Microalgae are a sustainable source of high-value bioactive compounds with significant nutritional and therapeutic benefits. They contain essential dietary components, including omega-3 polyunsaturated fatty acids, eicosapentaenoic acid (EPA), and docosahexaenoic acid (DHA). These fatty acids are known for their positive effect on mental, vision, and cardiovascular health in humans. In the present study, the stress-inducing effect of indole acetic acid (IAA), and abscisic acid (ABA), on Monoraphidium contortum SRR472 was investigated with a focus on omega-3 fatty acid biosynthesis. Our study highlighted, both phytohormones acted as biochemical stressors, significantly enhancing cellular proliferation, biomass accumulation, and omega-3 fatty acids biosynthesis. Among the treatments, ABA exerted the most stimulatory effect on overall cell growth and biomass. Notably, IAA induced stress significantly increased omega-3 polyunsaturated fatty acids, specifically linolenic acid, EPA, and DHA (9.10 ± 0.01, 3.75 ± 0.07, and 4.1 ± 0.1 %). Elevated levels of reactive oxygen species (ROS) and antioxidant enzymes under phytohormone further confirmed the cellular stress response. The gene expression analysis revealed a substantial upregulation of key fatty acid biosynthetic pathway genes, including FAD (fatty acid desaturase), D6E (elongase ∆6b1), and D6D (∆6-desaturase) were significantly enhanced by 5.3, 2.1, and 1.4-fold, respectively, relative to the control following IAA stress in the microalgae. These results support the role of stress-driven transcriptional regulation in enhancing PUFA production. These findings demonstrate that IAA and ABA-induced oxidative stress serve as a potent trigger for boosting omega-3 fatty acid accumulation in microalgae, offering a strategic approach for optimizing omega-3 PUFA yields in microalgal bioprocessing.
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