Optimization of Somatic Embryogenesis and Agrobacterium-mediated Transformation of Elite Wheat (Triticum aestivum) Cultivars of Pakistan

Immature embryos of twenty six wheat genotypes were regenerated on Murashige and Skoog (MS) medium supplemented with different levels of plant growth regulator (PGRs). 2 4-D at 2 mg L-1 level was found to be the most favorable callus inducer. Bobwhite exhibited maximum embryogenic callus production followed by Ufaq-2000' Punjab-2011' and Sehar-2006'. Regeneration potential of selected wheat genotypes was evaluated on two PGRs indole-3-acetic acid (IAA) and kinetin (N6-furfuryladenine) in 4 different combinations. The auxin-cytokinin combination in IK4 (1 mg L-1 IAA and 1 mg L-1 Kinetin) media was the most promising for regeneration as variety Bobwhite presented the highest plantlet frequency. Immature embryos of cultivar Sehar-2006' were utilized for Agrobacterium mediated transformation and GUS activity was observed in callus to determine the expression pattern. Transgenic plants of 'Sehar-2006' were confirmed by PCR and Southern blotting. Transformation efficiency of 2.45 % was achieved which reinstated the viability of optimized regeneration system and this particular wheat genotype for utilization in crop improvement program. Copyright 2014 Friends Science Publishers