Project Description

What is C.R.O.P.S?

The Current Situation

Canadian agriculture is one of the world’s most productive, significantly contributing to Alberta’s economy, with agriculture being the province's top renewable resource (Agriculture in Alberta, 2023). Our project addresses a critical issue affecting Canada’s number one oilseed plant, canola, which belongs to the Brassica family along with mustard, broccoli, brussel sprouts, and cauliflower (What is Canola?, 2024). Canola, known for its vibrant yellow flowers, produces seeds vital to its economic success. In 2021, Canada produced over 30% of the world’s canola and 60% of the global trade, with Alberta contributing $4 billion and representing 30% of Canada's canola revenue. Canola production adds $29.9 billion annually to the Canadian economy and supports 207,130 jobs (Canola Statistics 2024). However, Albertan canola crops have been threatened by Plasmodiophora brassicae, a soil-borne protist causing clubroot disease. This pathogen forms galls on plant roots, reducing nutrient uptake and causing 60-90% yield loss. P. brassicae can persist in soil for up to 20 years, with clubroot first reported in Alberta canola in 2003, and by 2020, over 3000 fields reported infestations (Clubroot disease of canola and mustard, 2024; Kageyama & Asano, 2009).

Current detection strategies for the pathogen involve sending soil samples to a remote lab location to undergo PCR/qPCR (Polymerase Chain Reactions) to determine the concentration of spores present in the field (About Canola 2023). These tests cost ~$95 per sample, leading to expensive detection costs for large fields. Scouting for the pathogen early in the growing season may allow farmers to detect the pathogen's presence enabling better crop planning and resource management. Last year the University of Lethbridge Collegiate iGEM team explored PbEL04, a small cysteine-rich protein unique to P. brassicae which is involved in causing the swelling of the cortical cells resulting in large masses/galls on the roots of the plant (Jiang, et al., 2022). They determined this protein shares structural similarities to that of Epidermal Growth Factor (EGF) protein that could be used for designing a complementary chimeric peptide that could be used for detecting the pathogen. Previously, they aimed to create a direct enzyme-linked immunosorbent assay (ELISA) to test the binding of our chimeric peptide with PbEL04 (iGEM ULethbridge 2023). This year we are advancing the project by designing a lateral flow assay to provide farmers with an easier detection device that can be utilized in the field. A lateral flow assay is the same type of test used in Covid-19 test strips and pregnancy tests (Higgins et al., 2021). Our approach utilizes the user-friendliness of the lateral flow assay, which does not require prior experience or training in biochemistry techniques. This also aligns with the needs of farmers and soil sellers, who require a quick method to determine whether their fields or soil are infected. Currently it takes around two to three weeks for a PCR lab result, while the lateral flow assay would give you an answer within an hour.

Current mitigation strategies for clubroot disease involve crop rotations and effective field hygiene to reduce its spread and impact. Although genetically engineered seeds have helped maintain yields in infested fields, the pathogen adapts quickly, making it an ongoing challenge (Alberta Clubroot Management Plan, 2022).

Our Proposal

We propose developing a biopesticide to help farmers increase their yields and reduce spore loads. Biopesticides, using natural microorganisms like Bacillus subtilis and Pseudomonas fluorescens, offer a promising approach for managing plant diseases. These bacteria work synergistically, enhancing plant health and disease resistance (Nquyen et al., 2020).Bacillus subtilis increases the production of enzymes that degrade pathogens, reducing the disease load in the soil (Guo et al. 2015). Meanwhile, Pseudomonas fluorescens boosts the plant's immune response by enhancing the synthesis of salicylic acid (SA), a key component in plant defense (Pelludat et al., 2003). In Canada, the regulation of genetically modified organisms (GMOs) ensures safety through stringent oversight by bodies like the Canadian Food Inspection Agency (CFIA) and Health Canada (Halvorsen et al., 2022). To further enhance biosafety, we plan to use inducible promoters and a toxin-antitoxin kill switch to control the engineered traits and ensure safe elimination from the environment if necessary (Regulation and Policy 2024).

Our approach involves creating an effective detection and mitigation strategy that complements one another and allows for the further flourishing of the canola industry in Alberta. By integrating the Clubroot Oversight and Prevention System (C.R.O.P.S) for early pathogen detection with innovative biopesticide solutions using microorganisms like Bacillus subtilis and Pseudomonas fluorescens, we offer a sustainable and effective method to combat clubroot disease. This strategy promotes the safe use of synthetic biology in agriculture, ensuring long-term crop health and productivity while supporting the economic stability of Alberta's canola farmers.

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References

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