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<p>Cannabinoids, the bioactive compounds unique to <i>Cannabis</i> plants, demonstrate significant therapeutic potential in treating epilepsy, pain management, and inflammatory conditions. Despite their clinical importance, the genetic mechanisms underlying cannabinoid biosynthesis in seed hemp remain poorly understood. Our study systematically identified 25 key genes involved in the cannabinoid synthesis pathway within the seed hemp genome. Comprehensive bioinformatics analyses were conducted to characterize their physicochemical properties, evolutionary relationships, gene structures, promoter cis-elements, and expression patterns. Transcriptomic profiling of ‘Yushe’ cultivar revealed tissue-specific expression patterns across roots, stems, leaves, and male/female flowers. These genes were organized into four distinct biosynthetic pathways: MEP (methylerythritol phosphate), GPP (geranyl diphosphate), acetic acid, and cannabinoid-specific pathways. Promoter analysis identified critical cis-elements associated with light responsiveness, phytohormone signaling, and stress adaptation. Notably, seven genes (<i>CsaDXS</i>, <i>CsaHDS</i>, <i>CsaHDR</i>, <i>CsaGPP(lsu)</i>, <i>CsaHPL</i>, <i>CsaTHCAS</i>, <i>CsaCBDAS</i>) exhibited higher expression in male leaves, while four genes (<i>CsaDXR</i>, <i>CsaGPP(lsu)</i>, <i>CsaHPL</i>, <i>CsaCBDAS</i>) were upregulated in female flowers. Comparative genomic analysis revealed that medicinal <i>cannabis</i> types possessed higher gene copy numbers for <i>CsaDXS</i>, <i>CsaHDS</i>, <i>CsaTHCAS</i>, and <i>CsaCBDAS</i>, which may account for their elevated cannabinoid production. Functional validation through VIGS-mediated silencing of <i>PT</i> gene demonstrated significant downregulation of <i>THCAS</i> and <i>CBDAS</i> expression. This study provides crucial genetic resources for functional characterization of cannabinoid synthase genes and facilitates the rational utilization of seed hemp germplasm.</p>

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Genome-Wide Identification and Expression Analysis of Genes Related to Cannabinoid Biosynthesis Pathways in Seed Hemp

  • 期刊论文

  • 发表日期：

    2025年10月15日

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  • Zuqing Yang,
  • Huawei Wei,
  • Xueqing Pan,
  • Jiahui Chen,
  • Huifan Kang,
  • Yuxin Lin,
  • Shaoqiang Jiang,
  • Xuping Feng,
  • Aifen Tao,
  • Jianmin Qi,
  • Jiantang Xu,
  • Hongmei Kang,
  • Liwu Zhang

摘要

Cannabinoids, the bioactive compounds unique to Cannabis plants, demonstrate significant therapeutic potential in treating epilepsy, pain management, and inflammatory conditions. Despite their clinical importance, the genetic mechanisms underlying cannabinoid biosynthesis in seed hemp remain poorly understood. Our study systematically identified 25 key genes involved in the cannabinoid synthesis pathway within the seed hemp genome. Comprehensive bioinformatics analyses were conducted to characterize their physicochemical properties, evolutionary relationships, gene structures, promoter cis-elements, and expression patterns. Transcriptomic profiling of ‘Yushe’ cultivar revealed tissue-specific expression patterns across roots, stems, leaves, and male/female flowers. These genes were organized into four distinct biosynthetic pathways: MEP (methylerythritol phosphate), GPP (geranyl diphosphate), acetic acid, and cannabinoid-specific pathways. Promoter analysis identified critical cis-elements associated with light responsiveness, phytohormone signaling, and stress adaptation. Notably, seven genes (CsaDXS, CsaHDS, CsaHDR, CsaGPP(lsu), CsaHPL, CsaTHCAS, CsaCBDAS) exhibited higher expression in male leaves, while four genes (CsaDXR, CsaGPP(lsu), CsaHPL, CsaCBDAS) were upregulated in female flowers. Comparative genomic analysis revealed that medicinal cannabis types possessed higher gene copy numbers for CsaDXS, CsaHDS, CsaTHCAS, and CsaCBDAS, which may account for their elevated cannabinoid production. Functional validation through VIGS-mediated silencing of PT gene demonstrated significant downregulation of THCAS and CBDAS expression. This study provides crucial genetic resources for functional characterization of cannabinoid synthase genes and facilitates the rational utilization of seed hemp germplasm.