A fast and reliable method for ploidy determination of regenerated Triticum aestivum from anther culture is described in the present work. The anthers of 47 hybrid combinations of winter wheat were cultured in vitro and the length of the guard cells was measured in regenerated plants. Thus, plants with a guard cell length of 32-46 mm were haploid, whilst plants with a guard cell length of 54-74 mm were diploid. Plants with a guard cell of 46-54 mm required further chromosome investigation for the exact determination of ploidy level.

The experiment was conducted to assess the effect of different concentration combination of 2,4D, a-NAA and kinetin on callus induction and regeneration of haploid in anther culture of wheat. Potato-2 liquid medium supplemented with different concentration combinations was used for this purpose. Among all the formulations, efficient callus induction and plant regeneration were observed in liquid potato medium containing following concentrations (mg l-1) of plant growth regulators: 2,4D-0.75; a-NAA-0.1 and kinetin 0.75. This formulation supported to produce haploid plantlets directly without transferring calli to regeneration medium.