doi: 10.15389/agrobiology.2021.1.146eng

UDC: 633.1:631.559.2:579.64



M.L. Sidorenko1, 2 ✉, N.A. Sleptsova1, A.N. Bykovskaya1,
V.V. Berezhnaya3, A.G. Klykov3

1Federal Scientific Center of the East Asia Terrestrial Biodiversity, Far Eastern Branch RAS, 159, Prosp. 100-letiya Vladivostoka, Vladivostok, 690022 Russia, e-mail,;
2Far Eastern Federal University, Campus, 10 Ajax Bay, Russky Island, Vladivostok, 690922 Russia, e-mail (✉ corresponding author);
3Chaika Federal Scientific Center of Agrobiotechnology in the Far East, Far Eastern Branch RAS, 30, ul. Volozhenina, Ussuriysk, 692539 Russia, e-mail,

Sidorenko M.L.
Berezhnaya V.V.
Sleptsova N.A.
Klykov A.G.
Bykovskaya A.N.

Received November 12, 2019


Mineral fertilizers which can significantly increase crop productivity have an adverse effect on the soil and the environment as a whole when used for a long time. Microorganisms, as an alternative to mineral fertilizers, stimulate plant growth and development due to their ability to fix nitrogen, produce siderophores, phytohormones and enzymes, dissolve inaccessible elements of mineral nutrition, suppress plant pathogens, and increase consumption of water and nutrients. However, the effectiveness of such preparations highly depends on factors of a new environment. We believe that bacteria from soils that have long been exposed to various agricultural practices may be good plant stimulants. In the presented study, for the first time, we have isolated local active strains of nitrogen-fixing and phosphate-mobilizing bacteria from soils subjected to 74-year stationary intensive farming in the conditions of the Russian Far East and revealed isolates and their combinations which stimulate wheat and barley seed germination and seedling growth. The aim of the work was to study the plant-stimulating properties of nitrogen-fixing and phosphate-mobilizing bacteria from soils that have been actively exposed to mineral fertilizers for a long time. Bacteria were isolated from soils sampled in October 2015 (the experimental field 8, Federal Research Center for Agrobiotechnology FEB RAS, Ussuriysk, Primorsky Territory, Russia). The seeds of wheat Triticum aestivum L. cultivar Primorskaya 50, and barley Hordeum vurlage L. cultivar Tikhookeanskii (collection of the Federal Research Center for Agrobiotechnology FEB RAS) were treated. Of 68 bacterial isolates with different cultural and morphological properties, three isolates, the Acinetobacter spp. N1, Azotobacter spp. N2, and Clostridium spp. N3 were nitrogen fixers, and four isolates, the Serratia spp. P6, Bacillus spp. P7), Arthrobacter spp. P8, and Pantoea spp. P19 were phosphate-mobilizing bacteria. Tests with the monocultures of nitrogen-fixing and phosphate-mobilizing isolates and their different binary compositions showed a 13-51 % increase (p ≤ 0.05) in wheat seed germination energy and 15-54 % increase (p ≤ 0.05) in barley seed germination energy compared to the untreated control. Laboratory germination of wheat seeds increased by 2-32 %, barley seeds by 7-30 % compared to untreated control. The barley seedlings were 1.8 times longer, and the roots were 2.7 times longer. Th binary combination N2P19, P6P19, and P8P19 caused the highest height of seedlings (120-140 mm, p ≤ 0.05), and with P6P7, N2P19, and P6P19 the roots were the longest (120-130 mm, p ≤ 0.05). These results allow us to conclude that short-term soaking seeds in the suspensions of the tested nitrogen-fixing and phosphate-mobilizing isolates improves seed germination energy and laboratory germination, and increases shoot and root length. Binary bacterial compositions have a greater effect on seed germination than monocultures. The strains N1 (Acinetobacter spp.), N2 (Azotobacter spp.), and P19 (Pantoea spp.) are the best stimulants. Species-specific differences in plant response to the treatment is probably due to lack of genetic, biochemical and physiological complementarities between specific plant species and the bacteria.

Keywords: biological fertilizers, diazotrophs, phosphate-mobilizing bacteria, soil, long-term chemicalization, Triticum aestivum L., wheat, Hordeum vurlage L., barley, seeds, germination energy. laboratory germination, seedlings.



  1. Lissaletta L., Billen G., Garnier J., Bouwman L., Velazquez E., Mueller N.D., Gerber J.S. Nitrogen use in the global food system: past trends and future trajectories of agronomic performance, pollution, trade, and dietary demand. Environmental Research Letters, 2016, 11(9): 1-15 CrossRef
  2. Aleshchenkova Z.M., Safronova G.V., Mel'nikov N.V., Esenbaeva A.E., Ten O.A. Vestnik Bashkirskogo universiteta, 2015, 20(1): 82-86 (in Russ.).
  3. Matin X.M., Sumathi C.S., Kannan V.R. Influence of agrochemical and Azotobacter spp. application on soil fertility in relation to maize growth under nursery conditions. Eurasian Journal of BioSciences, 2011, 5(1): 19-28 CrossRef
  4. Korshunova T.Yu., Silishchev N.N., Loginov O.N., Monakov Yu.B. Vestnik Bashkirskogo universiteta, 2007, 12(3): 34-35 (in Russ.).
  5. Savci S. An agricultural pollutant: chemical fertilizer. International Journal of Environmental Science and Development, 2012, 3(1): 77-79 CrossRef
  6. Gamzaeva R.S. Izvestiya Sankt-Peterburgskogo gosudarstvennogo agrarnogo universiteta, 2015, 40: 38-41 (in Russ.).
  7. Ahmemad M., Khan M.S. Alleviation of fungicide-induced phytotoxicity in greengram [Vigna radiata (L.) Wilczek] using fungicide-tolerant and plant growth promoting Pseudomonas strain. Saudi Journal of Biological Sciences, 2012, 19(4): 451-459 CrossRef
  8. Lukin S.M., Marchuk E.V. Dostizheniya nauki i tekhniki APK, 2011, 8: 18-21 (in Russ.).
  9. Nelson L.M. Plant growth promoting rhizobacteria (PGPR): prospects for new inoculants. Crop Management, 2004, 3(1): 301-305 CrossRef
  10. Shabaev V.P. Agrokhimiya, 2016, 8: 82-87 (in Russ.).
  11. Tirry N., Tahry Joutey N., Sayel H., Kouchou A., Bahafid W., Asri M., El Ghachtouli N. Screening of plant promoting traits in heavy metals resistant bacteria: prospects in phytoremediation. Journal of Genetic Engineering and Biotechnology, 2018, 16(2): 613-619 CrossRef
  12. Espidicar Z., Yarnia M., Ansari M., Mirshekari B., Asadi Rahmani H. Differences in nitrogen and phosphorus uptake and yield components between barley cultivars grown under arbuscular mycorrhizal fungus and pseudomonas strains co-inoculation in rainfed condition. Applied Ecology and Environmental Research, 2017, 15(4): 195-216 CrossRef
  13. Fukami J., Cerezini P., Hungria M. Azospirillum: benefits that go far beyond biological nitrogen fixation. AMB Express, 2018, 8(1): 73 CrossRef
  14. Martínez-Viveros O., Jorquera M.A., Crowley D.E., Gajardo G., Mora M.L. Mechanisms and practical considerations involved in plant growth promotion by Rhizobacteria. Jornal of Soil and Plant Nutrition, 2010, 10(3): 293-319 CrossRef
  15. Hazan R., Que Y.A., Maura D., Rahme L.G. A method for high throughput determination of viable bacteria cell counts in 96-well plates. BCM Microbiology, 2012, 12(1): 259 CrossRef
  16. Hanaka A., Ozimek E., Majewska M., Anna Rysiak A., Jaroszuk-Ściseł J. Physiological diversity of Spitzbergen soil microbial communities suggests their potential as Plant Growth-Promoting Bacteria. International Journal of Molecular Sciences, 2019, 20(5): 1207 CrossRef
  17. Pruntova O.V., Sakhno O.N. Laboratornyi praktikum po obshchei mikrobiologii [Laboratory course in general microbiology]. Vladimir, 2005 (in Russ.).
  18. Bome N.A., Belozerova A.A., Bome A.Ya. Biologicheskie svoistva semyan i fenogeneticheskii analiz kul'turnykh rastenii [Biological properties of seeds and phenogenetic analysis of cultivated plants]. Tyumen', 2007 (in Russ.).
  19. Zvyagintsev D.G. Pochva i mikroorganizmy [Soil and microorganisms]. Moscow, 1987 (in Russ.).
  20. Zolotarev V.N. Agrokhimiya, 2015, 7: 11-16 (in Russ.).
  21. Kifle M.H., Laing D.M. Effects of selected diazotrophs on maize growth. Frontiers in Plant Science, 2016, 7: 1429 CrossRef
  22. Hahn L., de Sá E.L.S., Osório Filho B.D., Machado R.G., Damasceno R.G., Giongo A. Rhizobial inoculation, alone or coinoculated with Azospirillum brasilense, promotes growth of wetland rice. Revista Brasileira de Ciência do Solo, 2016, 40: e0160006 CrossRef
  23. Myresiotis C.K., Vryzas Z., Papadopoulou-Mourkidou E. Biodegradation of soil applied pesticides by selected strains of plant growth promoting rhizobacteria (PGPR) and their effects on bacterial growth. Biodegradation, 2012, 23(2): 297-310 CrossRef
  24. Sochorec M.R., Knot P. The effect of fertilizer seed coating on the germinating capacity and initial development of some turf grass species and white clover. Acta Universitatus et Silviculturae Mendeliane Brunensis, 2012, 60(5): 199-204 CrossRef
  25. Batool S., Iqbal A. Phosphate solubilizing rhizobacteria as alternative of chemical fertilizer for growth and yield of Triticum aestivum (Var. Galaxy 2013). Saudi Journal of Biological Sciences, 2019, 26(7): 1400-1410 CrossRef
  26. Zavalin A.A. Biopreparaty, udobreniya i urozhai [Biopreparations, fertilizers and crop]. Moscow, 2005 (in Russ.).
  27. Xie H., Pasternak J.J., Glick B.R. Isolation and characterization of mutants of the plant growth-promoting rhizobacterium Pseudomonas putida CR12-2 that overproduce indoleacetic acid. Current Microbiology, 1996, 32(2): 67-71 CrossRef
  28. Mnasri N., Chennaoui C., Gargouri S., Mhamdi R., Hessini K., Elkahoui S., Djebali N. Efficacy of some rhizospheric and endophytic bacteria in vitro and as seed coating for the control of Fusarium culmorum infecting durum wheat in Tunisia. European Journal of Plant Pathology,2017, 147(3): 501-515 CrossRef
  29. Widawati S., Suliasih S. The effect of plant growth promoting rhizobacteria (PGPR) on germination and seedling growth of Sorghum bicolor L. Moench. IOP Conf. Series: Earth and Environmental Science, 2018, 166(1): 012022 CrossRef
  30. Afzal A., Bano A. Rhizobium and phosphate solubilizing bacteria improve the yield and phosphorus uptake in wheat (Triticum aestivum). International Journal of Agriculture and Biology, 2008, 10(1): 85-88.
  31. Kuz'min N.A., Seitova O.V. Vestnik Ryazanskogo gosudarstvennogo agrotekhnicheskogo universiteta im. P.A. Kostycheva, 2010, 2: 27-30 (in Russ.).
  32. Costa L.C., Tavanti R.F.R., Tavanti T.R., Pereira C.S., Desenvolvimento de cultivares de soja após inoculação de estirpes de Bacillus subtilis. Nativa, 2019, 7(2): 126-132 CrossRef






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