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HP-Questionnaire investigation

From the following data ,it can be concluded that in the perception of earthworms , "improving the ventilation and permeability of the soil, ability of water storage and protecting fertilizer and soil efficiency on water usage " and "some key processes such as influence of soil organic matter decomposition and nutrient cycling " gain the highest proportion, 79% and 88%, respectively, while the other understanding is not in the minority, the proportion of more than 45%. Thus it can be seen ,consumers' understanding of earthworms is relatively clear and our research is related to it------the earthworm intestinal bacterial species modification project.

Fig.1 People's congnition on functions of earthworms

From the perspective of earthworm product recognition and market demand, more than 60% of people know a lot about the effects of heavy metals in the soil on plants and the environment. However, the understanding of earthworms and environmental pollution related issues is less impressive. Data shows that the number of people who worry about the impact of environmental pollution on the survival of earthworms is only 7% more than the number who don’t think about it so far. It can be seen that a large number of people have not paid attention to this kind of problem. Because of the influence of heavy metals in soil, we want to modify and improve the intestinal bacterial species of earthworms.

Fig.2 People's congnition on affections of heavy metal

With regard to feeding earthworms, 72 percent of people are not aware of the existence of food that could feed earthworms.Moreover, in the respect of earthworm feed, more attention is being paid to the effects of drugs and their impact on the environment.

Fig.3 People's concerns on feed

About 65% of the people are satisfied with this microbial agent, which can degrade the heavy metal mercury in the soil, degrade the heavy metal mercury in the soil in a wide range, and enhance the resistance of earthworm to soil mercury Wu, while 32% of the people are still concerned about this. The uncertainty of biological pesticides in agriculture makes it impossible for human beings to accept the uncertainty caused by gene mutation. Most people also expect further improvements in its future biosafety. Our research is secure in terms of safety and cost, and efficiency and safety are our advantages.

Fig.4 About people's concerns about products Fig.5 People's acceptance of the product

Summary of Interviews-Scientific research institution

During the summer vacation, our HP teams visited the Chinese Academy of Sciences and the Shanghai Academy of Agricultural Sciences to interview relevant experts and explain our project idea to them. Through two interviews, we have a deeper understanding of the project and planning. BRIEF DESCRIPTION OF EXPERT: Zhang Peng, research fellow, Ph. D., center of Excellence in Plant Chinese Academy of Sciences Zheng Xianqing, associate research fellow, Shanghai Academy of Agricultural Sciences

1.About earthworms

Came to the Academy of Agricultural Sciences, met as an earthworm expert Zheng Xianqing teacher, learned that our project on earthworms, he led us to the room where he raised earthworms. The earthworms in the flowerpots were his test subjects. Teacher Zheng pointed to the earthworms moving in the pots: “It’s very valuable. We’ve done some research before, and that means that earthworms can generate more than two billion dollars a year in waste disposal alone. This is only a small part of the story, and the economic benefits of biomedicine are far greater. But there are not many people who study earthworms nowadays, so I’m glad that your experiment will involve earthworms.”

Fig.6 Earthworm culture basin Fig.7 Professor Zheng, Academy of Agricultural Sciences

2.Current technology and suggestion

In both interviews, we raised the question of “what are the existing methods for dealing with heavy metals in soil?”. Zhang Peng showed us the results of their research, “mizutani-soil probiotics”. To sum up, there are three main options:

1.Chemical Chelation

2.Low-accumulating plants

for example: The leaf stalks of food crops are rich in heavy metals, and the ears are used

3.Low-accumulating plants

for example: plants that do not absorb a heavy metal in soil where the metal exceeds the legal limit In addition,Professor Zheng put forward to us: mercury pollution in nature is not only mercury ions, but also organic mercury. In fact, organic mercury is more toxic and can be enriched in organisms, causing even more harm. We hope that our project can take organic mercury into account. After searching for data, we found that some bacteria in nature can express the organic mercury lyase Merb, which can transform organic mercury It is inorganic mercury, so we add Merb in the design, and decompose organic mercury into mercury ions through Merb protein. It is hoped that the organic mercury can be transformed into inorganic mercury and finally be fixed by the hydrogen sulfide produced by Atlcd. In communication with Professor Zhang, Professor Zhang pointed out that due to the extremely low content of mercury ions in soil, the output signal may be very low, and the protein content and fluorescence intensity may not meet the requirements. He proposed to us to design a signal amplification circuit. After consulting relevant information, we found that iGEM also had mercury ion detection in previous years [source: iGEM Gold Award in 2019] https://2019.igem.org/Team :BEAS_ Referring to the project, we decided to use rinp80a protein as our amplification element.

Fig.8 Soil probiotics Fig.9 Professor Zhang, Institute of plant physiology, Chinese Academy of Sciences

3.Legal Ethics and security

At the moment, there are some people who are not very receptive to GM, and feel that GM is something that goes against nature. To this end, Zhang Peng explained: The broad sense of genetically modified everywhere, like cucumbers. AH, before the wild cucumbers were so small (the teacher made a gesture, as big as the little finger) , and very bitter. Through the process of artificial constant selection, finally we have today’s cucumber. The state controls the technology so strictly that anyone who can get it on the market can be assured. On legal ethics, Zhang Peng told us: our country has very strict regulations on the release of genetically modified organisms from the real environment. The Department of Agriculture has regulations for that. Generally, if you’re doing a field experiment or a small experiment, you can apply. After the application, you are issued a permit, and then you can do the corresponding experiment. But for now, the state won’t allow it if GM managers do a lot of releasing. So it’s OK to experiment or build a method or a special case. I don’t remember exactly what it said, but you can check it out after the interview.

Fig.10 Relevant laws

4.Prospects for development

The two teachers generally approved of our project. On one hand, it has carried out this soil remediation. In some areas of our country, heavy metals in the soil are very serious, and using this technology may lead to a very good improvement. The earthworm, on the other hand, is a Chinese medicine -- a type of Earth Dragon. Therefore, it is a win-win result to use earthworms which can not only play the role of soil remediation but also play the role of cultivation of Chinese medicinal materials. Zhang Peng also suggested improvements that we could combine with earthworms. He said, “If the effect of using microorganisms that can't collect heavy metals can be doubled, then using microorganisms that can collect heavy metals to do this experiment can eventually reach 10 times. I think this project should be combined with the work for microbes.”

Summary of the interview-Chongming

On October 18,2020, our team went to Shanghai Chendao Agricultural Science and Technology Co. , Ltd. in Chongming District, Shanghai. Qian, who is in charge of the earthworm project, was interviewed to explain the current research and demands ofagricultural production companies for such products. At present, the earthworm as a pesticide is mainly red earthworm. Earthworms ferment their adapted feed in the intestine. The vermicompost excreted can be used as a good fertilizer to make the crops grow better and increase the crop yield greatly. The feed that earthworms eat comes from a nearby dairy farm. This is a win-win approach, both to help dairy farms deal with the manure of cattle and to provide fodder for earthworms. However, due to environmental pollution, cattle feed contains heavy metals, so their manure also contains heavy metals. Fortunately, earthworms contain heavy metal-degrading proteins that can degrade heavy metals. Unlike existing earthworm fertilizer applications, our project can more efficiently address the problem of heavy metals in the soil. That is, by feeding earthworms with genetically engineered Bacillus subtilis feed, to increase the efficiency of earthworms’ modification of heavy metal mercury, and on top of that, merRb protein can be used to detect whether the soil is contaminated with heavy metals. Compared with the existing methods of earthworm treatment, our product is more efficient and has the ability of identification. Manager Qian gave high recognition and said that if one day our “earthworm products” successful development, he is willing to cooperate with us.

Fig.11 Mr. Qian, the farmer Fig.12 Paddy field Fig.13 farmland Fig.14 Farmer Mr. Qian with us

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