Activity of nitrogen fixation and antioxidant enzymes in symbiotic systems Glycine max – Bradyrhizobium japonicum for complex treatment with lectin and fungicides

  • S. Y. Kots Institute of Plant Physiology and Genetics
  • T. P. Mamenko Institute of Plant Physiology and Genetics
  • A. V. Pavlyshche Institute of Plant Physiology and Genetics
Keywords: soybean; rhizobia; superoxide dismutase; guaiacol peroxidase; ascorbate peroxidase; symbiosis


The dynamics of the nitrogen fixation activity of the root nodules, the growth of the vegetative mass of plants and the change in the activity of antioxidant enzymes (superoxide dismutase, ascorbate and guaiacol peroxidase) in different soybean organs for treatment of seeds by rhizobia incubated with lectin, in combination with fungicides have been studied. The objects of the study were symbiotic systems formed with the participation of soybean (Glycine max (L.) Merr.) Almaz and Bradyrhizobium japonicum (standard strain 634b) incubated with lectin. As disinfectants of soybean seeds, the following preparations with fungicidal activity were used – Maxim XL 035 PS, Fever, Standak Top according to one rate of active substance consumption of each preparation specified by the manufacturer. One part of the seeds treated with fungicides was inoculated with pure culture of suspension of rhizobia for one hour (titre of suspension concentration was 108 cells/ml). Another part of the seeds treated with fungicides was inoculated with rhizobia suspension, which was previously incubated with a solution of commercial lectin soybean at a concentration of 100 μg/ml. The research was conducted in strictly controlled conditions of a model vegetative experiment using microbiological, physiological, biochemical methods, gas chromatography, spectrophotometry. It was found that processing of soybean seeds with fungicides (Fever and Maxim XL) together with rhizobium inoculation contributed to the preservation of the nitrogen fixation activity of the root nodules and the growth of vegetative mass of plants. Under these conditions, the intensification of the activity of superoxide dismutase and ascorbate peroxidase was observed, as well as inhibition of the activity of guaiacol peroxidase in soybean root nodules in the phase of three true leaves and increased activity of all investigated enzymes in the phase of mass flowering. It has been established that the use of complex treatment of seeds by soybean rhizobia incubated with lectin and fungicides leads to an increase in the activity of superoxide dismutase and guaiacol peroxidase in root nodules in the phase of three true leaves and the growth of the activity of ascorbate peroxidase in the phase of mass flowering. At the same time, the inhibition of the growth of vegetative mass of plants and their symbiotic properties occurred, as evidenced by the decrease in the nitrogen fixation activity of the root nodules for the joint treatment of seeds with fungicides and lectin. A specific reaction of investigated enzymes in the roots and leaves of soybean was shown, which was more pronounced in the phase of three true leaves, indicating the development of a typical antioxidant reaction to a complex treatment, as a kind of stress that is leveled to the phase of mass flowering. The degree of reaction of antioxidant enzymes in the studied symbiotic systems Glycine max – Bradyrhizobium japonicum depends on the nature of the active substance fungicides and the manifestation of their joint effect in a complex with rhizobia incubated with lectin.


Alscher, R. G., Erturk, N., & Heath, L. S. (2002). Role of superoxide dismutases (SODs) in controlling oxidative stress in plants. Journal of Experimental Botany, 53(372), 1331–1341.

Araujo, R. S., Cruz, S. P., Souchie, E. L., Martin, T. N., Nakatani, A. S., Nogueira, M. A., & Hungria, M. (2017). Preinoculation of soybean seeds treated with agrichemicals up to 30 days before sowing: Technological innovation for large-scale agriculture. International Journal of Microbiology, 1‒11.

Babosha, A. V. (2008). Inducible lectins and plant resistance to pathogens and abiotic stress. Biochemistry, 73(7), 812–825 (in Russian).

Bikrol, A., Saxena, N., & Singh, K. (2005). Response of Glycine max in relation to nitrogen fixation as influenced by fungicide seed treatment. African Journal of Biotechnology, 4(7), 667–671.

Bradford, M. A. (1976). Rapid and sensitive method for the quantitation of the microgram quantities of protein utilising: The principle of protein – dye binding. Analytical Biochemistry, 72, 248–254.

Chrispeels, M. J., & Raikhel, N. V. (1991). Lectins, lectin genes and their role in plant defence. Plant Cell, 3, 1–9.

Dalton, D. A., Baird, L. M., Langeber, L., Taugher, C. Y., Anyan, W. R., Vance, C. P., & Sarath, G. (1993). Subcellular localization of oxygen defense enzy mes in soybean (Glycine max (L.) Merr.) root nodules. Plant Physiology, 102(1), 481–489.

Dalton, D., Joyner, S. L., Becana, M., Iturbe-Ormaetxe, I., & Chatfield, J. M. (1998). Antioxidant defenses in the peripheral cell layers of legume root nodules. Plant Physiology, 116, 37–43.

Egley, G. H., Paul, R. N., Vaughn, K. C., & Duke, S. O. (1983). Role of peroxida se in the development of water impermeable seed coats in Sida sprinosa L. Planta, 157(1), 224–232.

Erofeeva, E. A. (2015). Dependence of guaiacol peroxidase activity and lipid peroxidation rate in drooping birch (Betula pendula Roth) and tillet (Tilia cordata Mill.) leaf on motor traffic pollution intensity. Dose-Response: An International Journal, 1‒6.

Evtushenko, M. D., Marutin, F. M., Turenko, V. P., Zherebko, V. M., & Sekun, M. P. (2004). Phytopharmacologia. Vusha Osvyta, Kyiv (in Ukrainian).

Fedorova, E. E., Zhivnevskaya, G. Y., Kalibernaya, Z. V., Artemenko, E. N., Iz mailov, S. F., & Gus'kov, A. V. (2000). Metabolism of IOA with the estab lishment of symbiosis between Phaseolus vulgaris and Rhizobium phaseoli. Russian Journal of Plant Physiology, 47(2), 231–235 (in Russian).

Fox, J. E., Gulledge, J., Engelhaupt, E., Burow, M. E., & McLachlan, J. A. (2007). Pecticides reduce symbiotic efficiency of nitrogen-fixing rhizobia and host plants. Proceeding of the National Academy of Sciences USA, 104(24), 10282–10287.

Hardy, R. W. F., Holsten, R. D., Jackson, E. K., & Burns, R. C. (1968). The acety lene-ethylene assay for nitrogen fixation: Laboratory and field evalution. Plant Physiology, 43(8), 1185–1207.

Hivrale, A. U., & Ingale, A. G. (2013). Plant as a plenteous reserve of lectin. Plant Signaling and Behavior, 8(12), 1–7.

Hoff, P. L. D., Brill, L. M., & Hirsch, A. M. (2009). Plant lectins: The ties that bind in root symbiosis and plant defense. Molecular Genetics Genomics, 282(1), 1–15.

Iturbe-Ormaetxe, J., Matamoros, M. A., Rubio, M. C., Dalton, D. A., & Becana, M. (2001). The antioxidant of legume nodule mitochondria. Molecular Plant-Microbe Interaction, 14(10), 1189–1196.

Kobak, S. Y., Kolisnik, S. I., & Serevetnyk, O. V. (2016). The most common diseases of soybean and the effectiveness of BASF products for their control. Agrobusiness Today, 10(329), 46–47 (in Ukrainian).

Kyrychenko, E. V. (2014). Phytolectines and diazotrophs – polyfunctional com ponents of complex biological compositions. Biotechnologia Acta, 7(1), 40–53 (in Ukrainian).

Lehotzky, R. E., Partch, C. L., Mukherjee, S., Cash, H. L., Goldman, W. E., Gard ner, K. H., & Hooper, L. V. (2010). Molecular basis for peptidoglycan recog nition by a bactericidal lectin. Proceeding of the National Academy of Sciences USA, 107(17), 7722–7727.

Marco, I. (2013). Integrated soybean protection against diseases. Agrobusiness Today, 11(258), 16–21 (in Ukrainian).

Matamoros, M. A., Dalton, D. A., Ramos, J., Clemente, M. R., Rubio, M. C., & Becana, M. (2003). Biochemistry and molecular biology of antioxidants in the rhizobia ‒ legume symbiosis. Plant Physiology, 133(2), 499–509.

Mittler, R. (2002). Oxidative stress, antioxidants and stress tolerance. Trends Plant Sciences, 7, 405–410.

Mohammadi, K., Sohrabi, Y., Heidari, G., Khalesro, S., & Majidi, M. (2012). Ef fective factors on biological fiation. Agricaltural Research, 7(12), 1782‒1788.

Moran, J. F., James, E. K., Rubio, M. C., Sarath, G., Klucas, R. V., & Becana, M. (2003). Functional characterization and expression of a cytosolic iron-super oxide dismutase from cowpea root nodules. Plant Physiology, 133(2), 773–782.

Moreira, R. A., Ainouz, I. L., Oliveira, J. T., & Cavada, B. S. (1991). Plant lectins, chemical and biological aspects. Memorias Instituto Oswaldo Cruz, 86(2).

Nakano, Y., & Asada, K. (1981). Hydrogen peroxidase is scavenged by ascor bate-specific peroxidase in spinach chloroplasts. Plant Cell Physiology, 22(5), 867–880.

Nikolaevsky, V., Sirenko, V., & Titova, L. (2017). Effect of pre-seed bacterializa tion of seeds on disease development and yield of soybean. Stiinta Agricola, 1, 55–59.

Owens, R. A., Blecburn, M., & Ding, B. (2001). Possible involvement of the phloem lectin in long-distance varied movement. Molecular Plant Microbe Interact, 14(7), 905–909.

Parsiavash, L., Saboora, A., & Nejad, S. Z. M. (2015). Investigating on the stability of peroxidase extracted from soybean (Glycine max var. williams) and effects of Na+ and K+ ions on its activity. Journal of Cell and Molecular Research, 7(2), 94–101.

Petrichenko, V. F., & Kots, S. Y. (2014). Symbiotic systems in modern agricul tural production. Bulletin of NAS of Ukraine, 3, 57–66 (in Ukrainian).

Raychauhuri, S. S., & Deng, X. W. (2000). The role of superxide dismutase in com bating oxidative stress in higher plants. The Botanical Review, 66(1), 89–98.

Rubio, M. C., James, E. K., Clemente, M. R., Bucciarelli, B., Fedorova, M., Vance, C. P., & Becana, M. (2004). Localization of superoxide dismutases and hydrogen peroxide in legume root nodules. The American Phytopatho logical Society, 17(12), 1294–1305.

Schmitz, N., Huystee, R., & Gijzen, M. (1997). Characterization of anionic soy bean (Glycine max) seed coat peroxidase. Canadian Journal of Botany, 75(8), 1336‒1341.

Sergienko, V. (2012). Diseases of soybeans and measures of its limitation. Agro business Today, 11(234), 28–30 (in Ukrainian).

Shao, H.-B., Chu, L.-Y., Lu, Z.-H., & Kang, C.-M. (2008). Primary antioxidant free radical scavenging and redox signaling pathways in higher plant cells. International Journal of Biological Sciences, 4(1), 8‒14.

Sharma, P., Jha, A. B., Dubey, R. S., & Pessarakli, M. (2012). Reactive oxygen species, oxidative damage, and antioxidative defense mechanism in plants under stressful conditions. Journal of Botany, 20(12), 1–26.

Zhiznevskaya, G. Y., Troitskaya, G. N., Borodenko, L. I., & Izmailov, S. F. (2001). Peroxidase and catalase in root nodules of fodder beans at an effective and ineffective symbiosis with rhizobia. Physiology and Biochemistry of Cultural Plants, 33(4), 285–290 (in Ukrainian).
How to Cite
Kots, S. Y., Mamenko, T. P., & Pavlyshche, A. V. (2018). Activity of nitrogen fixation and antioxidant enzymes in symbiotic systems Glycine max – Bradyrhizobium japonicum for complex treatment with lectin and fungicides. Regulatory Mechanisms in Biosystems, 9(2), 148-155.