Introduction: The B3 transcription factor has been identified in Arabidopsis thaliana, Oryza sativa, and Solanum lycopersicum, among other species. This family of transcription factors regulates seed growth, development, and stress. Cannabis is a valuable crop with numerous applications; however, no B3 transcription factors have been identified in this plant. Materials and Methods: The cannabis B3 gene family was identified and analyzed using bioinformatics analysis tools, such as the NCBI database, plantTFDB website, TBtools, and MEGA software. Quantitative real-time polymerase chain reaction (qRT-PCR) experiments were used to confirm its function. Results: The cannabis B3 family contains 65 members spread across 10 chromosomes. The isoelectric point ranged from 10.03 to 4.65, and the molecular weight ranged from 99,542.88 to 14,310.9 Da. Most of the members were found in the nucleus. The upstream promoter region of the gene contains a variety of cis-acting elements related to the stress response. RNA-seq data and qRT-PCR results showed that CsB3 genes were expressed differently in five organs of female Diku plants and in glandular hairs of nine distinct types of female cannabis inflorescences. Collinearity analysis revealed that there were more homologous genes between cannabis and dicotyledons than monocotyledonous plants, which was consistent with the evolutionary relationship. Conclusions: Hormones and external environmental factors might influence CsB3 expression. Furthermore, some genes such as CsB3-02, CsB3-07, CsB3-50, CsB3-62, and CsB3-65 may participate in cannabis growth and development and play a role in secondary metabolite synthesis. This study provides a solid foundation for further research into the gene function of the cannabis B3 family.
Keywords: B3 transcription factors; Cannabis sativa; evolutionary analysis; expression pattern.