Funded Projects

Agriculture and Farming

The Future of Fertilizer

Principal Investigator: Lawrence Wackett, Distinguished McKnight University Professor in the Department of Biochemistry, Molecular Biology, and Biophysics

Co-investigators: Carl Rosen, Professor and Department Head in the Department of Soil, Water, and Climate.

Industry Partners: The Office of Technology Commercialization has filed a provisional patent application. Two companies, one a Minnesota company and the other a multi-national, are currently evaluating the technology.

Problem: Urea is the primary source of nitrogen in agricultural fertilizer. However, due to high application rates, high levels of nitrate runoff into nearby waterways and threaten human health and local ecosystems.

Solution: Different urea-based compounds are used or found in fertilizers. The investigators plan to design and test these urea-based molecules for their positive and negative effects on plants. The optimal ratios of the compounds can produce better fertilizers.

Impact: By 2035, the amount of urea used as fertilizer is projected to increase from 165 billion tons to over 450 billion tons. A successful demonstration of optimally blended urea-based fertilizers could provide lower costs to farmers and offer a scalable solution to nitrate runoff and water impairment issues.

Mycoalgae Biofilm for Aquaculture Bioremediation and Animal-Feed Production

Principal Investigators: Bo Hu, Aravindan Rajendran

Lichen, a natural ecosystem with phototrophic algae and heterotrophic fungi symbiotically growing on the solid surface of rock or roof, is not readily applied in engineering field due to their low growth rate. A concept of “simulated lichen system” is recently developed by our UMN research group (UMN invention disclosure case # 20140274) that we can select different desired microalgae and fungal combinations that will be growing on the surface of some specific polymers to form the biofilm. Microalgae are naturally growing on the surface of the nutrient-rich water; however, biological treatment of polluted waters using microalgae is limited by problems associated with the settling and separation of algae downstream of the treatment site. The proposed methodology using bioaugment filamentous fungi in lichen biofilms overcomes this limitation, by efficiently retaining algae and recovering the nutrients and recycling of useful nutrients. We are extending this technology to aquaculture waste water bioremediation, and recycle the nutrients as proteinaceous microbial biomass feed for the aquatic animals. This process modification could make the aquafarming more efficient as commercial feeds may account for more than 50% of the total production costs. “Mycoalgae biofilm” also works on photosynthetic aeration to replace the conventional and energy intensive mechanical aeration for biological processes, and also the strains complement each other by the required respiratory gases. The biofilm composition can also be tailored based on the influent stream components, feasibility of the strains to grow together and its nutrition value as animal feed.

A Novel Way to Extract Resources from Minnesota Food Waste while Removing PFAS

PI: Bridget Ulrich
Industry Partners: Leigh Behrens, MPA, Planning Specialist, Ramsey/Washington Recycling and Energy
Postdoc: Shilai Hao

Research Problem

Over 35 million tons of food waste are disposed of in the US annually, representing substantial untapped potential for recovery of nutrients and energy. Food waste can be broken down by microorganisms to produce biofuels and biofertilizer through a process called anaerobic digestion (AD). However, poly- and perfluoroalkyl substances (PFAS) have been widely detected in food waste, and these persistent contaminants are highly resistant to degradation during AD and in the environment. Therefore, innovative treatment approaches are required to recover nutrients and energy from food waste without redirecting PFAS contamination to the environment.

Solution statement

The goal of this project is to evaluate the effectiveness of a treatment train approach incorporating AD and hydrothermal treatment (HT) to simultaneously remediate PFAS and recover nutrients and energy from food waste. HT decomposes food waste at high temperatures and pressures in an aqueous environment, producing energy-rich biofuel and nutrient- and carbon-rich biofertilizer. Moreover, HT has recently been demonstrated to rapidly degrade PFAS, representing a potential means of producing PFAS-free biofertilizers that can be safely applied to fields and gardens. This project will evaluate the effectiveness of sequential treatment by AD and HT for PFAS remediation and resource recovery from actual food waste collected by the Ramsey/Washington Energy and Recycling Board.

Impact statement

Development of innovative approaches to recover resources from waste streams in a way that minimizes environmental impact is of direct relevance to several of the United Nation’s Sustainable Development Goals. AD has been recognized as one of the most promising means of recovering resources from food waste while minimizing greenhouse gas emissions, but the potential for spreading PFAS contamination to the environment through fertilizer application is a substantial limitation of the approach. We seek to minimize the overall environmental impact of resource recovery from food waste by developing a treatment approach that simultaneously minimizes greenhouse gas emissions, produces renewable energy and biofertilizers, and remediates PFAS. This will lead to broad sustainability improvements across the Food-Energy-Water Nexus, and provide market opportunities for small businesses to locally collect, convert, and reutilize food waste

Putting the ‘Morph’ Back in Morphology for Fungal Remediation

Primary Investigator: Jonathan Schilling
Co-Investigators: Aiym Bakytbaikyzy (Postdoctoral Research Scholar)
Industry Partners: MycoWorks
Award Type: Seed Grant – Graduate Research Scholar

Problem: Filamentous ‘white rot’ type fungi produce enzymes known as Type II Peroxidases (PODs) that not only break down plant lignin, but also break down a vast range of pollutants. For this reason, a common bioremediation co-metabolism strategy involves spreading wood mulch on top of contaminated soils to attract or inoculate with white rot fungi to stimulate POD secretion and to inadvertently (but beneficially) degrade pollutants. This strategy works, it is cheap, and it has been widely adopted; however, when remediation timeframes are limited for diffusible pollutants or when concentrations of pollutants exceed POD titers achievable by typical inoculated fungi, this approach falls short. This is unfortunate, given that the alternatives are far more expensive, and it often leaves remediation alternatives out of reach, all together.

Solution: Two possible solutions are 1) to make the more effective alternatives cheaper, or 2) to make the cheap solution more effective (our approach). Our specific approach is to target fungal strains and growth conditions that force more hyphal septation and branching/binding in order to boost POD secretion per unit of fungal biomass. This is straightforward and it is logical, but it has been not been addressed, in part because of the limitations of the fungus, Pleurotus ostreatus (Oyster mushroom) most commonly used in these applications.

Impact: Improving efficiency of mycoremediation via peroxidase enzyme increases and hyphal densification could have tremendous potential impact on industrial clean-up of pollutants. These enzymes are selective in the reactions that they catalyze, but the cascading secondary reactions are non-selective, often targeting ring-structure compounds that resemble the intended target, lignin. These ring-structure compounds include many pollutants that are otherwise challenging to clean up, including hydrocarbons, PCBs, and dioxins, making these lignin-degrading white rot fungi have potentially great value for dispersed, low-cost bioremediation.

Phytoremediation of PFAS Using Nanomaterials

Primary Investigator: Christy Haynes
Co-Investigators: Riley Lewis (Postdoctoral Research Scholar)
Industry Partners: Ecolab, Connecticut Agricultural Experiment Station
Award Type: Seed Grant – Graduate Research Scholar (with NRRI Travel Grant)

Problem: Per- and polyfluoroalkyl substances (PFAS) are ubiquitous in the environment and represent a health threat worldwide. There are more than 7500 PFAS, and all have strong C-F bonds that render them persistent in the environment. In Minnesota, PFAS are emerging contaminants found in landfills, fire training facilities, and groundwater, so the MN Department of Health has been sampling within and tracking PFAS-affected communities. Methods for PFAS clean-up are urgently needed.

Solution: This work will develop novel nanomaterials (NNMs) that facilitate sorption and mobility of low-concentration PFAS into hemp plants for phytoremediation. While there are several reasonable sorbents available for water applications, this work will develop high-affinity NNMs for soil and sediment remediation.

Impact: The proposed project will develop NNMs for use in phytoremediation of PFAS and provide information on the mechanism and efficacy of the new remediation method, allowing transfer of PFAS from the diffuse and difficult-to-manage soil compartment to mobile, readily handled plants.

Reducing Nitrous Oxide from Agriculture

Primary Investigator: Timothy Griffis
Co-Investigators: Alexander Frie, Rodney Venterea
Industry Partners: Minnesota Farm Winery Association
Award Type: Seed Grant – Postdoctoral Research Scholar

Problem: Since the industrial revolution, the use of synthetic nitrogen (N) as fertilizer has driven increased agricultural yields. To meet growing demand, the use of N fertilizer has grown 40-times as large since 1940. Increased usage has led to a steep increase in emissions of nitrous oxide (N2O), a greenhouse gas roughly 300 times more potent than CO2. N2O has been labeled as the most important anthropogenic oxygen depleting substance. Minnesota’s 72 agricultural counties need new treatment methods to continue a sustainable approach to fertilizer use.

Solution: The majority of agriculturally-based N2O emissions come from denitrification, a biological process through which nitrate is converted to ozone-depleting nitrogenous gases. MnDRIVE Researchers have identified procyanidins, a compound produced by grapes and berries, as an inhibitor of denitrification. This project will use procyanidins from multiple sources and measure their ability to reduce N2O emissions. Researchers will also identify optimal levels of procyanidins to apply for the desired decrease in denitrification.

Impact: Climate change and O-Zone depletion are two pressing issues defining environmental protection in the 21st century. The potential of procyanidins to decrease nitrous oxide emissions is yet to be fully released, but their utilization could be key in mitigating climate change and stratospheric O-zone depletion. Procyanidin is produced by grapes and could increase the demand for juice and wine production, providing economic stimulation for Minnesota industries.

Reducing the Environmental Cost of Glyphosate

Primary Investigator: James Cotner
Co-Investigators: Brianna Loeks-Johnson (Graduate Scholar); Ben Fry (Undergraduate Scholar)
Industry Partners: Minnesota Department of Agriculture
Award Type: Seed Grant – Graduate Research Scholar & Undergraduate Research Scholar

Problem: Glyphosate is the most widely used herbicide on our planet. It’s extensive use in the Midwest, including Minnesota, has been extremely beneficial to agricultural production. However, this organophosphate continues to accumulate in soil and water, and is not easily degraded by microbes. Some recent publications have, however, suggested that bacteria like cyanobacteria or blue-green algae are more capable of using partially degraded glyphosate than other algae. This suggests that glyphosate may be contributing to the increased presence of blue-green algae in Minnesota water bodies and ecosystems, where glyphosate is used. The problem requires further investigation into how it might be influencing Minnesota lakes and rivers and the associated soils.

Solution: The Cotner Team will first examine the ability of harmful algal bloom (HAB) species of cyanobacteria to consume glyphosate. The cyanobacteria will also be analyzed for genes known to be involved in the consumption of glyphosate. MnDRIVE researchers will then screen a library of freshwater bacterial strains in the Cotner Lab for their ability to degrade glyphosate and other phosphonates under varying environmental conditions. Bacteria that are capable of degrading glyphosate and phosphonate will then be characterized for potential bioremediation use.

Impact: Identifying microbes capable of degrading glyphosate from areas of high glyphosate use will lead to a new understanding of bacterial species that may be contributing to harmful algal blooms (HABs) throughout Minnesota. Characterizing microbes from the Cotner bacterial library capable of glyphosate degradation will also demonstrate potential for future bioremediation technologies.

Repurposing Drainage Ditches to Remove Nitrogen

Primary Investigator: Jeffrey Strock
Co-Investigators: Satoshi Ishii, Hao Wang (Graduate Scholar)
Industry Partners: Minnesota Drainage Viewers Association, I&S Group, Inc. (ISG), Minnesota Department of Agriculture, Sand County Foundation, and USDA Natural Resources Conservation Service
Award Type: Seed Grant – Graduate Research Scholar

Problem: Drainage ditches on agricultural land improve field drainage efficiency by providing preferred flow paths for excess water. However, the increased flow rates do not allow dissolved nitrogen and phosphorus time to react, bind, or be absorbed by ditch sediment/soil or vegetation. These ditches transport high concentrations of nutrients and other contaminants into nearby streams and rivers, which degrades local and downstream ecosystems.

Solution: If drainage ditches behaved more like natural wetlands (chemically, biologically, and microbiologically), they could remove higher levels of nitrogen from field runoff, both through denitrification and plant uptake. Strock and his team will engineer and install low-grade weirs (aka – low dams) at a field site near Lamberton, with the goal to modestly raise upstream water levels, decrease flow velocity, and thus enhance nitrogen removal. The laboratory component of the project will analyze biological and microbial communities effective for nitrogen uptake within their low-grade weir design, with a focus on cold-climate denitrifiers.

Impact: Each year roughly 60 million pounds of nitrogen flows through drainage ditches leading to the Minnesota River. Improving the ability of drainage ditches to naturally remove more nitrate offers a low-cost solution to one of Minnesota’s most persistent environmental challenges. If successful, not only will this solution decrease the amount of dissolved nitrogen in Minnesota’s major waterways, but it would also decrease the amount of dissolved nitrogen leaving Minnesota via the Mississippi.

Using Nano-silica Materials to Remove Nitrate From Water

Primary Investigator: Kara Nell
Award Type: Seed Grant – Undergraduate Research Scholar

Problem: A majority of Minnesota’s waterways hold an elevated concentration of nitrate. Commonly emanating from runoff of agricultural fertilizers, high nitrate concentrations can cause algal blooms, eutrophication, and even direct human health issues. Yet, removing nitrate from these waters without influencing other molecules and drastically changing the water chemistry, possibly detrimentally, requires the ability to specifically target these molecules. Current nitrate filtration methods lack this specificity.

Solution: Early work by Nell’s lab showed that several novel silica materials selectively target specific anions like nitrate in competitive environments. They are currently working to determine their particular affinity with and selectivity for nitrate. This project will work to optimize nano-scale structures made of silica (aka – nano-silica materials) for the targeted binding and removal of nitrate from water, while minimizing the capture and removal of other water-soluble molecules.

Impact: The use of functionalized nano-silica materials would improve the ability of filtration techniques to be chemical selective for water remediation. This technology would not only be beneficial to multiple aspects of the water remediation process, but also may seed the way for direct application to soils – in a proactive implementation.

Green Alternatives to Toxic Fertilizer Components

Primary Investigator: Lawrence Wackett
Co-Investigators: Romas Kazlauskas, Mikael Elias, Carl Rosen, Lambros Tassoulas (Graduate Scholar)
Industry Partners: Minnepura Technologies
Award Type: Seed Grant – Graduate Research Scholar

Problem: Urea-based fertilizers contain biuret, which is a contaminant toxic to plants. Currently, an extraction process is used to reduce the amount of biuret present in these fertilizers, but the technology is expensive and doesn’t fully remove the toxin. Some plants such as citrus and avocado actually require low-biuret fertilizers, but this product is 2-2.5 times more expensive than untreated urea. More effective biuret removal and less expensive low-biuret fertilizers are in demand from the agriculture industry..

Solution: Researchers in the Wackett Lab have identified over 1000 biuret hydrolase enzymes from different bacteria. These microbial enzymes are effective at converting biuret into urea, thus taking the toxic contaminant and turning it into the valuable main ingredient in fertilizer, urea. Further work is needed, however, to identify biuret hydrolase enzymes that are most stable, purify them, and then test them in urea solutions. Crops sensitive to biuret will be sprayed with enzyme-treated fertilizer to test the efficacy of the enzymes-based removal process. These tests will provide important proof-of-concept data for the commercialization of enzyme-based biuret removal.

Impact: Efficient biuret removal should lower the cost of fertilizers for farmers and increase agricultural productivity, especially for biuret-sensitive crops. The research has global potential as urea-based fertilizers are used extensively, but the solution may prove to be impactful across Minnesota as well.

Bioaugmented Nitrogen Removal from Drainage System Water

Primary Investigator: Michael Sadowsky
Co-Investigators: Jeff Strock; Satoshi Ishii; Reda Abou-Shanab and Prince Mathai (Postdoctoral Scholars)
Industry Partners: Minnesota Department of Agriculture, Agricultural Utilization Research Institute, Minnesota Corn Growers Association
Award Type: Seed Grant – Postdoctoral Research Scholar

Problem: Roughly 60 million pounds of nitrogen flow through drainage systems in the Minnesota River basin each year. The nitrogen accumulates from agricultural runoff and enters the Mississippi River, resulting in environmental damage to local ecosystems as well as those downstream of the Mississippi, such as hypoxic zones in the Gulf of Mexico. Most drainage systems are unequipped to remove nitrogen before runoff enters groundwater systems. Recently developed bioreactors only extract about 30% of nitrogen from water and are not optimized for early spring cold temperatures – when runoff volume peaks.

Solution: MnDRIVE researchers will begin by modifying existing, above-ground bioreactors in a field setting. The bioreactors will be inoculated with cold-adapted denitrifying bacteria (bioaugmentation) and observed at temperature settings between 5 and 15 degrees celsius. Ethanol will then be applied to microbial communities as a source of carbon, a process called biostimulation that will maximize the ability of microbes to remove nitrogen. Nitrogen removal capabilities will be recorded and genetic analysis will also be used to determine the abundance of microbes at the various settings used. Once the ideal conditions have been identified, current bioreactors at the Southwest Research and Outreach Center will be modified for in-field testing where similar measurements will be made to test the effectiveness of the above-ground biostimulation technology.

Impact: Agriculture is one of Minnesota’s largest industries, but also contributes large amounts of nitrogen to runoff water. Applying biostimulation strategies to existing bioreactors will help increase nitrogen removal. Removing nitrogen from water before it enters rivers and streams will improve conditions of local ecosystems and reduce contribution to hypoxic zones downstream.

Using Fungi to Protect Soybean Crop from Nematodes

Primary Investigator:Christine Salomon
Co-Investigators: Sophia Powells, Elaine Kappel
Industry Partners: NA
Award Type: Seed Grant – Undergraduate Research Scholar

Problem: Soybean cyst nematode (SCN, Heterodera glycines) is a pathogen that causes crop losses of over 30% for soybean farmers globally. Although some control measures have shown success in preventing the disease, there is no treatment for SCN once a field has been infected.

Solution: Some species of fungi produce linoleic acid, which is toxic to nematodes. The Salomon Lab plans to develop a treatment for SCN using these linoleic acid-producing fungi. The Lab will test various species to determine which is most effective in inhibiting the nematodes. The initial experiments were focused on developing methods to efficiently quantify the amount of linoleic acid and several structurally related fatty acids from crude extracts obtained for each environmental fungal isolate. Pure fatty acids were chemically modified so that they could more easily be detected and quantified. Once this method was optimized with the pure compounds, the crude fungal extracts were subjected to the same derivatizations and analyzed to detect and measure any naturally occurring fatty acids. The strains with the highest levels of inhibitory fatty acids will be used for additional testing with soybean cyst nematodes and nematode eggs.

Impact: Minnesota produces more soybeans than any other state in the U.S. Developing a treatment for SCN could prevent financial losses for farmers in our state who grow the crop for food and energy.

Fueled By Food Waste

Primary Investigator: Bo Hu
Co-Investigators: Tim LaPara
Industry Partners: Second Harvest Heartland
Award Type: Demonstration Grant

Problem: In the United States, roughly 40% of all food goes to waste, including food donated to non-profit food banks like Minnesota’s Second Harvest Heartland. Unusable or spoiled food is an unpredictable and fluctuating cost for foods banks who need to pay for the removal of the wasted food.

Solution: MnDRIVE Environment researchers designed an anaerobic bioreactor to digest food waste on site. Not only could this approach address the issue of excess food waste and disposal, but the bioreactor also produces methane, the principal component of natural gas. This offers a new energy source or revenue stream. In addition, a second bio-electrochemical system removes hydrogen sulfide, a toxic and odorous component, from the biogas so that the digester can be operated more safely in the neighborhood.

Impact: The demonstration project will help Second Harvest evaluate the feasibility of a commercial-scale anaerobic bioreactor installation that could one day heat their facility and/or create new revenue streams to stabilize the non-profit organization. This type of digestion technology is of significant interest from public solid waste disposal sites.

Fungi for Phosphate Removal

Primary Investigator: Chris Lenhart
Co-Investigators: Laura Bender (Graduate Scholar); Kirsten Haus (Northstar STEM Alliance Researcher), Alli Graper (undergraduate honors thesis)
Industry Partners: Legvold Farms; Agricultural Drainage Management Coalition
Award Type: Seed Grant – Graduate Research Scholar

Problem: Streambank vegetative buffers (also known as riparian strips) are required by Minnesota State Law to be maintained along all waterways, to filter out sediment and nutrients such as nitrogen and phosphorus. While riparian buffers effectively remove particulate phosphorus, they don’t do as well removing dissolved phosphorus. Soil fungi within buffer strips can also contribute to removal of dissolved phosphorus by enhancing plant uptake as most native grasses have symbiotic relationships with mycorrhizal fungi to uptake nutrients. Fungal communities are greatly impacted by decades of farm tillage so when buffers are installed the microbial community is not optimized for phosphorus uptake, nor for native plant establishment, which are both key to buffer function.. Therefore, removal of dissolved phosphorus from buffers and edge-of-field practices to prevent transport into water bodies remains a challenge.

Solution: The removal of soluble phosphorus in riparian buffers may be improved by enhancing the symbiotic relationship between plant roots and fungal communities. To test this, Lenhart and Bender have compared multiple riparian vegetative plots – one with and one without mycorrhizal fungi added and with different types of native grass species – within a riparian buffer along a stream in Minnesota. Phosphorus levels in the plants, soil, and water within each plot have been measured and analyzed over the 2019-2020 period to determine whether addition of mycorrhizal fungal to riparian buffers plots improves phosphorus uptake and removal from tile drainage flow routed through the buffer. Additionally, this in situ work is being compared to controlled environment studies being conducted using 100 gallon tanks on the St. Paul campus.

Impact: Improving the removal of dissolved phosphorus in vegetative buffers would help achieve Minnesota’s State Nutrient Reduction Strategy goals. These goals aim to preserve water ecosystem health by lowering excess nutrient levels that can cause eutrophication and algal blooms. Buffers in sandy, compacted or degraded soils are most likely to benefit from fungal amendments. Findings from the studies are being

Algal Biofertilizer to Improve Soil Health

Primary Investigator: Rob Gardner**
Co-Investigators: Adriana Alvarez De la Hoz (Graduate Scholar)
Industry Partners: Chippewa Valley Ethanol Company; USDA
Award Type: Seed Grant – Graduate Research Scholar

Problem: To meet the growing global food demands, agriculture needs to increase productivity. However, growth of traditional farming practices degrade overall soil fertility, which leads to over application of fertilizers and the subsequent leaching and runoff of excess nutrients into groundwater and surface water bodies. With the predominant use of synthetic fertilizers, researchers observe elevated levels of nitrogen (N), phosphorus (P), and carbon dioxide (CO2) in agricultural runoff. This is a direct contributor to poor water quality, and often causes eutrophication of surface water bodies across Minnesota. Eutrophication removes oxygen from the water which promotes the growth of unwanted and often toxic algal growth. These algae are detrimental to the surrounding ecosystem and potentially harmful to humans.

Solution: Sustainable agricultural practices have emerged as a set of tools for farmers, to help them better manage aspects like soil fertility, erosion control, water quality, and nutrient cycling. The Gardner lab has early results indicating that the use of microalgae (single-cellular) as a component in natural bio-based fertilizer improves soil fertility, while also replacing the need for synthetic fertilizers. This lab-based study reports that traditional synthetic urea fertilizer harms soil microorganisms while algal biofertilizer enhances soil health in multiple ways. Unlike synthetic fertilizers, algae supply organic matter and favor soil microbial dynamics. Algae stabilize soil particles and structure, thereby reducing erosion rates, and they release organic nutrients at a slower rate than synthetic fertilizers. This results in longer nutrient availability for crops and less nutrient loss to runoff. And lastly, the remaining live algae capture atmospheric N2 and CO2 in the soil surface, offering a natural nitrogen input and carbon capture potential.

Impact: Many aspects of this proactive approach for improving soil health offer beneficial outcomes for arable MN soils compared to traditional farming techniques. The use of algal biofertilizer as a replacement of synthetic urea fertilizer can contribute to long-term soil fertility and lower carbon emissions from crop production by reducing synthetic fertilizer demand. Stabilizing soil particles is also critical for reducing erosion and nutrient runoff rates, which should result in lower nutrient levels in waterways that receive agricultural runoff.

** MnDRIVE regrets to report that Dr. Gardner passed away in October 2019.

Identifying the Best Genetic Sequences for Removing Nitrate

Primary Investigator: Jeffrey Gralnick
Co-Investigators: NA
Industry Partners: MPCA (additionally Barr Engineering, Geosyntech Consultants)
Award Type: Research Scholar – Postdoctoral Research Scholar

Problem: Fertilizer use in Minnesota’s agricultural industry has led to environmental nitrate contamination. High levels of nitrate cause methemoglobinemia (“blue baby syndrome”) and increase the risk of non-Hodgkin’s lymphoma. Nitrates from Minnesota also contribute to the “dead zone” in the Gulf of Mexico. To combat this, certain bacteria are used to break down nitrates into nitrogen-containing gases; however, these bacteria prefer to breathe oxygen instead of nitrate when oxygen is present. If bacteria breathe oxygen, they won’t remove nitrates, restricting the practicality of using bacteria to solve the problem.

Solution: Researchers in the Gralnick Lab will test a new genomic technology that may allow them to deregulate nitrate respiration in bacteria. This would allow bacteria to break down nitrate even while oxygen is present. The lab will use a method called 2D-TnSeq to identify sets of genetic mutations that improve denitrification rates. They will test 2D-TnSeq on Escherichia coli first as a proof-of-concept before targeting Shewanella species, an efficient denitrifying group of bacteria.

Impact: Shewanella species can efficiently convert nitrate to innocuous nitrogen gas in anaerobic environments. Genetic modification using 2D-TnSeq could make the denitrification process viable in aerobic environments as well. This would provide more opportunities for nitrate bioremediation using Shewanella species in the environment.

Optimizing the Treatment and Use of Manure to Reduce Nitrogen Runoff

Primary Investigator: Kechun Zhang
Co-Investigators: NA
Industry Partners: Ascenix
Award Type: Seed Grant – Postdoctoral Research Scholar

Problem: There is no shortage of manure in Minnesota. In fact, the over 18,000 feedlots in Minnesota alone generate an amount of manure equivalent to 50 million people. Much of this manure is repurposed for use as a source of organic nitrogen fertilizer. However, most of the nitrogen in manure comes from protein which plants cannot use directly. After application, this proteinaceous nitrogen may runoff into local water systems, damaging human and ecosystem health.

Solution: MnDRIVE researchers will engineer bacteria to degrade the amino acids, a key component of proteins, from manure. Degraded amino acids will then act as a source of carbon for the bacteria to use in producing biofuels and other marketable chemicals. Nitrogen will also be separated during the amino acid degradation, and will serve as a source of fertilizer.

Impact: A novel approach to manure management would result in more useful fertilizer sources without the negative effects of nitrogen runoff. Optimizing the engineered bacteria could lead to a fully functional management system and create economic incentive to reduce the environmental impact of manure application.

On-site Fungal Treatment to Remove Phosphorus from Animal Manure

Primary Investigator: Bo Hu
Co-Investigators: NA
Industry Partners: Dennis Haubenschild Farms
Award Type: Demonstration Grant – Postdoctoral Research Scholar

Problem: Manure is often applied as a fertilizer to agricultural fields to supplement nitrogen and phosphorus in the soil. As the livestock industry trends towards more concentrated farming, higher levels of manure must be disposed of per acre of field. This leads to phosphorus accumulation in the soil because manure has a higher ratio of phosphorus to nitrogen than what is needed for plant growth. Excessive phosphorus causes eutrophication in lakes and rivers, which kills organisms living in the water.

Solution: Fungal treatment can recover phosphorus in manure, which would lower the phosphorus to nitrogen ratio to a more suitable level for field application. The Hu Lab will demonstrate this process at Haubenschild Farm to determine its cost-effectiveness at a commercial scale. MnDRIVE researchers will also investigate inexpensive methods to sterilize the manure, so the fungi can grow in place of pathogenic bacteria.

Impact: Fungal treatment of manure would mitigate the environmental impacts of agriculture by reducing phosphorus accumulation in fields. In addition, partially removing phosphorus from manure could produce a phosphorus-specific fertilizer byproduct and result in an additional form of revenue for agricultural facilities.

Lowering Copper Concentrations in Agricultural Pesticides

Primary Investigator: Christine Salomon
Co-Investigators: NA
Industry Partners: NA
Award Type: Seed Grant – Undergraduate Research Scholar

Problem: Copper is regularly used to treat fungal and bacterial diseases in the agriculture industry. In Minnesota, pesticides that contain copper can leach into local water systems. While low levels of copper do not impact humans, the metal can be toxic for microbial communities, plants, invertebrates, and fish. New pesticides with lower concentrations of copper are needed to reduce the environmental impact of treating fungal and bacterial diseases in the agricultural industry.

Solution: The Salomon Lab recently identified a Streptomyces bacterial isolate from the Soudan Iron Mine that produces antifungal metabolites. These metabolites inhibit the growth of pathogenic yeasts and show increased antifungal activity with the addition of low concentrations of copper. MnDRIVE researchers will determine which agricultural pathogens the isolates work against and optimal copper ratios that increase antifungal activity the most.

Impact: Using a naturally produced antifungal will reduce the need for toxic levels of copper in agricultural pesticides. The project will include application of the low-copper products on plants in a greenhouse setting to demonstrate proof-of-concept. A new, low-copper pesticide will help protect Minnesota’s environment from copper contamination.

Using Bacteria to Recycle Wasted Phosphorus

Primary Investigator: Mikael Elias
Co-Investigators: Bo Hu (Co-PI)
Industry Partners: MN Department of Agriculture and Metropolitan Council Environmental Services
Award Type: Seed Grant – Postdoctoral Research Scholar

Problem: Phosphorus (P) fertilizer is essential for large-scale agriculture. The element will reach peak production by 2040 and soon after will begin to decline, negatively impacting global food production. Meanwhile, agricultural runoff of P fertilizer in the Midwest is a major polluter of the Great Lakes and Mississippi River. Excess P in freshwater creates an overproduction of algae that reduces biodiversity and can be toxic to water ecosystems.

Solution: The Elias Lab is engineering a bacterial-based bioscavenger that will remove P from agricultural waste materials and recycle it as a fertilizer. The Lab’s novel approach uses laboratory molecular engineering to increase the phosphate uptake system in bacteria. The project will result in an efficient “phosphate-scavenging bug” to be applied in a biofilter system for agricultural sources.

Impact: A phosphorus recycling system could both reduce water pollution and relieve P shortages. The solution will be of high interest to environmental protection agencies, P-producing industries (livestock, chemical producers), and companies invested in sustainable food production. The system will address Minnesota’s phosphorus runoff concerns and a global challenge in food production.

Using Beneficial Microbes to Protect Minnesota’s Potato Crops from Disease

Primary Investigator: Linda Kinkel
Co-Investigators: Scott Bates (Postdoctoral Research Scholar)
Industry Partners: NA
Award Type: Seed Grant – Postdoctoral Research Scholar

Problem: Soilborne plant pathogens limit Minnesota potato production every year. To combat the pathogens, chemicals are applied to the soil in a process called fumigation. However, soil fumigation is expensive and kills a large proportion of beneficial microbes that counteract potato plant pathogens, resulting in an increase in the intensity of disease effects.

Solution: The Kinkel lab is researching how to apply certain groups of microbes, coupled with soil nutrient amendments, to support existing soil microbiomes in potato plants before or after soil fumigation. Researchers will apply microbes to 10 different potato growing sites across Minnesota to assess the disease rate and microbial composition of the soil after application.