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Project Description

Overview​

Project Fungilyzer represents an innovative approach to addressing phosphate depletion in agricultural soils. Our primary objective is to engineer a genetically modified fungus with the unique capability to efficiently absorb and store phosphate from the soil environment. In times of phosphate scarcity, this fungus will act as a dynamic bio-fertilizer, recognizing the deficiency and facilitating the transfer of essential phosphate nutrients to plants by triggering a programmed cell death process known as apoptosis.

Our Inspiration​

Our inspiration for the Fungilyzer project stems from the pressing global challenges of phosphate depletion and eutrophication. The urgency of these issues, coupled with the finite nature of phosphate resources, fuels our determination to find a novel, sustainable solution. We are driven by the belief that we can reshape the future of agriculture, not just by increasing yields but by doing so in a manner that conserves the environment and safeguards the planet's health. We are motivated to provide the intelligence that can offer an innovative alternative to mitigate these challenges and revolutionize phosphate resource management.

This project is a testament to our belief in the potential of SynBio to offer groundbreaking solutions to global challenges. The prospect of reengineering a fungus to function as a dynamic bio-fertilizer that can sense and respond to soil conditions fuels our passion for this work.

Background​

The world faces critical challenges related to limited phosphate resources and eutrophication, both of which have global ramifications. Phosphorus (P) stands as a vital macronutrient for plants, comprising approximately 0.2% of a plant's dry weight. It assumes a fundamental role in the constitution of essential molecules, including nucleic acids, phospholipids, and ATP (adenosine triphosphate). As a result, a consistent and sufficient provision of this nutrient is indispensable for the growth and development of plants. Furthermore, phosphate ions (Pi), derived from phosphorus, play a critical role in governing crucial enzyme reactions and the intricate regulation of metabolic pathways.1 The sustainability of modern agricultural systems largely relies on continuous applications of phosphate-based fertilizers, primarily derived from finite phosphate rock reserves. Consequently, there exists an imminent threat of resource scarcity that could profoundly impact global food security.2 To address these pressing concerns, scientists have sought innovative solutions to mitigate the depletion of natural resources while enhancing agricultural productivity. Phosphate runoff presents a dual dilemma, as it not only hampers crop development but also constitutes a major source of phosphorus (P) pollution in aquatic ecosystems, thus contributing significantly to the widespread eutrophication of streams and lakes.3 In light of these challenges, our research endeavors have culminated in the development of a genetically modified fungus, underpinned by a sophisticated genetic switch, tailored to effectively address the issues of eutrophication and phosphate availability. This bioengineered fungal agent functions as a biological fertilizer, strategically augmenting phosphate uptake and delivering it to plants precisely when needed. Consequently, our innovative approach curtails phosphate runoff, simultaneously fostering increasing plant growth rates.

Problem​

Phosphate, a vital yet finite natural resource, faces ongoing depletion as its demand in the modern world escalates exponentially, driven by the increasing need for food production. Phosphorus (P) plays a pivotal role in ensuring optimal conditions for seed and root development, as well as conferring strength and quality to cereal crops, among other critical functions.4 Remarkably, even with substantial chemical fertilizer application, global soils continue to experience phosphorus depletion, primarily due to the runoff caused by precipitation and other climatic shifts.

Given this challenging scenario, there arises a compelling need for a biological fertilizer solution that possesses the unique capacity to efficiently capture and store phosphate, thereby minimizing wastage to the greatest extent possible.

Our Solution​

In response to the escalating demand for a phosphate management system that not only conserves this limited resource but also delivers it to plants in a controlled manner when the soil's phosphate levels are insufficient, the iGEM team Duesseldorf decided to develop a pioneering biological fertilizer named "Fungilyzer". Fungilyzer is founded on the intricate mechanism of apoptosis, a genetically regulated cell death process that can be harnessed to release phosphate on demand, making it available for plant growth and nourishment. The pressing challenge we aim to address is the dual-fold issue of depleting phosphate reserves and the detrimental consequences of phosphate runoff caused by precipitation and changing climatic conditions. Our innovative approach not only optimizes phosphate uptake by plants but also provides a safeguard against the wasteful runoff of this critical nutrient. To achieve this, we have selected the fungus S. cerevisiae as our model organism, owing to its well-established symbiotic relationship with plants. Through meticulous genetic engineering, we have introduced a sophisticated genetic switch within S. cerevisiae. This genetic switch serves a dual purpose: It actively sequesters available phosphate from the soil and possesses the ability to sense phosphate deficiency. When phosphate levels in the soil fall below the required threshold, our genetically modified fungus promptly triggers the apoptotic process, culminating in the release of stored phosphate. This precisely timed delivery mechanism ensures that plants receive the necessary phosphate precisely when they need it for growth and development.

High Phosphate conditions

In high phosphate condition the Fungilyzer will take up excess phosphate for later use by the plant.

Low Phosphate conditions

In low phosphate conditions the Fungilyzer will release previously strored phosphate by lysis to ensure plant survival.

To regulate the cell lysis in our Fungilyzer, we employ a native Phosphate-repressed promoter, PHO5 (BBa_K4706002), sourced from S. cerevisiae. This promoter enhances the expression of downstream coding sequences in the absence of phosphate.5 Our plan is to use it in conjunction with a fluorescent reporter, meffRFP (BBa_K4706005), to enable the measurement of phosphate levels through fluorescence. For inducing cell death as part of our project to release stored phosphate, we initially had only one choice in the registry, BAX. However, given our need for controlled cell death, we sought an alternative protein. Our search led us to AtBAG6 (BBa_K4706007), a protein from Arabidopsis thaliana known in literature for its role in regulating controlled cell death.6 This protein will play a central role in our project by facilitating the final release of phosphate. The significance of this protein in our project lies in its pivotal role in enabling the ultimate release of phosphate. It will be located downstream of PHO5(BBa_K4706002), serving the purpose of inducing cell death under conditions of limited phosphate availability.

Future Prospects in the Real World​

The Fungilyzer project has the potential to redefine the future of agriculture and environmental sustainability in the real world. If successfully implemented, this innovative approach could address the critical issues of phosphate depletion and eutrophication, with far-reaching implications.

In the agricultural sector, the introduction of Fungilyzer as a biological fertilizer could lead to increased crop yields and quality. Farmers would benefit from a sustainable and efficient phosphate management system, reducing their reliance on finite phosphate rock reserves and chemical fertilizers. This, in turn, could contribute to improved food security on a global scale and reduced environmental harm caused by excessive phosphate runoff.

Moreover, the Fungilyzer project offers a promising example of harnessing genetic engineering for environmental conservation. By curbing phosphate runoff, it could mitigate the widespread eutrophication of water bodies, preserving aquatic ecosystems and water quality. This could have positive effects on industries such as fisheries and tourism, which rely on healthy aquatic environments.

In summary, the real-world prospects of the Fungilyzer project are multifaceted, encompassing enhanced agricultural sustainability, global food security, and improved environmental health. As we look ahead, the successful implementation of this innovative solution has the potential to transform the way we manage phosphate resources, leaving a lasting impact on the world we live in.

  1. D. P. Schachtman, R. J. Reid, and S. M. Ayling, ‘Phosphorus Uptake by Plants: From Soil to Cell’, Plant Physiol, vol. 116, no. 2, pp. 447–453, Feb. 1998, doi: 10.1104/PP.116.2.447.↩
  2. M. Hijri, H. J. Biofertil Biopestici, and J. Biofertil Biopestici, ‘The Use of Mycorrhizae to Enhance Phosphorus Uptake: A Way Out the Phosphorus Crisis Arbuscular Mycorrhizal Fungi Genomics View project African Soil Microbiome View project Biofertilizers & Biopesticides’, 2011, doi: 10.4172/2155-6202.1000104.↩
  3. Y.-Y. Yang, M. M. Tfaily, J. L. Wilmoth, and G. S. Toor, ‘Molecular Characterization of Dissolved Organic Nitrogen and Phosphorus in Agricultural Runoff and Surface Waters 2 3 4’, 2022.↩
  4. M. R. Hart, B. F. Quin, and M. L. Nguyen, ‘Phosphorus runoff from agricultural land and direct fertilizer effects: a review’, J Environ Qual, vol. 33, no. 6, pp. 1954–1972, Nov. 2004, doi: 10.2134/JEQ2004.1954.↩
  5. Korber P, Barbaric S. The yeast PHO5 promoter: from single locus to systems biology of a paradigm for gene regulation through chromatin. Nucleic Acids Res. 2014;42(17):10888-902. doi: 10.1093/nar/gku784. Epub 2014 Sep 4. PMID: 25190457; PMCID: PMC4176169.↩
  6. Kang CH, Jung WY, Kang YH, Kim JY, Kim DG, Jeong JC, Baek DW, Jin JB, Lee JY, Kim MO, Chung WS, Mengiste T, Koiwa H, Kwak SS, Bahk JD, Lee SY, Nam JS, Yun DJ, Cho MJ. AtBAG6, a novel calmodulin-binding protein, induces programmed cell death in yeast and plants. Cell Death Differ. 2006 Jan;13(1):84-95. doi: 10.1038/sj.cdd.4401712. PMID: 16003391.↩