Integrated Human Practices
Overview
Food security has always been a top concern for humanity. However, large-scale food production often leads to various infections and diseases, significantly reducing crop yields. Among these, fungal infections caused by Rhizoctonia solani, a root-nodule fungus, are particularly severe. To address this challenge, our project is dedicated to developing a double-stranded RNA (dsRNA) solution specifically targeting Rhizoctonia solani, ensuring its harmlessness to the environment and safety for human health. These efforts aim to mitigate the impact of diseases on the yield of corn and other crops. Our project has gained public recognition and demonstrates a sense of responsibility towards community development. Despite skepticism about the reliability and safety of synthetic products, this indicates that there is still much work to be done in the practical application of this technology. We need to rigorously test and improve our dsRNA solution to ensure its effectiveness and safety. Our approach is based on extensive research, expert consultation, education, and communication, which are interconnected and provide valuable contributions to the improvement of our project. All activities conducted throughout the project are also based on solid research and collaboration with experts. The advice and feedback from all professors, agricultural experts, and professionals in related fields have made a valuable impact on the improvement, evaluation, and development of our project. Their knowledge and suggestions have helped us deepen our understanding of the objectives and shape our final product.
Through the efforts of HP, we can accomplish the following:
Development of a feasible dsRNA solution: By extensively engaging with professionals associated with the project, we aim to develop an effective dsRNA-based treatment for rhizobia to prevent significant reductions in food crop yields. This approach is eco-friendly, safe for human health, and economically viable. Public Influence: Through comprehensive research and the gathering of various information, we have gained a better understanding of the critical role that synthetic biology plays in agricultural production. However, for the public, a significant portion lacks knowledge about this area. We are launching extensive public awareness campaigns in the agricultural and health sectors to increase awareness and support for our project.
The following mind map detail our pathway towards integrated human practices and activities related to our project.
// IHP Structure //
Section I
To have a better setting for public information collection and design of our project, we start with brainstorming and background research, such we are setting different aspects for us to investigate, both through statistical analysis and technological methods. Also, we conduct surveys and interviews to have a deeper understanding of our project, with an analysis of SWOT.
Literature Research
Background
Corn as an important food crop and feed crop in our country, and in our team, we were discussing the topic when we are thinking about food, since everyone in our team seems love eating, and talking about on the topic of food, the crops are an important part in our diet. In the world, almost all of the countries use the corns as one of their main sources for food. According to the US Department of Agriculture, China’s corn production will account for 23.02% in 2020, ranking second in the world. China is also one of the world’s largest corn consumers, with consumption reaching 289 million tons in 2020.
![Global maize production, consumption, and trade: trends and R&D implications | Food [1]](https://static.igem.wiki/teams/5474/i2.png)
The large production of food will always contribute to some infections and diseases, and by having such pests or diseases, the yield of production of corns drops rapidly. So, through research and finding, we found one important infection that caused the massive decrease in corn production – Corn sheath. It is a fungal disease caused by Rhizoctonia Solani. In recent years, with the dense production of corns, continuous cropping in main corn producing areas for many years and the lack of application of high-quality and high-resistance corn varieties, the incidence of corn sheath blight has accelerated, with the incidence reaching 40% in general years and 100% in severe cases, resulting in a rapid decline in corn yield and quality.
RNA interference
![Geographical distribution of Rhizoctonia solani [3]](https://static.igem.wiki/teams/5474/i3.png)
RNA interference (RNAi) is an endogenous-specific gene post-transcriptional silencing mechanism triggered by double-stranded RNA (dsRNA). After dsRNA is introduced into the organism, it is broken down into 21~23 bp small interfering RNA (siRNA) by RNase III called Dicer in the cell. The siRNA binds to the target mRNA under the action of the silencing complex (RISC) and is sequence-specifically degraded. Target mRNA prevents the synthesis of corresponding protein products, causing the loss of function of the target gene. RNA interference (RNAi) is a gene expression regulation mechanism conserved in eukaryotes such as animals, plants, fungi, and nematodes. Inhibiting target gene expression mainly through nucleic acid sequence-specific interactions is an important defense mechanism for eukaryotes to prevent viral infection, prevent the invasion of foreign nucleic acids such as transposons, and regulate gene expression. In recent years, it has become possible to control plant diseases, insect pests and fungal diseases by inhibiting functional gene expression through RNAi, which has also opened up new ways to prevent and control postharvest fungal diseases of fruits and vegetables. Spray-induced gene silencing (SIGS) is an RNA interference-based approach that involves synthesizing target double-stranded RNA (dsRNA) in vitro and spraying it onto the surface of fruits and vegetables that are infested with pathogens. This method inhibits the target gene’s transcription and translation, providing a means for the prevention and treatment of pathogens. SIGS is characterized by its simplicity of operation and does not rely on the host’s gene transformation system. The primary component is dsRNA, a biomacromolecule that is non-toxic and non-residual. In the field of post-harvest disease control for fruits and vegetables, SIGS holds great promise for application. Our project employs this method.
How to prepare the dsRNA?
The method of preparing dsRNA is usually to separately transcribe and synthesize sense and antisense RNA sequences, and then anneal to generate dsRNA fragments. This method has a complicated process, is expensive, and the quality of the dsRNA obtained is not high. Commercial kits emerged as the times require, but the high price of dsRNA synthesis has limited the large-scale application of SIGS in the field of fruit and vegetable disease control. Low-cost microbial reactors (such as E. coli) are easy to operate and can be rapidly propagated on cheap media, making mass production of dsRNA possible.
Ethics, Value, Safety of using dsRNA
The use of dsRNA technology to control the fungal pathogen Rhizoctonia solani presents both opportunities and challenges. Using this innovative solution, we investigated ethical, potential benefits and safety measures.
(1) Ethics
Environmental Impact: The use of dsRNA technology in agriculture may have environmental implications. Concerns include the potential for non-target effects if dsRNA’s specificity is not strong, which could affect beneficial fungi and microbes. Additionally, there are worries about the long-term persistence and spread of dsRNA in the environment. Researchers and policymakers must carefully evaluate these environmental impacts and develop strategies to ensure that dsRNA-based solutions do not disrupt the natural balance of agricultural ecosystems.
Food Safety and Consumer Trust: The application of dsRNA technology to food crops raises significant concerns about food safety and consumer trust. The use of this technology must be transparent, and any potential risks to human or animal health through the food chain must be thoroughly tested and proven safe. Consumers should have the right to make informed choices, which may require clear labeling requirements for crops treated with dsRNA entering the food supply. Establishing consumer trust is crucial because public perceptions of the safety and ethical standards of this technology will greatly influence its broader acceptance and adoption.
Responsible Innovation: Deploying dsRNA technology for agricultural applications requires a responsible approach to innovation. This includes conducting thorough risk assessments and implementing robust mitigation strategies to minimize unintended consequences. It is also important to ensure equitable access to the technology to prevent monopolistic control, as this could restrict the benefits it brings to farmers and communities. Involving various stakeholders, including farmers, local communities, and relevant experts, in the development and deployment of dsRNA solutions helps address issues and promotes inclusive and responsible innovation.
Ethical Research Practices: Research and development of solutions based on dsRNA must adhere to the highest ethical standards. This includes following established biosafety protocols, obtaining necessary regulatory approvals, and adhering to principles of scientific integrity and responsible research conduct. Researchers must also navigate complex intellectual property issues to balance innovation needs with ensuring the widespread accessibility of the technology. Maintaining ethical research practices is essential for building trust and ensuring responsible advancement of this powerful biotechnology.
(2) Value
Targeted Disease Control: Traditional fungicides have limited effectiveness and can lead to chemical residues and resistance. In contrast, dsRNA technology provides more effective disease control by precisely silencing genes essential to the pathogen. Compared to broad-spectrum fungicides, dsRNA technology is more precise and has less impact on the environment. It targets specific pathogens and reduces the impact on non-target organisms, helping to maintain healthy and balanced agro-ecosystems.
Sustainable management: dsRNA technology can help develop more sustainable disease management strategies by targeting genetic weaknesses of pathogens to provide long-lasting protection, reducing the reuse of chemical fungicides, lowering input costs for farmers, and reducing reliance on harmful synthetic compounds.
Adaptability: Rhizobia adapt to a wide range of fungicides and develop resistance. dsRNA technology’s flexibility allows rapid development of new sequences to cope with change and maintain effective disease control.
Yield and quality: Effective control of rhizobia can significantly improve crop yield and quality, protect plants from the destructive effects of fungal pathogens, increase farmers’ economic returns, and improve food security.
(3) Safety
Target specificity: Ensuring the target specificity of dsRNA is one of the key safety considerations in using this technology. Rigorous testing and validation must be conducted to demonstrate that dsRNA does not have unintended effects on non-target organisms, including beneficial fungi, microorganisms, or other organisms in the agroecosystem.
Environmental fate and persistence: Studying the degradation and accumulation of dsRNAs in the environment, such as soil and water bodies, is critical to assessing their long-term safety. Understanding their fate and persistence in the environment can help minimize ecological risks.
Animal Health: A comprehensive food safety assessment is needed to ensure that dsRNA-based solutions do not pose a risk to human or animal health through the food chain. This includes assessing the uptake, metabolism and bioaccumulation potential of dsRNAs and conducting toxicology studies to validate their safety.
Regulatory Compliance: The development and deployment of dsRNA-based solutions for agricultural applications must comply with the relevant regulatory framework and be approved by the authorities. This ensures that the technology meets safety standards and potential risks are addressed prior to commercial application.
Monitoring and stewardship: Ongoing monitoring and stewardship programs are essential to identify and respond to any safety issues that may arise during commercial use. Establishing effective post-deployment monitoring and reporting mechanisms helps to maintain the safety of the technology over time. Safe and responsible use of dsRNA for Rhizoctonia solani control can be achieved by prioritizing target specificity, environmental fate, food safety, regulatory compliance, and monitoring systems. Continued research, stakeholder engagement and commitment to safety are essential to realize the full potential of this innovative biotechnology.
SWOT Analysis
Survey
Purpose
The purpose of this public survey is to gather community feedback on the agricultural product corns. All the insights will help us understand public perceptions, consumption behaviors, and preferences regarding the taste, pesticides residue and newly developed dsRNA product. All the responses will be invaluable in guiding future developments and regulatory practices related to this product and helping us better develop a solution towards the corns sheath blight. We don’t just want to understand people’s view and preferences but also popularizing our own products and requesting for opinions and consumers expectations.
Result Analysis
We investigated age as well as their work occupation and whether they had expertise in agriculture. It shows that the survey population is well distributed in terms of age and covers a wide range of occupations. The next question was about the frequency of consumption of maize in a week, which we hoped to understand consumer concerns and preferences for maize as a major agricultural product. The survey showed that most consumers purchase maize at least once a week, with about 80% of the respondents opting for 1-4 times, indicating the importance of maize in the agricultural and food industry.
In a survey analyzing consumer behavior and preferences for purchasing corn, participants were asked to rank the factors they consider when purchasing corn. The results showed that taste and the safety of agricultural products are each considered equally important by consumers, with the origin of the corn being the least influential factor in their purchasing decisions. Taste and food safety are the top priorities for consumers.
The following question concerns the daily price of corn, which consumers can generally accept at around 6 yuan per unit. This result aids in understanding the current market price of corn and in adjusting the pricing for our dsRNA pesticide products. We need to manage the cost and selling price of our products. Consequently, we strive to keep our synthetic pesticide prices affordable, aligning with the current market conditions. Next, we surveyed consumers’ preferences for different types of corn. From the data tables and charts, it is evident that consumers have no clear preference for the variety of corn. Yellow and white corn each have 33% of supporters, and about 38% of consumers like both colors.
The next question addressed consumers’ concerns about pesticide residues, with approximately 48% expressing concern and 52% showing no concern. This suggests that about half of the population is concerned about pesticide residues, while the other half seems to believe they are harmless and not a concern. We may need to educate and raise awareness about the harms and health risks associated with pesticide residues, which also helps us understand the public’s knowledge of pesticides.
We posed a direct question about our product: “Would you purchase a biopesticide that is effective, environmentally friendly, and an alternative to traditional pesticides, even if it is slightly more expensive?” The results indicate that our product has gained public recognition, with many expressing a positive attitude towards our biopesticide, with approximately 70.86% indicating they might buy it. This positive feedback suggests a promising future for our product and indicates potential for greater development.
The last question explored broader applications for our dsRNA products, such as applications in shampoos to fight fungus. According to our survey results, more than half of the participants would consider purchasing such synthetic biology products. This shows that our products have the potential for a wide range of applications in several fields. This feedback helps to improve our product understanding and future strategic planning.
Conclusion
The results of the survey indicate that maize is an agricultural product that is in high demand by the public. Although price sensitivity is an important consideration, consumers prefer quality, suggesting that the use of synthetic biology to improve crop quality could be successful in the marketplace. Public perception of pesticide residues is mixed, suggesting the need for better health education. However, there is a positive attitude towards biotechnology, as an alternative to traditional pesticides, which shows the promising future of innovative agricultural solutions in the market. In addition, the survey explored the possibility of applying biotechnology to other products, such as shampoos, and the results were encouraging. Overall, the results of these surveys will guide our dsRNA pesticide development, focusing on quality and safety to meet consumer expectations.
Interview: Farmers from ChongQing Village
To understand consumer demands and preferences for pesticide product types and the existing issues with current pesticide products, our team conducted two interviews in a corn-growing village in Chongqing. We randomly interviewed some farmers.
During interviews with a number of farmers, we found that the seedling stage is a sensitive period during corn cultivation that is susceptible to a number of diseases. During this stage, farmers use pesticides such as herbicides and insecticides, but not very frequently. At the same time, they are also very concerned about environmental protection during the planting process and hope that the use of pesticides will reduce the pollution of the environment. The farmers held a cautious attitude towards new biopesticides, and their main concerns included the safety of use, effectiveness, and price. These comments guided us to pay great attention to designing highly targeted gene sequences in the subsequent experimental design, and to adopt lower-cost methods in the experimental process to control the cost.
Section II
Through research and literature review, we established our basic approach: the RsCAT enzyme encoded by the rhizobia can clear hydrogen peroxide, thereby reducing the plant’s immune response and facilitating Rhizoctonia Solani infection. Building on this discovery, we designed a dsRNA sequence that can target and silence RsCAT, and constructed its recombinant expression vector. We then transformed this expression vector into E. coli HT115(DE3), successfully expressing the targeted dsRNA, which we named RsCAT-dsRNA. As we began our laboratory work, we conducted more interviews with professionals. These in-depth discussions provided us with valuable insights and guidance for our experiments. Throughout the project implementation, we will continue to receive support and assistance from these experts and professionals to make necessary adjustments and improvements.
Interview: Professor Zhou Tan
Prof. Tan is an associate professor at the College of Life and Environmental Sciences, Hangzhou Normal University. Expert in the field of molecular biology.
Key points
In the interview, Prof. Tan said that it is very important to understand the mechanism of action of the fungus, such as whether there are genes that affect the growth and health of the leaves, or whether toxins are released. In addition, the professor mentioned that by changing the concentration of dsRNA and controlling the length of DNA, we can obtain stable dsRNA and reduce the manufacturing cost of products. In addition, Prof. Tan talked about how targeting specific genes or DNA can increase the effectiveness of a product and reduce the harm caused to the environment or humans.
Interview : Dr. William Mills
William (Billy) Mills completed his Ph.D. in Biochemistry at the Johns Hopkins University School of Medicine in Baltimore, Maryland, and began serving as an Assistant Professor in the Department of Science at Mount Saint Mary’s University in the fall of 2023.
Key points
When staple crops are damaged or infected, their yields can decrease significantly. The use of this dsRNA product can effectively prevent large-scale crop yield loss, making its development and application crucial. The key to the experiment lies in selecting appropriate target gene sequences to minimize off-target effects. When dsRNA technology is commercialized, cost control becomes a significant issue. When considering cost accounting, it is important to first invest in the laboratory and then conduct further research in the external market. Negotiating and discussing with large companies can increase the chances of successful product launch. Through conversations with Professor Mills, we gained valuable insights and adjusted our experiments accordingly to explore methods for reducing costs.
Academical Meet Up
The academic gathering between high school and university iGEM teams provides a valuable platform for learning, exchange, and collaboration. This event aims to promote the exchange of ideas, feedback, and suggestions to foster the common growth and development of all participants. Each team introduces its project, outlining its objectives, methods, and preliminary results, many of which have provided inspiration in terms of both topic selection and experimental methods. Following the project presentations is a Q&A session to facilitate in-depth discussion and exchange. Teams receive detailed feedback from peers and experts, which helps identify strengths, weaknesses, and areas for improvement. Furthermore, experts provide practical advice on technical aspects, project management, and potential future directions.
Team Q&A
Q：How efficient is the product?
A：Current experiments indicate that E. coli can efficiently express dsRNA, and we have extracted dsRNA with a relatively high concentration. The disease resistance experiments are still ongoing.
Q: How do you prove that the product targets a specific gene?
A: That’s a great question. By measuring the content of hydrogen peroxide in tobacco leaves and comparing it with tobacco dsRNA. The results show that the tobacco leaves sprayed with dsRNA have more brown DAB staining (indicating the presence of hydrogen peroxide) than those sprayed with water. Therefore, it indicates that the dsRNA is functional. We will also design qRT-PCR experiments later to test the expression of the CAT gene.
Q: How do you solve the stability issue of dsRNA?
A: By conducting experiments to place dsRNA for different durations and then detecting its stability through gel electrophoresis, which is still ongoing. We have found in literature that encapsulating dsRNA in nanomaterials can make it more stable. We will continue to advance the experiments to see how to improve this aspect.
The academic meeting between high school and university iGEM teams and industry experts has been a great success. It provides a unique opportunity for knowledge sharing, feedback, and collaboration, significantly improving project quality and fostering a supportive community of practice. This event fully demonstrates the benefits of collective learning and cooperation, achieving a win-win for all participants. We will also revise the experiments and improve them based on the advice of the experts and professors.
Section III
As the project progresses into the later stages, we continue to actively seek feedback and suggestions in order to better evaluate our work and plan the next steps. Through extensive interviews and exchanges with industry professionals, we have clearly identified our strengths and future development strategies. More importantly, through this process of evaluation and communication, we have also gained insights into our weaknesses and shortcomings, providing valuable opportunities for further optimization of the product.
Interview: Professor Chaofeng Li
Professor Li, a Ph.D. from the University of Chinese Academy of Sciences, has been working at the College of Agronomy and Biotechnology at Southwest University as an Associate Professor since 2022. His research focuses on maize lodging resistance, stress tolerance, and maize nutrition.
Key points
Professor Li expressed support for our strategy and during the conversation, we shared the experimental results that the project has achieved. He acknowledged our preliminary work, pointing out that while traditional pesticides have a significant impact on crop diseases, they often pose greater harm to the environment and can lead to the development of resistance in pests. He emphasized that dsRNA technology has advantages that traditional insecticides do not possess. At the same time, Professor Li also raised several points that our project needs to pay attention to. First, given the characteristics of dsRNA, the timing and location of applying the product to crops are crucial, as this will directly affect the efficiency and effectiveness of dsRNA. Therefore, we must pay attention to the conditions and location of application. The concentration of dsRNA used is also critical; we need to determine the optimal concentration because using a concentration that is too low may have minimal impact on disease resistance, while a concentration that is too high will result in increased costs. Another issue to consider is the versatility of the product’s application. We need to know whether this dsRNA technology can also be effective on other crops, such as wheat, rice, and potatoes. Through this interview, we have gained a deeper understanding of the project and the product, which will help us to improve the product and enhance its application value.
Interview: Researcher Chunfang Zhao
Dr. Zhao works at the Grain Crops Research Institute of the Jiangsu Academy of Agricultural Sciences. She has long been engaged in plant functional gene research and is deeply involved in the genetic research of high-quality rice varieties.
Key points
The final application of the product will be in the field, so we need to communicate with professionals in the agricultural industry, which will help us further improve the product. Dr. Zhao, as a frontline worker who has worked in the field for many years, has provided us with a lot of guidance. Dr. Zhao mentioned that in order to accurately evaluate the efficacy of bio-pesticides, large-scale field trials must be conducted. These trials will provide us with key data on the effectiveness of bio-pesticides in actual agricultural conditions. By observing the performance of the product in large-scale applications, we can assess its impact on crop yield and quality, and thereby determine its practical feasibility and potential benefits for widespread use in agriculture. While laboratory results can prove the effect of dsRNA, external environmental factors, such as temperature, humidity, and other conditions, as well as the presence of fertilizers, may affect the function of dsRNA, and these are uncertain. Another issue that needs to be considered is the widespread applicability of the product, including the functional verification on other food crops and vegetables. In summary, in our discussion with Dr. Zhao Chunfang, she emphasized the importance of conducting large-scale field trials and made important suggestions on applying the product to more crops to enhance market competitiveness.
Interview: Dr. Roger Worthington
Dr. Roger Worthington, he is a consultant in the fields of ethics, global health and sustainable development. He is co-chair of the Sustainable Development Goals Publishers’ Compact Fellows. He regularly speaks at United Nations conferences on health, ethics and sustainable development, runs professional development courses for NHS doctors and mentors young academics from around the world.
Key points
The issues related to public health and ethics in the project should not only be investigated at the initial stage of the project but should be continuously addressed throughout the project’s progression. Through interviews and discussions with Dr. Worsington, we have been able to identify the ethical aspects that require particular attention during the project. Whether the crops are used for feeding livestock or for human consumption, it is essential to clearly define the target crops, as this has a significant impact on economic benefits and crop quality. Considering factors such as climate conditions, population density, religious beliefs, and lifestyle habits in different regions of the world, all of these will influence the application of dsRNA bio-pesticides. The project needs to focus not only on issues during the pesticide application phase but also on the potential environmental pollution that may occur during the production process. Dr. Worsington has provided insights from multiple perspectives, which has deepened our understanding of the project and offered valuable guidance for future planning.
Interview 8: Professor Weimin Hu
Professor Weimin Hu, from Zhejiang University from the College of Agricultural and Biology Technology, the department of Agriculture. Now working towards the field of crop germplasm innovation and seed engineering, also genetics and breeding of maize.
Key points
Our conversation with Dr. Hu provided valuable guidance for the future development and practical application of the dsRNA-based pesticide targeting rhizobia. Dr. Hu emphasized that our product must align with government environmental protection regulations to ensure it meets the strict standards of green agriculture. He stressed that our pesticide should demonstrate significant efficacy, ideally preventing at least 75% of the target diseases. Additionally, Dr. Hu mentioned that reliance solely on laboratory assessments is insufficient; field trials are crucial for validating the effectiveness and adaptability of our product in different agricultural environments. This feedback will guide the next steps of our project, ensuring that our innovative solution meets environmental requirements and is effective in field practice.
Summary
Agriculture is not only the foundation of society but also the lifeline of the economy. It is not only about food production but also involves environmental protection and resource conservation. In our preliminary research, we noted the significant harm that corn sheath rot poses to corn production, which became the starting point for our project. Through literature research, we learned about the enormous potential of using SIGS technology to spray dsRNA onto plants as a new type of pesticide for disease control. Through SWOT analysis, questionnaires, and in-depth interviews with farmers in rural areas, we gradually clarified the needs and knew the focus of our experiments.
In designing the experiment, we paid special attention to safety, product effectiveness, and cost. During the experimental process, in order to meet the demands for product quality and price, we specifically designed experimental plans to find a production method that is low in cost and meets quality standards. Additionally, in our conversation with Professor Tan, he guided us to optimize the selection of target fragments to enhance product effectiveness. At the meetup session, the stability of dsRNA was a common concern. Through more effective target fragment design and optimization of experimental methods, our experimental results showed that RsCAT-dsRNA has good disease resistance and relatively low cost.
In later discussions with experts, we also received valuable advice and clarified the direction for our next steps. For products that are about to be applied in the field, field experiments are indispensable, and safety is always the top priority in production and use.
The image above shows our visit to the Ecological Agriculture Experimental Station of Zhejiang University, where we witnessed the spectacular large glass greenhouses and field experimental scenes. We deeply felt the cutting-edge research and innovative technologies that researchers are conducting, which are gradually transforming traditional agricultural methods. These research results have not only significantly increased crop yields and quality but also contributed to environmental protection. In the face of global challenges related to food security, climate change, and sustainable development, these achievements are particularly important. Our work will continue!
Reference
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