Team:CAU China/Integrated Human Pratice/Researchers

In the process of our experiment, we got the help of many professionals. For example, Professor Huiqiang Lou and Professor Long Zhang provided us with yeast plasmids PAGDT7 and PABKT7 for vector construction; Professor Wangpeng Shi provided many wheat seedlings and wheat seeds for feeding locusts for our experiments. Of course, we had more in-depth communication and discussions with many experts. We would like to express our sincere gratitude to the professors and teachers who gave us help and advice.

    Introduction: Mr. Wangpeng Shi is a professor of Entomology Department in the College of Plant Protection, China Agricultural University. His main research directions are pest biological control, insect behavior, and safety evaluation of genetically modified crops. Involving the use of microspores, Beauveria bassiana, Metarhizium anisopliae and other beneficial organisms to carry out research on the biological control theory and technology of locusts and other major pests; the regulation of host behavior by insect pathogenic microorganisms and biochemical molecular mechanisms; the effect of genetically modified crops on non-target organisms Indoor and field ecological safety evaluation standards and the impact on the behavior of non-target organisms, etc. We contacted him, hoping to learn more about the feasibility of our design and what needs to be improved.

    Communication content: We explained to Professor Shi the idea of designing the project, that is, to kill young locusts using RNAi technology against several specific genes as targets. Professor Shi said that our topic selection is of strategic significance, and suggested that we can use nanoparticle materials to achieve the penetration of RNA into the epidermis of locusts. Professor Shi also pointed out that due to the safety of genetically modified organisms, it is not realistic to promote it in the short term. At the same time, the professor denied our preset to kill locusts through trapping. In addition, Professor Shi provided great support for our experimental materials.

    Perfection: Through the communication with Professor Shi, we gave up the idea of using traps to kill locusts, and decided to use RNAi technology to further improve and screen more suitable genes.

    Introduction:Shuo Yan is a professor of Entomology Department in the College of Plant Protection, China Agricultural University. His main research direction is functional genomics of agricultural insects: identifying a series of key regulatory genes for insect growth and development, using RNAi and transgenic technology to destroy the key gene functions of pests ; Research on the mechanism and application of nanomaterials in entomology: use nanomaterials to carry exogenous nucleic acids, toxic proteins, and drug molecules into insects or plant cells, and carry out research on the mechanism of nanocarriers to improve the efficiency of exogenous insecticidal factors. Interfere with development and behavior, and explore new strategies for pest genetic control. We communicated with Professor Yan Shuo, hoping to learn more about the feasibility of RNAi technology and information about nanomaterials.

    Communication content: Professor Shuo Yan told us that in terms of RNAi, our project has no technical difficulties in theory and can be achieved. Regarding nanocarrier delivery of dsRNA, we can currently achieve nanocarrier-mediated The penetrate of dsRNA through body wall, and direct spraying can interfere with the expression of target genes, which has a good lethal effect. The establishment of a locust poisoning system for a batch of key lethal genes can theoretically be realized.

    Perfection: Through the communication with Professor Shuo Yan, we further searched for relevant literature and materials, hoping to realize the delivery process through nanomaterials, but unfortunately, due to the difficulty of design and high cost of nanomaterials, it is not suitable for production. In the end, we did not choose nanomaterials for delivery.

    Introduction:Miss. Cao Chuan is an associate professor of Entomology Department in the College of Plant Protection, China Agricultural University. Her main research direction is to use Drosophila as a model, using molecular biology, genomics and evolutionary genetics to explore the molecular and genetics mechanism of insect immunity , genetic diversity and evolution of resistance and pathogenicity, the interaction and co-evolution of insects and microorganisms. In addition, it also involves the biological control of pests, that is, studying the use of insect viruses as a means of biological control of agricultural pests, exploring the pathogenic characteristics and mechanisms of viruses, and the immune response of host insects.We communicated with Associate Professor Chuan Cao about the modification and safety of Beauveria bassiana, Metarhizium anisopliae.

    Communication content: Associate Professor Chuan Cao provided us with suggestions on safety. She said that 1. Enhancing strain resistance will also increase its adaptability in the environment and its competitiveness against wild-type strains. If the transformed strains are used as biological pesticides to effectively prevent insects in a short period of time by spraying them in large quantities, consideration should be given to reducing their adaptability compared to the wild type. 2. Following the above ideas, genetic modification can be used to reduce the spore production of Beauveria bassiana or Metarhizium anisopliae, reduce the probability of its lateral spread and the number of spores in the environment.

    Introduction:Mr. Xinzheng Huang is a lecturer of Entomology Department in the College of Plant Protection, China Agricultural University. His main research direction involves identification of plant volatiles, insect pheromone (sex pheromone, aggregation pheromone) active components and control target insects’ behavior, etc. We communicated with Dr. Huang, hoping to get suggestions on selection of specific target genes for locust when locust plague breaks out.

    Communication content: Dr. Huang explained to us that there are some phenotypic differences between the desert locust's dispersal type and the social type. For example, a large number of volatile compounds are released by mature male locusts of the social type, including phenylacetonitrile, benzaldehyde, guaiacol and phenol. While they are not detected in newly emergence or dispersal type males. In addition, according to a literature report in Science, the transformation of locusts from living alone to living in groups begins when they attract each other and gather together, discover and smell each other's scent, or touch each other with their hind limbs. This touch forms a physiological stimulus in the body and stimulates the serotonin chemical substance in the locust's brain nerves. Serotonin is a "necessary and sufficient condition" for stimulating this transformation. The author pointed out that the development of drugs targeting the serotonin signaling pathway may provide a new method for preventing and controlling locust disasters. However, some people believe that the serotonin signaling pathway is widespread in the animal kingdom, so other species may suffer significant collateral damage.

    Introduction:Mr. Tan Shugan is a lecturer of Entomology Department in the College of Plant Protection, China Agricultural University. His main research direction is insect pathology and behavior. We also consulted Dr. Tan about target gene selection.

    Communication content: Dr. Tan said that in terms of genetic selection for genetic modification of engineered bacteria (RNAi and foreign gene insertion), he suggested: 1. Genes related to the growth and development of migratory locusts (molding and mating); 2. With Beauveria bassiana Genes related to body wall invasion (chitinase, etc.); 3. Genes related to changes in two ecotypes of migratory locusts. Dr. Tan thinks that these three directions are all good, but for a project, it is recommended to fix one or two, and that the genes that control the growth and development of migratory locusts are the best choice.

    Perfection: We consulted the recommendations of Dr. Huang and Dr. Tan, and finally selected three specific genes for the fifth instar locust using ATPase, CHS, and TSP as our target genes for designing RNAi.

    Introduction:Researcher Zehua Zhang is an expert in the agricultural industry technology system, a member of the expert group of the Chinese Association for Science and Technology "Prediction and Prevention of Diseases and Pests", a member of the "Pollution-free Agricultural Products Certification" review committee of the Agricultural Products Quality and Safety Center of the Ministry of Agriculture, and an expert in grassland protection and monitoring of the Ministry of Agriculture Committee, member of the Biological Control Professional Committee of the Chinese Society of Plant Protection, and secretary general of the Grassland Plant Protection Committee of the Chinese Society of Grassland. Mainly engaged in locust disaster research and the use of fungal preparations to control locusts and other target pests. The characteristics of the biodiversity of the ecosystem and the sustainable supporting technologies for the prevention and control of locusts such as natural enemies, Microsporidium and Metarhizium anisopliae. We communicated with researcher Zehua Zhang, hoping to understand the current status of locust control and some of his supplementary suggestions for our project.

    Communication content: For the project plan, Mr. Zhang suggested that we need to pay attention to the interference rate of RNAi on locusts, that is, the research on whether RNAi can effectively inhibit the corresponding genes of locusts, so that we can judge whether our RNAi technology is really effective. Teacher Zhang also told us that the current low promotion of biological pesticides is mainly due to the low cost of chemical pesticides and quick results. Farmers pursue economic benefits and have weak ecological awareness, so they are dependent on chemical pesticides. However, with the advancement of technology and people's health and ecological awareness, biological pesticides will have great application value. At the same time, Mr. Zhang pointed out that in terms of policy, a reliable supervision agency is currently needed.

    Perfection: Under the suggestion of Teacher Zhang, we supplemented the design of using RNAi technology to transform the biotechnology Metarhizium anisopliae to enhance its virulence and resistance, and designed an vitro injection experiment. Through communication with Teacher Zhang, we learned that the people's understanding of locust disasters and prevention has limitations, so we designed a questionnaire to investigate related issues (see the questionnaire survey for details).