show that the genotypes displayed different sensitivities to CuSO4, indicating that the transformation efficiency was highly genotype-dependent. The integration of transgenes in the T0 transformants was demonstrated by polymerase chain reaction (PCR) analysis of the obtained resistant plantlets with primers specific to the SINA gene. Among the three genotypes studied, ‘Isly’ showed the highest efficiency of 9.75%, followed by ‘Amria’ with 1.25% and ‘Chaoui’ with 1%.
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