doi: 10.15389/agrobiology.2022.3.476eng

UDC: 631.4/.6:631.95:57.042



E.Ya. Rizhiya1 , L.V. Boitsova1, V.E. Vertebniy1, J. Horak2, M.A. Moskvin1, V.I. Dubovitskaya1, Yu.V. Khomyakov1

1Agrophysical Research Institute, 14, Grazhdanskii prosp., St. Petersburg, 195220 Russia, e-mail (✉ corresponding author),,,,,;
2Slovak University of Agriculture in Nitra, Hospodárska 7, 949 76 Nitra, Slovakia, e-mail

Rizhiya E.Ya.
Moskvin M.A.
Boitsova L.V.
Dubovitskaya V.I.
Vertebniy V.E.
Khomyakov Yu.V.
Horak J.

Received February 17, 2022

Biochars are considered as an attractive tool in agriculture for carbon sequestration and improvement of soil functions. Biochar addition to soils can raise the pH, increase the organic carbon content, enhance nutrient retention, and increase microbial biomass. The introduction of biochar to different soils is an irreversible action. After entering the soil environment, the so called “aging” of biochar begins, due to the water-physical processes occurring in the soil, e.g., moistening, drying, freezing and thawing. Therefore, it is necessary to understand the directions of various changes occurring in the soil when using this ameliorant. The two-year field experiment to study the effect of biochar on the dynamics of some soil enzymes during of biochar aging was performed with the aim to reveal mechanisms of interaction between soil and biochar and to justify the sensitiveness of enzymes on a biochar amendment to the soil. The studied loamy sand Spodosol soil had medium and high soil quality. The experimental design included the soil (control) and the soil with 20 t/ha biochar introduced in the top arable layer (0-10 cm) of 4 m2 plots in 3 replicates. The impact of birch (Betula spp.) biochar produced by fast pyrolysis at 600 °C was studied. Chemical characteristics of the biochar were as follows: Corg. — 88.9 %, Ntot. — 0.43 %, H — 3.2%, O — 5.1 %, pHН2О 8.3, water content — 3.1 %, ash content — 1.8 %. In 2019, a seed mixture of oat (Avеna satíva L.) cv. Borrus and common vetch (Vicia sativa L.) L’govskii cv. was cultivated on the plots at the rate of 200 kg/ha (or 85 g per 4 m2). In 2020, white lupine (Lupinus albus L.) cv. Dega was cultivated as green manure for winter wheat at the rate of 200 kg/ha. Soil and biochar samples were collected from a 0-10 cm layer of the humus horizon in May to August (at 14-day intervals) with an Endelman soil drill (Royal Eijkelkamp B.V., the Netherlands). The activity of peroxidase (EC and polyphenoloxidase (EC was assessed by the photocolorimetric method (l = 440 nm and l = 590 nm, respectively), and the assessment of temporal changes in the oxidation of the surface of biochar was studied by IR spectrometry (an FSM 2202 Michelson spectrometer, Infraspek, Russia). The biochar was found to increase the activity of the studied enzymes, on average by 12-13 %, as compared to the activity in soils without biochar. The peroxidase activity was on average 1.5 times higher than that of the polyphenoloxidase and significantly (p < 0.05) depended on the degree of soil quality. The ratio of polyphenoloxidase to peroxidase in the soil with medium soil quality was approximately 20 % lower than in the soil with high soil quality, where the conditions (temperature, humidity, amount of organic matter) were optimal for humus synthesis. It was found that all treatments showed the soil humification factor less than 1, which indicates the predominance of the processes of mineralization of humic substances in the soil over their immobilization. The biochar increased the mineralization of organic matter by 11.5 % compared to soils without biochar. Over the two-year experiment, IR spectroscopy revealed a tendency to an increase in the amount of hydroxyl, carbonyl, and carboxylate groups compared to the initial biochar, which is consistent with the data on the increase in the activity of polyphenoloxidase and peroxidase. Our findings confirm that biochar introduced into the loamy sand Spodosol remained stable during two years and did not significantly affect the enzymatic activity of soils.

Keywords: soddy-podzolic sandy loam soil, biochar, peroxidase, EC, polyphenol oxidase, EC, IR spectra of biochar.



  1. Lehmann J., Rillig M.C., Thies J., Masiello C.A., Hockaday W.C., Crowley D. Biochar effects on soil biota — a review. Soil Biology and Biochemistry, 2011, 43(9): 1812-1836 CrossRef
  2. Huang Z., Hu L, Dai J. Effects of ageing on the surface characteristics and Cu(II) adsorption behaviour of rice husk biochar in soil. Open Chemistry, 2020, 18(1): 1421-1432 CrossRef
  3. Beusch C. Biochar as a soil ameliorant: how biochar properties benefit soil fertility — a review. Journal of Geoscience and Environment Protection, 2021, 9(10): 28-46 CrossRef
  4. Nie T., Hao P., Zhao Z., Zhou W., Zhu L. Effect of oxidation-induced aging on the adsorption and co-adsorption of tetracycline and Cu2+ onto biochar. Science of The Total Environment, 2019, 673: 522-532 CrossRef
  5. Zheng H., Wang Z., Deng X., Herbert S., Xing B. Impacts of adding biochar on nitrogen retention and bioavailability in agricultural soil. Geoderma, 2013, 206: 32-39 CrossRef
  6. Rechberger M.V., Kloss S., Rennhofer H., Titner J., Watzinger A., Soja G., Lichtenegger H., Zehetner F. Changes in biochar physical and chemical properties: accelerated biochar aging in an acidic soil. Carbon, 2017, 115: 209-219 CrossRef
  7. Li H., Lu X., Xu Y., Liu H. How close is artificial biochar aging to natural biochar aging in fields? A meta-analysis. Geoderma, 2019, 352: 96-103 CrossRef
  8. Gámiz B., Velarde P., Spokas K.A., Celis R., Cox L. Changes in sorption and bioavailability of herbicides in soil amended with fresh and aged biochar. Geoderma, 2019, 337: 341-349 CrossRef
  9. Huang Z., Hu L., Zhou Q., Guo Y., Tang W., Dai J. Effect of aging on surface chemistry of rice husk‐derived biochar. Environmental Progress & Sustainable Energy, 2018, 37(1): 410-417 CrossRef
  10. Fuertes A.B., Camps Arbestain M., Sevilla M., Masiá-Agullo J.A., Fiol S., López R., Smernik R.J., Aikenhead W.P., Arce F., Macías F. Chemical and structural properties of carbonaceous products obtained by pyrolysis and hydrothermal carbonization of corn stover. Australian Journal of Soil Research, 2010, 48(7): 618-626 CrossRef
  11. Batista E.M.C.C., Shultz J., Matos T.T.S., Fornari M.R., Ferreira T.M., Szpoganicz B., de Freitas R.A., Mangrich A.S. Effect of surface and porosity of biochar on water holding capacity aiming indirectly at preservation of the Amazon biome. Scientific Reports, 2018, 8: 10677 CrossRef
  12. Marshall J., Muhlack R., Morton B.J., Dunnigan L., Chittleborough D., Kwong C.W. Pyrolysis temperature effects on biochar—water interactions and application for improved water holding capacity in vineyard soils. Soil Syst., 2019, 3(2): 27 CrossRef
  13. Mukherjee A., Zimmerman1 A.R., Hamdan R., Cooper W.T. Physicochemical changes in pyrogenic organic matter (biochar) after 15 months field aging. Solid Earth Discuss, 2014, 6: 731-760 CrossRef
  14. Shakhnazarova V.Yu., Orlova N.E., Orlova E.E., Bankina T.A., Yakkonen K.L., Rizhiya E.Ya., Kichko A.A. Influence of biochar on the taxonomic composition and structure of prokaryotic communities in agro soddy-podzolic soil. Sel'skokhozyaistvennaya biologiya [Agricultural Biology], 2020, 55(1): 163-173 CrossRef
  15. Gonçalves Lopes E.M., Mendes Reis M., Leidivan Almeida Frazão, da Mata Terra L.M., Lopes E.F., dos Santos M.M., Fernandes L.A. Biochar increases enzyme activity and total microbial quality of soil grown with sugarcane. Environmental Technology & Innovation, 2021, 21: 101270 CrossRef
  16. Wang D., Li C., Parikh S.J., Scow K.M. Impact of biochar on water retention of two agricultural soils — a multi-scale analysis. Geoderma, 2019, 340: 185-191 CrossRef
  17. Sánchez-García M., Sánchez-Monedero M. A., Roig A., López-Cano I., Moreno B., Benitez E., Cayuela M.L. Compost vs biochar amendment: a two-year field study evaluating soil C build-up and N dynamics in an organically managed olive crop. Plant and Soil, 2016, 408(1/2): 1-14 CrossRef
  18. Khaziev F.Kh. Metody pochvennoy еnzimologii [Methods of soil enzymology]. Moscow, 2005 (in Russ.).
  19. Boytsova L.V., Rizhiya E.Ya., Dubovitskaya V.I. Agrokhimiya, 2021, 9: 23-31 CrossRef (in Russ.).
  20. Gul’ko A.E., Khaziev F.Kh. Pochvovedenie, 1992, 11: 55-67 (in Russ.).
  21. Metody pochvennoy mikrobiologii i biokhimii /Pod obshchey redaktsiey D.G. Zvyagintseva [Methods of soil microbiology and biochemistry. D.G. Zvyagintsev (ed.)]. Moscow, 1991 (in Russ.).
  22. KeiluweIt M., Nico P.S., Johnson M.G., Kleber M. Dynamic molecular structure of plant biomass-derived black carbon (biochar). Environmental Science & Technology, 2010, 44(4): 1247-1253 CrossRef
  23. Njuma O.J., Davis I., Ndontsa E.N., Krewall J.R., Liu A., Goodwin D.C. Mutual synergy between catalase and peroxidase activities of the bifunctional enzyme KatG is facilitated by electron hole-hopping within the enzyme. Journal of Biological Chemistry, 2017, 292(45): 8408-18421 CrossRef
  24. Libra J. A., Ro K.S., Kammann C., Funke, Berge N.D., Neubauer Y., Titirici M.M., Fühner C., Bens O., Kern J., Emmerich K.-H. Hydrothermal carbonization of biomass residuals: a comparative review of the chemistry, processes and applications of wet and dry pyrolysis. Biofuels, 2011, 2(1): 71-106 CrossRef
  25. Chunderova A.I. Pochvovedenie, 1970, 7: 22-26 (in Russ.).
  26. Lammirato C., Miltner A., Kaestner M. Effects of wood char and activated carbon on the hydrolysis of cellobiose by b-glucosidase from Aspergillus niger. Soil Biology and Biochemistry, 2011, 43(9): 1936-1942 CrossRef
  27. Ahmad M., Rajapaksha A.U., Lim E.U., Zhang M., Bolan N., Mohan D., Vithanage M., Lee S.S., Ok Y.S. Biochar as a sorbent for contaminant management in soil and water: a review. Chemosphere, 2014, 99: 19-33 CrossRef
  28. Ibrahim M.M., Tong C., Hu K., Zhou B., Xing S., Mao Y. Biochar-fertilizer interaction modifies N-sorption, enzyme activities and microbial functional abundance regulating nitrogen retention in rhizosphere. Science of The Total Environment 2020, 739: 140065 CrossRef
  29. Ameloot N., De Neve S., Jegajeevagan K., Yildiz G., Buchan D., Funkuin Y.N., Prins W., Bouckaert L., Sleutel S. Short-term CO2 and N2O emissions and microbial properties of biochar amended sandy loam soils. Soil Biology and Biochemistry, 2013, 57: 401-410 CrossRef
  30. Bailey V.L., Fansler S.J., Smith J.L., Bolton H. Reconciling apparent variability in effects of biochar amendment on soil enzyme activities by assay optimization. Soil Biology and Biochemistry, 2011 43(2): 296-301 CrossRef
  31. Park J., Choppala G., Bolan N, Chung J., Chuasavathi T. Biochar reduces the bioavailability and phytotoxicity of heavy metals. Plant Soil, 2011, 348: 439-451 CrossRef
  32. Kumar S., Mastro R., Ram L., Sarkar P., George J., Selvi V. Biochar preparation from Parthenium hysterophorus and its potential use in soil application. Journal of Ecological Engineering, 2013, 55: 67-72 CrossRef
  33. Paz-Ferreiro J., Gasco G., Gutierrez B., Mendez A. Soil biochemical activities and the geometric mean of enzyme activities after application of sewage sludge and sewage sludge biochar to soil. Biology and Fertility of Soils, 2012, 48: 511-517 CrossRef
  34. Wu F., Jia Z., Wang S., Chang S., Startsev A. Contrasting effects of wheat straw and its biochar on greenhouse gas emissions and enzyme activities in a Chernozemic soil. Biology and Fertility of Soils, 2013, 49: 555-565 CrossRef
  35. Kuzyakov Y., Bogomolova I., Glaser B. Biochar stability in soil: decomposition during eight years and transformation as assessed by compound-specific 14C analysis. Soil Biology and Biochemistry, 2014, 70: 229-236 CrossRef
  36. Tozzi F.V.D.N., Coscione A.R., Puga A.P., Carvalho C.S., Cerri C.E.P., de Andrade C.A. Carbon stability and biochar aging process after soil application. Horticult. Int. J., 2019, 3(6): 320-329 CrossRef
  37. Nosenko T.N., Sitnikova V.E., Strel’nikova I.E., Fokina M.I. Praktikum po kolebatel’noy spektroskopii [Workshop on vibrational spectroscopy]. St. Petersburg, 2021 (in Russ.).
  38. Ahmad M., Lee S.S., Dou X., Mohan D., Sung J.-K., Yang J.E, Ok Y.S. Effects of pyrolysis temperature on soybean stover and peanut shell-derived biochar properties and TCE adsorption in water. Bioresource Technology, 2012, 118: 536-544 CrossRef
  39. Naisse C., Girardin C., Lefevre R., Pozzi A., Maas R., Stark A., Rumpel K. Effect of physical weathering on the carbon sequestration potential of biochars and hydrochars in soil. GCB Bioenergy, 2015, 7: 488-496 CrossRef
  40. Ghaffar A., Ghosh A., Li F., Dong X., Zhang D., Wu M., Li H., Pan B. Effect of biochar ageing on surface characteristics and adsorption behavior of dialkyl phthalates. Environmental Pollution, 2015, 206: 502-509 CrossRef







Full article PDF (Rus)

Full article PDF (Eng)