doi: 10.15389/agrobiology.2015.3.369eng

UDC 631.559.2:631.847.21:579.64

Supported by EurAsEC program «Innovative biotechnologies», State Contract № 14.М04.12.0011.


A.P. Kozhemyakov1, Yu.V. Laktionov1, T.A. Popova1, A.G. Orlova1,
A.L. Kokorina2, O.B. Vaishlya3, E.V. Agafonov4, S.A. Guzhvin4,
A.A. Churakov5, M.T. Yakovleva6

1All-Russian Research Institute for Agricultural Microbiology, Federal Agency of Scientific Organizations, 3, sh. Podbel’skogo, St. Petersburg, 196608 Russia,

2Saint Petersburg State Agrarian University, 2/1, Sankt-Peterburgskoe sh., St. Petersburg, 196608 Russia,

3Tomsk State University, 36, pr. Lenina, Tomsk, 634050 Russia,

4Donskoi State Agrarian University, pos.Persianovskii, Oktyabr’skii Region, Rostov Province, 346493 Russia,

5Krasnoyarsk State Agrarian University, 66, pr. Svobodnyi, Krasnoyarsk, 660041 Russia,

6Yakutsk Research Institute of Agriculture, Federal Agency of Scientific Organizations, 23/1, ul. Bestuzheva-Marlinskogo, Yakutsk, 677001 Russia,

Received March 30, 2015


Herein we summarize the results of a comprehensive study aimed on the creation of liquid form of biological products for symbiotic and associative rhizobacteria. The objects of study were nodule bacteria from the rhizosphere of galegae (Rhizobium galegae), soybean (Bradyrhizobium japonicum) and associative rhizobacteria (Arthrobacter mysorens, Azospirillum brasilense, Agrobacterium radiobacter), as well as soybean (Glycine max), corn (Zea) and wheat (Trticum) plants. As carriers for bacteria, we used gamma-sterile peat, vermiculite and improved sterile liquid media on the basis of components of vegetable or synthetic origin, with the introduction of nutrients and stabilizers (humates, glycerine, potassium sorbate, carboxymethylcellulose, etc.). The efficacy of tested products and forms were compared in a biological survey carried out in different regions of the Russian Federation. Under the conditions of Leningrad Region the effectiveness was evaluated for the preparations based on recently used and perspective strains of alfalfa nodule bacteria Sinorhizobium meliloti. An optimized combination of stabilizers, nutrients and protective substrates provided a long-term storage and high efficiency of biological products. Nevertheless, plant nutrient media (bean broth) did not provide the required quality. Using bean medium for cultivation it is possible to obtain the desired titer of bacteria in the product, but their numbers are rapidly decreasing. Furthermore, preparations obtained with use of bean broth lose quality after 2-3 months after the beginning of storage because of extraneous microflora therein. The new forms of biochemicals designed on the basis of a synthetic nutrient medium can be stored at room temperature for at least 6 months, when sterile-packed in plastic bottles. High bacteria titer (3.6-4.2 billion CFU/ml of liquid formulation) was obtained after cultivation. It is shown that an improved liquid form has a high efficiency. Its application on leguminous plants increased grain yield by 20-40 % in soy, and by 15-25 % in peas. Productivity of cereal crops (wheat, barley) increased by 15-25 %, and the productivity of alfalfa was 20-45 % higher. A significant increase in crop quality was found to be due to higher protein content. The effectiveness of biological products considerably depended on agro-ecological conditions and biological features of the crops. Using different strains of nodule bacteria Sinorhizobium meliloti in the Leningrad Region allowed us to create the effective plant-microbe systems. Thus on the sod-podzolic soil the strain A-4 was most effective providing yield increase by more than 25 %, while on sod-carbonate soil it was the strain A-5 which increased the productivity of alfalfa by 140 %.

Keywords: biological product, nodule bacteria, rhizobacteria association, forms of biological products.


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  1. Zavalin A.A. Biopreparaty, udobreniya i urozhai[Biologicals, fertilizers and crop yield]. Moscow, 2005.
  2. Tikhonovich I.A., Kruglov Yu.V., Kozhemyakov A.P., Chebotar' V.K., Kandybin N.V., Laptev G.Yu. Biopreparaty v sel'skom khozyaistve [Biologicals in agriculture]. Moscow, 2005.
  3. Blagoveshchenskii G.V., Voitovich I.V., Shtyrkhunov V.D., Ol'khovyi V.E. Kormoproizvodstvo, 2003, 4: 20-23.
  4. Emel'yanova A.G., Yakovleva M.T., Kozhemyakov A.P. Priem uluchsheniya plodorodiya merzlotnykh pochv s ispol'zovaniem inokulyatsii semyan lyutserny zheltoi shtammami kluben'kovykh i assotsiativnykh bakterii. Metodicheskoe posobie [Improving permafrost soil fertility by inoculation of yellow alfalfa seeds with nodule and associative bacteria]. Yakutsk, 2014.
  5. Kozhemyakov A.P., Timofeeva S.V., Popova T.A. Zashchita i karantin rastenii,  2008, 2: 42-43.
  6. Mishustin E.N. V sbornike: Mineral'nyi i biologicheskii azot v zemledelii SSSR [In: Mineral and biological nitrogen in agriculture of the USSR]. Moscow 1985: 3-11.
  7. Kozhemyakov A.P. Mikrobiologicheskii zhurnal, 1997, 59(4): 22-28.
  8. Beveridge C.A., Mathesius U., Rose R.J., Gresshoff P. Common regulatory themes in meristem development and whole-plant homeostasis. Curr. Opin. Plant Biol., 2007, 10: 44-51 CrossRef
  9. Marra L.M., Fonseca Sousa Soares C.R., Oliveira S.M., Avelar Ferreira P.A., Soares B.L. Biological nitrogen fixation and phosphate solubilization by bacteria isolated from tropical soils. Plant Soil, 2012, 357: 289-307 CrossRef
  10. Khotyanovich A.V. Metody kul'tivirovaniya azotfiksiruyushchikh bakterii, sposoby polucheniya i primenenie preparatov na ikh osnove (metodicheskie rekomendatsii) [Nitrogen-fixing bacteria cultivation, biologicals' production and application: recommendations]. Leningrad, 1991.
  11. Tittabutr P., Payakapong W., Teaumroong N., Singleton P.W., Boon-
    kerd N. Growth, survival and field performance of bradyrhizobial liquid inoculant formulations with polymeric additives. Science Asia, 2007, 33: 69-77.
  12. Albareda M., Rodriguez-Navarro D.N., Camacho M., Temprano F.J.  Alternatives to peat as a carrier for rhizobia inoculants: solid and liquid formulations. Soil Biol. Biochem., 2008, 40: 2771-2779 CrossRef
  13. Covarrubias S.A., de-Bashan L.E., Moreno M., Bashan Y. Alginate beads provide a beneficial physical barrier against native microorganisms in wastewater treated with immobilized bacteria and microalgae. Appl. Microbiol. Biotechnol., 2012, 93(6): 2669-2680 CossRef
  14. Denton M.D., Pearce D.J., Ballard R.A., Hannah M.C., Mutch L.A., Norng S. A multi-site field evaluation of granular inoculants for legume nodulation. Soil Biol. Biochem., 2009, 41: 2508-2516 CrossRef
  15. Diaz-Zorita M., Fernandez-Canigia M.V. Field performance of a liquid formulation of Azospirillum brasilense on dry land wheat productivity. Eur. J. Soil Biol., 2009, 45: 3-11 CrossRef
  16. John R.P., Tyagi R.D., Brar S.K., Prevost D. Development of emulsion from rhizobial fermented starch industry wastewater for application as Medicago sativa seed coat. Eng. Life Sci., 2010, 10: 248-256.
  17. Laktionov Yu.V. Bakterial'nye preparaty [Bacterial biologicals]. LAP LAMBERT Academic Publishing, 2011 (ISBN 978-3-8433-1137-3).
  18. Laktionov Yu.V., Popova T.A., Andreev O.A., Ibatullina R.P., Ko-
    zhemyakov A.P. Sel'skokhozyaistvennaya biologiya [Agricultural Biology], 2011, 3: 116-118 (
  19. Tikhonovich  I.A.,  Kozhemyakov  A.P.,  Provorov  N.A. Genetic potential of plants for improving the beneficial microbe interactions. NATO ASI series. In: Biological fixation of nitrogen for ecology and sustainable agriculture. Berlin, 1997: 191-194.
  20. Belimov A.A., Vorob'ev N.I., Kozhemyakov A.P. Byulleten' VNIISKhM, 1989, 52: 6-12.
  21. Laktionov Yu.V., Kozhemyakov A.P. Materialy dokladov 7-i konferentsii «Perspektivy ispol'zovaniya novykh form udobrenii, sredstv zashchity i regulyatorov rosta rastenii v agrotekhnologiyakh sel'skokhozyaistvennykh kul'tur» [Proc. 7th Conf. «Prospects for the use of new forms of fertilizers, plant protection products and plant growth regulators in agrotechnology».]. Anapa,  2012: 86-87.
  22. Laktionov Yu.V., Popova T.A., Andreev O.A., Ibatullina R.P., Kozhemyakov A.P. V sbornike: Sovremennye podkhody v biotekhnologii Respubliki Tatarstan [In: Recent approach to biotechnology in the Republic of Tatarstan]. Kazan', 2013: 34-38.
  23. Laktionov Yu.V., Belobrova S.N., Kozhemyakov A.P., Vorob'ev N.I., Sergaliev N.Kh., Amenova R.K. Plodorodie, 2013, 5: 24-25.
  24. Berestetskii O.A., Dorosinskii L.M., Kozhemyakov A.P. Izvestiya AN SSSR, seriya biologicheskaya, 1987, 5: 670-679.