Abstract
Apis mellifera L. includes several recognized subspecies that differ in their biological properties and agricultural characteristics. Distinguishing between honey bee subspecies is complicated. We analyzed the Folmer region of the COX1 gene in honey bee subspecies cultivated at bee farms in Russia and identified subspecies-specific SNPs. DNA analysis revealed two clearly distinct haplogroups in A. mellifera mellifera. The first one was characterized by multiple cytosine-thymine (thymine–cytosine) transitions, one adenine-guanine substitution, and one thymine–adenine substitution. The nucleotide sequence of the second haplogroup coincided with sequences from other subspecies, except the unique C/A SNP at position 421 of the 658-bp Folmer region. A. mellifera carnica and A. mellifera carpatica could be distinguished from A. mellifera mellifera and A. mellifera caucasica by the presence of the A/G SNP at position 99 of the 658-bp Folmer region. The G/A SNP at position 448 was typical for A. mellifera carnica. A. mellifera caucasica COX1 sequence lacked all the above-mentioned sites. We developed a procedure for rapid identification of honey bee subspecies by PCR with restriction fragment length polymorphism (RFLP) using mutagenic primers. The developed molecular method for honey bee subspecies identification is fast and inexpensive.
Highlights
The honey bee (Apis mellifera L.) is a major producer of honey
We found that the highest intrabreed variability of the COX1 gene was in A. mellifera mellifera and analyzed nucleotide substitutions typical of this subspecies
We found that the COX1 gene can be used as a marker for distinguishing honey bee subspecies
Summary
The honey bee (Apis mellifera L.) is a major producer of honey. Honey bees are important pollinators of entomophilic crops [1]. Several bee breeds (subspecies) are recommended for breeding within the territory of the Russian Federation. One of the important areas of bee breeding is the preservation of bee subspecies genotypes diversity, which can be used as a starting material for breeding and crossbreeding. Breed purity is an important factor in these processes. In Russia, there are more than 20 breeding bee farms, and there are Apis mellifera populations from the Primorye region. It is believed that these bees are more resilient to parasitic mites [2,3]. All of the above factors determine the importance of correctly identifying honey bee subspecies in Russia
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