Fayaz Mohd, Angmo Tsering, Katoch Kajal, Majeed Aasim, Kundan Maridul, Wajid Mir Abdul, Pal Koushik, Misra Prashant
Plant Sciences and Agrotechnology Division, CSIR-Indian Institute of Integrative Medicine, Canal Road, Jammu, 180001, India.
Academy of Scientific and Innovative Research (AcSIR), Ghaziabad, 201002, India.
Planta. 2025 May 11;261(6):135. doi: 10.1007/s00425-025-04709-x.
The functional characterization of promoter regions of CBDAS and PT genes of cannabinoids biosynthesis suggests that multiple factors including tissue-specific, phytohormones, and stress-related signals modulate their activity. Cannabis sativa L. has tremendous potential as a future crop for producing clinically important cannabinoid metabolites. While the cannabinoid biosynthetic pathway is largely known, the mechanistic details about its regulation are less understood. Decrypting the environmental and developmental factors regulating cannabinoid biosynthesis pathway may prove beneficial in pathway engineering and molecular breeding programs. Functional characterization of the promoter regions of key cannabinoid biosynthesis genes can provide useful insights into their transcriptional regulation. This study, therefore, is focused to uncover the role of different phytohormones and abiotic factors in influencing the activity of CsCBDAS and CsPT1 promoters through the development of promoter-GUS fusion expressing transgenic lines of Nicotiana tabacum. Spatial analysis across different tissues revealed that CsCBDAS and CsPT1 promoters drive a high level of GUS staining in leaf and flowers of the transgenic lines. A strong GUS staining was detected in the glandular trichomes of both tobacco transgenic lines. The results showed that out of the five hormones, three (IAA, GA, and SA) and four (IAA, GA, SA, and ABA) caused significant activation of CsCBDAS and CsPT1 promoters, respectively. While the light, heat, cold, salt, and wound stress induced promoter activity of both CsCBDAS and CsPT1, the drought stress was found to induce the activity of CsCBDAS promoter only. Validation of the expression patterns of these genes under different conditions in C. sativa through qRT-PCR suggested that phytohormones and abiotic factors may influence the cannabinoid biosynthesis in C. sativa by modulating their promoter activity.
大麻素生物合成中CBDAS和PT基因启动子区域的功能表征表明，包括组织特异性、植物激素和胁迫相关信号在内的多种因素调节它们的活性。大麻作为一种未来作物，在生产具有临床重要性的大麻素代谢产物方面具有巨大潜力。虽然大麻素生物合成途径在很大程度上已为人所知，但其调控的机制细节仍了解较少。解密调节大麻素生物合成途径的环境和发育因素可能对途径工程和分子育种计划有益。关键大麻素生物合成基因启动子区域的功能表征可为其转录调控提供有用见解。因此，本研究旨在通过构建烟草启动子-GUS融合表达转基因株系，揭示不同植物激素和非生物因素对CsCBDAS和CsPT1启动子活性的影响。对不同组织的空间分析表明，CsCBDAS和CsPT1启动子在转基因株系的叶片和花中驱动高水平的GUS染色。在两种烟草转基因株系的腺毛中均检测到强烈的GUS染色。结果表明，在五种激素中，三种（IAA、GA和SA）和四种（IAA、GA、SA和ABA）分别显著激活了CsCBDAS和CsPT1启动子。虽然光、热、冷、盐和伤口胁迫诱导了CsCBDAS和CsPT1的启动子活性，但发现干旱胁迫仅诱导CsCBDAS启动子的活性。通过qRT-PCR验证这些基因在大麻不同条件下的表达模式表明，植物激素和非生物因素可能通过调节其启动子活性来影响大麻中的大麻素生物合成。
