Genetic Diversity in Candidate Single-Nucleotide Polymorphisms Associated with Resistance in Honeybees in the Czech Republic Using the Novel SNaPshot Genotyping Panel

<b>Background/Objectives</b>: The increasing pressure from pathogens and parasites on <i>Apis mellifera</i> populations is resulting in significant colony losses. It is desirable to identify resistance-associated single-nucleotide polymorphisms (SNPs) and their variability for the purpose of breeding resilient honeybee lines. This study examined the genetic diversity of 13 SNPs previously studied for associations with various resistance-providing traits, including six linked to Varroa-specific hygiene, five linked to suppressed mite reproduction, one linked to immune response, and one linked to chalkbrood resistance. <b>Methods</b>: Genotyping was performed using a novel SNaPshot genotyping panel designed for this study. The sample pool consisted of 308 honeybee samples in total, covering all 77 administrative districts of the Czech Republic. <b>Results</b>: All examined loci were polymorphic. The frequency of positive alleles in our population is medium to low, depending on the specific SNP. An analysis of genotype frequencies revealed that most loci exhibited the Hardy-Weinberg equilibrium. A comparison of the allele and genotype frequencies of the same locus between samples from hives and samples from flowers revealed no significant differences. The genetic diversity, as indicated by the heterozygosity values, ranged from 0.05 to 0.50. The fixation index (<i>F</i>) was, on average, close to zero, indicating minimal influence of inbreeding or non-random mating on the genetic structure of the analyzed samples. <b>Conclusions</b>: The obtained results provide further insights into the genetic variation of SNPs associated with the immune response and resistance to pathogens in honeybee populations in the Czech Republic. This research provides a valuable foundation for future studies of honeybee diversity and breeding.