applied sciences  
The Potential of Complementary Sex-Determiner Gene Allelic Diversity for Studying the Number of Patrilines within Honeybee Colonies [PERSON], [PERSON], [PERSON], [PERSON], [PERSON] and [PERSON],*  
1 Laboratory of Molecular and Cellular Immunology, Ludwik Hirszfeld Institute of Immunology and Experimental Therapy Polish Academy of Sciences, Weigla 12, 53-114 Wroclaw, Poland; [EMAIL_ADDRESS] (R.M. ); [PERSON] (J.-N.-T.); [EMAIL_ADDRESS] (K.A.-W.); [PERSON]) Animal Production Department, agricultural Institute of Slovenia, Haquetova Ulica 17, 1000 Ljubljana, Slovenia; [EMAIL_ADDRESS] Correspondence: [EMAIL_ADDRESS]; Tel. : +48-071-3709980 (ext. 333)  
Footnote 1: [[https://www.mdpi.com/journal/applsci](https://www.mdpi.com/journal/applsci)]([https://www.mdpi.com/journal/applsci](https://www.mdpi.com/journal/applsci))  
Footnotediversity, enhancing the colony's ability to adapt to changing environmental conditions, resist diseases, and improve its overall fitness [6]. Studying patrilines provides insights into genetic diversity within honeybee colonies, the reproductive success of different drones, the effects of genetic diversity on colony health and productivity, and social dynamics among worker bees. Understanding the genetic diversity within colonies is also important from a practical standpoint, as it informs decisions regarding improving the desired traits in breeding programs such as disease resistance, gentleness, and productivity [7]. One of the important components of a successful breeding program is mating control, in which genetically superior individuals are identified and used in the selection of breeding lines to generate genetic progress [8]. Due to peculiarities in the reproductive biology of honeybees, the implementation of mating control is demanding and needs a proper evaluation of its efficiency. The assessment of mating control success is measured by how many drones (partilines) the queen mated with came from known (installed) drone-producing colonies and how many from unknown colonies (originating from a large area around the mating site). Current methods employ the indirect genotyping of the worker bee brood using several polymorphic microsatellite _loci_ to estimate the number and the origin of the patrilines in each colony and the share of the patrilines that belong to the known drones--the ones that belong to the drone-producing colonies that were installed [9]. Microsatellite markers, also known as simple sequence repeats (SSRs) or short tandem repeats (STRs), are widely used tools to study inter- and intra-colonial genetic diversity in honeybees [10; 11]. These markers consist of short repeated units of 1-6 base pairs in length, mainly located in the non-coding regions of the genome [12]. Microsatellite markers are highly polymorphic, meaning that the number of repeats can vary significantly among individuals in a population. The analysis of microsatellite markers involves the PCR amplification of specific _loci_ and determining their length variation in the amplified fragments using capillary electrophoresis or, less frequently, DNA sequencing techniques. Microsatellite analysis has been successfully used in honeybee research for studying social organization, population structure, genome mapping, and evolutionary history [6; 13; 14]. Numerous microsatellite _loci_ in the honeybee genome have been described, and different sets of markers are used for specific studies. However, microsatellite marker analysis has limitations, including the occurrence of homoplasy (alleles identical in length but with different sequences), null alleles (alleles that fail to yield PCR products, leading to false homozygosity), and a limited number of alleles for a given microsatellite marker in a population, resulting in the low resolution of the analysis.