Clock genes regulate sex pheromone production and male mating ability in <i>Bactrocera dorsalis</i>

Many animals display physiological and behavioral activities limited to specific times of the day. Certain insects exhibit clear daily rhythms in their mating activities that are regulated by an internal biological clock. However, the specific genetic mechanisms underlying this regulation remain largely unexplored. Mating in the fruit fly Bactrocera dorsalis exhibits a daily rhythm and is dependent on sex pheromones produced in the male rectum. We used transcriptome sequencing and clustered regularly interspaced short palindromic repeats (CRISPR) / CRISPR-associated nuclease 9 techniques to understand whether the daily rhythmicity of mating in B. dorsalis and sex pheromone production in the rectum are regulated by clock genes. The results showed that the production of sex pheromones by B. dorsalis males is rhythmic (low during the day and high at night) and is influenced by clock genes. Knockout of the clock genes cryptochrome 1 (cry1) and timeless (tim) reduced the production of sex pheromones and significantly impaired mating ability in males. In addition, quantitative polymerase chain reaction results from 5 different tissues showed cry1 was highly expressed in the head, whereas tim was highly expressed in both the head and rectum (a key site for male sex pheromone production). Transcriptome analysis confirmed that cry1 (head) and tim (head and rectum) exhibit rhythmic expressions consistent with sex pheromone rhythmicity. These results suggest that cry1 may be related to a central clock neuron (like the suprachiasmatic nucleus), whereas the rhythmic expression of tim in the rectum indicates the potential presence of peripheral oscillators. Our study reveals new targets and ideas for improved control of the fruit fly.