Team:UNILausanne/Human Practices

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Human Practices

Interview with Danilo Christen

To gage the use and feasibility of our project, we discussed it at length with Dr. Danilo Christen, the head of a research group on fruit crops in the Alpine region working at Agroscope, the Swiss centre of excellence for agricultural research. Trained as an agricultural engineer, employed by the Swiss Confederation and an expert in fruit crops, he was the perfect choice to get a first opinion on our project idea.

Dr. Christen informed us that, although drastic freezing events like the one we experienced this spring are usually quite rare, this is the second occurrence in the past 5 years. Both times, these extreme freezing spells caused devastating losses in the apricot crop in our region, making this problem an important one to address.

Our interviewee first described the current methods used by farmers to protect their crops. In the plains, farmers spray the trees with water that then freezes on the young fruits, lending a protective effect. Although this method works quite well, about half of the apricot trees are grown on the mountainside, which does not boast the same irrigation systems. During freezing nights in spring, farmers light candles under the trees on the hillside, generating heat. However, during this year’s freeze, violent northern winds blew away the heat generated by the few candles that managed to be lit, or blew the candles out entirely. The young fruits were therefore completely unprotected and phenomenal amounts froze and fell to the ground that day.

Our interviewee concluded that the use of a protein solution to protect the young fruit was a great idea, especially for the hillside trees. The candles generally used are very costly, reaching around 3,000 US$ per hectare and per night. Furthermore, they are made of paraffin, so they pollute the environment, and don’t protect the sensitive tissues enough during strong freezing spells.
Products to protect crops from frost are in high demand, not only in Switzerland but in neighboring countries. In France, vineyards were ravaged by frost in multiple regions this spring. According to our interviewee, our product could be broadly marketed, as the demand is there.

We then explained our idea to combat Pseudomonas syringae pv. syringae with the goal of reducing its role in the heightening of frost damage. Our interviewee was emphatic that attacking this pathogen was a marvelous idea. This pathogen not only exacerbates the problems due to frost, but also causes plant damage in general. Our interviewee viewed our attack of this pathogen as doubly beneficial - not only would we be diminishing the losses during freezing spells, but we would also be ridding the trees of an infestation that generally deteriorates their health.

P. syringae have such an impact on apricot production that, according to Dr. Christen, their important infestation of the Swiss German region is the reason why apricot cultivation is not possible in this area. In Romandy, the Swiss French region, P. syringae infestations have a particularly high impact as well, as these bacteria are used to generate snow in the ice canons employed on the sides of ski slopes. Apricots are cultivated in the same area, causing trees grown on the mountainside to have a higher exposure to these bacteria.

We finally discussed with our interviewee the steps necessary to get such a product recognized and approved. To approve a product, we would first need to prove its effectiveness. The second factor we would need to check is its impact on the environment: its toxicity on insects and auxiliary fauna, as well as its toxicity in water. Finally, we would need to check the toxicity of our product on humans.

At the end of our lengthy discussion, Dr. Christen assured us that our project design seemed excellent and that addressing this problem was a great idea.

Integrated Human Practices

Interview with Simon Gasser

Our first thought when approaching the human practices portion of our project was to reach out to apricot farmers and get their input on our project. However, each person we contacted was unavailable. Farmers in our region are overworked and underpaid, and the strain put on them due to the loss of apricot crops, one of the most highly valued crops in our region, is immense.

Instead, we searched for advice on a way to adapt our product to their needs from phytosanitary product manufacturers in Switzerland. We first contacted a representative from Stähler, a company that develops products hand in hand with farmers, adapting them to their needs. We reached out to Simon Gasser, who is in charge of developing and registering products for the company.
The main conclusion from our interview was that the product and implementation of our ideas was theoretically feasible. The antifreeze proteins (AFP) and tailocin solution contain no living organisms, which is key to allowing their use, as dispersing GMOs is currently illegal in Switzerland. Their production on a large scale would be pricey, but apricots are a very profitable crop and Simon assured us that farmers would be ready to pay around 500 francs per hectare to save them from the frost. Our interviewee suggested that our product should be sprayed or pulverized on the plant, as this would be the most efficient delivery method. Based on his advice, we decided to formulate our product to be in liquid form, deliverable as a spray to be applied directly to the plant. Simon Gasser assured us that this was feasible, as such products are usually pulverized by a tractor working its way through the rows of crops.
During this interview, we also learned that, to register our product, we would need to split the two solutions. The AFP solution would be registered as a fertilizer, which entails a streamlined version of the registration process. On the other hand, the tailocin solution would be registered as a phytosanitary product, which takes much more time and effort to accomplish.

Interview with Anne-Gabrielle Wüst Saucy and Silvain Aubrey

To deepen our understanding of the legalities of GMO production in our country and their impact on our project, we contacted Silvain Aubrey from the Swiss Federal Office for Agriculture (FOAG) and Anne-Gabrielle Wüst Saucy from the Federal Office for the Environment (FOEN). In our lengthy interview with them, they emphasized that even if the use of GMOs in agriculture is currently illegal in Switzerland, it is still important to do research on the subject. Even Swiss companies specialized in the production of products used in agriculture do so, and then target the European Union’s market to sell their products as the rules regarding GMO use are more lenient. However, as apricot losses due to frost is a local problem, navigating through the legal restrictions in Switzerland was important to us. With the input of our interviewees, we decided to split our product into three different parts. The first product is a totally non-GMO AFP solution that could be implemented in Switzerland currently. The second one is the tailocin solution, containing only the protein complex, and used to combat the plant pathogen P. syringae syringae – implementing it in Switzerland would be legal. Phages containing the CRISPR/Cas9 construct are not considered GMOs, but they modify our pathogenic bacteria to deactivate the gene coding for the Ice Nucleation Protein, and therefore produce a GMO. The use of this mechanism would currently be illegal in our country, but our interviewees urged us to pursue this idea regardless. This is exactly what we did. Since the phage is an alternative to the tailocin treatment, we would still have a solution to provide to our local farmers, whilst exploring another novel idea.

Meetings influencing phage design

We had several meetings with Dr. Gregory Resch, head of bacteriophage research at CHUV, the university hospital in Lausanne, in the context of the phage subsection of our project. He explained that the lytic strategy we had envisioned to delete a gene in our pathogen was ill-advised. Our initial idea had been to use a lytic phage whose lysin genes and genes responsible for replication had been deleted, in an attempt to avoid killing the pathogen. Dr. Resch suggested we use a lysogenic phage instead, since using our initial design, the infected bacterium would die regardless of the deletion of the targeted genes.

Our initial idea for the phage project was to delete the ice nucleation protein gene in Pseudomonas syringae pv. syringae using homologous recombination. Dr. Resch suggested we implement a double stranded cut induced by CRISPR/Cas9 technology instead, which would then be resolved via homologous recombination. This method would increase the odds of gene deletion.

After tweaking our design according to Dr. Resch’s recommendations and doing further research, we sought out the help of Dr. Astri Kusumawardhani to explore the use of a phagemid instead of a phage. She allowed us to better understand phagemids and confirmed that using a phagemid was more adapted to our project. Working with Dr. Kusumawardhani allowed us to get expert feedback and chose the phagemid best suited to our design.

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