Proposed Implementation

We have done extensive research on implementation of our project. This section covers aspects of implementation that are built into our work, and things that still need to close the gap between in-lab and the real world!

Introduction

The greenhouse industry and pesticides in Canada are heavily regulated to protect the growers, greenhouse workers, and Canadian buyers and as such, for our project to be commercialized, we must meet regulatory body requirements. The regulatory body in our case would be Health Canada, specifically the Pesticide Management Regulatory Agency (PMRA) (Government of Canada, 2004). The PMRA requires all pesticides to undergo a pesticide registration process that involves the following:

The Environmental Risk assessment has 2 components: what happens when the pesticide is in the environment and whether or not the pesticide harms non-target organisms. The Health Evaluation is various studies to determine if the pesticide poses any threat to human health. The Laboratory Services component is the PMRA’s analysis of the chemistry behind the pesticide. The Value Assessment component determines if the pesticide is effective at managing pests.

The PMRA does presubmision consultations so we will submit an application before we start the pesticide registration process to ensure we have a smooth application (Government of Canada, 2016). Also, we would need to be approved as a Minor Use Pesticide as we our target buyer are greenhouse growers and greenhouse application falls under minor use (Government of Canada, 2022).
To successfully meet these requirements, we can either have our biopesticide undergo the tests required at the University of Guelph’s own Agriculture and Food Laboratory, which has over 17 years of experience in performing tests for the Pesticide Registration process (University of Guelph Agriculture and Food Laboratory Services, n.d.).
We would also need to explore US regulations for pesticides as growers in Canada export their produce to the US and the US exports their produce to Canada, thanks to our trade agreements and close proximity to the US.

Pesticide Application

We have considered 2 methods of application to give growers choice. The first application method is a powdered form of our biopesticide. This method would allow for growers to incorporate our bioinsecticide into their soil, which would be extremely beneficial for flower growers. Flower growers would only need to induce our biopesticide once they choose to use it and it can remain inactive in their soil for as long as they choose. A consideration that must be made is how to make the powdered form applicable to water based systems like hydroponics. From our HP work, we have learned that most growers use hydroponics. To solve this question we must investigate how we can induce B. subtilis to form biofilms in plant roots in a hydroponic system as is further discussed on the Entrepreneurship page. The second application method is a coated seed, which uses gelatin and the sporulation capacity of B. subtilis to form seed coatings around tomato seeds. We will also need to finalize work in regards to the concentration that our product must be applied by growers. This information will be present on our biopesticide label, which is a legal document that makes growers responsible for the correct application of pesticides.

Safety

Our project was designed with safety as its baseframe. We initially looked into using Cry proteins instead of Cyt proteins, however Cry proteins have a broad target range of insects which can be detrimental to the beneficial insects and wildlife (Valtierra-de-Luis et al., 2020). For instance, many major pollinators are in the Lepidoptera order such as butterflies, so using Cry proteins would harm these beneficial insects and not pass the pesticide registration process. Cyt proteins however are very specific and can target specific insects within a small range. Due to their specificity Cyt proteins were chosen for our product.

Another important consideration of our project was the acknowledgement of our project as a genetically modified organism (GMO). Society has differing views on GMOs, but we wanted to negate concern of our GMO in the environment through the development of our project. Firstly, we chose greenhouses as our preferred growing environment since greenhouses are more contained than fields, however we must acknowledge that they are not 100% contained as a lab would be. To close this gap in containment of our biopesticide, we excluded the fertility factor in our plasmid to prevent its genome transmission via horizontal gene transfer (HGT). HGT is when two related bacteria, for instance our modified B. subtilis and the native population of B. subtilis, share genetic information. However as our B. subtilis lacks the factor that enables HGT it cannot share its DNA. This exclusion prevents our plasmid from giving other bacteria engineered antibiotic resistance, specifically ampicillin and chloramphenicol resistance, which is majorly frowned upon in the scientific community.
We also looked at the inclusion of a kill switch in our plasmid for the easy disposal of our biopesticide. Please read the Novel Kill Switches section under Best Supporting Entrepreneurship for a longer explanation of the benefits of a Kill Switch. A potential kill switch is PBSX, a phage like bacteriocin that has been shown to lyse B. subtilis (7). The gene encoding this suicide gene can be put under the control of an inducible promoter such as PMcl1, a hemolymph inducible promoter (Poon, 2016). Due to time constraint development of this kill switch will be a future plan.
For more information about our safety procedures please read the Safety page on our wiki.

Works Cited

  • Government of Canada. (2004). Pesticide Registration Process. Government of Canada.
  • Government of Canada. (2016). Pre-submission Consultations. Government of Canada.
  • McDonnell, G. E., Wood, H., Devine, K. M., & Mcconnell, D. J. (1994). Genetic control of bacterial suicide: regulation of the induction of PBSX in Bacillus subtilis. Journal of Bacteriology, 176(18), 5820–5830.
  • Government of Canada. (2022). Minor-use pesticides. Government of Canada.
  • McDonnell, G. E., Wood, H., Devine, K. M., & Mcconnell, D. J. (1994). Genetic control of bacterial suicide: regulation of the induction of PBSX in Bacillus subtilis. Journal of Bacteriology, 176(18), 5820–5830.
  • Poon, C. (2016). Part:BBa_K2040100. iGEM. Accessed at: http://parts.igem.org/Part:BBa_K2040100
  • University of Guelph Agriculture and Food Laboratory Services. (n.d.). GLP Services at the AFL. University of Guelph Agriculture and Food Laboratory Services.
  • Valtierra-de-Luis, D., Villanueva, M., Berry, C., & Caballero, P. (2020). Potential for Bacillus thuringiensis and Other Bacterial Toxins as Biological Control Agents to Combat Dipteran Pests of Medical and Agronomic Importance. Toxins, 12(12), 773–.
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