Reconsidering the direct cross approach for gene transfer from Aegilops tauschii to hexaploid wheat using diversified D genome donors

The direct cross-hybridization approach between 16 Aegilops tauschii (AT) and 10 bread wheat (W) lines was carried out in reciprocal directions. W (♀) x AT (♂) yielded no embryo formation whereas embryo formation ranging from 34.09 to 43.33% was obtained in AT (♀) x W (♂) crosses in all 160 combinations. The major bottleneck of low plant regeneration in AT (♀) x W (♂) crosses was cleared by adopting detached tiller culture and embryo rescue medium adjuvants such as casein hydrolysate (200 mg/L) and activated charcoal (2 g/L). The regeneration frequency as a result of these two interventions increased from 3.61 to 19.29 per cent. Two innovative methods of chromosome doubling included in situ colchicine (0.05%) treatment with cotton swab method and pre-regeneration colchicine treatment (0.1 and 0.2%) to pollinated ovaries were found to be effective. The findings of the study make successful exploitation of direct cross approach (AT (♀) x W (♂)) possible in wheat improvement.