Difference between revisions of "Team:Calgary/Human Practices"

 
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           </div>
 
           <p>
 
           <p>
             <b>Providing a solution to the green seed problem</b> could be the key to
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             Providing a solution to the green seed problem could be the key to
 
             alleviating losses felt by those in the canola industry. Our team’s
 
             alleviating losses felt by those in the canola industry. Our team’s
 
             vision was to make yOIL that solution. In order to do so, we
 
             vision was to make yOIL that solution. In order to do so, we
             had to ensure that we created something that was <b>effective,
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             had to ensure that we created something that was effective,
 
             usable,
 
             usable,
             and targeted</b> areas which impacted people the most. The only
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             and targeted areas which impacted people the most. The only
 
             way to achieve this goal, was to <b>integrate our end-users in  
 
             way to achieve this goal, was to <b>integrate our end-users in  
 
             creating our solutions</b>.
 
             creating our solutions</b>.
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             throughout the project lifecycle, and ultimately the all-encompassing solutions found in yOIL.
 
             throughout the project lifecycle, and ultimately the all-encompassing solutions found in yOIL.
 
           </p>
 
           </p>
 +
 +
          <p>yOIL started off small in scope. Nothing more than a process to remove chlorophyll from canola oil.
 +
            It wasn't until we spoke to everyone involved in the industry that we realized we could do so much more.
 +
            <br><br>
 +
            <b>This is the story of how yOIL was born from a desire to help oil producers, and grew into an all-encompassing solution through
 +
          iterative meetings with contacts from all over the industry.</b></p>
  
  
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             speak with. Our primary research indicated that there are two main entities
 
             speak with. Our primary research indicated that there are two main entities
 
             impacted by the green seed issue: farmers and oil producers. In addition to
 
             impacted by the green seed issue: farmers and oil producers. In addition to
             those two, we also identified <dfn>agronomists</dfn> and organizational bodies working
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             those two, we also identified agronomists and organizational bodies working
 
             within the canola oil industry. From here, we initiated conversations
 
             within the canola oil industry. From here, we initiated conversations
 
             about the green seed issue and discussed how we could utilize synthetic biology
 
             about the green seed issue and discussed how we could utilize synthetic biology
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                 <div class="modal-body">
 
                 <div class="modal-body">
                   <p>Craig Shand is a local Southern Albertan Farmer and <dfn>Agronomist</dfn>. It was essential for us to understand the exact effects green seed has on Canadian agriculture from someone directly in the industry who is experiencing those losses.
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                   <p>Craig Shand is a local Southern Albertan Farmer and Agronomist. It was essential for us to understand the exact effects green seed has on Canadian agriculture from someone directly in the industry who is experiencing those losses.
 
                     <br><br>The green seed problem creates losses at the canola crushers, and those losses are passed on to farmers like Craig. When farmers are already operating on a sliver of a profit margin, the downgrade of crops from $10 per bushel to $5-6 per bushel can make or break a growing season. Or as Craig succinctly puts it: <b>“Green is the difference between profit” </b>,
 
                     <br><br>The green seed problem creates losses at the canola crushers, and those losses are passed on to farmers like Craig. When farmers are already operating on a sliver of a profit margin, the downgrade of crops from $10 per bushel to $5-6 per bushel can make or break a growing season. Or as Craig succinctly puts it: <b>“Green is the difference between profit” </b>,
 
                     <br><br>Our project strived to keep money in the pockets of farmers. Talking to Craig informed our team how we could leverage our scientific advances to help Albertan farmers. Our solution would have to go beyond simply purifying chlorophyll from oil. We would need to create a market for green canola seed.
 
                     <br><br>Our project strived to keep money in the pockets of farmers. Talking to Craig informed our team how we could leverage our scientific advances to help Albertan farmers. Our solution would have to go beyond simply purifying chlorophyll from oil. We would need to create a market for green canola seed.
                     <br><br>As an Agronomist, Craig is heavily involved in <dfn>field scouting</dfn> and trying to help as many farmers as possible manage green seed. He identified two major problems:First, the Albertan growing season is relatively short and is dictated by the frost-free time period. Canola seeds must mature before frost hits, otherwise green seeds will form. Frost is an ephemeral boogeyman to farmers, you do not know when it is coming until it is too late.
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                     <br><br>As an Agronomist, Craig is heavily involved in field scouting and trying to help as many farmers as possible manage green seed. He identified two major problems:First, the Albertan growing season is relatively short and is dictated by the frost-free time period. Canola seeds must mature before frost hits, otherwise green seeds will form. Frost is an ephemeral boogeyman to farmers, you do not know when it is coming until it is too late.
 
                     <br><br>Secondly, when farmers take their harvest to grain elevators for grading, that grading process is done manually. It is inefficient, time consuming and worse, highly subjective. Craig highlighted the fact that you could take your harvest to two different grain elevators and get two different gradings. Gradings being the difference between selling at a premium or selling at a loss, Craig opened our eyes to another problem that can be addressed.
 
                     <br><br>Secondly, when farmers take their harvest to grain elevators for grading, that grading process is done manually. It is inefficient, time consuming and worse, highly subjective. Craig highlighted the fact that you could take your harvest to two different grain elevators and get two different gradings. Gradings being the difference between selling at a premium or selling at a loss, Craig opened our eyes to another problem that can be addressed.
 
                     <br><br></p>
 
                     <br><br></p>
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                 <div class="modal-body">
 
                 <div class="modal-body">
 
                   <p>Ward Toma is the General Manager of the <a class="info-link" href="https://albertacanola.com/about/"> Alberta Canola Producers Commission (ACPC),</a>an organization which supports the “long-term success of canola farmers in Alberta through research, extension, consumer engagement, and advocacy for canola farmers”(https://albertacanola.com/about/). As General Manager, Ward is responsible for overseeing the ACPC and is often the public face advocating for Albertan canola. His background in Agricultural Economics was a perfect jumping point into a long, detailed conversation about where Albertan agriculture needs help.
 
                   <p>Ward Toma is the General Manager of the <a class="info-link" href="https://albertacanola.com/about/"> Alberta Canola Producers Commission (ACPC),</a>an organization which supports the “long-term success of canola farmers in Alberta through research, extension, consumer engagement, and advocacy for canola farmers”(https://albertacanola.com/about/). As General Manager, Ward is responsible for overseeing the ACPC and is often the public face advocating for Albertan canola. His background in Agricultural Economics was a perfect jumping point into a long, detailed conversation about where Albertan agriculture needs help.
                     <br><br>Starting with Green Seed, Ward confirmed what we started to fear after talking to Craig: the success of a farmer’s growing season is largely defined by green seed content. By his account, 6-7% of an annual crop consists of green seed that is blended into together into the bulk of the crop, reducing the value of the crop. He suggested that potentially recovering the chlorophyll and making it into something valuable would be unique and profitable in comparison to the <dfn>acid-activated clays</dfn> method. Creating a market for green seed and increasing demand would give leverage to the farmer when negotiating prices at the grain elevators.  
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                     <br><br>Starting with Green Seed, Ward confirmed what we started to fear after talking to Craig: the success of a farmer’s growing season is largely defined by green seed content. By his account, 6-7% of an annual crop consists of green seed that is blended into together into the bulk of the crop, reducing the value of the crop. He suggested that potentially recovering the chlorophyll and making it into something valuable would be unique and profitable in comparison to the acid-activated clays method. Creating a market for green seed and increasing demand would give leverage to the farmer when negotiating prices at the grain elevators.  
 
                     Ward confirmed that green seed is the main factor affecting canola crop value throughout a harvest cycle.
 
                     Ward confirmed that green seed is the main factor affecting canola crop value throughout a harvest cycle.
                     <br><br>Ward was also kind enough to give us a lead on another excellent resource, pointing us towards Dr. Veronique Barthet, a researcher at the <dfn>Canadian Grain Commission</dfn>.
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                     <br><br>Ward was also kind enough to give us a lead on another excellent resource, pointing us towards Dr. Veronique Barthet, a researcher at the Canadian Grain Commission.
 
                     <br><br></p>
 
                     <br><br></p>
 
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                 </div>
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                 <div class="modal-body">
 
                 <div class="modal-body">
                   <p>Dr. Veronique Barthet is a Research Scientist and Program Manager for Oilseeds Monitoring and Research at the <dfn>Canadian Grain Commission</dfn>'s Grain Research Laboratory. Her areas of expertise lie in researching grain quality factors, seed oxidative stability and developing analytical quality measurement methods. She shared with us how the Grain Research Laboratory quantifies the quality of crops, such as canola, in an objective manner.
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                   <p>Dr. Veronique Barthet is a Research Scientist and Program Manager for Oilseeds Monitoring and Research at the Canadian Grain Commission's Grain Research Laboratory. Her areas of expertise lie in researching grain quality factors, seed oxidative stability and developing analytical quality measurement methods. She shared with us how the Grain Research Laboratory quantifies the quality of crops, such as canola, in an objective manner.
                     <br><br>The Canadian Grain Commission relies mostly on <dfn>spectroscopy</dfn> and high performance liquid chromatography (<dfn>HPLC</dfn>) to determine the chlorophyll content of green seeds. She also told us that chlorophyll a is in the highest abundance in green seed oil, a consideration which would affect our project design.
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                     <br><br>The Canadian Grain Commission relies mostly on spectroscopy and high performance liquid chromatography (HPLC) to determine the chlorophyll content of green seeds. She also told us that chlorophyll a is in the highest abundance in green seed oil, a consideration which would affect our project design.
 
                     <br><br>Dr. Barthet’s knowledge of the current method for purifying chlorophyll from canola oil, the acid-activated clay method, also proved to be invaluable to our project development. According to her, the clay used in the purification process is disposed following use, highlighting that creating a re-usable method would bring economic value to the oil processors. The acid-activated clay method is the only purification method that she is aware of meaning that is the only technique we would need to outperform.
 
                     <br><br>Dr. Barthet’s knowledge of the current method for purifying chlorophyll from canola oil, the acid-activated clay method, also proved to be invaluable to our project development. According to her, the clay used in the purification process is disposed following use, highlighting that creating a re-usable method would bring economic value to the oil processors. The acid-activated clay method is the only purification method that she is aware of meaning that is the only technique we would need to outperform.
 
                     <br><br>Finally, Dr. Barthet emphasized that any solution we propose would have to be integrated into current oil processing plants, and would ultimately need to be commercially and economically viable. She believes that as long it is commercially viable, an alternative to acid-activated clays would be welcome in the canola oil processing community.</p>
 
                     <br><br>Finally, Dr. Barthet emphasized that any solution we propose would have to be integrated into current oil processing plants, and would ultimately need to be commercially and economically viable. She believes that as long it is commercially viable, an alternative to acid-activated clays would be welcome in the canola oil processing community.</p>
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                 <div class="modal-body">
 
                 <div class="modal-body">
                   <p>Angela Brackenreed is an Agronomy Specialist with the <dfn>Canola Council of Canada</dfn>.  
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                   <p>Angela Brackenreed is an Agronomy Specialist with the Canola Council of Canada.  
 
                     We spoke with Angela to assess the viability of pursuing a project handling the rampant green seed problem.  
 
                     We spoke with Angela to assess the viability of pursuing a project handling the rampant green seed problem.  
 
                     Since we spoke to her at the relative onset of our project, we valued her ability to give us a generalized outlook on the canola industry  
 
                     Since we spoke to her at the relative onset of our project, we valued her ability to give us a generalized outlook on the canola industry  
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                   <p>Dallas Gade is a project manager at Richardson Oilseed, one of the biggest producers of oilseed in Western Canada.  
 
                   <p>Dallas Gade is a project manager at Richardson Oilseed, one of the biggest producers of oilseed in Western Canada.  
 
                     Earlier in the summer we had an excellent conversation with Dallas regarding the process flow in an industrial sized  
 
                     Earlier in the summer we had an excellent conversation with Dallas regarding the process flow in an industrial sized  
                     oil processing facility. He walked us through the oil production process step by step, even providing us with <dfn>process flow diagrams</dfn>.
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                     oil processing facility. He walked us through the oil production process step by step, even providing us with process flow diagrams.
 
                     Our lengthy conversation could have kept going for days, so Dallas was kind enough to invite us to visit  
 
                     Our lengthy conversation could have kept going for days, so Dallas was kind enough to invite us to visit  
 
                     Richardson’s refinery in Lethbridge, Alberta to continue the conversation there.  
 
                     Richardson’s refinery in Lethbridge, Alberta to continue the conversation there.  
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               <div class="col-sm text-center">
 
               <div class="col-sm text-center">
                 <button onclick="location.href='https://2019.igem.org/Team:Calgary/Modelling/StandardizedSeedGrading'" class="projButton">Mean Green Machine</button>
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                 <button onclick="location.href='https://2019.igem.org/Team:Calgary/Modelling/StandardizedSeedGrading'" class="projButton">Mean Green Machine<p style="text-align: center">standardize seed grading</p></button>
 
               </div>
 
               </div>
 
               <div class="col-sm text-center">
 
               <div class="col-sm text-center">
                 <button onclick="location.href='https://2019.igem.org/Team:Calgary/SunnyDays'" class="projButton">Sunny Days</button>
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                 <button onclick="location.href='https://2019.igem.org/Team:Calgary/SunnyDays'" class="projButton">Sunny Days<p style="text-align: center">weather prediction</p></button>
 
               </div>           
 
               </div>           
 
             </div>
 
             </div>
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           <p>
 
           <p>
 
             Midway through the summer, we were lucky enough to be able to attend canolaPALOOZA.  
 
             Midway through the summer, we were lucky enough to be able to attend canolaPALOOZA.  
             An annual event held each year in Lacombe, Alberta, canolaPALOOZA is the “agronomy  
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             An annual event held in Lacombe, Alberta, canolaPALOOZA is the “agronomy  
 
             event of the summer” where canola experts from all over the prairies come together  
 
             event of the summer” where canola experts from all over the prairies come together  
 
             to discuss all things canola. Everyone from farmers, agronomists, mycologists and  
 
             to discuss all things canola. Everyone from farmers, agronomists, mycologists and  
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             It was an excellent opportunity for us to propose our  
 
             It was an excellent opportunity for us to propose our  
 
             ideas to a wide cross-section of the canola industry, and get feedback on the commercial viability of our  
 
             ideas to a wide cross-section of the canola industry, and get feedback on the commercial viability of our  
             solutions. The event was extremely valuable because this is where we about the fungal infestations  
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             solutions. The event was extremely valuable because this is where we learned about the fungal infestations  
             currently affecting canola crops in Alberta. A fortuitous meeting which would give us the inspiration to repurpose  
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             currently affecting canola crops in Alberta. A fortuitous meeting which would <b>give us the inspiration to repurpose  
             chlorophyll into an anti-fungal agent. CLOSE THE LOOP
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             chlorophyll into an anti-fungal agent.</b>
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             <br><br>
 
             <br><br>
 
             This event lead to a connection with the Canadian Grain Commission, who expressed their <b>interest in the standard  
 
             This event lead to a connection with the Canadian Grain Commission, who expressed their <b>interest in the standard  
 
             seed grading system</b>. They informed us that there had been a similar project in the past that was successful, but got
 
             seed grading system</b>. They informed us that there had been a similar project in the past that was successful, but got
 
             scrapped due to its cost. This inspired us to make our standardized grading machine as inexpensive as possible.
 
             scrapped due to its cost. This inspired us to make our standardized grading machine as inexpensive as possible.
            <br><br>
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            Having already decided to convert captured chlorophyll to pheophorbide, we then dedicated additional efforts to
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            researching the affect of pheophorbide on fungal infestations affecting canola.
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             <br><br>
 
             <br><br>
 
             To learn about this process, click the button below.
 
             To learn about this process, click the button below.
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               <div class="col-sm text-center">
                 <button onclick="location.href='https://2019.igem.org/Team:Calgary/Anti-Fungal'" class="projButton">Chlorophyll Extraction System</button>
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                 <button onclick="location.href='https://2019.igem.org/Team:Calgary/Anti-Fungal'" class="projButton">Repurposing Chlorophyll</button>
 
               </div>
 
               </div>
 
             </div>
 
             </div>
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             With the need for our solutions verified, it was time for us to put shovels in the ground  
 
             With the need for our solutions verified, it was time for us to put shovels in the ground  
 
             ( or bacteria on the agar)  
 
             ( or bacteria on the agar)  
             and begin designing yOIL. Our development would be agile and adaptable.  
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             and begin designing yOIL. Our development would be <b>agile and adaptable</b>.  
 
             Continually meeting with stakeholders and integrating feedback throughout the  
 
             Continually meeting with stakeholders and integrating feedback throughout the  
 
             design process to ensure yOIL holistically embodies the needs of everyone  
 
             design process to ensure yOIL holistically embodies the needs of everyone  
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             The clay method is not reusable, environmentally friendly, or selective to chlorophyll, thus,  
 
             The clay method is not reusable, environmentally friendly, or selective to chlorophyll, thus,  
 
             we needed to come up with an improved solution. Proteins are organic and are  
 
             we needed to come up with an improved solution. Proteins are organic and are  
             made to be highly specific to their substrate, so we thought to design a system that utilized a protein to capture chlorophyll, but we weren’t
+
             made to be highly specific to their substrate, so we thought to design a system that utilized a protein to <a class = "abody" href = "https://2019.igem.org/Team:Calgary/ChlorophyllExtraction">capture chlorophyll</a>, but we weren’t
 
             confident as to which chlorophyll-binding protein to use, how it could be used in an oil environment, and what other
 
             confident as to which chlorophyll-binding protein to use, how it could be used in an oil environment, and what other
             considerations we needed to be aware of.
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             considerations to be aware of.
 
             <br><br>
 
             <br><br>
 
             To answer these questions, we spoke to protein bio-chemists, micro-biologists, plant biologists, and chemical engineers who gave us insight
 
             To answer these questions, we spoke to protein bio-chemists, micro-biologists, plant biologists, and chemical engineers who gave us insight
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                 </div>
 
                 </div>
 
                 <div class="modal-body">
 
                 <div class="modal-body">
                   <p>Angela Brackenreed is an Agronomy Specialist with the Canola Council of Canada. We spoke with Angela to assess the viability of pursuing a project handling the rampant green seed problem. Since we spoke to her at the relative onset of our project, we valued her ability to give us a generalized outlook on the canola industry and give us an idea of the movers and shakers in the bustling Canadian Canola industry.
+
                   <p>Angela Brackenreed is an Agronomy Specialist with the Canola Council of Canada.  
                     <br><br>We learned that farmers in particular can use a few agronomic techniques to mitigate the impact of green seed on their crops. If a farmer can get an idea of when frost will occur, they can then accommodate the weather when choosing when to seed and when to swath. However, Angela made us realize that the industry follows a production pipeline of canola oil, from farmer to family tables, and the green seed problem effects every person along the way. We were surprised to learn from Angela that the canola industry is very accepting of genetically modified organisms, and we would just need to manage public perception if we designed such an approach. However, it was relieving to learn that 100% of canola grown is genetically modified and that the industry itself would not have ethical qualms to integrate a synthetic biology solution.
+
                    We spoke with Angela to assess the viability of pursuing a project handling the rampant green seed problem.  
 +
                    Since we spoke to her at the relative onset of our project,  
 +
                    we valued her ability to give us a generalized outlook on the canola industry and  
 +
                    give us an idea of the movers and shakers in the bustling Canola industry.
 +
                     <br><br>We learned that farmers in particular can use a few agronomic techniques to  
 +
                    mitigate the impact of green seed on their crops. If a farmer can get an idea of when frost will occur,
 +
                    they can then accommodate the weather when choosing when to seed and when to swath.  
 +
                    However, Angela made us realize that the industry follows a production pipeline of canola oil,  
 +
                    from farmer to family tables, and the green seed problem effects every person along the way.  
 +
                    We were surprised to learn from Angela that the canola industry is very accepting of genetically modified organisms,
 +
                      and we would just need to manage public perception if we designed such an approach.  
 +
                      However, it was relieving to learn that 100% of canola grown is genetically modified and that the industry itself  
 +
                      would not have ethical qualms to integrate a synthetic biology solution.
 
                     <br><br></p>
 
                     <br><br></p>
 
                 </div>
 
                 </div>
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                     review of other tags that could be used in its place in an industrial system, such as a  
 
                     review of other tags that could be used in its place in an industrial system, such as a  
 
                     galactose-binding protein tag or biotin-carboxy carrier protein. Additionally, he suggested  
 
                     galactose-binding protein tag or biotin-carboxy carrier protein. Additionally, he suggested  
                     that secreted the protein could also minimize the costs of purification, and that trying  
+
                     that secreting the protein could minimize the costs of purification, and that trying  
 
                     different signal peptides with our protein could be a consideration.
 
                     different signal peptides with our protein could be a consideration.
 
                     <br><br></p>
 
                     <br><br></p>
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           <p>
 
           <p>
             Chlorophyll cannot be removed from the acid-activated clay chlorophyll after it is bound; however, by using chlorophyll binding proteins the chlorophyll can be released after.
+
             Chlorophyll cannot be removed from the acid-activated clay chlorophyll after it is bound;  
             In our initial discussions with Ward Toma, he suggested that repurposing chlorophyll could bring new revenue into the canola industry and mitigate some of the losses caused by  
+
            however, by using chlorophyll binding proteins the chlorophyll can be released after.
             green seed. Dallas Gade further confirmed that valuable byproducts could be useful to offset expensive input costs. Pheophorbide a is a natural catabolite of chlorophyll that  
+
             In our initial discussions with Ward Toma, he suggested that <a class = "abody" href = "https://2019.igem.org/Team:Calgary/RepurposingChlorophyll"> repurposing chlorophyll</a>
             has been investigated recently for its photosensitizing abilities in experimental anti-cancer and anti-fungal treatments. Seeing how lucrative the product is, we decided to  
+
            could bring new revenue into the canola industry and mitigate some of the losses caused by  
 +
             green seed. Dallas Gade further confirmed that valuable byproducts could be useful to offset expensive input costs.
 +
            Pheophorbide a is a natural catabolite of chlorophyll that  
 +
             has been investigated recently for its photosensitizing abilities in experimental anti-cancer (Xodo et al., 2012) and anti-fungal treatments (Liang et al.,2016).
 +
            Seeing how valuable the product could be, we decided to  
 
             genetically engineer part of the chlorophyll degradation pathway in <i> E. coli </i> to produce pheophorbide a.
 
             genetically engineer part of the chlorophyll degradation pathway in <i> E. coli </i> to produce pheophorbide a.
 
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                   <p>Dr. Gijs van Rooijen is the current Chief Scientific Officer of Genome Alberta. He obtained his MSc in Molecular Sciences from the Agricultural University in Wageningen and a PhD in Plant Molecular Biology from the University of Calgary. Dr. Gijs worked for SemBioSys Genetics and (co)authored and co(invented) 25 issued patents in the US.
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                   <p>Dr. Gijs van Rooijen is currently the Chief Scientific Officer of Genome Alberta. He obtained his MSc in Molecular Sciences from the Agricultural University in Wageningen and a PhD in Plant Molecular Biology from the University of Calgary. Dr. Gijs worked for SemBioSys Genetics and (co)authored and co(invented) 25 issued patents in the US.
 
                     <br><br> The goal of the meeting was to obtain a professional opinion regarding the value and feasibility of the project, and the issues we would face in terms of its execution. We also wanted to talk more about enzyme essays for our specific proteins.
 
                     <br><br> The goal of the meeting was to obtain a professional opinion regarding the value and feasibility of the project, and the issues we would face in terms of its execution. We also wanted to talk more about enzyme essays for our specific proteins.
                     <br><br> Dr. Gijs expressed concerns about scaling up our project idea, especially since we are using proteins and purifying them with His tags which would be very expensive in the long run. He had some reservations with binding affinity to chlorophyll and yielding enough CBP from E.coli to make it worthwhile. Additionally, loss of product at each step would be inevitable.  He stated that 10% of the work is the proof of concept but 90% is the optimization. In terms of biotechnology, a project would have more value if the technology is something “people are screaming for”. We should also consider whether the stakeholders are looking for innovation or whether they are happy with the status quo. To make a successful startup, “Helping farmers is a noble goal” but at the end of the day you need to deal with people looking at spreadsheets. Dr. Gijs suggested testing pheophorbide on breast cancer cell lines, or other potential applications to increase the value of the project. Being a plant biotechnologist he thinks that genetic modification at the crop level would be the best solution because it is best to tackle a problem at the base level. Ultimately, Dr. Gijs thought we would face many hurdles to develop the project into a commercial system.
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                     <br><br> Dr. Gijs expressed concerns about scaling up our project idea, especially since we are using proteins and purifying them with His tags which would be very expensive in the long run. He had some reservations with binding affinity to chlorophyll and yielding enough chlorophyll binding protein from E.coli to make it worthwhile. Additionally, loss of product at each step would be inevitable.  He stated that 10% of the work is the proof of concept but 90% is the optimization.
 +
                    In terms of biotechnology, a project would have more value if the technology is something “people are screaming for”.  
 +
                    We should also consider whether the stakeholders are looking for innovation or whether they are happy with the status quo.
 +
                      To make a successful startup, “Helping farmers is a noble goal” but at the end of the day you need to deal with people looking at spreadsheets.  
 +
                      Dr. Gijs suggested testing pheophorbide on breast cancer cell lines, or other potential applications to increase the value of the project.
 +
                      Being a plant biotechnologist he thinks that genetic modification at the crop level would be the best solution because it is best to tackle a problem at the base level.
 +
                        Ultimately, Dr. Gijs thought we would face many hurdles to develop the project into a commercial system.
 
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             Craig, Angela, and Ward, stressed two major hurdles farmers face when they produce and sell their seeds to oil processors;
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             Craig, Angela, and Ward, stressed two major hurdles farmers face when they produce and sell their seeds to oil processors:
             the unpredictable weather and the unstandardized seed grading system. We turned to computation as a solution to address  
+
             unpredictable weather and unstandardized seed grading practices. We turned to computation as a solution to address  
             these problems. Current weather forecasts cannot predict weather 3-4 months into the future, preventing farmers from  
+
             both problems. Current weather forecasts cannot predict weather 3-4 months into the future, preventing farmers from  
             employing agronomic techniques that could save their crops. Therefore, we wanted to see if we could design a long-term  
+
             employing agronomic techniques that could save their crops. This drove us to design a long-term weather prediction system  
            weather prediction system for farmers to use.
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            to give farmers the tools they need to make the right choices.
 
             <br><br>
 
             <br><br>
             The current grading system involves a worker manually crushing a batch of 500 seeds and identifying each seed's colour based on a colour chip.
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             The current seed grading system involves a worker manually crushing a batch of 500 seeds and identifying each seed's colour based on a colour chip.
 
             The percentage of green seeds found denotes the grade of the batch, with a lower grade indicating higher green seed content.  
 
             The percentage of green seeds found denotes the grade of the batch, with a lower grade indicating higher green seed content.  
 +
            Higher green seed percentage means that a farmer's harvest is sold at a discount to grain elevators, effectively taking money out of their pockets.
 
             This process is very subjective, as it relies on the human eye to determine colour, and therefore grade.  
 
             This process is very subjective, as it relies on the human eye to determine colour, and therefore grade.  
 
             Farmers need a standardized system that will produce the same result every time, so they do not incur loss due to human subjectivity.
 
             Farmers need a standardized system that will produce the same result every time, so they do not incur loss due to human subjectivity.
             We sought expert advice to build a machine that can solve this problem.
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             We sought expert advice to build a machine that can solve this problem. Not just any machine. A <a class = "abody" href = "https://2019.igem.org/Team:Calgary/Modelling/StandardizedSeedGrading"> mean green machine </a>.
 
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                   <p>               1. Do you believe there is value to standardize % Distinctly Green Seed with a tool?
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                   <p>
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                    Jeff Danielson is the Manager of Grain Inspection for Viterra Inc. Viterra is Canada's largest grain handler, so it was essential to get feedback
 +
                    on our proposed seed grading platform, as it would directly affect their workflow. Jeff was kind enough to respond to our questions over email.
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                    <br><br><b>1. Do you believe there is value to standardize % Distinctly Green Seed with a tool?</b>
 
                     <br><br>Yes, a more uniform result that removes the variability of individuals perception of color would be beneficial.
 
                     <br><br>Yes, a more uniform result that removes the variability of individuals perception of color would be beneficial.
  
                     <br><br>2. How is % Distinctly Green Seed currently evaluated at your elevator?
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                     <br><br><b>2. How is % Distinctly Green Seed currently evaluated at your elevator?</b>>
 
                     <br><br>The process is done by counting 500 seeds with a canola “ruler”, then the seeds are transferred to a piece of masking tape, the seeds are crushed flat using a hand roller. Seeds are crushed flat in order to expose the inside of the seed.  The seeds that qualify as distinctly green (DGR) are then counted and a percentage is calculated. The Canadian Grain Commission provides a visual color reference guide to help determine what is distinctly green and what is not distinctly green. This whole process can take anywhere from 30 seconds to a couple of minutes depending on the graders ability and the canola quality.
 
                     <br><br>The process is done by counting 500 seeds with a canola “ruler”, then the seeds are transferred to a piece of masking tape, the seeds are crushed flat using a hand roller. Seeds are crushed flat in order to expose the inside of the seed.  The seeds that qualify as distinctly green (DGR) are then counted and a percentage is calculated. The Canadian Grain Commission provides a visual color reference guide to help determine what is distinctly green and what is not distinctly green. This whole process can take anywhere from 30 seconds to a couple of minutes depending on the graders ability and the canola quality.
  
                     <br><br>3. Do you know how % Distinctly Green Seed gets converted into a dollar amount?
+
                     <br><br><b>3. Do you know how % Distinctly Green Seed gets converted into a dollar amount?</b>>
 
                     <br><br>The Canadian Grain Commission dictates the allowable tolerance for distinctly green in all three grades of Canola: Canola 1 Canada can have a maximum of 2% DGR, Canola 2 Canada can have a maximum of 6% DGR, Canola 3 Canada can have a maximum of 20% DGR. The price for these different grades or qualities of Canola is set by  standard market forces of supply and demand. The problem with DGR is that it takes more time and resources to process and purify the finished oil.  So this time and resources means increased expenses which means canola with DGR is less valuable. Canola crushing facilities would know the actual cost to process higher levels of DGR.
 
                     <br><br>The Canadian Grain Commission dictates the allowable tolerance for distinctly green in all three grades of Canola: Canola 1 Canada can have a maximum of 2% DGR, Canola 2 Canada can have a maximum of 6% DGR, Canola 3 Canada can have a maximum of 20% DGR. The price for these different grades or qualities of Canola is set by  standard market forces of supply and demand. The problem with DGR is that it takes more time and resources to process and purify the finished oil.  So this time and resources means increased expenses which means canola with DGR is less valuable. Canola crushing facilities would know the actual cost to process higher levels of DGR.
  
                     <br><br>4. Who do you think will use a tool to standardize % Distinctly Green Seed?
+
                     <br><br><b>4. Who do you think will use a tool to standardize % Distinctly Green Seed?</b>
 
                     <br><br>Primary elevators, port terminals, crushing facilities, Canadian Grain Commission, independent service providers, some producers
 
                     <br><br>Primary elevators, port terminals, crushing facilities, Canadian Grain Commission, independent service providers, some producers
  
                     <br><br>5. What is the frequency at which suppliers and yourselves disagree on the grade of the canola?
+
                     <br><br><b>5. What is the frequency at which suppliers and yourselves disagree on the grade of the canola?</b>
 
                     <br><br>The tolerance for DGR is quite wide within each grade, so there tends to be limited disagreement.  The variability of the human eye and interpretation of color, can lead to disagreements around samples that are on the margins of the grade tolerances. Keep in mind that DGR canola is quite uncommon.  Canola is a very hardy plant and tends to reach full maturity in most crop years.  There can be years where there is essentially no DGR canola in the harvest.
 
                     <br><br>The tolerance for DGR is quite wide within each grade, so there tends to be limited disagreement.  The variability of the human eye and interpretation of color, can lead to disagreements around samples that are on the margins of the grade tolerances. Keep in mind that DGR canola is quite uncommon.  Canola is a very hardy plant and tends to reach full maturity in most crop years.  There can be years where there is essentially no DGR canola in the harvest.
  
                     <br><br>6. Would you rather use a physical device or an app to standardize Canola Grading?
+
                     <br><br><b>6. Would you rather use a physical device or an app to standardize Canola Grading?</b>
                     <br><br> As long as it works consistently, is accurate and easy to use it wouldn’t matter.
+
                     <br><br>As long as it works consistently, is accurate and easy to use it wouldn’t matter.
  
                     <br><br>7. Do you know anyone that has access to images of counts/quality of canola seeds? We can use this information to accurately build our device.
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                     <br><br><b>6. Would you rather use a physical device or an app to standardize Canola Grading?</b>
                     <br><br> The Canadian Grain Commission would be able to provide you with the Canola Color Reference material.  There is a GCG office located in Calgary.  I can provide you with samples of green canola and referrals of where to buy supplies for crushing canola for grading.
+
                     <br><br>The Canadian Grain Commission would be able to provide you with the Canola Color Reference material.  There is a GCG office located in Calgary.  I can provide you with samples of green canola and referrals of where to buy supplies for crushing canola for grading.
 
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                   <p> The team talked with Dr. Alim to gain some more context on certain aspects of the seed grading project. Namely, we wished to learn about methods of analyzing digital colour information and relating it to cut-off points based on a colour chip. This is due to the fact that current seed grading is done by comparing seed smears to a physical colour chip. As Dr. Alim’s areas of interest include applications of signal processing to computer graphics and visualization, he was quite helpful in pointing us in the right direction.
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                   <p> Dr. Usman Alim is an Associate Professor and Director of the Data Science program at the University of Calgary.
                     <br><br>Dr. Alim initially brought up deep learning approaches like with convolutional neural networks, but the team explained that a lack of labelled seed data precluded this methodology from being used. As an alternative, Dr. Alim suggested using clustering methodologies to identify pixel clusters for yellow seeds and distinctly green seeds, even if arbitrary cut-off surfaces needed to be applied. These clusters would utilize different colour spaces, as colour spaces are essentially data transforms. Additionally, Dr. Alim brought up the Lab colourspace, as it designed so that even small differences in values correspond with human-visible differences. Unfortunately, when exploring the colour spaces of the seeds, there was too much data and too much overlap in pixel values between seeds for clustering methods to be applied. However, examination of different colour spaces revealed that they could be represented as geometric volumes, leading to the current approach of using colour space distance calculations for pixel analysis.
+
                    The team talked with Dr. Alim to gain some more context on certain aspects of the seed grading project.
 +
                    Namely, we wished to learn about methods of analyzing digital colour information and relating it to cut-off points based on a colour chip.  
 +
                   
 +
                    As Dr. Alim’s research background into applications of signal processing to computer graphics and visualization made him an ideal contact to influence Mean Green Machine.
 +
                     <br><br>Dr. Alim initially brought up deep learning approaches like with convolutional neural networks,
 +
                    but the team explained that a lack of labelled seed data precluded this methodology from being used.  
 +
                    As an alternative, Dr. Alim suggested using clustering methodologies to identify pixel clusters for yellow seeds and distinctly green seeds,
 +
                      even if arbitrary cut-off surfaces needed to be applied. These clusters would utilize different colour spaces, as colour spaces are essentially data transforms.
 +
                      Unfortunately, when exploring the colour spaces of the seeds, there was too much data and too much overlap in pixel values between seeds for clustering methods to be applied.  
 +
                      However, examination of different colour spaces revealed that they could be represented as geometric volumes,
 +
                      leading to the current approach of using colour space distance calculations for pixel analysis.
 
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                   <p>Sean Mason was a mentor for the hardware of a 3rd year mechanical engineering student  
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                   <p>Sean Mason is a journeyman machinist and
                    at UofC with years of experience as a machinist, was critical in concretizing mean green
+
                    mechanical engineering student. Our team had no previous experience with CADs or working with
                    machine. Our team had no previous experience with <b>Definition</b>CADs or working with
+
 
                     machine shops. Sean Mason acted as a mentor for designing parts in Solidworks,  
 
                     machine shops. Sean Mason acted as a mentor for designing parts in Solidworks,  
                     design considerations, measurements, tolerances, who to talk to for sourcing the parts,  
+
                     design considerations, measurements, tolerances, sourcing parts,  
 
                     and choosing the right materials for the job. Whilst it remained an iGEM project, Sean  
 
                     and choosing the right materials for the job. Whilst it remained an iGEM project, Sean  
 
                     Mason stood right behind us and made sure we were doing things correctly in Solidworks  
 
                     Mason stood right behind us and made sure we were doing things correctly in Solidworks  
                     by providing tutorials on different aspects of Solidworks and more than once made us  
+
                     by providing tutorials on different aspects of Solidworks and made us  
                     consider significant portions of the design to add functionality or ease of use. Ideas  
+
                     re-consider significant portions of the design to add functionality or ease of use. Ideas  
                     such as <b>Definition</b>chamfers, which were never considered were added in due to his  
+
                     such as chamfers, were implemented based on his expertise in bringing designs to life.  
                    expertise in creating real-life projects that work. He also oversaw safety for drilling,  
+
                    He also oversaw safety for drilling,  
                     sanding, and glueing for the first assembly of the machine. His ideas on how to create an  
+
                     sanding, and gluing for the first assembly of the machine. His masterful insight on creating an
                     easy-to-assemble standardization tool helped us create the way it is now. </p>
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                     easy-to-assemble standardization tool helped us make the meanest, greenest, seed grader in the prairies. </p>
 
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             <h1>4. Evaluate</h1>
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             <h2>4) Pheophorbide</h2>
            <h2>Pheophorbide- A unique solution to an industry problem</h2>
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           <p>
 
           <p>
             Attending CanolaPALOOZA allowed us to evaluate our project <b>to see if we were effectively  
+
             Attending CanolaPALOOZA allowed us to evaluate the need for our project to see if we were effectively  
             accomplishing our goal</b> of helping the canola industry. Overall, we got very positive feedback  
+
             accomplishing our goal of helping the canola industry.
 +
            Overall, we got very positive feedback  
 
             on our projects, but we did learn about other problems the industry faces aside from green seed.  
 
             on our projects, but we did learn about other problems the industry faces aside from green seed.  
             Canola suffers from a <b>fungal disease</b> caused by <i>Sclerotinia sclerotiorum</i> leading to
+
             Fungal species such as <i>Sclerotinia sclerotiorum</i>  
             huge losses in crop yield annually <b> (REFERENCE???) </b>. This was interesting to us because our
+
             is "one of the most serious and damaging diseases of [canola seed]" (Garg et al., 2010). We saw an opportunity here.
             product, pheophorbide, is an anti-fungal agent that could be used to address this problem.
+
            An opportunity to <a class = "abody" href = "https://2019.igem.org/Team:Calgary/RepurposingChlorophyll">
 +
             repurpose the chlorophyll</a> captured from canola oil and convert it into pheophorbide for
 +
            use as an anti-fungal agent. But first, we needed to investigate how pheophorbide can work as an anti-fungal.
 
             <br><br>
 
             <br><br>
             We decided to <b>further investigate</b> the effect <i>S. sclerotiorum</i> has on the canola industry  
+
             We decided to further investigate the effect <i>S. sclerotiorum</i> has on the canola industry  
 
             (particularly on the farmers) and how we could use pheophorbide as a unique solution to combat it.
 
             (particularly on the farmers) and how we could use pheophorbide as a unique solution to combat it.
 
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                 <div class="modal-body">
                   <p>Our early background research identified pheophorbide as a candidate for use as a therapeutic agent for photodynamic therapy. Photodynamic therapy (PDT) is a medical treatment which utilizes photosensitizing compounds, such as Pheophorbide, in the presence of specific wavelengths of light in order to produce a directed cytotoxic effect. PDT is currently being investigated for applications in cancer treatment and there already exist several commercial photosensitizers such as Photofirin. We were interested in Pheophorbide’s potential as a PDT photosensitizing agent, but we had to talk to someone to learn more about it.
+
                   <p>Our early background research identified pheophorbide as a candidate for use as a therapeutic agent for photodynamic therapy.
                     <br><br>Dr. Ronald Moore is a professor of Surgery and Oncology at the University of Alberta and currently serves as the Mr. Lube Chair in Uro-Oncology Research. He has over 25 years of experience in researching novel therapies for genitourinary malignancies.
+
                    Photodynamic therapy (PDT) is a medical treatment which utilizes photosensitizing compounds, such as Pheophorbide,  
                     <br><br>Our discussion with Dr. Moore gave us an excellent overview of PDT’s current usage as well as what considerations doctors make when choosing a photosensitizing agent for PDT. Generally, the world of experimental cancer research is constantly in flux, changing with new developments and discoveries. While the current consensus is positive, tumorous cells substantially retain photosensitizers, the future's uncertain and the demand for photosensitizing agents in the future is shrouded in darkness.
+
                    in the presence of specific wavelengths of light in order to produce a directed cytotoxic effect (Xodo et al. 2016).
                     <br><br>Following our conversation with Dr. Moore, it was immediately apparent that we could not put all our eggs into one PDT-basket. We needed to explore other ways. During our literature review, we identified PDT as a potential method to treat fungal infestations. After our meeting with Dr. Moore we realized that pheophorbide has more potential as an anti-fungal agent than as an experimental cancer treatment drug.
+
                    PDT is currently being investigated for applications in cancer treatment and there already exist several commercial photosensitizers  
 +
                    such as Photofirin. We were interested in Pheophorbide’s potential as a PDT photosensitizing agent,  
 +
                    but we had to talk to someone to learn more about it.
 +
                     <br><br>Dr. Ronald Moore is a professor of Surgery and Oncology at the University of Alberta and currently serves as the  
 +
                    Mr. Lube Chair in Uro-Oncology Research. He has over 25 years of experience in researching novel therapies for genitourinary malignancies.
 +
                     <br><br>Our discussion with Dr. Moore gave us an excellent overview of PDT’s current usage as well as what considerations doctors make when  
 +
                    choosing a photosensitizing agent for PDT. Generally, the world of experimental cancer research is constantly in flux, changing with  
 +
                    new developments and discoveries. While the current consensus is positive, tumorous cells substantially retain photosensitizers,  
 +
                    the future's uncertain and the demand for photosensitizing agents in the future is shrouded in darkness.
 +
                     <br><br>Following our conversation with Dr. Moore, it was immediately apparent that we could not put all our eggs into one PDT-basket.
 +
                    We needed to explore other ways. During our literature review, we identified PDT as a potential method to treat fungal infestations.
 +
                    After our meeting with Dr. Moore we realized that pheophorbide has more potential as an anti-fungal agent than as an experimental cancer treatment drug.
 
                   </p>
 
                   </p>
 
                 </div>
 
                 </div>
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                 </div>
 
                 </div>
 
                 <div class="modal-body">
 
                 <div class="modal-body">
                   <p>After the meeting with Dr. Moore, we immediately had to learn more about working with fungi and anti-fungal assays. Through background research, we had the idea to use the “drop method”, applying drops of pheophorbide to different locations on an agar plate, with varying concentrations to see if it inhibited fungal growth. However, we needed to validate our experimental design. So we met with Dr. Heather Addy.
+
                   <p>After the meeting with Dr. Moore, we immediately had to learn more about working with fungi and anti-fungal assays.  
                     <br><br>Dr. Heather Addy is a mycologist and plant biologist who specializes in plant-fungal interactions at the University of Calgary. Her expertise in the field meant we absolutely had to talk to her to inform the design of our anti-fungal assays.
+
                    Through background research, we had the idea to use the “drop method”, applying drops of pheophorbide to different locations on an agar plate,
                     <br><br>Dr. Addy gave us the idea to use the “disc method”, immersing paper discs in solubilized pheophorbide, placing them around a fungal culture placed in the centre of a potato dextrose agar plate. The fungal colony would grow and eventually come into contact with the pheophorbide discs. Over the course of the experiment, we would measure the distance from the centre of the plate to the edge of the fungal colony’s growth. If pheophorbide does in fact have an inhibitory effect on fungal growth, then there would be a decreased rate of growth for the portions of the colony interacting with pheophorbide.
+
                    with varying concentrations to see if it inhibited fungal growth. However, we needed to validate our experimental design. So we met with Dr. Heather Addy.
                     <br><br>Dr. Addy generously put us in touch with Fran Cusack, a Biological Sciences Technician who prepares fungal samples for classes. Fran was kind enough to provide us with samples of <i>Pestalotiopsis microspora</i> and <i>S. sclerotiorum</i>, the same fungus which commonly afflicts canola crops.
+
                     <br><br>Dr. Heather Addy is a mycologist and plant biologist who specializes in plant-fungal interactions at the University of Calgary.  
                     <br><br>We now had the information and tools at our disposal, to begin testing pheophorbide’s application as an anti-fungal agent. However, there was still a gap in our knowledge regarding the what the average Albertan farmer goes through when faced with fungi, like <i>S. sclerotiorum</i>
+
                    Her expertise in the field meant we absolutely had to talk to her to inform the design of our anti-fungal assays.
 +
                     <br><br>Dr. Addy gave us the idea to use the “disc method”, immersing paper discs in solubilized pheophorbide, placing them around a  
 +
                    fungal culture placed in the centre of a potato dextrose agar plate. The fungal colony would grow and eventually come into contact with the pheophorbide discs.
 +
                    Over the course of the experiment, we would measure the distance from the centre of the plate to the edge of the fungal colony’s growth.  
 +
                    If pheophorbide does in fact have an inhibitory effect on fungal growth, then there would be a decreased rate of growth for the portions of the colony interacting with pheophorbide.
 +
                     <br><br>Dr. Addy generously put us in touch with <a class ="abody" href ="https://2019.igem.org/Team:Calgary/Attributions#ChlorophyllRepurposing">Fran Cusack</a>, a Biological Sciences Technician who prepares fungal samples for classes.  
 +
                    Fran was kind enough to provide us with samples of <i>Pestalotiopsis microspora</i> and <i>S. sclerotiorum</i>, the same fungus which commonly afflicts canola crops.
 +
                     <br><br>We now had the information and tools at our disposal, to begin testing pheophorbide’s application as an anti-fungal agent.  
 +
                    However, there was still a gap in our knowledge regarding the what the average Albertan farmer goes through when faced with fungi, like <i>S. sclerotiorum</i>
 
                   </p>
 
                   </p>
 
                 </div>
 
                 </div>
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                 </div>
 
                 </div>
 
                 <div class="modal-body">
 
                 <div class="modal-body">
                   <p>To say John Mayko is merely an Agronomy Specialist from Mundare, Northern Alberta would vastly undersell his involvement with the Alberta Canola industry and his contribution to yOIL.
+
                   <p>To say John Mayko is merely an Agronomy Specialist from Mundare, Northern Alberta would vastly undersell his involvement  
                     <br><br>Receiving his Masters degree in Engineering Agrology from the University of Alberta, John is the president of Apex Agrology Services and currently sits on the Board of Directors for the Alberta Canola Producers Commission. His many years as a Senior Agri-Coach meant he could give us a clear indication of how fungi affect Albertan farmers.
+
                    with the Alberta Canola industry and his contribution to yOIL.
                     <br><br>John himself has had to deal with fungus. According to him, “Anybody growing canola in Alberta will have to deal with it”. There is no question that fungus is an issue for canola farmers, but what is being done about it?
+
                     <br><br>Receiving his Masters degree in Engineering Agrology from the University of Alberta,  
                     <br><br>Unfortunately, there is no fix for fungus. Once a crop has been afflicted by fungal blight, it must be discarded, there is no turning back the clock. At the early bloom stage, every farmer must make a decision whether or not to apply anti-fungal treatments to their crop. It is a costly proposition ($20-$30 per acre according to Dr. Kelly Turkington) which does not give a 100% guarantee.
+
                    John is the president of Apex Agrology Services and currently sits on the Board of Directors for the Alberta Canola Producers Commission.  
                     <br><br>From John, we learnt that our pheophorbide application would have to be preventative not prescriptive. He also gave us the indication that for our product to be viable, it would have to be cheaper to apply than current methods.
+
                    His many years as a Senior Agri-Coach meant he could give us a clear indication of how fungi affect Albertan farmers.
 +
                     <br><br>John himself has had to deal with fungus. According to him, <b>“Anybody growing canola in Alberta will have to deal with it”</b> .  
 +
                    There is no question that fungus is an issue for canola farmers, but what is being done about it?
 +
                     <br><br>Unfortunately, there is no fix for fungus. Once a crop has been afflicted by fungal blight,  
 +
                    it must be discarded, there is no turning back the clock. At the early bloom stage, every farmer must make a decision whether or not to apply anti-fungal treatments to their crop.  
 +
                    It is a costly proposition ($20-$30 per acre according to Dr. Kelly Turkington) which does not give a 100% guarantee.
 +
                     <br><br>From John, we learnt that our pheophorbide application would have to be preventative not prescriptive.
 +
                    He also gave us the indication that for our product to be viable, it would have to be cheaper to apply than current methods.
 
                     <br><br>Now that we had an idea of the farmer’s perspective towards fungi, it was time for us to learn the pathologist’s perspective.
 
                     <br><br>Now that we had an idea of the farmer’s perspective towards fungi, it was time for us to learn the pathologist’s perspective.
 
                   </p>
 
                   </p>
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                 </div>
 
                 </div>
 
                 <div class="modal-body">
 
                 <div class="modal-body">
                   <p>Dr. Turkington holds a Masters Degree and Ph.D in Plant Pathology, focusing on the epidemiology of <i>S. sclerotiorum</i> stem rot in canola. For the last 23 years, Kelly has been working as a Research Scientist for Agriculture and Agri-Food Canada at the Lacombe Research Centre in Alberta.
+
                   <p>Dr. Turkington holds a Masters Degree and Ph.D in Plant Pathology, focusing on the epidemiology of <i>S. sclerotiorum</i> stem rot in canola.
 +
                    For the last 23 years, Kelly has been working as a Research Scientist for Agriculture and Agri-Food Canada at the Lacombe Research Centre in Alberta.
 
                     <br><br>We consulted him to learn more about the progression of <i>S. sclerotiorum</i> fungus, its impact on farmers and preventative and prescriptive measures to combat it.
 
                     <br><br>We consulted him to learn more about the progression of <i>S. sclerotiorum</i> fungus, its impact on farmers and preventative and prescriptive measures to combat it.
                     <br><br>According to Kelly, the cost to treat fungi like <i>S. sclerotiorum</i> can be around $20-$30 per acre, severely reducing a farmer’s bottom line. To assess risk of fungal growth, farmers employ a checklist by assigning point values to certain factors including the plant, the host and the environment. This is a very broad indication of risk and is not an exact science as there is ambiguity in the checklist. Some companies have started using DNA-based chips to quantitatively determine the percentage infestation of a plant.
+
                     <br><br>According to Kelly, the cost to treat fungi like <i>S. sclerotiorum</i> can be around $20-$30 per acre,  
                     <br><br>In determining if Pheophorbide would be an ideal anti-fungal agent, he directed us to consider the full chemical profile of pheophorbide. Not just it’s effects on fungi but also on non-target organisms. Another consideration is the societal aspect of such a product. Members of the farming community and industry only care if the product is cheap and effective, but society as a whole tends to support products which are “organic” or come from the environment in a responsible manner.
+
                    severely reducing a farmer’s bottom line. To assess risk of fungal growth,  
 +
                    farmers employ a checklist by assigning point values to certain factors including the plant, the host and the environment.  
 +
                    This is a very broad indication of risk and is not an exact science as there is ambiguity in the checklist.  
 +
                    Some companies have started using DNA-based chips to quantitatively determine the percentage infestation of a plant.
 +
                     <br><br>In determining if Pheophorbide would be an ideal anti-fungal agent, he directed us to consider the full chemical profile of pheophorbide.
 +
                    Not just it’s effects on fungi but also on non-target organisms. Another consideration is the societal aspect of such a product.
 +
                    Members of the farming community and industry only care if the product is cheap and effective, but society as a whole tends to support products which are
 +
                    “organic” or come from the environment in a responsible manner.
 
                   </p>
 
                   </p>
 
                 </div>
 
                 </div>
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           <div class="header-area">
 
           <div class="header-area">
             <h1 id="designIterate">5. Iterate on Design</h1>
+
             <h1 id="designIterate">4. Evaluate and Iterate on Design</h1>
 
             <h2>Further improving our project design</h2>
 
             <h2>Further improving our project design</h2>
 
           </div>
 
           </div>
 
           <p>
 
           <p>
             <b>We are dedicated to improving our solutions to better serve the people in our community.</b> We have
+
             We are dedicated to improving our solutions to serve the people in our community. We involved
             made it our mission to involve our stakeholders and academic advisors in the <b>ideation, design,
+
             our stakeholders and academic advisors in every stage of our solutions. From investigation and ideation, to
             and testing of our solutions.</b>
+
             prototyping and testing, yOIL's design process was purely human-centered.
 +
       
 
             <br><br>
 
             <br><br>
             To do this, we hosted a mid-summer faculty talk, inviting the experts we consultated with  
+
             We hosted a mid-summer faculty talk, inviting experts we consultated with  
             earlier in the project. Additionally, with our tools for the farmers, we met with the <b>Canadian Grain  
+
             earlier in the project, to <b>close the loop</b> with them. Additionally, with our tools, we met with the Canadian Grain Commission
             Commission</b> to <b>perform testing of our standardized seed grading machine</b>. Our weather prediction model  
+
             to perform <b>testing of our standardized seed grading machine</b>. Our weather prediction model, <a class = "abody" href ="https://2019.igem.org/Team:Calgary/SunnyDays">Sunny Days</a>
 
             is <b>undergoing peer-review at Alberta Academic Review</b> with the goal of being published and made available to  
 
             is <b>undergoing peer-review at Alberta Academic Review</b> with the goal of being published and made available to  
             the academic community for other researchers to improve.  
+
             the academic community for improvement.  
 
           </p>
 
           </p>
  
 
           <div class="fullInterview">
 
           <div class="fullInterview">
 
             <h2>Faculty Talk</h2>
 
             <h2>Faculty Talk</h2>
 +
            <p>
 
             Once we consolidated and incorporated the advice from our expert consultations with professors,  
 
             Once we consolidated and incorporated the advice from our expert consultations with professors,  
 
             and canola industry members, we were able to design and define the various wet and dry lab projects
 
             and canola industry members, we were able to design and define the various wet and dry lab projects
             encompassing yOIL. From here, we decided to host a mid-summer faculty talk to get further feedback  
+
             encompassing yOIL. From here, we decided to host a mid-summer faculty talk to get iterative feedback  
             on our experimental design and current progress. Supervisors, past iGEM Calgary members, and our business contacts, and the aforementioned professor were also invited to the presentation to give us feedback on the feasibility of our projects.
+
             on our experimental design and current progress. Supervisors, past iGEM Calgary members, and business contacts,  
 +
            were also invited to the presentation to provide their unique perspectives.
 
             <br><br>
 
             <br><br>
             Overall, we received positive feedback on our dry and wet lab projects. The biochemistry and  
+
 
             microbiology professors we invited confirmed that our T7-inducible genetic circuit with BL21 was simple
+
             Overall, we recieved positive feedback and constructive criticism on all aspects of yOIL. Biochemists and  
             and well characterized for protein production. However, we were informed that our signal peptides may not
+
             microbiologists in attendance confirmed that our T7-inducible circuits in BL21(DE3) <i>E.coli</i> was the way to go.
            be as efficient as extracting the protein from the cell lysate, but would be a good future direction to
+
             However, we were informed that using signal peptides won't be as efficient as extracting proteins from cell lysate, but is a  
            consider. Additionally, we got a few specific comments about the emulsion system on how to improve it where
+
            good consideration for future scale-up. The idea to use an emulsion system was well recieved as a workaround for preventing
            to get more information about how to design efficient emulsions. However, the general use of an  
+
            our protein from denaturing in oil, and we were commended for considering future industrial scale-up in that regard.
            emulsion-system was well received and we were commended for considering the steps necessary for its industrial scale-up  
+
             Initially, we thought to add ethanol to the chlorophyll binding proteins to release chlorophyll, but we were given an alternative
             (we have this right). A few professors also gave suggestions on how to improve the lab experiments we planned
+
            which would instead temporarily weaken the protein's structure, thus releasing chlorophyll.
            for chlorophyll repurposing. We initially thought to add ethanol to the chlorophyll binding proteins to release chlorophyll,  
+
            but we were given alternative solutions that would instead temporarily weaken the protein’s structure to release
+
            chlorophyll. Additionally, the use of a magnesium utilizing protein was an idea given to characterize an enzyme in the chlorophyll degradation pathway.
+
 
             <br><br>  
 
             <br><br>  
             In conclusion, we were able to take the advice we received from our meetings at the beginning  
+
             We were able to take the advice we received from our meetings at the beginning  
 
             of our project to create clear wet lab and dry lab components each with a defined experimental workflow.  
 
             of our project to create clear wet lab and dry lab components each with a defined experimental workflow.  
 
             The faculty talk was an opportunity for us to concisely present our plans to recieve expert feedback  
 
             The faculty talk was an opportunity for us to concisely present our plans to recieve expert feedback  
 
             again, and iteratively improve our project design.
 
             again, and iteratively improve our project design.
 +
            </p>
  
 
             <br><br><br>
 
             <br><br><br>
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               To verify field implementation of the standardized seed grading project into industry, we went to visit seed grading experts
 
               To verify field implementation of the standardized seed grading project into industry, we went to visit seed grading experts
 
               <a href="https://2019.igem.org/Team:Calgary/Attributions" target="_blank">Romeo Honorio and Scott Kippin</a>  
 
               <a href="https://2019.igem.org/Team:Calgary/Attributions" target="_blank">Romeo Honorio and Scott Kippin</a>  
               from the Canadian Grain Commission (CGC). We met Romeo at where they explained current
+
               from the Canadian Grain Commission (CGC). We brought along our first prototype for Mean Green Machine (MGM)
               attempts to standardize seed grading. Our visit allowed us to have large graded seed samples from the experts,
+
               to get feedback on our hardward implementation.
              that way we can base our grading algorithm on their counts. They also provided significant feedback on
+
              <br><br>
               standardizing the grading conditions. At the CGC office all the walls were a specific grey, and the grading
+
 
              tables another specific grey, and the light fixtures were at very controlled intensities and highly diffused.
+
              We got large graded seed samples from the experts themselves to see how our grading algorithm stacks up.
               These design considerations were embraced by standardized grading machine to allow for a better prototype.  
+
               Seeing the highly standardized environment where seeds are graded, muted grey walls and tables to control light diffusion,
               Another piece of learned information is that farmers do not care about specific percentages of DGR, only the  
+
               gave us more design considerations regarding <a class = "abody" href = "https://2019.igem.org/Team:Calgary/Modelling/StandardizedSeedGrading">
              final grade as that is their paycheck. Grain elevators, oil refineries, and crushers are significantly invested  
+
               Mean Green Machine</a>'s first prototype. More importantly we learned that farmers do not care about specific percentages of  
 +
              distinctly green seed (DGR), only the final grading which also indicates how green their paycheque will be.
 +
             
 +
              <br><br>
 +
             
 +
              Grain elevators, oil refineries, and crushers are significantly invested  
 
               in knowing the exact percentage of the grade as it directly affects the final product canola oil. Even if we  
 
               in knowing the exact percentage of the grade as it directly affects the final product canola oil. Even if we  
 
               started this project by hearing about a farmer's grievance with the grading system, our project would have serious effects  
 
               started this project by hearing about a farmer's grievance with the grading system, our project would have serious effects  
               throughout the canola pipeline. So we learned that whilst farmers would benefit from our invention,  
+
               throughout the canola pipeline. Whilst farmers would benefit from our invention,  
 
               the other cogs in the canola industry would be more significantly affected by the standardization of DGR grading.
 
               the other cogs in the canola industry would be more significantly affected by the standardization of DGR grading.
 
               <br><br>
 
               <br><br>
 
               Another piece of information that we learned is that there exists some machines that can grade chlorophyll  
 
               Another piece of information that we learned is that there exists some machines that can grade chlorophyll  
               successfully on the market, but they are usually slow, and cost upwards of 40,000-60,000 dollars which is more than what  
+
               successfully on the market, but they are usually slow, and cost upwards of $40,000-$60,000 which is more than what  
 
               grain elevators are willing to pay. Romeo liked our machine but wishes it will be taken further both in algorithm  
 
               grain elevators are willing to pay. Romeo liked our machine but wishes it will be taken further both in algorithm  
 
               quality and user-oriented hardware design, especially in terms of the speed at which it is built. He also emphasized buying the  
 
               quality and user-oriented hardware design, especially in terms of the speed at which it is built. He also emphasized buying the  
               best camera possible as the camera affects the entire project, hopefully an infrared camera to help us have better  
+
               best camera possible as the camera affects the entire project, hopefully an infrared camera to give better readings.
               readings, they also suggested a future direction of grading wheat as the current methods are even more prone to human error than canola.
+
               The also gave us the idea to apply MGM to grading wheat, an excellent future direction to reduce the human error present in that sector.
 
             </p>
 
             </p>
 
           </div>
 
           </div>
 +
 +
          <div class="header-area">
 +
              <h1>References</h1>
 +
              <ol type="1">
 +
                <li><p class ="hangingindent">E Xodo, L., Rapozzi, V., Zacchigna, M., Drioli, S., & Zorzet, S. (2012).
 +
                  The chlorophyll catabolite pheophorbide a as a photosensitizer for the photodynamic therapy. Current medicinal chemistry, 19(6), 799-807.</p></li>
 +
                  <li><p class ="hangingindent">Liang, Y. I., Lu, L. M., Chen, Y., & Lin, Y. K. (2016).
 +
                    Photodynamic therapy as an antifungal treatment. Experimental and therapeutic medicine, 12(1), 23-27.</p></li>
 +
                    <li><p class ="hangingindent">Garg, H., Atri, C., Sandhu, P. S., Kaur, B., Renton, M., Banga, S. K., ... & Banga, S. S. (2010).
 +
                      High level of resistance to Sclerotinia sclerotiorum in introgression lines derived from hybridization between wild crucifers and
 +
                      the crop Brassica species B. napus and B. juncea. Field Crops Research, 117(1), 51-58.</p></li>
 +
                </ol>
 +
            </div>
 +
            <p>
 +
 +
            </p>
 
         </div>
 
         </div>
 
       </div>
 
       </div>

Latest revision as of 19:58, 16 November 2019

Human Practices

Solutions for Real People

Human Centered Design

How can we solve problems that affect people?

Providing a solution to the green seed problem could be the key to alleviating losses felt by those in the canola industry. Our team’s vision was to make yOIL that solution. In order to do so, we had to ensure that we created something that was effective, usable, and targeted areas which impacted people the most. The only way to achieve this goal, was to integrate our end-users in creating our solutions.

We utilized a human-centered design process and identified a set of steps to guide our work. This lead to integration with key stake holders throughout the project lifecycle, and ultimately the all-encompassing solutions found in yOIL.

yOIL started off small in scope. Nothing more than a process to remove chlorophyll from canola oil. It wasn't until we spoke to everyone involved in the industry that we realized we could do so much more.

This is the story of how yOIL was born from a desire to help oil producers, and grew into an all-encompassing solution through iterative meetings with contacts from all over the industry.

1. Understand the Problem

Key Stakeholder Interviews

Before BioBrick design and lab work began, there were questions we needed answered. We needed to know what exactly the green seed issue entails. How large it is? Who does it impact? What is being done to stop it?

To answer these questions, we identified four groups of people that we needed to speak with. Our primary research indicated that there are two main entities impacted by the green seed issue: farmers and oil producers. In addition to those two, we also identified agronomists and organizational bodies working within the canola oil industry. From here, we initiated conversations about the green seed issue and discussed how we could utilize synthetic biology as a solution.



What did we discover?

Through our discussions with these industry members, we learned that every stage of canola oil production faces tremendous losses due to the green seed problem. We can think of these stages as part of a pipeline.



With a consumer market desiring pure oil, the green seed problem forces the players in the production pipeline to somehow deal with the green oil produced.


Farmers have no tools to plan for unexpected frost or drought, and gamble their harvest every year on unpredictable whether.

Leading to farmers inevitably producing green seeds, which get downgraded and are sold at a lower price.

The process to grade green seeds is not standardized or objective, making the system less fair.

Oil processors are forced to spend more time and money to purify the oil produced from the green seeds they receive, the archaic acid-activated clay purification method is unsustainable.


Every person we talked to asserted the impact green seed has had on them and emphasized the need for new solutions that would mitigate the losses incurred.

2. Ideate

Propose solutions and verify their need

Having identified and understood the multiple adverse aspects of the green seed problem, our team began brainstorming solutions.

After discussing and analyzing our stakeholders' needs, we decided on two synthetic biology solutions and one engineering based solution. These are all aimed at the oil processing stage where current methods are expensive, result in oil loss, and are not environmentally friendly.

To discover more about these solutions, click on the buttons below.




The above solutions focused on the oil processing stage. However, after hearing stories from farmers like Craig about their losses due to green seed, we were determined to help them as well.

Preventing green seed entirely would be the most beneficial solution for farmers, but we knew that was infeasible in our time frame. We determined that we could help them by creating software and hardware solutions that help with crop management and seed grading.

To learn about these software and hardware solutions, click on the buttons below.




Verifying need - Canolapalooza



Midway through the summer, we were lucky enough to be able to attend canolaPALOOZA. An annual event held in Lacombe, Alberta, canolaPALOOZA is the “agronomy event of the summer” where canola experts from all over the prairies come together to discuss all things canola. Everyone from farmers, agronomists, mycologists and even an undergraduate iGEM team from the University of Calgary descended on a lone canola field in Leduc to talk shop.

Many of our initial contacts, including Ward Toma and Dr. Barthet, were in attendance, in addition to many others. It was an excellent opportunity for us to propose our ideas to a wide cross-section of the canola industry, and get feedback on the commercial viability of our solutions. The event was extremely valuable because this is where we learned about the fungal infestations currently affecting canola crops in Alberta. A fortuitous meeting which would give us the inspiration to repurpose chlorophyll into an anti-fungal agent.

This event lead to a connection with the Canadian Grain Commission, who expressed their interest in the standard seed grading system. They informed us that there had been a similar project in the past that was successful, but got scrapped due to its cost. This inspired us to make our standardized grading machine as inexpensive as possible.

To learn about this process, click the button below.




3. Design Solutions

Expert consultations

With the need for our solutions verified, it was time for us to put shovels in the ground ( or bacteria on the agar) and begin designing yOIL. Our development would be agile and adaptable. Continually meeting with stakeholders and integrating feedback throughout the design process to ensure yOIL holistically embodies the needs of everyone involved in the canola oil pipeline.

1) Chlorophyll Extraction System

The clay method is not reusable, environmentally friendly, or selective to chlorophyll, thus, we needed to come up with an improved solution. Proteins are organic and are made to be highly specific to their substrate, so we thought to design a system that utilized a protein to capture chlorophyll, but we weren’t confident as to which chlorophyll-binding protein to use, how it could be used in an oil environment, and what other considerations to be aware of.

To answer these questions, we spoke to protein bio-chemists, micro-biologists, plant biologists, and chemical engineers who gave us insight into how to design our system.

2) Chlorophyll Repurposing

Chlorophyll cannot be removed from the acid-activated clay chlorophyll after it is bound; however, by using chlorophyll binding proteins the chlorophyll can be released after. In our initial discussions with Ward Toma, he suggested that repurposing chlorophyll could bring new revenue into the canola industry and mitigate some of the losses caused by green seed. Dallas Gade further confirmed that valuable byproducts could be useful to offset expensive input costs. Pheophorbide a is a natural catabolite of chlorophyll that has been investigated recently for its photosensitizing abilities in experimental anti-cancer (Xodo et al., 2012) and anti-fungal treatments (Liang et al.,2016). Seeing how valuable the product could be, we decided to genetically engineer part of the chlorophyll degradation pathway in E. coli to produce pheophorbide a.

Producing functional eukaryotic enzymes in bacteria can be a challenge, especially those involved in a specific pathway. We decided to talk to a microbial biochemistry and a plant biotechnologist about the feasibility of our idea.



3) Tools for the farmers

Craig, Angela, and Ward, stressed two major hurdles farmers face when they produce and sell their seeds to oil processors: unpredictable weather and unstandardized seed grading practices. We turned to computation as a solution to address both problems. Current weather forecasts cannot predict weather 3-4 months into the future, preventing farmers from employing agronomic techniques that could save their crops. This drove us to design a long-term weather prediction system to give farmers the tools they need to make the right choices.

The current seed grading system involves a worker manually crushing a batch of 500 seeds and identifying each seed's colour based on a colour chip. The percentage of green seeds found denotes the grade of the batch, with a lower grade indicating higher green seed content. Higher green seed percentage means that a farmer's harvest is sold at a discount to grain elevators, effectively taking money out of their pockets. This process is very subjective, as it relies on the human eye to determine colour, and therefore grade. Farmers need a standardized system that will produce the same result every time, so they do not incur loss due to human subjectivity. We sought expert advice to build a machine that can solve this problem. Not just any machine. A mean green machine .

4) Pheophorbide

Attending CanolaPALOOZA allowed us to evaluate the need for our project to see if we were effectively accomplishing our goal of helping the canola industry. Overall, we got very positive feedback on our projects, but we did learn about other problems the industry faces aside from green seed. Fungal species such as Sclerotinia sclerotiorum is "one of the most serious and damaging diseases of [canola seed]" (Garg et al., 2010). We saw an opportunity here. An opportunity to repurpose the chlorophyll captured from canola oil and convert it into pheophorbide for use as an anti-fungal agent. But first, we needed to investigate how pheophorbide can work as an anti-fungal.

We decided to further investigate the effect S. sclerotiorum has on the canola industry (particularly on the farmers) and how we could use pheophorbide as a unique solution to combat it.

4. Evaluate and Iterate on Design

Further improving our project design

We are dedicated to improving our solutions to serve the people in our community. We involved our stakeholders and academic advisors in every stage of our solutions. From investigation and ideation, to prototyping and testing, yOIL's design process was purely human-centered.

We hosted a mid-summer faculty talk, inviting experts we consultated with earlier in the project, to close the loop with them. Additionally, with our tools, we met with the Canadian Grain Commission to perform testing of our standardized seed grading machine. Our weather prediction model, Sunny Days is undergoing peer-review at Alberta Academic Review with the goal of being published and made available to the academic community for improvement.

Faculty Talk

Once we consolidated and incorporated the advice from our expert consultations with professors, and canola industry members, we were able to design and define the various wet and dry lab projects encompassing yOIL. From here, we decided to host a mid-summer faculty talk to get iterative feedback on our experimental design and current progress. Supervisors, past iGEM Calgary members, and business contacts, were also invited to the presentation to provide their unique perspectives.

Overall, we recieved positive feedback and constructive criticism on all aspects of yOIL. Biochemists and microbiologists in attendance confirmed that our T7-inducible circuits in BL21(DE3) E.coli was the way to go. However, we were informed that using signal peptides won't be as efficient as extracting proteins from cell lysate, but is a good consideration for future scale-up. The idea to use an emulsion system was well recieved as a workaround for preventing our protein from denaturing in oil, and we were commended for considering future industrial scale-up in that regard. Initially, we thought to add ethanol to the chlorophyll binding proteins to release chlorophyll, but we were given an alternative which would instead temporarily weaken the protein's structure, thus releasing chlorophyll.

We were able to take the advice we received from our meetings at the beginning of our project to create clear wet lab and dry lab components each with a defined experimental workflow. The faculty talk was an opportunity for us to concisely present our plans to recieve expert feedback again, and iteratively improve our project design.




Canadian Grain Commission

To verify field implementation of the standardized seed grading project into industry, we went to visit seed grading experts Romeo Honorio and Scott Kippin from the Canadian Grain Commission (CGC). We brought along our first prototype for Mean Green Machine (MGM) to get feedback on our hardward implementation.

We got large graded seed samples from the experts themselves to see how our grading algorithm stacks up. Seeing the highly standardized environment where seeds are graded, muted grey walls and tables to control light diffusion, gave us more design considerations regarding Mean Green Machine's first prototype. More importantly we learned that farmers do not care about specific percentages of distinctly green seed (DGR), only the final grading which also indicates how green their paycheque will be.

Grain elevators, oil refineries, and crushers are significantly invested in knowing the exact percentage of the grade as it directly affects the final product canola oil. Even if we started this project by hearing about a farmer's grievance with the grading system, our project would have serious effects throughout the canola pipeline. Whilst farmers would benefit from our invention, the other cogs in the canola industry would be more significantly affected by the standardization of DGR grading.

Another piece of information that we learned is that there exists some machines that can grade chlorophyll successfully on the market, but they are usually slow, and cost upwards of $40,000-$60,000 which is more than what grain elevators are willing to pay. Romeo liked our machine but wishes it will be taken further both in algorithm quality and user-oriented hardware design, especially in terms of the speed at which it is built. He also emphasized buying the best camera possible as the camera affects the entire project, hopefully an infrared camera to give better readings. The also gave us the idea to apply MGM to grading wheat, an excellent future direction to reduce the human error present in that sector.

References

  1. E Xodo, L., Rapozzi, V., Zacchigna, M., Drioli, S., & Zorzet, S. (2012). The chlorophyll catabolite pheophorbide a as a photosensitizer for the photodynamic therapy. Current medicinal chemistry, 19(6), 799-807.

  2. Liang, Y. I., Lu, L. M., Chen, Y., & Lin, Y. K. (2016). Photodynamic therapy as an antifungal treatment. Experimental and therapeutic medicine, 12(1), 23-27.

  3. Garg, H., Atri, C., Sandhu, P. S., Kaur, B., Renton, M., Banga, S. K., ... & Banga, S. S. (2010). High level of resistance to Sclerotinia sclerotiorum in introgression lines derived from hybridization between wild crucifers and the crop Brassica species B. napus and B. juncea. Field Crops Research, 117(1), 51-58.