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Implementation of our projects' scientific research and advice requires us to apply this information into the real world in order to serve as beneficial to our society and ecosystems. Wheat and barley are the second most produced food commodities in Australia (Australian Bureau of Statistics, 2022). Although Australia is in the top 10 countries for food availability the inability to export wheat for the rest of the world can affect the global supply chain and ultimately lead the Australian economy to plummet. Majority of products involving wheat are dependent on Australian farmers. Cereal rust has caused major losses to wheat crops, and continues to be a significant threat (Park, 2008). Due to Australian cereal crop yields being low, public attitudes towards synthetic biology solutions have become more accepted (Park, 2008). Nevertheless, there are multiple concerns that must be addressed when proposing to implement a genetically modified solution into a natural environment. Our team believes it is essential that our proposed implementation carefully considers the technical, safety, ethical and socio-cultural concerns.
During our conversation with Professor Matthew Kearnes, we discussed the environmental and ethical concerns of our project, specifically discussing how our project could affect genetic diversity. He informed us to contemplate how the use of our project could potentially perpetuate the practice of monoculture agriculture. This form of agriculture is known to contain negative environmental effects including loss of genetic diversity, reliance on non-renewable inputs and soil degradation. As a team, we fully acknowledge the importance of being cautious and considerate when navigating how to implement our project into the environment. We plan to implement our solution in a four-step phase, starting with the long term observation of our modified organism in a controlled and contained simulated soil environment. Using this approach, our team will be able to determine the most promising delivery of our project in regards to the environment and society.
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The UNSW iGEM team believes that proposed end users are parties that will assist our team in introducing our project into the real world. Our elected end users are government and non-government organisations whose incentives are to conserve and protect the soil environment.
Consulting with Professor Matthew Kearnes, an ethicist and social scientist allowed us to explore and eliminate who we wanted our proposed end users to be. During our conversation with him we questioned what our end users incentives were and what issues the general public and stakeholders could have with our synthetic biology solution.
Asking ourselves these questions led us to consider whether our product would be commercially profitable to our proposed end-users. Through this our team has decided that although profit can understandably be our proposed end-users motive they should have a stronger desire to solve food shortages and be less dependent on toxic pesticides. Next, our stakeholders must feel secure in that they are confident that their voices will be heard and integrated into our solution. Finally, our proposed end users must have enough scientific knowledge and control to utilise our solutions for the benefit of food security and freedom from pesticides.
The government is one of our ideal end users as they have an obligation to act upon the community’s best interest. The Grains Research And Development Corporation (GRDC) is a government agency that specialises in ensuring profitability for Australian growers. They invest in projects that deliver advanced and refined farming technology to the Australian grains industry (GRDC, 2021). The iGEM team believes cooperation with GRDC will be beneficial to both parties as both fill each other's criterion.
Another proposed end user are non-government organisations (NGOs) who develop solutions that are valued by their customers and seek improvement for their company, not only in the commercial but also in the environmental and social sector. Companies such as AWB limited who work with international non-government food corporations can affect our solution beyond Australian borders. This global reach would allow this solution to benefit wheat farmers and communities around the world. Working with NGOs gives them independence from government regulations, additionally they are less likely to be pressured by political values.
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Conversing with Microbiologist, Professor Torsten Thomas allowed our team to gain greater insight into how to implement our project into the real world. In our discussion he talked to us about the complexities when dealing with plant diseases, and how one must test their project on the wheat crops' different disease stages. It is impossible to consider the greater impact of our investigation on the soil environment unless we are able to observe our product in a simulated soil environment.
The first stage of our project’s proposed implementation involves the ex-situ testing of our project in a controlled soil environment. Our project will be tested on three different pathogens including Puccinia spp.. Our next step would be to transfer our GMO into an in-situ soil environment that has low biodiversity levels, once again testing on all three pathogens. The last step which will be undergone if the two previous steps are successful is releasing our project into a natural environment with normal to harsh biodiversity conditions.
Our team places public opinion as a priority. Because of this we wanted to ensure our proposed implementation was responsible and beneficial to the world. Our team created a proposed implementation that was intended to be as safe and free from risk as possible. Our three stage method was designed to eliminate risk to the soil and surrounding microbial life that exists within it.
Performing a long-term observation of our project on the pathogens Puccinia spp. within a controlled and contained environment simulating soil conditions.
Performing a long term observation of our project on the pathogens Puccinia spp. in a depleted natural soil environment.
If both stages 1 and 2 are successful, then our GMO can be released into the natural environment in normal to hard biodiversity conditions.
Continual observation of our project is essential during this time.
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One aspect that all of our stakeholders urged us to complete was the need for extensive testing before releasing our genetically engineered project into an in-situ environment. In order to test our project in-situ, several applications for the physical containment facility certification must be made. This includes receiving a PC1, PC2 and PC3 certification. It is also important for our team to comply with the Office of the Gene Technology (‘OGTR’) standards. They believe that, “Genetically modified (GM)… crops grown in Australia were approved for commercial release only when the regulator found that the GM crops were safe for human health and the environment as non-GM versions” (Office of Gene Technology Regulator, 2020).
Social Licensing
Successfully implementing our project into the environment is highly dependent on social licensing. The four stakeholder groups we chose to concentrate on were academics, government officials, Traditional Owners and businesses which correlated with our cause. Listening to each stakeholders social values ensures that our solution will be free of discontent. The concept of social licensing was discussed with social scientist Doctor Lucy Carter. She informed us that there is often a disconnect between scientists and people, as many scientists begin their experimentation neglecting their stakeholders' concerns.
Engaging with our stakeholders and the public was highly encouraged by Doctor Carter. In response to this, the UNSW iGEM team consulted with our four stakeholders often, listening to their commentary. Connecting with the public is also essential as they are the major class of individuals which determine whether our product is accepted into society. Additionally, engaging with the public allows us to educate the wider population about the benefits of synthetic biology on creating a permanent solution to cereal rust.
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Through our discussion with Indigenous environmental and climate advocate, Joshua Gilbert we realised the importance of scientists communicating thoroughly with Indigenous communities at the early stages of research. Joshua stated “engaging with Mob and those affected is very important,” thus contacting Indigenous non-government organisations such as black duck foods allows our team to return our economic benefits directly back to Indigenous people. The team understands that Traditional Owners have a deep spiritual connection to the land and we need to be mindful and respect their opinions on our project and how we implement it into nature. It is vital that we engage early with Traditional Owners of the land we seek to implement our work on in order to hear their voices on our solution.
An important factor that plays a role in the success of our proposed implementation is the level of acceptance the public shows towards the GMO being implemented. Therefore, overcoming this barrier would mean a greater involvement in educating and receiving feedback from the public is needed. Creating surveys asking for the public’s opinion on our project and their concerns about it ensured that we also took the public's concerns seriously. Feedback from the survey displayed people’s worry that genetic engineering of food crops could lead to loss of genetic diversity and that there were many ethical concerns regarding the use of GMOs.
Science communication would also need to be utilised in elucidating the stigma surrounding the use of GMOs on the natural environment.
Conversing with social scientists and the public allowed us to realise there were multiple topics of debate regarding the ethics of introducing a genetically modified species into the environment. Alinta Furnell, co-founder of synthetic biology company Synbiote urged us to concentrate on the biosafety concerns and to consistently talk and educate the public about our project. Additionally, communicating their concerns within our project ensures that the public feels heard.
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Considering that our proposed solution to crop rust involves the release of genetically modified organisms into the environment, the potential for unintentional invasion by genetically modified species outside their intended environment is a major environmental concern. If this is not prevented, it may lead to changes in soil ecosystem dynamics and possibly to a loss of biodiversity. To limit the likelihood of this happening, biosafety considerations must be integrated into the design of our projects. Furthermore, transgenic microorganisms can be designed with bio-containment systems such as kill switches or natural/synthetic malnutrition. These systems will result in cell death upon exposure to specific conditions, such as the presence of specific molecules or a change in temperature or pH value. Thus, the fusion of our modified e.coli with other species can be prevented by designing a kill switch.
In order to avoid unintentional invasion by our engineered E. coli, we plan to include a kill switch section in our design. Kill switch is an efficient mechanism that can cause synthetic bacteria to die without the presence of certain chemicals (Chan et al. 2016) . MazEF is a candidate toxin-antitoxin kill switch system for our project. MazF encodes a stable toxin that has the ability to cleave mRNA at a specific site. MazE is a labile anti-MazF protein that can prevent death of e.coli from MazF toxin (Zhang et al. 2003). Moreover, this mechanism has been proved to be effective in controlling the death of individual E. coli in the literature, this indicates its potential for limiting engineered E. coli growth (Hazan et al. 2004).
Additionally, rust infection can cause increasing H2O2 accumulation in wheat as the rust effector (PstGSRE1) has the ability to defeat ROS-Induced Defense (Qi et al. 2019). Thus, we planned to use H2O2 as a signaling molecule to control the expression of MazE. Furthermore, our last year team verified that higher concentrations of H2O2 advance the expression of the TrxC promoter, so TrxCp is a candidate promoter for our project. In this case, if a large amount of H2O2 shows up means that there is rust at the site, which can be a signal to induce TrxCp and MazE antitoxin can be expressed. This will result in preventing the death of the E. coli we designed, and the rust will be killed after our designed peptide expression. On the another hand, if not enough H2O2 is monitored, then the MazE antitoxin will not be expressed and the MazF toxin will cause the death of the E. coli, thus avoiding the unintentional invasion.
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Maintaining the stability of the peptide in transit may be a challenge for us. Special transport methods, such as cold chain transport may be required. Further experiments are required to verify optimal conditions to store engineered e. coli that can make sure its stability.
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