In 2022, shortly after Russian forces launched a full-scale invasion of his country, Oleh Prylutskyi was studying fungal communities in soil. He was assessing how such communities vary in both space and time as part of a global project conducted by the Society for the Protection of Underground Networks (SPUN). Much to his dismay, Prylutskyi realized Ukraine didn’t have a single data point to feed into the analysis.
“There are a lot of fungal researchers in Ukraine with a long history, but Ukraine was a blank spot, nothing on the map,” Prylutskyi said.
Documenting Fungal Biodiversity
By July 2023, Prylutskyi, an associate professor at Kharkiv National University, and his colleagues at SPUN had secured funding from the Human Frontier Science Program (HFSP), allowing small multidisciplinary teams across the country to begin to gather soil samples that could be analyzed for their environmental DNA (eDNA). The aim was to reveal the country’s fungal species diversity, which is important because only between 92% and 95% of fungi worldwide have been scientifically described. Preserving this fungal biodiversity is critical because once they are gone, any economic, health, or ecosystem benefits these species might have provided are lost, too.
The project involved 34 contributors in 17 teams, yielding 225 soil samples across 21 of Ukraine’s 24 oblasts, or administrative regions, including areas close to the front lines.
“Ukrainian scientists are determined to fully utilize every opportunity to study the astonishing diversity and complexity of life, from the mighty ancient trees to microscopic soil fungi.”
Prylutskyi and his wife, biologist and founder of the Ukrainian Bat Rehabilitation Center Alona Prylutska, conducted soil sampling and bat surveys throughout central and western Ukraine, at one point collecting soil samples by lamplight.
“It was challenging to measure the study plot correctly and identify plants around, but we did it successfully, listening to the owl calls above our heads,” Prylutska said.
Another team, comprising botanists Dariia Borovyk at the M.G. Kholodny Institute of Botany and Kateryna Lavrinenko at Bohdan Khmelnytsky National University, visited the steppe slopes, floodplain meadows, and forests of the ancient Ukrainian Crystalline Shield formation.
“We tried to choose locations farthest from settlements with minimal anthropogenic impact on the soil microflora,” Lavrinenko said, and as a result, the only large animals they “noticed along the way were wild birds or startled roe deer.”
Dariia Borovyk samples the alluvial sandy grasslands of Khortytsia, an island in the Dnieper River, in September 2023. Credit: Volodymyr Shyriaiev
“Ukrainian scientists,” Lavrinenko said, “are determined to fully utilize every opportunity to study the astonishing diversity and complexity of life, from the mighty ancient trees to microscopic soil fungi.”
Prylutskyi explained that the project’s most dangerous sampling point was the bottom of Kakhovka Reservoir, which had been drained when its dam was destroyed in the conflict; the sample point was just 4 kilometers from Russian-controlled firing positions.
“All returned safe and sound, though Russian drones were flying around,” he said, adding that the project never encouraged any of its collaborators to take unnecessary risks.
“Staying in Ukraine is a risk on a daily basis, thousands have died for no particular reason—sleeping at home or being in public places—so I doubt any Ukrainian would say that pushing the scientific frontier is not worth taking a slightly higher risk than average.”
DNA Sleuthing
At each sampling site, the teams mapped out a 30-meter by 30-meter quadrant and took nine soil samples. “From several kilograms of soil, we got 20–50 grams of soil; then in the lab, we mixed it again very carefully and took 200 nanograms (0.2 g of soil),” Prylutskyi said. “We are hoping that this careful mixing ensures a representative sample.”
Most fungi are microscopic and do not produce mushrooms, noted Mark Anthony, an assistant professor of fungal ecology at the University of Vienna who isn’t involved in the project. To identify these microscopic organisms, the scientists use DNA metabarcoding, a technique that uses genetic sequencing techniques to determine which fungi species are present in a sample and their relative abundance.
“It is currently the best tool available to study fungal biodiversity when people do it right,” Anthony said. “This [project’s] data is also good for the international community because earlier research shows that Ukraine is an area where we know very little about fungal biodiversity.”
The sequences from the soil samples are now being analyzed in labs in the United States, and results are expected by the end of the year.
“These Organisms Are a Black Box”
“These organisms are a black box: We don’t know where they are distributed, nor what environmental conditions they can tolerate—Oleh Prylutskyi is closing this gap,” said Michael Van Nuland, lead data scientist at SPUN.
Julia Köninger, a soil ecologist and postdoc at the Universidade de Vigo in Spain who isn’t involved in the project, explained that documenting soil health was important because war itself introduces pollutants to soils. Human activity at battle sites can leave behind oil and heavy metals; heavy tanks compact soils, and craters from missiles may erode topsoil.
“I have read about scientists that lived during World War II…and I used to wonder why didn’t they just go running and screaming. Why were they describing fungi species in the middle of a war? Now I understand them: Today is the perfect time to try to make the most brave ideas happen.”
“Understanding the diversity and health of soil fungi before the conflict provides a reference point to assess the damage caused by the war,” Köninger said. “This knowledge is vital for documenting environmental war atrocities and understanding the long-term ecological impact.”
Meanwhile, Prylutskyi has kept working.
“I have read about scientists that lived during World War II in Poland and Ukraine, and I used to wonder why they didn’t just go running and screaming. Why were they describing fungi species in the middle of a war?” he said. “Now I understand them: Today is the perfect time to try to make the most brave ideas happen.”
—Andrew J. Wight, Science Writer
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Citation: Wight, A. J. (2024), Ukrainian scientists race to document soil fungi, Eos, 105, https://doi.org/10.1029/2024EO240366. Published on 15 August 2024.
Text © 2024. The authors. CC BY-NC-ND 3.0Except where otherwise noted, images are subject to copyright. Any reuse without express permission from the copyright owner is prohibited.
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