Non-Destructive Genotyping of Honeybee Queens to Support Selection and Breeding

SIMPLE SUMMARY: Genotyping of an individual usually requires tissue samples obtained by methods that may cause an injury. In the case of honeybee queens, most of such injuries cause supersedure of the queen. We have investigated the possibility of using two non-destructive sources to obtain genetic information from an individual queen: feces and exuviae. Both sources turned out to be acceptable in terms of quality of DNA and usefulness in genotyping. In practical use, any of the two could be used to shorten generation interval in queen breeding and decrease costs of the selection process. ABSTRACT: In traditional bee breeding, the honeybee queen is chosen for breeding based on the performance of the colony produced by its mother. However, we cannot be entirely certain that a specific queen will produce offspring with desirable traits until we observe the young queen’s new colony. Collecting the queen’s genetic material enables quick and reliable determination of the relevant information. We sampled exuviae, feces, and wingtips for DNA extraction to avoid fatally injuring the queen when using tissue samples. Quantity and purity of extracted DNA were measured. Two mitochondrial markers were used to determine the lineage affiliation and exclude possible contamination of DNA extracts with non-honeybee DNA. dCAPS (derived Cleaved Amplified Polymorphic Sequences) markers allowed detection of single nucleotide polymorphisms (SNPs) in nuclear DNA regions presumably associated with Varroa sensitive hygiene and set the example of successful development of genotyping protocol from non-destructive DNA sources. One of the logical future steps in honeybee breeding is introducing genomic selection and non-destructive sampling methods of genetic material may be the prerequisite for successful genotyping. Our results demonstrate that the extraction of DNA from feces and exuviae can be introduced into practice. The advantage of these two sources over wingtips is reducing the time window for processing the samples, thus enabling genotyping directly after the queen’s emergence.