Contribution

Synthetic biology involves creating biological systems from basic life elements, using engineering principles to design organisms with specific functions. By combining biology, genetic engineering, and engineering principles, researchers construct systems that function like circuits. Bioluminescence, especially from organisms like fireflies or fungi, offers useful biological components for creating synthetic systems. These systems have applications in environmental monitoring, scientific research, and creating bioluminescent plants. The introduction of foreign genes into plants enables them to glow, demonstrating the practical applications of synthetic biology.

Our project focuses on creating bioluminescent plants that not only serve as emotional therapy tools but also act as environmental sensors. By integrating fungal bioluminescence pathways with optimized plant pathways, we significantly improved the plants' ability to emit light. We also introduced a formaldehyde-sensing system that triggers increased bioluminescence when formaldehyde levels rise. These plants are designed to be visible in the dark, providing therapeutic effects while also acting as biosensors for pollutants.

1. Enhanced Bioluminescent Pathway:

• TAL and C3H Genes: By introducing TAL (Tyrosine Ammonia-Lyase) and C3H (Cinnamate-4-Hydroxylase) genes, we improved caffeic acid biosynthesis, a key precursor in the bioluminescence pathway. This significantly increased the brightness of the plant's glow.
• Codon Optimization: To ensure the efficient expression of these genes in plants, we performed codon optimization of, increasing the expression levels of key enzymes involved in the bioluminescent pathway.

2. Formaldehyde Response System:

• Formaldehyde Sensor: We incorporated formaldehyde-responsive elements that activate when exposed to elevated levels of formaldehyde, triggering increased production of caffeic acid. This results in enhanced bioluminescence, transforming the plant into a living sensor for harmful environmental pollutants.

3. Innovative Experimental Methods:

• Transient Transformation System: Using a transient transformation system, we tested the functionality of various genetic constructs in tobacco seedlings, allowing us to quickly validate the key catalytic enzymes involved in the bioluminescent and caffeic acid synthesis pathways. This method significantly shortened experimental cycles.
• Molecular Biology Tools: We applied advanced molecular biology tools to rapidly characterize the function of enzymes involved in the synthetic pathway, providing new methods for characterizing metabolic functions in synthetic biology.

4. Gene Expression Optimization:

• We used transient expression of TAL and C3H to successfully increase the concentration of caffeic acid in the plant's metabolic pathway. This strategy represents a novel application in synthetic biology, enhancing natural bioluminescence mechanisms and laying the groundwork for future biotechnological applications.

5. Detailed Experimental Protocol and Tool Sharing:

• Experimental Protocol: We meticulously documented our entire experimental process and created an experimental manual to help future iGEM teams and researchers replicate or build on our work. This manual is openly accessible and provides step-by-step instructions for constructing bioluminescent plants.

English Version:

Chinese Version:

• Quantitative Tools: We used tools like ImageJ for quantitative analysis of bioluminescence, measuring grayscale values to objectively assess luminescence intensity. This standardized method ensures reproducibility and provides a reliable way to compare results across different experiments.

6. Component and Biological System Improvements:

• Enhanced Caffeic Acid Pathway: By overexpressing genes related to caffeic acid production, we optimized the overall bioluminescence of the plant. This offers a new approach for optimizing other biological systems within synthetic biology.
• Environmental Sensing Module: Our integration of a formaldehyde-responsive module not only enhances the plant's bioluminescent function but also contributes to synthetic biology by providing a new environmental monitoring tool.

• Horticultural Therapy and Bioluminescence: Our bioluminescent plants are designed to improve mental health by combining the calming effects of light with the therapeutic potential of horticulture. These plants offer a form of natural dark therapy, helping alleviate stress, anxiety, and potentially even depression.
• Bioluminescent Environmental Sensors: By integrating formaldehyde-responsive elements, our plants can serve as biosensors for pollutants like formaldehyde, providing a visual indication of toxic environments. This represents a significant step toward the application of synthetic biology in environmental science.

• Outreach and Education: Through community and school outreach, we are raising awareness about the potential of synthetic biology and its applications. We aim to educate the public not only about bioluminescence but also about mental health and the potential therapeutic benefits of integrating biotechnologies with traditional wellness practices.

• Biosafety and Ethical Guidelines: Throughout the project, we adhered to strict biosafety standards, ensuring the responsible development and application of genetically modified plants. We considered the environmental impact, making sure that the modified plants do not pose risks to ecosystems.
• Ethical Responsibility in Synthetic Biology: We take responsibility for evaluating the potential societal impacts of our project, ensuring that it aligns with ethical principles and contributes positively to both science and society.