The purpose of the present work was to evaluate carrot plants obtained from anther cultures with respect to their ploidy and homozygosity. Ploidy was determined using flow cytometry. Homozygosity of analyzed plants was determined using isoenzymes, glucose-6-phosphate isomerase (PGI) and aspartate aminotransferase (AAT). The cytometric tests revealed that more than 90% of the carrot plants obtained from anther cultures were doubled haploid. In the initial assessment of polymorphism of the two enzymatic systems in selected androgenetic carrot plants of the cultivars: Berjo, Kazan F₁, and Splendid F₁, it was proven that 100% of those plants were homozygotes in respect to PGI and also with respect to AAT. In the second experiment, the obtained androgenetic progeny of the heterozygous donor plant of cultivar Narbonne F₁ was found to be 94% homozygotic with respect to PGI and 100% homozygotic in the case of AAT. For the androgenetic plants of the cultivar Kazan F₁, 89% of them were homozygotic with respect to AAT but PGI enzyme system did not differentiate the homozygotic and heterozygotic androgenetic progeny. These results indicated that enzyme polymorphism depends on carrot genotype, therefore, analysis of some (more than one) isozymes was necessary to confirm homozygosity of plants. PGI and AAT can be a useful tool for determining homozygosity in androgenetic carrot plants.