RNA-sequencing elucidates the regulation of delayed nuptial flight behavior in honey bee queens

Controlling queen bees to mate with selected drones is a challenging aspect of honeybee breeding.Comparing to the traditional, spatial, mating isolation (mating station) temporal isolation as a method for honey bee mating has fewer restrictions and shows significant potential for application. This approach aims to regulate mating by delaying the mating time of queens and drones. In this study, we examined the nuptial flight behavior of queens subjected to an human-induced delay in mating. Based on the timing of the queens' nuptial flights, we divided them into three groups: the regular flight group (RF), the delayed flight group (DF), and the delayed no-flying group (DNF). Transcriptome sequencing and bioinformatics analysis were performed on the brain tissues of the queens. Core biological clock genes Cry2 and Per respond to light, time and temperature cues, regulating the expression of related genes (e.g., HSPs, cytochrome P450) that directly influence nuptial mating behavior transmission. Other significant changes in the expression of genes related to the biological clock, immunity, metabolism, odor, and pheromones were observed. Further study of these genes may be key to advancing methods for regulating mating time and achieving controlled mating.