To tackle the limitations of chemical pesticides in citrus aphid control^[1],we developed the APHiGO green management system.We have designed and characterized multiple new parts for RNAi,and developed the open-source dsRNA design platform Aphigem and the based off-target analysis database PostgreSQL to support future RNAi teams.Based on these core components,we have built the CitrusShield intelligent platform to achieve real-time monitoring and precise intervention in a closed-loop management of pest conditions.In terms of entrepreneurial promotion,we have written the RNA pesticide regulation blue book and business plan,actively promoting technology transformation to contribute to a greener,more precise,and sustainable future for agriculture.

At the beginning of the project, we conducted an in-depth survey of existing dsRNA design tools.We found that not only does the iGEM community lack a complete and ready-to-use integrated dsRNA computer-aided design system,but also various design systems available in society have a series of problems such as the absence of reasonable off-target species and the non-public source code,which prevents further functional expansion according to needs.To address these issues,we developed an integrated dsRNA design platform that combines automation,selectability, off-target analysis, and visualization.This platform aims to solve the design problems of siRNA and multi-integrated dsRNA for the iGEM community and other teams.It can automatically complete the design based on the algorithm process using the sequence information provided by users. More importantly,we adhere to the principles of open source and open collaboration,making the platform's source code completely public and transparent.We look forward to future iGEM teams building on this foundation to seamlessly integrate advanced functions such as artificial intelligence,and jointly promote the continuous evolution of this tool,making it a cornerstone in the application of synthetic biology in agriculture.To ensure that this tool is widely and correctly used, we have specially written a detailed user guide, which you can view in its entirety on our Software.The guide provides complete instructions from installation to use,significantly reducing the barrier to user adoption. Learn more in Software.

Fig 1. The figure illustrates the various components of our backend code and the complete operation process.

During the design and development of the project,we noticed that the iGEM community currently lacks an independent and usable off-target analysis database.While some relevant databases exist in the public domain,they are often limited by incomplete species coverage and restricted user access. To address this issue,we independently constructed a PostgreSQL database structured on a "Class-Species-Gene" structure,aimed at integrating with our developed fusion dsRNA design platform.At the same time, the abundant species types and related transcriptome information stored in the database can also provide data support for other iGEM teams.In addition,the database has excellent scalability,allowing future teams to further develop it based on the existing structure. We hope that in the future,it will gradually break through its current role of data input and retrieval,integrating more functionalities to become an independent data processing center.Learn more in Software.

Fig 2. The figure illustrates the three core tables in the database architecture and their specific structures and contents.

In the current field of agricultural pest control, particularly for Toxoptera citricida , traditional methods often rely on chemical pesticides, which pose issues such as environmental pollution, non-target damage, and increased pest resistance^[2]. While some teams in the iGEM community have explored biological control approaches, there is still a lack of a comprehensive closed-loop management system that integrates early detection, precise intervention, and intelligent decision-making. To fill this gap, we have developed the CitrusShield smart pest control platform, aiming to create a scalable and adaptable new paradigm for pest management. This platform not only targets the brown citrus aphid but can also be quickly transferred to the management of other pests by replacing specific components, providing the iGEM community with a standardized method framework from sensing to action.

To achieve this goal, we have constructed a closed-loop system where three core modules work in synergy:
·Early Detection System: Based on engineered Bacillus subtilis sensing of honeydew signals, it releases MeSA gas, with pest monitoring achieved through IoT sensors.
·Trunk Injection RNAi System:Using four generations of RNAi molecules and MS2 virus-like particle-based targeted delivery, it establishes systemic protection in plants.
·Engineered Metarhizium anisopliae System: During the emergency response phase, it performs dual pest-killing actions of “infection + RNAi,” with a light-controlled safety switch to ensure environmental control.

In terms of functional verification, we have experimentally confirmed the feasibility of the system's full chain from signal detection to precise intervention. The engineered Bacillus subtilis successfully synthesized and released methyl salicylate (MeSA) under sucrose induction, and our independently developed gas sensor showed high sensitivity and specificity to this signal, enabling real-time pest detection and wireless transmission^[3]. Additionally, the RNAi molecules we designed effectively silenced aphid target genes, leading to aphid death. The engineered Metarhizium anisopliae can not only stably express RNAi molecules but also successfully infect the body walls of aphids, achieving a dual killing effect of "fungal infection + gene silencing".

In the future, we expect CitrusShield to become a reusable and iterative pest control platform within the iGEM community. By replacing sensor components, RNAi targets, or pathogenic fungal chassis, this framework can quickly adapt to other crop pests, truly realizing "one platform, multiple scenarios" for precise pest management. We also hope that this will help drive the evolution of iGEM projects from isolated innovations to systematic and paradigmatic solutions, promoting a smarter, greener, and more sustainable future for agricultural biological control. Learn more in Citrus Shield.

Fig 3. Overview of the APHiGO Project.

At the Nanjing Synthetic Biology Industry Expo, we identified key issues within China's RNA pesticide regulatory system, such as the lack of classification standards and unclear registration data requirements. These challenges have caused the industrialization process to lag behind international advanced levels. As a result, we wrote Analysis of China's RNA Pesticide Regulations, systematically outlining the relevant clauses and approval processes within the existing pesticide regulatory framework in China. This provides directly applicable regulatory references for future iGEM teams undertaking agricultural synthetic biology projects, helping to reduce the costs of starting from scratch.Learn more in Entrepreneurship.

Building on this foundation, we further compiled the China's RNA Pesticide Industry Development Blue Book: Analysis of International RNA Pesticide Regulations and Considerations Based on China's National Conditions. The goal is to go beyond the perspective of individual projects and, from an industry, policy, and international standpoint, provide a systematic framework and recommendations for the healthy and orderly development of RNA pesticides in China. This blue paper compares the regulatory policies for RNA pesticides in major countries and regions such as the United States, European Union, and Japan, while also proposing localized development strategies based on China's specific context. It offers a reusable framework for future teams to conduct policy analysis or project planning with an international perspective. Learn more in Integrated Human Practices.

Fig.4 Analysis of China's RNA Pesticide Regulations(left) and China's RNA Pesticide Industry Development Blue Book(right)

In terms of technology transfer and intellectual property, we have submitted multiple patent applications related to RNAi core technologies, including A Template for in vitro Transcription Synthesis of dsRNA and Its Application, A dsRNA Automatic Design System and Method Based on Multi-Rule Screening and Off-Target Effect Analysis, and A Multi-Target Double-Stranded RNA Molecule Targeting Toxoptera citricida and Its Application. These patents not only protect our core technological solutions but also provide reusable experience for future iGEM teams in areas such as patent search, application processes, and technology transfer.To assist with this, we have compiled a Patent Application Process Manual to help future iGEM teams reduce the time spent on independent exploration.

In terms of community collaboration, we have actively participated in the iGEM UAM-initiated project The Do-Not List for the iGEM Competition, sharing practical challenges and strategies encountered during the iGEM season. This initiative helps global iGEM teams avoid common obstacles and build more robust and successful synthetic biology projects.

In terms of regional communication, we have summarized the planning and execution experience of the iGEM exchange meeting in South China, forming a detailed exchange meeting plan and points for attention to provide references for future iGEM team Meetups.

In terms of the iHP concept, we innovatively created iHP-themed comics. Using a vivid and intuitive visual format, we explain the concept and practice methods of Integrated Human Practices (iHP), share our understanding and experience of HP, and help global iGEMers better understand and implement responsibility and innovation. Learn more in Integrated Human Practices.

Additionally, we were invited to participate in the iGEM China Team Experience Series interviews, where we shared our practical experience in team building, regional exchanges, and student development. This not only provided a Chinese perspective to the global community but also offered a replicable "growth guide" for new domestic teams.

We have developed a comprehensive business plan for the RNA pesticide product of this project, covering market analysis, cost structure, product iteration path, and investment prospects. This plan provides a clear commercialization roadmap for the APHiGO project and aims to serve as a complete, detailed and reusable reference template for future iGEM teams exploring the commercialization of synthetic biology outcomes. To maximize its community value, we have shared the business plan as open-source on Wiki, helping teams systematically consider the possibility of industrializing scientific research achievements at an early stage of the project and guiding them from lab research to practical application, further strengthening the educational significance of iGEM in "responsible innovation" and "entrepreneurial spirit." Learn more in Entrepreneurship.

Fig.7 Business plan