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In a selection of honeybees from autochthonous ecotypes, different lines must be identified. Honeybee lineages are usually distinguished by classical morphometrics and molecular markers, but these approaches are both costly and time-consuming to implement. Recognition of the purity of races is very important for regional and country regulations to allow a sustainable conservation of the huge variety of local honeybees. A geometric morphometric approach has been frequently used. In this work, honeybee samples were collected from stationary apiaries (belonging to the centers for honeybee queen selection) from two different Serbian areas: Vršac (northeastern Serbia, mostly flatland) and Vranje (southern Serbia, mostly mountainous), and two different Montenegrin areas: Bijelo Polje (northern Montenegro, mountainous region) and Sutomore (southern Montenegro, coastal region). Each sample consisted of 150 honeybee workers, collected from 10 hives (15 specimens each). On the honeybee left forewings, a total of 19 vein intersections were used to determine the differences among the honeybees using MorphoJ 1.4a software. Canonical variate analysis (CVA) slightly separated the honeybee lines into one overlapping cloud of specimens at the individual level. The first canonical variable (60.57% of the total variability) discriminated mainly between Bijelo Polje and Sutomore honeybee lines. Therefore, on the colony level, CVA separated all four groups of breeding honeybee lines. The results show that geometric morphometrics are reliable in the discrimination of honeybee lines within subspecies only at the colony level. [ABSTRACT FROM AUTHOR]