Proposed Implementation

It takes more than having a solution work in the lab to make it a success. That’s why we went out and thought about how it would be feasible to implement our Outer Membrane Vesicle based solution on a scale large enough to make it work in day to day agriculture.

How does BOOM V work?

The modular nature of our design makes it very simple to adapt a wide range of scenarios. Traditionally, pesticides that combat specific pathogens have to be adapted on a regular basis, since these pathogens adapt themselves to these pesticides in a constant race between pathogen and plant. BOOM V has a significant advantage here, as we use pathogen-associated molecular patterns (PAMPs) to trigger the pattern-triggered immunity (PTI) in plants.
In our lab, we were able to show that a treatment with OMVs induced an immune response in plants.

Important Questions

Who are the end users?
How should it be distributed and used?
How is our solution produced efficiently?
What does the market look like?
What are the challenges and limitations of an implementation?
What safety aspects are there to be considered?

The individual steps to produce OMVs aren’t overly complicated. The main steps to produce them are operating the bioreactor, the centrifuge and making sure that safety aspects are adhered to. This is why we think it would be best to hand over the production to professionals.
Here, you can find our OMV isolation protocol.

What makes BOOM V so good?

Our current synthetic pesticide use isn’t sustainable. With our project, we enable broad treatment to various plants, since the design of our OMV based solution isn't developed for certain plant varieties but rather for general use. Additionally, our OMVs are biodegradable, so they don’t decrease the soil quality over time as seen with synthetic pesticides in a study conducted by the Swiss Institute of Agricultural Research Agroscope [1].

Who wants to use BOOM V?

After talking to both conventional as well as organic farmers, we learned that both kinds of producers would be interested in trying out our solution (see our interviews with farmer Mozzini or winegrower Studer).
In our collaboration with the Swiss university EPFL, we observed that the concerns of the Swiss population were mostly directed towards the use of pesticides, rather than GMOs. However, these results need to be considered with care, since most people who answered the survey tended to be young and came from an academic background.
Interestingly, in our collaboration with the Indian iGEM team MIT_MAHE and the Singaporean team NUS, we observed that specifically farmers prefer biopesticides, rather than the conventional synthetic pesticides. Still, among both farmers and consumers, GMOs are not seen as a preferred solution for the future, as many participants felt that too little is known about their effects on human health and the environment [2].

Who can use BOOM V?

In Switzerland, we have a temporary ban on the commercial use of genetically engineered plants. The decision whether Switzerland will allow their cultivation will be made in four years only, in 2025 [3].
To understand more about our political situation, please visit our handbook, especially the chapter "Will the temporary ban on GEOs in Switzerland ever see an end?".
This means that currently, the only possibility to apply our OMVs in Swiss agriculture is to spray the vesicles onto the plants. The other option would be to release genetically engineered bacteria in the soil. Though Swiss regulations don't allow the second option, we still decided to follow both possible application paths because in many countries in the world, GM-derived plants are standard in agricultural use. In an interview with Council of States and entrepreneur Ruedi Noser, he recommended that if we wanted to turn our iGEM project into a start-up, we should go abroad: “In Switzerland, the admission procedure for a GEO-product takes very long and is extremely expensive. This could quickly become fatal for a small start-up. But the market isn’t bound to national borders, and I think your project could have huge potential.” [4]

It was during a visit to the Swiss Federal Institute for Agricultural Research Agroscope that we saw the consequences of this division in opinions on the matter of GMOs. After crops had been destroyed through an act of vandalism a few years back, the field trials there are now protected with barbed wire fences:

This is in stark contrast to an anecdote that Council of States Ruedi Noser told us during our interview when remembering a trip to California: In a research institute at the University of Berkeley, GMO plants just stood in some hallway, without any protective measures, so in theory, everyone could just take them away [4].

How to distribute and use BOOM V?

Nowadays, farmers heavily rely on synthetic pesticides to protect their crops. This leads to ecological as well as economical problems: On one hand, the chemical compounds from synthetic pesticides stay in the soil for a long time, decreasing the water quality [1] and leading to losses in biodiversity. And on the other hand, farmers often depend on big transnational seed and pesticide conglomerates, all while prices have increased in the last years, as Swiss farmer Roberto Mozzini confirmed to us [5]. Our OMVs are degradable and would allow a faster, cheaper and more broad application since they don’t target one pathogen specifically.
For more details on our contributions to sustainability, please visit our Sustainability page.

Following feedback that we got from talking to farmers and representatives of farming associations, we understood that our solution has to be above all cheap, safe to use, and easy to apply. This is why we believe that the best solution for the implementation of the spray option our OMVs would be to have local hubs with bioreactors that are handled by professionals. The produced OMVs could then locally be distributed according to the current needs of farmers.
Swiss Council of States Ruedi Noser encouraged us to consider going abroad, where the laws concerning GMOs and their use are less strict than in Switzerland [4].

An Example for Application

One potential application example is the case of powdery mildew, a fungal disease strongly reducing crop yields worldwide. We know that powdery mildew loves humid environments. After heavy rainfall, there is a high chance of a prospective fungal attack, so we could prepare the plants by spraying OMVs onto their leaves. This will induce an immune response, thus priming the plant for the upcoming pathogen attack instead of only reacting after the pathogen-caused damage has already happened. This is only one of various possible application examples. Our technology could also work for other diseases like leaf rust et cetera.

A global pathogen: Leaf rust on corn leaves

Of course, there are pathogens against which our OMV based solution would not work, like insects. The defence against an insect attack includes very different mechanisms which our project doesn’t solve.

Cost and Efficiency

After talking to an expert in the industry, we realised that efficiency will be key if we are to produce OMVs on a large enough scale.
While it is true that you can heighten production efficiency by working with larger bioreactors, from a certain scale onwards efficiency goes down again. We were told that industrial size productions can be less efficient than lab based productions by a factor of 3-4 in some cases.
To improve efficiency, producing locally and therefore reducing the transportation costs and emissions is another key towards making the whole solution sustainable.

[6]

The Market for BOOM V

Since a multitude of pesticide solutions are commercially available, we decided to narrow down our analysis to fungicides within the Swiss market. Taking as a reference point the current fungicidal solutions, we estimated under which price our solution could be commercially competitive. We evaluated the current annual market potential for our alternative in Switzerland.
Our market potential is around 11.7 Mio. CHF (ca. 13 Mio. USD) which shows the potential our solution has. For more information on the market for BOOM V in Switzerland, please visit our Entrepreneurship page.

Challenges and Limitations for Implementation

In order to make sure that our OMVs can be implemented in the most efficient way and across different agricultural backgrounds, further analysis concerning the minimal quantities of OMVs for field application will need to be conducted. Furthermore, during our lab work we worked with the plant A. thaliana only. If the OMV solution is to be used in a real world context, experiments with other plant species will need to be done first in the lab, and then in field trials. At the same time, A. thaliana is a good indicator since many other plants in agriculture belong to the same brassicaceae family. If a crop is not from this particular family, our modular design will allow optimal stimulation of the immune response through adjustment of the elicitor on the OMV membrane.
We would like our OMV solution to be cheap, but the initial investment would be quite high since lab equipment like centrifuges are very expensive.

Further Illustration of the Challenges for Implementation

Implementation of Safety Features

To keep the ecosystem and the people around it safe, our solution needs a few safety precautions, with which we believe that a successful implementation could take place.
We want to stress that it would be best if the centrifuge and bioreactors were to be handled by professionals.
On a technical level, an important safety point is that we follow two approaches, one where we release a GMO into the soil, and one where we spray OMVs. However, if bacteria were to be released into the soil, we would have to find ways to biologically contain the bacterium, for example kill switches, genetic parts that cannot work on other organisms, non-canonic AAs, etc [7].
For more about the design of our project, please visit our Design page.

Summary

It keeps the costs as low as possible, making the most of bioreactor and distribution efficiency.
We can ensure that only the GMO derived products, and not the GMOs themselves leave the production facility.
Our OMVs are more sustainable than synthetic pesticides as they are biodegradable.

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[1] Widespread Occurrence of Pesticides in Organically Managed Agricultural Soils — The Ghost of a Conventional Agricultural Past?, by Judith Riedo, Felix E. Wettstein, Andrea Rösch, Chantal Herzog, Samiran Banerjee, Lucie Büchi, Raphaël Charles, Daniel Wächter, Fabrice Martin-Laurent, Thomas D. Bucheli, Florian Walder, and Marcel G. A. van der Heijden, Environmental Science & Technology, 2021, 55 (5), 2919-2928, DOI: 10.1021/acs.est.0c06405, 27. September 2021
[2] Consolidated Survey Insights from the collaboration with NUS and MIT_MAHE
[3] Swiss Federal Council: About the temporary ban, 20. October 2021
[4] Ständerat, Self-Sufficiency and Start-ups: An Interview with Ruedi Noser by iGEM UZH 2021, 21. October 2021
[5] Greenhouses, Growth and G...Tomatoes: An Interview with Roberto Mozzini by iGEM UZH 2021, 21. October 2021
[6] Picture created with Biorender.com on 18. October 2021
[7] Synthetic biology approaches to biological containment: pre-emptively tackling potential risks. Essays in biochemistry, by Torres, L., Krüger, A., Csibra, E., Gianni, E., & Pinheiro, V. B. (2016). , 60(4), 393–410. https://doi.org/10.1042/EBC20160013

Team:UZurich/Implementation