Social Research

Questionnaire (I)

The general public is an important part of the stakeholders and their opinions and needs should be fully considered. Therefore, we launched the first round of online research for the general public.

Objectives:

To assess the health needs of the general public, their knowledge of natural plant metabolites, and their awareness of the relationship between cucumber and health, and to provide information to support further research on the function and wide range of applications of synthesizing flavonoid substances using cucumber epidermal hairs.

In the questionnaire, we collected 657 valid data.
Summary

The results of the questionnaire reflect the increased concern and health awareness of health issues, less awareness that cucumbers are rich in flavonoids, and the main reasons for choosing to eat cucumbers are taste and price. Most people are willing to increase the frequency of consumption if they are aware of the health benefits of cucumber. At the same time, there is a greater demand for knowledge about the natural plant metabolite flavonoids, and people would like to know more about their intake and effects.

People want to be able to maintain their health in a simple, affordable and safe way. Therefore, the promotion of cucumber, which is rich in nutrients high in flavonoids, is in line with public demand. We will further improve the popularization of flavonoids and other related knowledge to meet the public's demand for health knowledge and promote more scientific dietary choices and the maintenance of a healthy lifestyle.

Expert interviews
Technical feasibility interviews

Firstly for the technical feasibility of the research proposal, we interviewed Dr. Sun of China Agricultural University, who said that the modification of the key genes of cucumber epidermal hair initiation, CsTBH, and flavonoid metabolism genes (PAL) is feasible from the current research.

Highlights:
Highlights

Key gene for flavonoid synthesis in cucumber:

The CsTBH gene plays an important regulatory role in cucumber, and mutations in this gene will cause the entire cucumber plant to no longer produce hairs, as well as increase the percentage of female flowers in cucumber, and make the plant weaker and more susceptible to disease. In addition, the subsequent metabolic pathway regulated by this gene, the synthesis of flavonoids mainly occurs in the glandular epidermal hairs, which affects the proportion of female and male flowers, as well as plant growth and disease resistance of cucumber fruits. The PAL gene is mainly involved in the phenylpropane metabolic pathway and regulates the synthesis of flavonoid precursors. Although the stabilization of its editing in cucumber is not clear at present, by analogy with the function of this gene in other species, it can be hypothesized that its modification may have an impact on the content of flavonoids in cucumber.

Technical aspects:

There are some mainstream modification methods, such as RNA interference technology, vector transformation, overexpression, and CRISPR knockout gene. These techniques can be used to achieve the regulation of flavonoid synthesis in cucumber by altering the expression or function of genes.

Challenges of technical feasibility:

There are technical problems such as low transformation efficiency, stable inheritance and difficulty in obtaining the desired phenotype, for example, the transformation efficiency of gene modification is low and a large number of plants may need to be modified to obtain the desired results. In addition, even if the transformed plants are obtained, there is uncertainty as to whether their phenotypes are as expected, which requires continuous exploration and optimization.

Other considerations:

Social acceptance and ethical issues are also factors to be considered. There are concerns about the safety and potential effects of GM products, so adequate clinicological experiments need to be conducted to ensure that the modified cucumber is harmless to humans before it can be promoted in the market.

During genetic modification, the increase in flavonoids may lead to a strong defense state in the plant and affect the growth, so it is important to keep it within a manageable range and follow the natural growth of the plant while increasing the flavonoid content.

Summary

Through the interview, we have gained a better understanding of the modification of key genes in cucumber and the technical feasibility of using synthetic biology to increase flavonoid substances, and we will also utilize genetic transformation to stably overexpress the double genes after obtaining a positive strain. The content of flavonoids was determined using the transformed empty vector as a control. The effect of the CsTBH-CsPAL system on flavonoid synthesis will be determined through the analysis of the significance of differences in the content of the product. Further experiments were completed to actively explore and address the challenges faced.

Field Research - Plantation Visit

Our team members visited the Piovan planting base in Beijing to directly observe the growth of cucumbers: including the morphology of the plant, the growth stage, the characteristics of the epidermal hairs, etc., to understand the environmental factors of the growth of cucumbers, and to collect samples of cucumbers.

Interviewed Director Song who is the plantation to learn from practical experience and understand the views of frontline farmers on GM cucumbers and their actual needs for cucumber quality and characteristics. It helps us to better transform our research results into practical agricultural production applications, so that our project can be better adapted to the market demand.

Highlights
Highlights

1. Findings from observation

🥒In each section of the cucumber plant, flowers open, and the male and female plants are heterogamous.

🥒Epidermal trichome are distributed differently in different parts of the cucumber plant. On the leaves, epidermal trichome are usually denser, and on the stem, epidermal trichome are also distributed and may be slightly less dense than on the leaves. On the fruit, the distribution of epidermal trichome also varies from variety to variety.

🥒There are already very easy to observe epidermal trichome in the early growth stage of cucumber fruits.

2. Recognition in practice

🥒The modern agricultural plantation greenhouse adopts fully automated control technology, which can precisely regulate the temperature, humidity, and light in the greenhouse. This advanced facility creates the most suitable environment for the growth of cucumbers to ensure that they can thrive, and from the point of view of modern agricultural technology, it is believed that it can also provide stable and controllable conditions for our research.

3. Interview Summary

🥒Director Song said that in agricultural cultivation, although there are different varieties of cucumbers, the long cucumber with thorns still occupies the vast majority of the planting scale and market scale.

🥒Cucumber in the planting of the general do not leave seeds, because it is a hybrid selection of varieties, a generation of planting when the traits are excellent, but leave the seeds to plant again will be due to gene separation and reorganization of the quality and yield decline; leave the seeds to be strict conditions to ensure that the purity of the seed and vitality of the ordinary growers are difficult to have the relevant knowledge and equipment to operate.

🥒It is willing to accept cucumbers that can produce more flavonoids after genetic modification. Although the application of genetic modification technology in agriculture has great potential, they will still prioritize the market demand. Relevant national and regional laws and regulations must be strictly followed to ensure that its safety and legality have been fully evaluated and approved before planting.

Summary

Through our visit to the Beijing Paiwang planting base, we have gained a deeper understanding of cucumber growth, environmental factors, and the needs of frontline agriculturalists. This information will provide an important reference for our research. At the same time, we recognize that our research needs to take into account factors such as market demand, safety and legality to ensure that it can bring real value to agricultural production.

Questionnaire Survey (II)

Based on the analyzed results of the first round of questionnaire survey, and in combination with the suggestions of experts who gave us food for thought about safety and ethics during the interviews, we carried out a second round of combined online and offline survey on the public acceptance of the research project.

Objectives:

We collected information on the public's acceptance of genetic modification to increase the production of flavonoids in cucumbers and analyzed the influencing factors. We expect to lay a solid public foundation for the smooth progress of the research project and future market application.

We received a total of 546 valid data from this questionnaire:
Summary

Analysis of the survey results and data shows that the solution of using synthetic biology technology to enhance the yield of flavonoid substances in cucumber epidermal hairs has some potential, but also faces many challenges. On the one hand, the general public has a relatively positive attitude towards the technology, which provides a certain social base for product development and promotion. On the other hand, concerns such as health risks, costs, and technical complexity need to be addressed in subsequent studies. At the same time, diversified product types should be developed according to the needs and expectations of people of different ages, occupations and education levels, and the promotion process should focus on issues such as cost control, fulfillment of the needs of specific groups of people, health and safety protection as well as the balance of natural ecology, so as to enhance public acceptance and satisfaction. In addition, strengthening the popularization of synthetic biology technology and increasing the level of understanding of the general public will help to form a clearer and more positive attitude and promote the application and development of our projects in various fields.

Plant Secondary Metabolites Faculty Forum Learning Exchange
Objectives:

In order to gain a deeper understanding of flavonoid synthesis and provide valuable reference and inspiration for our project research. We attended the conference “Mining and analysis of genes related to citrus characteristic flavonoid synthesis” presented by Dr. Chenning Zhao (College of Food and Health, Zhejiang Agriculture and Forestry University).

Highlights:

RNA-seq technology was utilized to find the transcription factors involved in cucumber flavonoid synthesis and their regulatory mechanisms. By analyzing the transcriptome data, the upstream regulatory sequences of genes related to flavonoid synthesis and potential transcription factor binding sites can be found to understand the transcriptional regulatory network.HPLC-MS/MS technology can be used to accurately detect the contents and compositions of flavonoids in cucumber, which can help us to evaluate the effect of co-epitaxial modules, and provide a more accurate basis for optimization of the synthesis pathway.

Image source: Plant Secondary Metabolism Virtual Teaching and Research Section Forum--Zhao Chenning: Mining and analysis of genes related to the synthesis of citrus characteristic flavonoids (class screenshot)
Summary

Citrus research provides a molecular basis for derivative product development, which also reminds us to consider the direction of derivative product development in cucumber flavonoids research, and to utilize the multiple bioactivities of flavonoids for the development of products such as cosmetics, nutraceuticals, or pharmaceuticals with high added value.

Flavonoids of Interest - Directions for Pharmaceutical Applications Interviews
Objectives:

Flavonoids have certain pharmacological activities, so enhancing flavonoid production in cucumber could provide a new, relatively abundant source of pharmaceutical ingredients. In order to understand the current status of the application of flavonoids in the field of pharmaceuticals, the value of the application, and to obtain professional advice and recommendations. We connected with Dr. Li Wei, a researcher of the pharmaceutical company.

Highlights:
Highlights

Through interviews, we learned that.

🥒Currently in the field of medicine, flavonoid substances are of medicinal value, for example, flavonoids and flavonols have antioxidant, anti-inflammatory and antitumor effects, anthocyanins have antioxidant and cardiovascular health-protecting effects, and flavanones have antimicrobial, antiviral, and immunomodulatory effects.

🥒The absorption, metabolism and bioavailability of flavonoid substances from different sources may differ in the human body;

🥒Flavonoid ingredients are widely used in the development of nutraceuticals, such as grape seed extract, propolis, blueberry extract, and other nutraceuticals, and the flavonoid substances in cucumber would also be a good choice.

🥒Cucumbers are relatively inexpensive to grow and have high yields. Compared with the extraction of flavonoids from some expensive plants, the use of cucumber to extract or increase its flavonoid yield through synthetic biology technology is a cost advantage, which is conducive to reducing the cost of related pharmaceutical products.

Summary

Through the communication with Dr. Li, we have a better understanding of the practical needs and related problems of flavonoids in the application of pharmaceuticals and nutraceuticals. Some cases and market data he shared with us have made us realize the potentials of flavonoids in the prevention and treatment of a variety of diseases, as well as the prospects of their application in the field of pharmaceuticals and nutraceuticals, which have made the direction of our research more determined and clarified the practicality of our research.