The (1,3)-β-D-glucan also referred to as callose is a main component of cell walls of higher plants. Many physiological processes are associated with the changes in callose deposition. Callose is synthesised by the callose synthase complex while its degradation is regulated by the hydrolytic enzymes β-1,3-glucanases. The latter one specifically degrade (1,3)-β-D-glucans. This work is aimed to study β-1,3-glucanase activities in the leaves of plants at two leaf stage in two diploids (Agilops tauschii, Triticum monococcum L.), four tetraploids (Ae. cylindrica, Ae. triuncialis, T. araraticum, T. dicoccum) and two hexaploids (T. aestivum L, T. spelta L.). The leaves were subjected to qualitative and quantitative β-1,3-glucanase activity assays. Our results showed that the total β-1,3-glucanase activities were variable and genotype dependent. No significant correlation between β-1,3-glucanase activities and ploidy level was observed. The gel activity assays revealed a single fraction of ~52 kDa Glu1 that was found in all genotypes. The Glu1 fraction corresponds to a single or two acidic Glu isoforms in dependence on genotype. However, none of the acidic Glu fractions can be assigned as a specific for di-, tetra- or hexaploid genotypes. A single basic GluF isoform was detected and found as present in all genotypes.