4. Conclusion

A. mellifera subspecies are three parameters of the wing namely the cubital index, the hantel index, and the discoidal shift. These parameters, together with the data on the variability of the COI–COII mtDNA locus, make it possible to differentiate the dark-colored forest bee and bees of

**Microsatellite analysis (loci А043, А088, Ар081 and others)**

(10–30 bees from each bee colony)

**Establishment of races that participated in bee hybridization**

**Variants PQQ or PQQQ Variant Q**

**Analysis of variability of the COI-COII mtDNA locus**  (minimum 5 bees from each bee colony)

**Morphometric analysis**  (parameters of wing, including the cubital index, the hantel index, and the discoidal shift; minimum 30 bees from each bee colony)

> **Q and discrepancy of morphometric parameters with the standard of southern races**

**Q and the consistency of morphometric parameters with the standard of southern races**

**More precise definition (clarification) of genotypic composition, identification of ecotypes**

**PQQ/PQQQ and discrepancy of morphometric parameters with the dark forest bee standard**

Our data also indicate that only the exterior or just genetic traits may be insufficient to determine the origin of bees and only the simultaneous analysis of morphometric parameters and data on the variability of locus COI–COII of mtDNA allow to evaluate the breed and cases

Finally, a microsatellite analysis should be conducted to study genetic diversity of bee colonies and to clarify their origin (possibly ecotypes) and/or the origin of the hybrids. As the research on the variability of the nuclear DNA markers in different bee subspecies

southern breeds, as well as hybrids (see details in Refs. [17–19]).

of hybridization objectively.

**PQQ/PQQQ and the consistency of morphometric parameters with the dark forest bee standard**

174 Selected Studies in Biodiversity

**Regi (clarification) of More precise definition** 

**genotypic composition, identification of ecotypes**

Figure 2. Algorithm of the study of the bee colonies.

A screening study of bee colonies in Siberia made it possible to identify two populations of the dark-colored forest bee in the Krasnoyarsk Territory and the Tomsk Region. These A. m. mellifera populations identified in Siberia were described by a complex of morphometric and molecular genetic markers. According to the mtDNA analysis, all studied bee colonies were of the dark-colored forest bee origin in the maternal line (the bees had a variant PQQ of the COI–COII locus). According to the basic morphometric parameters, most bee colonies fully corresponded to the A. m. mellifera standard. As possible potential DNA markers, microsatellite loci specific for determining of the bee subspecies (A043, mrjp3) and/or ecotypes (A008) of the dark-colored forest bee have been identified from 18 analyzed microsatellites.

Thus, to identify and preserve dark-colored forest bee populations in Siberia, we studied the genetic diversity of local native bees, described the specific polymorphic variants of loci of mtDNA and nuclear genome, and proposed an algorithm for the search and a comprehensive study of the dark-colored forest bee.

As a result of our research, we can draw the following conclusions:


Development of diagnostic DNA markers is a scientific basis for the evaluation of quality of bee colonies in the dark-colored forest bee farm, created by Tomsk State University. In addition, a complex approach to the analysis of bee colonies (morphometric and molecular genetic analysis) allows obtaining genetic certification of bees, identifying the valuable line (ecotypes) of local bees, and protecting and making rational use of genetic resources of aboriginal bee subspecies.

This is one of the first attempts to introduce molecular genetic markers in the practice of beekeeping in Russia as the real possibility of the definition of bee subspecies (bee breeds). In the future, a similar comprehensive approach, including analysis of molecular genetic and morphometric markers, will be used for the selection of bee colonies with high economically significant indicators, disease resistance, and other parameters based on genotypic features of honeybees.

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