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http://dx.doi.org/10.5010/JPB.2021.48.4.246

Biolistic transformation of Moroccan durum wheat varieties by using mature embryo-derived calli  

Senhaji, Chaimae (Biotechnology Research Unit, Institut National de la Recherche Agronomique (INRA))
Gaboun, Fatima (Biotechnology Research Unit, Institut National de la Recherche Agronomique (INRA))
Abdelwahd, Rabha (Biotechnology Research Unit, Institut National de la Recherche Agronomique (INRA))
Diria, Ghizlane (Biotechnology Research Unit, Institut National de la Recherche Agronomique (INRA))
Udupa, Sripada (ICARDA–INRA Cooperative Research Project, International Center for Agricultural Research in the Dry Areas)
Douira, Allal (Laboratory of Botany, Biotechnology and Plant Protection, Faculty of Sciences-University Ibn Tofail, University campus)
Iraqi, Driss (Biotechnology Research Unit, Institut National de la Recherche Agronomique (INRA))
Publication Information
Journal of Plant Biotechnology / v.48, no.4, 2021 , pp. 246-254 More about this Journal
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 employed for obtaining transgenic wheat. However, most of these procedures employ immature embryos, which are not available throughout the year. Therefore, the present investigation 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 transformation. To enhance the regeneration capacity and transformation efficiency of the tested genotypes, the study compared the effect of copper supplementation in the induction 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%.
Keywords
Biolistic transformation; Copper sulphate; Durum wheat; Matures embryos; Plasmid pANIC-5E; SINA gene;
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