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<p>This study investigates the impact of polyethylene glycol (PEG)-induced osmotic stress in <i>Artemisia annua</i>, focusing on morpho-physiological changes, secondary metabolite synthesis, and gene expression. The finding reveals mild osmotic stress positively influenced glandular trichome density, which is a storage site for bioactive metabolites. However, higher levels of osmotic stress led to a significant decrease in the density of these trichomes. Under 8% PEG exposure proline, sugar, and anthocyanin levels increased, whereas decrease in growth, relative water content, chlorophyll content, and photosynthetic efficiency (Fv/Fm) was observed. Gas chromatography analysis showed a 1.49-fold increase in total essential oil compounds compared to controls, identifying seven major compounds: 1,8-cineole, camphor, β-caryophyllene, β-farnesene, β-coapane, selina-4,11-diene, and aristolone. Biosynthetic genes (<i>PAL</i>, <i>CHS</i>, <i>ADS</i>, <i>DBR2</i>, and <i>CYP71AV1</i>) exhibited upregulation by 2, 4.7, 4.1, 1.2, and 1.5-fold, respectively, when subjected to 8% PEG-induced stress, thereby enhancing artemisinin and other secondary metabolite production. To explore the role of AaAP2/ERF transcription factors under osmotic stress, a comprehensive genome-wide analysis identified 111 members across the DREB, ERF, AP2, RAV, and Soloists subfamilies. Promoter analysis revealed five homologues, with <i>AaDREB-09</i> showing the highest upregulation (2.8-fold) in shoot tissues under osmotic stress. Further protein docking analysis demonstrated that AaDREB-09 binds to the promoters of <i>AaDBR2</i> and <i>AaCYP71AV1</i>, enhancing the biosynthesis of artemisinin and dihydroartemisinic acid, thereby improving stress tolerance. These findings depict <i>AaDREB-09</i> as a promising target for genetic engineering to enhance secondary metabolite production and stress resilience in <i>A. annua</i>.</p>

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Functional identification of AaDREB-9 transcription factor in Artemisia annua L. and deciphering its role in secondary metabolism under PEG-induced osmotic stress

  • 期刊论文

  • 发表日期：

    2025年4月10日

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  • Sabitri Kumari,
  • Nidhi Rai,
  • Sneha Singh,
  • Pajeb Saha,
  • Mansi Singh Bisen,
  • Shashi Pandey-Rai

摘要

This study investigates the impact of polyethylene glycol (PEG)-induced osmotic stress in Artemisia annua, focusing on morpho-physiological changes, secondary metabolite synthesis, and gene expression. The finding reveals mild osmotic stress positively influenced glandular trichome density, which is a storage site for bioactive metabolites. However, higher levels of osmotic stress led to a significant decrease in the density of these trichomes. Under 8% PEG exposure proline, sugar, and anthocyanin levels increased, whereas decrease in growth, relative water content, chlorophyll content, and photosynthetic efficiency (Fv/Fm) was observed. Gas chromatography analysis showed a 1.49-fold increase in total essential oil compounds compared to controls, identifying seven major compounds: 1,8-cineole, camphor, β-caryophyllene, β-farnesene, β-coapane, selina-4,11-diene, and aristolone. Biosynthetic genes (PAL, CHS, ADS, DBR2, and CYP71AV1) exhibited upregulation by 2, 4.7, 4.1, 1.2, and 1.5-fold, respectively, when subjected to 8% PEG-induced stress, thereby enhancing artemisinin and other secondary metabolite production. To explore the role of AaAP2/ERF transcription factors under osmotic stress, a comprehensive genome-wide analysis identified 111 members across the DREB, ERF, AP2, RAV, and Soloists subfamilies. Promoter analysis revealed five homologues, with AaDREB-09 showing the highest upregulation (2.8-fold) in shoot tissues under osmotic stress. Further protein docking analysis demonstrated that AaDREB-09 binds to the promoters of AaDBR2 and AaCYP71AV1, enhancing the biosynthesis of artemisinin and dihydroartemisinic acid, thereby improving stress tolerance. These findings depict AaDREB-09 as a promising target for genetic engineering to enhance secondary metabolite production and stress resilience in A. annua.