Figure 1. Power-Interest Matrix.
Farmers are located in the lower reaches of the pesticide value chain, and as victims of nematode diseases as well as consumers of pesticides, communication with farmers is necessary.
By knowing the extent of farmers' understanding of nematode diseases, we are able to provide them with targeted knowledge of agricultural production.
By understanding farmers' priorities when choosing pesticides, we identified the project's product pain points and the core needs of consumers.
Director of the Village Committee of Baxiandong Village, Huilongxu Town
We set our sights on one of our team members' hometown, Yongzhou, Hunan Province, where the citrus industry is well developed and variety and quantity of biopesticides are used.
We hope to establish exchanges with farmers here and learn first-hand information about biopesticides and nematode diseases, starting from the downstream of the pesticide value chain. We selected the director of the village committee to conduct interviews for high-reliability and high-efficiency information.
Content:
(1)The region is primarily cultivated with citrus and other cash crops, with biopesticides being used frequently. The director noted that the main reason for selecting pesticides is based on actual needs. Additionally, they prioritize biological pesticides and will resort to chemical pesticides only when absolutely necessary.
(2)Farmers in the area often apply biopesticides in the same manner as chemical pesticides, such as through spraying. However, if our products require improvement through methods like seed dressing, farmers are willing to adopt them as long as the results are satisfactory.
(3)The director mentions the major drawbacks of biopesticides and chemical pesticides: chemical pesticides are prone to drug resistance, biopesticides are slightly more expensive, and the efficacy of both is unstable in adverse weather such as high temperatures.
(4)Farmers are generally aware of the increasing restrictions on chemical pesticides and the increasing support for biopesticides. And they can perceive that the current pesticides are becoming more and more low-toxicity and environmentally friendly.
(5)Important vegetation and crops in the area have been affected by nematodes, raising concerns among farmers regarding nematode disease management. However, the effectiveness of chemical pesticides has been limited, and common physical methods can easily damage citrus roots, resulting in poor treatment outcomes.
Reflection:
(1)This area is primarily cultivated with cash crops. Although the cost of biopesticides is slightly higher than that of chemical pesticides, their overall cost-effectiveness remains high. Consequently, farmers choose pesticides based on actual needs, and as long as the products are effective, they are willing to adopt more advanced techniques.
(2)During our discussions, we discovered that the spread of another rampant disease, Citrus Yellow Shoot, in the region may also be linked to nematodes. This connection was later confirmed through the team's further communication with the academic community.
(3)The director has high hopes for our success in managing diseases and contributing to the development of our hometown. The farmers' expectations deeply moved our team members, reinforcing our understanding of the project's significant relevance.
Reflection
We found that plant nematode diseases are indeed widespread, serious, and have far-reaching implications; however, there are significant differences in farmers' understanding of these diseases across various regions.
For instance, in Changle County, Shandong Province—a major vegetable cultivation area—local farmers possess a deeper understanding of nematode diseases and have developed a set of effective, environmentally friendly physical control methods.
Conversely, we discovered that farmers in some areas lack awareness of the characteristics and transmission mechanisms of plant nematode diseases, as well as how to implement targeted prevention and treatment strategies.
In response, we summarized and analyzed methods for the prevention, identification, and control of nematodes, sharing this agricultural knowledge with regions where awareness is low. (click here for details)
Moreover, we recognized that farmers' simple wishes center around good crops, happy families, and respect. Therefore, in our future interactions with farmers, we will uphold the spirit of seeking truth from facts, demonstrate genuine concern, and promote equality and respect.
COLLEGE OF PLANT SCIENCE & TECHNOLOGY OF HUA ZHONG AGRICULTURAL UNIVERSITY
Research Interests: The molecular mechanism of the nematode infection and interaction between plants and nematodes, mainly focusing on soybean cyst nematodes and rice-grasses root-knot nematodes.
The primary goal of our communication was to investigate the feasibility of utilizing salicylic acid as an input signal, confirming that our genetic circuit was being activated as anticipated. Besides, we are also going to ask her about the parasitic process of root-knot nematode to help improve the model.
Content:
(1) She pointed out that our engineered bacteria must be endophytic, or be able to exchange substances with plant roots, in order to sense the upregulation of plant salicylic acid, and let the VLP produced by cleavage enter the plant root system, and finally achieve contact with nematodes.
(2) When root-knot nematodes invade plants, they try to avoid damaging the cells, crossing from the meristem to the elongation zone until they reach the nutrient-rich vascular bundles and settling [1], which takes about a month. Therefore, it may take a long time for our engineered bacteria to feel the upregulation of salicylic acid and initiate the subsequent circuit, and it is difficult to achieve a timely response.
(3) The distribution of plant nematodes was hierarchical in soil depth, and its number and diversity were most abundant in the soil surface layer (0-30 cm), and the number gradually decreased with the increase of depth.
(4) Caenorhabditis elegans, a commonly used model animal, uses microorganisms as a source of nutrition and is widely distributed in soil. However, many plant nematodes can hardly use the nutrients of microorganisms, so they will only multiply in large numbers after parasitic plants.
(5) In addition to harmful plant parasitic nematodes, there are also various beneficial nematodes that live freely in the soil. We need to take beneficial nematodes into account in the scope of the project's products, and be aware that our products have the potential to affect other organisms that maintain the ecological balance of the soil.
(6) She pointed out that the main reason for the slow development of biopesticides is due to poor effectiveness. In addition to this, she also raised an aspect that we had not considered – the protection of patent rights. For example, some biopesticides are at risk of being stolen by others in the environment after they are applied. In addition, after the expiration of the patent term of biopesticides, there will be some imitations with poor effect. These factors will affect the enthusiasm for biopesticide research and development.
(7) The amount of SA in different plants and different tissues of the same plant is different, and we need to consider whether the circuit of the project can be applied to different plants. This provided the inspiration for our salicylic acid biosensor model. (click here for details)
Figure 2. Communication with Xiaoli Guo.
Reflection:
(1) After reviewing the literature, we learned that Bacillus velezensis is endophytic, which can not only achieve the expected goals of the circuit, but also that the bacterium itself has beneficial properties that promote plant growth and antifungal, which can complement the expected effects of the project [2].
(2) We need to consider the impact of the project on other organisms in the environment, such as microorganisms and beneficial nematodes. We found ourselves needing more information about this, so we decided to seek help from Dr. Gaofeng Wang, who studies plant-nematode interactions.
(3) For how to deal with the differences in salicylic acid in different plants, we need to seek more professional advice. We decided to consult with Professor Shunping Yan, who studies plant hormones and plant injury response mechanisms.
(4) Our project has designed a suicide circuit, which contributes to the protection of property rights in addition to environmental protection considerations.
COLLEGE OF PLANT SCIENCE & TECHNOLOGY OF HUA ZHONG AGRICULTURAL UNIVERSITY
Research Interests: Molecular mechanism of plant-nematode interaction and green prevention and control technology
The primary purpose of our communication was to assess the potential impact of the project on other organisms in the environment, gather more information about plant nematodes and free-living nematodes, and engage in an in-depth discussion on green control technologies for nematodes.
Content:
(1) This group of nematodes includes not only parasitic nematodes that are highly detrimental to plants but also beneficial, free-living nematodes that are essential components of the soil community and occupy key trophic levels in the soil food web. Therefore, we must consider the impact of our project on both beneficial nematodes and their predators.
(2) Professor Wang suggested selecting an insecticidal protein with a low molecular weight and mutating its degradation site to enhance its stability, thereby preventing it from being affected by the complex soil environment, such as decomposition by microorganisms. Additionally, we can utilize models to predict and evaluate its stability, ensuring it maintains a more stable spatial structure.
(3) In a complex soil environment, the primary environmental factors to consider include ultraviolet light and temperature (above ground), as well as pH and salinity (below ground).
(4) Nematodes possess a dormant mechanism in the soil; for instance, cyst nematodes can survive for up to ten years after forming cysts. Flooding can effectively reduce nematode populations, as evidenced by the lower numbers found in paddy fields compared to drylands. This reduction may be due to flooding-induced increases in reactive oxygen species in plants, which can inhibit nematode parasitism by creating oxidative stress.
(5) Mr. Wang outlined several common methods for controlling nematodes. The first method involves the use of highly toxic pesticides which were banned in China. The second method utilizes the new nematicidal molecule fluopyramide, but its high cost makes it viable only for cash crops. Furthermore, widespread use has led to nematode resistance against this chemical.Another approach is to develop crop varieties with enhanced stress resistance and improved plant immunity, achieving a balance between plant growth and nutrient absorption rates by nematodes. Lastly, employing nematode-resistant rootstocks during grafting is an option, though this method may result in root rot.
(6) Mr. Wang noted that some farmers may not fully understand plant nematode diseases. Many farmers lacking professional knowledge often observe stunted or abnormally developed plants and instinctively attribute these issues to insufficient water and fertilizer. While supplementing these resources can yield temporary improvements, it does not address the root cause of the problem, as discussed earlier. Consequently, many farmers remain unaware of the impact of plant nematodes. This highlights the importance of our dialogue with farmers to enhance their understanding. (click here for details)
(7) Mr. Wang also shared his insights on nematode control, suggesting that since plants have a certain level of tolerance to nematode parasitism, it may not be necessary to completely eliminate or block nematodes. Instead, we can focus on enhancing the disease resistance of plants. This can be achieved by screening for varieties with greater stress resistance or by inhibiting the formation of nematode feeding sites. Such strategies can effectively help plants combat nematode infestations.
(8) Drawing from his experience in actual production, Mr. Wang emphasized that effectiveness and simplicity are the primary factors farmers consider when selecting pesticides. 'Simplicity' encompasses aspects such as tolerant application timing, minimal application frequency, and lower accuracy requirements. 'Effectiveness' refers to the speed of action and duration of effect.
(9) Mr. Wang also reminded us to consider cost; otherwise, only farmers cultivating high-value crops would be able to afford these products. Finally, he pointed out the importance of transportation and storage for biopesticides. In many countries, agricultural capitalists work directly with manufacturers to arrange timely deliveries of pesticides in large quantities, reducing the need for prolonged storage. However, Chinese farmers often require pesticides to be stored for extended periods, so our products must be designed for long-term storage without significant loss of effectiveness over time.
Figure 3. Communication with Gaofeng Wang.
Reflection:
(1) The safety and efficacy of trans-aconitic acid, an important insecticide substance in our project, has been widely verified in the Chinese biopesticide market, proving that our project will not cause adverse environmental effects due to killing too many beneficial nematodes.
(2) The salicylate content fluctuates during the growth of plants, and there may also be other abiotic or biotic factors that activate the damage response of plants, resulting in the upregulation of salicylate and thus the activation of our circuit. But because the main role of our Plant Guard is to activate plant immunity, it can also play a protective effect on plants.
(3) We selected the smallest possible Cry6Aa protein according to the recommendations, and at the same time, several modules predicted the structures of various proteins, which played a guiding role in the project [3].
(4) The nematicidal methods on the market have great defects, and there is an urgent need for a universal green biopesticide, so our broad-spectrum project products have quite bright market prospects.
(5) Based on the information provided by the teacher, we improved the simulation of nematode distribution by modeling.
(6) Communication with experts confirms our earlier findings from farmers. In Hunan, we discovered that Citrus Yellow Shoot disease is severe, with nematode disease significantly contributing to its spread. Nematodes act as carriers of pathogenic bacteria and exacerbate plant abnormalities, indicating a close relationship between Citrus Yellow Shoot and nematodes. Additionally, plant nematodes can cause dysplasia, leading farmers to mistakenly uproot healthy plants, resulting in economic losses[4].
COLLEGE OF LIFE SCIENCE AND TECHNOLOGY OF HUAZHONG AGRICULTURAL UNIVERSITY
Research Interests: Response mechanism of plant DNA damage, mechanism of plant disease resistance, and plant hormone signal transduction
The primary purpose of our communication was to address the differences in salicylic acid content among various plants. Following inquiries from Professor Xiaoli Guo, we decided to seek further professional advice.
Content:
The amount of salicylic acid varies between different species of plants and different parts of the same plant, for example, in Arabidopsis thaliana, the background level of salicylic acid is relatively low, about between 5 and 30 μg/g fresh weights [5]. The content of SA in rice is between 5 and 30 μg/g fresh weights[6]. If we want to increase the range of applications of our products, we need to take into account the sensitivity of salicylic acid receptors.
Figure 4. Communication with Shunping Yan.
Reflection:
To enhance NahR's sensitivity to SA in order to address potential variations in SA concentration, we performed saturation mutagenesis on residues near the docking site based on NahR's affinity for SA. (click here for details)
Reflection
During our discussions with experts from various fields, they offered questions and suggestions for our project from a professional perspective, effectively promoting its optimization and iteration.
Firstly, they provided numerous recommendations for circuit optimization and modeling improvements based on relevant literature and actual production data, bringing our project closer to real-world production indicators.
Secondly, through the experts' insights, we gained a deeper understanding of plant nematode diseases and biopesticides, enhancing our comprehension of the project.
Finally, the experts affirmed the innovative and exploratory nature of our project design, emphasizing its significant relevance to actual production.
During this process, we received both criticism and recognition from the manufacturing sector, which helped us identify the differences between actual production conditions and the laboratory environment. This insight will enable our projects to better address the needs of our stakeholders.
A high-tech company that develops, produces and sells biopesticides, biofertilizers, microbial feed additives and their preparations.
The company's main products are closely related to our project, and we aim to gather more information about biopesticides and nematode pesticides through this exchange.
Additionally, we visited their production facility in Xiantao, Wuhan, with the goal of making our project more practical.
Content:
(1) Geographical and laboratory environments differ significantly, with regional soil conditions varying in terms of microorganisms, nutrients, pH, humidity, and other factors.
(2) The efficacy is often concentration-dependent. On one hand, we aim for bacteria to colonize leaf surfaces or rhizospheres and form biofilms, so it's important to ensure that the engineered bacteria reach the necessary concentrations. On the other hand, trans-aconitic acid, as a small organic molecule, may be degraded or utilized by other microorganisms, leading to reduced efficacy. Thus, its concentration must also be considered.
(3) We currently plan to apply our products in the form of powder, spray, or seed dressing. If designed as a spray, we must account for environmental factors that could affect protein expression, potentially reducing effectiveness compared to laboratory conditions.
(4) Plant roots grow continuously, especially during periods of rapid growth. During this time, nematodes can invade from the root tips, but our bacterial growth rate may not be fast enough to protect these areas, leaving some zones more vulnerable to nematode attack.
(5) Two major questions in the biopesticide market are: How many bacteria remain? Where are they located? While we can measure bacterial concentration, tracking their distribution in plant roots is more challenging.
Figure 5. Communication with experts(left). visiting the production facility(right).
Reflection:
(1) While the development of biopesticides has been encouraged by the government in recent years, the actual progress of biopesticide development in China may not be as rapid as anticipated.
(2) For our products to be commercially viable, their effectiveness must be carefully considered. This is influenced by various factors, such as gene expression levels and protein stability.
(3) We also need to evaluate the concentration and diffusion of trans-aconitic acid to better confirm our product's effectiveness, which will guide our modeling. (click here for details)
(4) The final form of the product remains a consideration. Spray formulations may lack effectiveness, leading us to explore seed dressing as an alternative. We will continue assessing its potential in future HP activities.
(5) The colonization module in our circuit enhances the expression of GgaA and TapA, which supports biofilm formation and strengthens Bacillus viability in external environments, ultimately improving product efficacy.
(6) Chemical pesticides possess systemic activity, allowing them to spread quickly with root growth and cover all parts of the plant. Biopesticides, however, usually lack this property, which is why their effectiveness can sometimes fall short compared to chemical pesticides.
(7)We aimed to understand the growth and specific distribution of engineered bacteria in the soil, so we planned to produce the corresponding colloidal gold test strips and conceive a device design to help quantify the content of engineered bacteria in soil samples. (click here for details)
The company's primary business involves the research, development, production, and sales of chemical pesticides.
The primary purpose of our communication was to understand the pros and cons of chemical pesticides and biopesticides by engaging with the marketing, R&D, and other departments within the industry. As a chemical pesticide company, they provided an objective and relevant evaluation of our product. Additionally, we aimed to explore the challenges the company faces in transitioning to biopesticide research, in order to inspire our project and enhance its practicality.
Content:
(1) From the perspective of the market, Minister Zhang of the Marketing Department introduced the detailed classification of pesticides and the current sales status of each pesticide. He carefully analyzed the performance of different types of pesticides in the market, including the sales trends of traditional chemical pesticides and biological pesticides.
(2) At the same time, Minister Zhang further revealed the four major links of pesticide sales - manufacturers, agents, retailers and farmers, so that we have a further understanding of the sales channels.
(3) In addition, he emphasized that in the current pesticide market environment, the pursuit of cost performance is the key to the development of pesticides, which is not only related to the economic benefits of enterprises, but also directly affects the use experience of farmers and the sustainability of agricultural production.
(4) Professor Jialong Sun from Qingdao Agricultural University highlighted the challenges of our project after gaining a detailed understanding of it. Currently, chemical pesticides dominate the market due to their high cost-effectiveness, rapid action, and broad application. In contrast, biopesticides remain at a competitive disadvantage due to their slower efficacy and relatively higher costs. This market reality presents a significant challenge for our project.
Figure 6. Communication with Jialong Sun.
(5) Pei Wu, the chief expert of the pesticide quality inspection and evaluation team at the Provincial Institute of Pesticide Science, shared valuable industry insights and professional suggestions that greatly broadened our perspective and offered new development directions for our project. Additionally, she introduced the key evaluation criteria for pesticide registration, which encompass five aspects: physical and chemical properties, pharmacological efficacy, residue levels, toxicological properties, and economic viability.
Figure 7. Communication with Pei Wu.
(6) Finally, we were fortunate to have an in-depth and productive exchange with General Manager Honghua Ren. Mr. Ren offered numerous practical suggestions for our project, providing a strong boost to its progress. Under his professional guidance, we also gained a clearer understanding of the project's future development path.
Figure 8. Communication with Honghua Ren.
Reflection:
(1) Following this exchange, we revisited our previous assumptions. For instance, we once believed that chemical pesticides were inherently more harmful. However, we learned that, with increasingly strict regulations in the pesticide industry, highly toxic or environmentally harmful chemical pesticides are now heavily restricted. There is also a rigorous regulatory system overseeing the production and use of other chemical pesticides.
(2) The number of registered biopesticides using Bacillus velezensis and Bacillus subtilis is considerable, with mature technologies and fierce market competition. This highlights the need for greater innovation in our products to enhance their market competitiveness.
(3) Like many biopesticides, our products may face the challenge of performing well in laboratory settings but being less effective in complex soil environments. Therefore, increased real-world application and continuous optimization are necessary to improve product performance.
(4) We gained a more comprehensive understanding of the pesticide industry from a market perspective. Stakeholders such as manufacturers, agents, retailers, and farmers provided valuable insights for improving our project. Moving forward, we are focused on enhancing cost-effectiveness and improving prevention and control measures, steadily advancing toward higher project goals.
(5) In the '14th Five-Year Plan' for the development of the national pesticide industry, most chemical pesticides are not prioritized. However, many chemical pesticide companies are not planning a rapid transition to the biopesticide industry, and the outlook for biopesticides remains uncertain due to their limited effectiveness.
Reflection
Our communication with the industry reflects our inclusiveness and sense of responsibility. Throughout this process, we integrated diverse perspectives and opinions, engaging with various business entities, including those that may be critical of our project. We addressed some of this feedback in subsequent project optimizations.
This exchange allowed us to gain insights into the project from previously unexplored perspectives, such as market dynamics and corporate R&D. Interactions between the chemical pesticide and biopesticide industries have enriched our background knowledge, deepening our understanding of the differences between these two types of products.
Through discussions with industry experts, we reflected on the effectiveness of our products and incorporated this feedback into our projects. Communication with companies has also enhanced the social responsibility of our initiatives.
Changle County, Weifang City, Shandong Province
We spoke with Ziyun Liu, Deputy Director of the Environmental Protection Bureau, and Xiangyang Feng, Chief of the Agricultural Management Section of the Agriculture and Rural Affairs Bureau, regarding the use of pesticides and the management of nematode diseases in today's agricultural production. During this conversation, team members also introduced Bacillus Gemini from HZAU-China 2024, which received high praise from both officials.
Content:
(1) Before the exchange, we had a solid understanding of China’s recent policies, such as the '14th Five-Year Plan' for Ecological Protection and Pesticide Industry Development, and had gathered cases of ecological degradation caused by excessive pesticide use. Our team raised concerns about environmental monitoring and protection given the widespread use of pesticides.
(2) We emphasized that traditional agricultural practices struggle to meet the demands for green and sustainable development, making ecological protection urgent. Ziyun Liu explained that enforcement against highly toxic pesticides has increased, with restrictions on low-toxicity pesticides and encouragement for eco-friendly alternatives. The government also sets production quotas for pesticide manufacturers to manage at the source.
(3) We also discussed nematode disease management, with Section Chief Feng highlighting the impact on greenhouse crops in Weifang, Shandong Province. He noted the limited effectiveness of current nematode control methods and the challenges of using machinery in greenhouses, making manual labor essential.
(4) In response, we introduced Bacillus Gemini and its mechanism of action. Leaders from the Environmental Protection Bureau and Agriculture and Rural Affairs Bureau praised the project for aligning with agricultural green transformation goals and providing scientific support for Changle County and beyond.
(5) Both sides discussed future R&D, cooperation, and promotion plans, agreeing that industry-academia collaboration and the integration of agricultural technology and environmental protection are crucial for sustainable agriculture. The Agriculture and Rural Affairs Bureau expressed willingness to provide a test base for product trials to support the successful implementation of Bacillus Gemini.
Figure 9. Communication with Ziyun Liu and Xiangyang Feng.
Reflection:
(1) The government continues to restrict the use of chemical pesticides while encouraging the development of environmentally friendly biopesticides. This aligns with the R&D direction and values of our project. In fact, during our exchange, we received encouragement, support, and commitment from government officials.
(2) Current methods for controlling nematode diseases have significant flaws, and there is no universally effective, environmentally friendly solution. Therefore, our product is groundbreaking, and our exploration of plant nematode pesticides holds great significance.
(3) Technicians highlighted the need to consider the depth of plant nematode distribution, offering a new perspective for the modeling phase of our project.
(4) Moving forward, we will prioritize green, lightweight, simple, and efficient solutions as core values in our project development.
Reflection
Through our communication with the government, we gained a deeper understanding of pesticide policies and the enhanced supervision measures in place. We confirmed that our project is encouraged and supported by the state, which boosts our confidence in promoting the project and affirms its potential to address practical problems. This alignment is significant for the green development of the pesticide industry.
Additionally, we discussed the current situation and management of nematode diseases with government officials, further enhancing our understanding of the disease. This knowledge will help us effectively conduct educational initiatives and respond more accurately to societal needs in the future.
Conclusion
By engaging with stakeholders in an equal, honest, and bidirectional manner, we discovered and addressed shortcomings in the project while also uncovering its unique value. With the guidance and assistance of stakeholders, we expanded the project's potential and achieved results beyond expectations.
For example, during discussions with chemical pesticide companies, we learned that current biopesticides face challenges in terms of efficacy and stability. In response to this issue, we considered improving the project by simulating the distribution of nematodes and the diffusion of TAA. Through communication with experts, we found that salicylic acid levels vary across different plant tissues, so we addressed this by simulating the sensitivity of salicylic acid receptors.
Additionally, through exchanges with various parties, we recognized the significant value of Bacillus Gemini in agricultural production, offering new hope to farmers in combating nematode-related diseases.
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