vetpressАграрная наукаAgrarian science0869-81552686-701XРедакция журнала "Аграрная наука"10.32634/0869-8155-2024-384-7-166-169vetpress-3183Research ArticleАГРОИНЖЕНЕРИЯ И ПИЩЕВЫЕ ТЕХНОЛОГИИAGROENGINEERING AND FOOD TECHNOLOGIESКультурально-физиологическая характеристика штаммов Sinorhizobium fredii селекции ВНИИ сои и их способность продуцировать витамины В9 и В12Cultural and physiological characteristics of Sinorhizobium fredii strains selected by the All-Russian Scientific Research Institute of Soybean and their ability to produce vitamins В9 and В12https://orcid.org/0000-0002-1141-1900ЯкименкоМ. В.YakimenkoM. V.

Мария Владимировна Якименко - ведущий научный сотрудник, заведующая лабораторией биологических исследований, кандидат биологических наук 

Игнатьевское шоссе, 19, Благовещенск, Амурская обл., 675000

Maria Vladimirovna Yakimenko - Leading researcher, Head of the Laboratory of Biological Research, Candidate of Biological Sciences 

19 Ignatievskoe highway, Blagoveshchensk, Amur region, 675000

yamv@vniisoi.ruhttps://orcid.org/0000-0003-0098-3484ТатаренкоИ. Ю.TatarenkoI. Yu.

Игорь Юрьевич Татаренко - старший научный сотрудник лаборатории биологических исследований, кандидат сельскохозяйственных наук 

Игнатьевское шоссе, 19, Благовещенск, Амурская обл., 675000

Igor Yurievich Tatarenko - Senior Researcher at the Laboratory of Biological Research, Candidate of Agricultural Sciences 

19 Ignatievskoe highway, Blagoveshchensk, Amur region, 675000

tigy@vniisoi.ruhttps://orcid.org/0000-0003-4611-767XСорокинаА. И.SorokinaA. I.

Арина Игоревна Сорокина - ведущий научный сотрудник лаборатории биологических исследований, кандидат ветеринарных наук 

Игнатьевское шоссе, 19, Благовещенск, Амурская обл., 675000

Arina Igorevna Sorokina - Leading Researcher at the Laboratory of Biological Research, Candidate of Veterinary Sciences 

19 Ignatievskoe highway, Blagoveshchensk, Amur region, 675000

aziradot@mail.ruВсероссийский научно-исследовательский институт сои (ФБГНУ ФНЦ ВНИИ сои)РоссияAll-Russian Scientific Research Institute of Soybean (FSBSI FRC ARSRI of Soybean)Russian Federation20243007202407166169Copyright © Якименко М.В., Татаренко И.Ю., Сорокина А.И., 20242024Якименко М.В., Татаренко И.Ю., Сорокина А.И.Yakimenko M.V., Tatarenko I.Y., Sorokina A.I.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.vetpress.ru/jour/article/view/3183

Актуальность. Микроорганизмы, стимулирующие рост растений (PGPB), в том числе и ризобии, с помощью различного физиологического, молекулярного и биохимического воздействий улучшают продуктивность растений. Из вышеперечисленных факторов наименее изучена роль витаминов. Для оценки способности ризобий продуцировать витамины В9 и В12 были отобраны штаммы вида Sinorhizobium fredii, которые представляют собой однородную группу и имели хороший и обильный рост биомассы на различных питательных средах. Методы. Культурально-физиологические свойства штаммов изучали общепринятыми лабораторными микробиологическими методами. Пересевы коллекционных штаммов ризобий, изучение различных свойств этих бактерий проводили на питательной среде МДА и МРС, а также на производственных питательных средах RM и TY производства фирмы HIMEDIA (Индия). Определение чувствительности штаммов S. fredii к антибиотикам проводили диско-диффузным методом. Содержание в бактериальной массе ризобий витаминов В9 и В12 определяли хемилюминесцентным иммунным методом с использованием иммунохимических систем Access на хемилюминесцентном анализаторе Access2. Результаты. Установлено, что изучаемые штаммы S. fredii имеют хороший или обильный рост бактериальной массы на питательных средах МРС, МДА, RM и TY. Они каталазоположительные, обладают высокой и средней устойчивостью к АБП. Наибольшую концентрацию витамина В9 в биомассе синтезировали штаммы СБ-39 (75,0 пг/мл), ББ-49 (66,6 пг/мл) и ТБ-488 (48,9 пг/мл), а витамина В12 — штаммы ББ-49, СБ-39 (1500 пг/мл).

Relevance. Microorganisms that stimulate plant growth (PGPB), including rhizobia, improve plant productivity through various physiological, molecular and biochemical effects. Of the above factors, the role of vitamins has been studied to a lesser extent. For assessing the ability of rhizobia to produce vitamins В9 и В12, Sinorhizobium fredii strains were selected, which represent a homogeneous group and had good and abundant biomass growth on various nutrient media. Methods. The cultural and physiological properties of the strains were studied using generally accepted laboratory microbiological methods. Passages of collection strains of rhizobia and the study of various properties of these bacteria were carried out on nutrient media MDA and MRS, and on production nutrient media RM and TY produced by HIMEDIA company (India). Determination of the sensitivity of S. fredii strains to antibiotics was carried out using the disc diffusion method. The content of vitamins B9 and B12 in the bacterial mass of rhizobia was determined by the chemiluminescent immune method with the use of paramagnetic particles, and by applying immunochemical Access systems, on the chemiluminescence analyzer Access2. Results. It was found that the studied strains S. fredii have good or abundant growth of bacterial mass on nutrient media MRS, MDA, RM and TY. They are catalase-positive, have high and medium resistance to antibiotics. The highest concentration of vitamin B9 in biomass was synthesized by strains SB-39 (75.0 pg/ml), BB-49 (66.6 pg/ml) and TB-488 (48.9 pg/ml), and vitamin B12 — by strains BB-49 and SB-39 (1500 pg/ml).

ризобиивитаминыкультуральные свойствапитательные средыантибиотикиrhizobiavitamin В9vitamin В12cultural propertiesnutrient mediaantibioticscatalaseReferencesSingh D., Ghosh P., Kumar J., Kumar A. Plant Growth-Promoting Rhizobacteria (PGPRs): Functions and Benefits. Singh D., Gupta V., Prabha R. (eds.). Microbial Interventions in Agriculture and Environment. Singapore: Springer. 2019; 2: 205–227. https://doi.org/10.1007/978-981-13-8383-0_7Singh D., Ghosh P., Kumar J., Kumar A. Plant Growth-Promoting Rhizobacteria (PGPRs): Functions and Benefits. Singh D., Gupta V., Prabha R. (eds.). Microbial Interventions in Agriculture and Environment. Singapore: Springer. 2019; 2: 205–227. https://doi.org/10.1007/978-981-13-8383-0_7Palacios O.A., Bashan Y., de-Bashan L.E. Proven and potential involvement of vitamins in interactions of plants with plant growth-promoting bacteria — an overview. Biology and Fertility of Soils. 2013; 50(1): 140–158. https://doi.org/10.1007/s00374-013-0894-3Palacios O.A., Bashan Y., de-Bashan L.E. Proven and potential involvement of vitamins in interactions of plants with plant growth-promoting bacteria — an overview. Biology and Fertility of Soils. 2013; 50(1): 140–158. https://doi.org/10.1007/s00374-013-0894-3Cronan J.E., Littel K.J., Jackowski S. Genetic and biochemical analyses of pantothenate biosynthesis in Escherichia coli and Salmonella typhimurium. Journal of Bacteriology. 1982; 149(3): 916–922. https://doi.org/10.1128/jb.149.3.916-922.1982Cronan J.E., Littel K.J., Jackowski S. Genetic and biochemical analyses of pantothenate biosynthesis in Escherichia coli and Salmonella typhimurium. Journal of Bacteriology. 1982; 149(3): 916–922. https://doi.org/10.1128/jb.149.3.916-922.1982Balabanova L., Averianova L., Marchenok M., Son O., Tekutyeva L. Microbial and Genetic Resources for Cobalamin (Vitamin B12) Biosynthesis: From Ecosystems to Industrial Biotechnology. International Journal of Molecular Sciences. 2021; 22(9): 4522. https://doi.org/10.3390/ijms22094522Balabanova L., Averianova L., Marchenok M., Son O., Tekutyeva L. Microbial and Genetic Resources for Cobalamin (Vitamin B12) Biosynthesis: From Ecosystems to Industrial Biotechnology. International Journal of Molecular Sciences. 2021; 22(9): 4522. https://doi.org/10.3390/ijms22094522Bernhardt C., Zhu X., Schütz D., Fischer M., Bisping B. Cobalamin is produced by Acetobacter pasteurianus DSM 35099. Applied Microbiology and Biotechnology. 2019; 103(9): 3875–3885. https://doi.org/10.1007/s00253-019-09704-3Bernhardt C., Zhu X., Schütz D., Fischer M., Bisping B. Cobalamin is produced by Acetobacter pasteurianus DSM 35099. Applied Microbiology and Biotechnology. 2019; 103(9): 3875–3885. https://doi.org/10.1007/s00253-019-09704-3Acevedo-Rocha C.G., Gronenberg L.S., Mack M., Commichau F.M., Genee H.J. Microbial cell factories for the sustainable manufacturing of B vitamins. Current Opinion in Biotechnology. 2019; 56: 18–29. https://doi.org/10.1016/j.copbio.2018.07.006Acevedo-Rocha C.G., Gronenberg L.S., Mack M., Commichau F.M., Genee H.J. Microbial cell factories for the sustainable manufacturing of B vitamins. Current Opinion in Biotechnology. 2019; 56: 18–29. https://doi.org/10.1016/j.copbio.2018.07.006Zhang Y., Li X., Wang Z., Wang Y., Ma Y., Su Z. Metabolic Flux Analysis of Simultaneous Production of Vitamin B12 and Propionic Acid in a Coupled Fermentation Process by Propionibacterium freudenreichii. Applied Biochemistry and Biotechnology. 2021; 193(10): 3045–3061. https://doi.org/10.1007/s12010-021-03584-yZhang Y., Li X., Wang Z., Wang Y., Ma Y., Su Z. Metabolic Flux Analysis of Simultaneous Production of Vitamin B12 and Propionic Acid in a Coupled Fermentation Process by Propionibacterium freudenreichii. Applied Biochemistry and Biotechnology. 2021; 193(10): 3045–3061. https://doi.org/10.1007/s12010-021-03584-yOh S., Cave G., Lu C. Vitamin B12 (Cobalamin) and Micronutrient Fortification in Food Crops Using Nanoparticle Technology. Frontiers in Plant Science. 2021; 12: 668819. https://doi.org/10.3389/fpls.2021.668819Oh S., Cave G., Lu C. Vitamin B12 (Cobalamin) and Micronutrient Fortification in Food Crops Using Nanoparticle Technology. Frontiers in Plant Science. 2021; 12: 668819. https://doi.org/10.3389/fpls.2021.668819de-Bashan L.E., Bashan Y. Joint Immobilization of Plant Growth-Promoting Bacteria and Green Microalgae in Alginate Beads as an Experimental Model for Studying Plant-Bacterium Interactions. Applied and Environmental Microbiology. 2008; 74(21): 6797–6802. https://doi.org/10.1128/AEM.00518-08de-Bashan L.E., Bashan Y. Joint Immobilization of Plant Growth-Promoting Bacteria and Green Microalgae in Alginate Beads as an Experimental Model for Studying Plant-Bacterium Interactions. Applied and Environmental Microbiology. 2008; 74(21): 6797–6802. https://doi.org/10.1128/AEM.00518-08de-Bashan L.E., Antoun H., Bashan Y. Cultivation factors and population size control uptake of nitrogen by the microalgae Chlorella vulgaris when interacting with the microalgae growth-promoting bacterium Azospirillum brasilense. FEMS Microbiology Ecology. 2005; 54(2): 197–203. https://doi.org/10.1016/j.femsec.2005.03.014de-Bashan L.E., Antoun H., Bashan Y. Cultivation factors and population size control uptake of nitrogen by the microalgae Chlorella vulgaris when interacting with the microalgae growth-promoting bacterium Azospirillum brasilense. FEMS Microbiology Ecology. 2005; 54(2): 197–203. https://doi.org/10.1016/j.femsec.2005.03.014Marek-Kozaczuk M., Skorupska A. Production of B-group vitamins by plant growth-promoting Pseudomonas fluorescens strain 267 and the importance of vitamins in the colonization and nodulation of red clover. Biology and Fertility of Soils. 2001; 33(2): 146–151. https://doi.org/10.1007/s003740000304Marek-Kozaczuk M., Skorupska A. Production of B-group vitamins by plant growth-promoting Pseudomonas fluorescens strain 267 and the importance of vitamins in the colonization and nodulation of red clover. Biology and Fertility of Soils. 2001; 33(2): 146–151. https://doi.org/10.1007/s003740000304Matamoros M.A. et al. Biosynthesis of Ascorbic Acid in Legume Root Nodules. Plant Physiology. 2006; 141(3): 1068–1077. https://doi.org/10.1104/pp.106.081463Matamoros M.A. et al. Biosynthesis of Ascorbic Acid in Legume Root Nodules. Plant Physiology. 2006; 141(3): 1068–1077. https://doi.org/10.1104/pp.106.081463Kang B.G., Kim W.T., Yun H.S., Chang S.C. Use of plant growth-promoting rhizobacteria to control stress responses of plant roots. Plant Biotechnology Reports. 2010; 4(3): 179–183. https://doi.org/10.1007/s11816-010-0136-1Kang B.G., Kim W.T., Yun H.S., Chang S.C. Use of plant growth-promoting rhizobacteria to control stress responses of plant roots. Plant Biotechnology Reports. 2010; 4(3): 179–183. https://doi.org/10.1007/s11816-010-0136-1Tazoe M., Ichikawa K., Hoshino T. Purification and Characterization of Pyridoxine 5′-Phosphate Phosphatase from Sinorhizobium meliloti. Bioscience, Biotechnology, and Biochemistry. 2005; 69(12): 2277–2284. https://doi.org/10.1271/bbb.69.2277Tazoe M., Ichikawa K., Hoshino T. Purification and Characterization of Pyridoxine 5′-Phosphate Phosphatase from Sinorhizobium meliloti. Bioscience, Biotechnology, and Biochemistry. 2005; 69(12): 2277–2284. https://doi.org/10.1271/bbb.69.2277Revillas J.J., Rodelas B., Pozo C., Martínez-Toledo M.V., González-López J. Production of B-group vitamins by two Azotobacter strains with phenolic compounds as sole carbon source under diazotrophic and adiazotrophic conditions. Journal of Applied Microbiology. 2000; 89(3): 486–493. https://doi.org/10.1046/j.1365-2672.2000.01139.xRevillas J.J., Rodelas B., Pozo C., Martínez-Toledo M.V., González-López J. Production of B-group vitamins by two Azotobacter strains with phenolic compounds as sole carbon source under diazotrophic and adiazotrophic conditions. Journal of Applied Microbiology. 2000; 89(3): 486–493. https://doi.org/10.1046/j.1365-2672.2000.01139.xMartínez-Viveros O., Jorquera M.A., Crowley D.E., Gajardo G., Mora M.L. Mechanisms and practical considerations involved in plant growth promotion by rhizobacteria. Journal of soil science and plant nutrition. 2010; 10(3): 293–319. https://doi.org/10.4067/S0718-95162010000100006Martínez-Viveros O., Jorquera M.A., Crowley D.E., Gajardo G., Mora M.L. Mechanisms and practical considerations involved in plant growth promotion by rhizobacteria. Journal of soil science and plant nutrition. 2010; 10(3): 293–319. https://doi.org/10.4067/S0718-95162010000100006Nakei M.D., Venkataramana P.B., Ndakidemi P.A. Soybean-Nodulating Rhizobia: Ecology, Characterization, Diversity, and Growth Promoting Functions. Frontiers in Sustainable Food Systems. 2022; 6: 824444. https://doi.org/10.3389/fsufs.2022.824444Nakei M.D., Venkataramana P.B., Ndakidemi P.A. Soybean-Nodulating Rhizobia: Ecology, Characterization, Diversity, and Growth Promoting Functions. Frontiers in Sustainable Food Systems. 2022; 6: 824444. https://doi.org/10.3389/fsufs.2022.824444Jaiswal S.K., Mohammed M., Ibny F.Y.I., Dakora F.D. Rhizobia as a Source of Plant Growth-Promoting Molecules: Potential Applications and Possible Operational Mechanisms. Frontiers in Sustainable Food Systems. 2021; 4: 619676. https://doi.org/10.3389/fsufs.2020.619676Jaiswal S.K., Mohammed M., Ibny F.Y.I., Dakora F.D. Rhizobia as a Source of Plant Growth-Promoting Molecules: Potential Applications and Possible Operational Mechanisms. Frontiers in Sustainable Food Systems. 2021; 4: 619676. https://doi.org/10.3389/fsufs.2020.619676Moore S.J., Warren M.J. The anaerobic biosynthesis of vitamin B12. Biochemical Society Transactions. 2012; 40(3): 581–586. https://doi.org/10.1042/BST20120066Moore S.J., Warren M.J. The anaerobic biosynthesis of vitamin B12. Biochemical Society Transactions. 2012; 40(3): 581–586. https://doi.org/10.1042/BST20120066Mastella L. et al. First report on Vitamin B9 production including quantitative analysis of its vitamers in the yeast Scheffersomyces stipitis. Biotechnology for Biofuels and Bioproducts. 2022; 15: 98. https://doi.org/10.1186/s13068-022-02194-yMastella L. et al. First report on Vitamin B9 production including quantitative analysis of its vitamers in the yeast Scheffersomyces stipitis. Biotechnology for Biofuels and Bioproducts. 2022; 15: 98. https://doi.org/10.1186/s13068-022-02194-yYadav A.N. et al. Biodiversity, and biotechnological contribution of beneficial soil microbiomes for nutrient cycling, plant growth improvement and nutrient uptake. Biocatalysis and Agricultural Biotechnology. 2021; 33: 102009. https://doi.org/10.1016/j.bcab.2021.102009Yadav A.N. et al. Biodiversity, and biotechnological contribution of beneficial soil microbiomes for nutrient cycling, plant growth improvement and nutrient uptake. Biocatalysis and Agricultural Biotechnology. 2021; 33: 102009. https://doi.org/10.1016/j.bcab.2021.102009Babalola O.O. Beneficial bacteria of agricultural importance. Biotechnology Letters. 2010; 32(11): 1559–1570. https://doi.org/10.1007/s10529-010-0347-0Babalola O.O. Beneficial bacteria of agricultural importance. Biotechnology Letters. 2010; 32(11): 1559–1570. https://doi.org/10.1007/s10529-010-0347-0Cesco S. et al. Plant-borne flavonoids released into the rhizosphere: impact on soil bio-activities related to plant nutrition. A review. Biology and Fertility of Soils. 2012; 48(2): 123–149. https://doi.org/10.1007/s00374-011-0653-2Cesco S. et al. Plant-borne flavonoids released into the rhizosphere: impact on soil bio-activities related to plant nutrition. A review. Biology and Fertility of Soils. 2012; 48(2): 123–149. https://doi.org/10.1007/s00374-011-0653-2Mukherjee T., Hanes J., Tews I., Ealick S.E., Begley T.P. Pyridoxal phosphate: Biosynthesis and catabolism. Biochimica et Biophysica Acta (BBA) — Proteins and Proteomics. 2011; 1814(11): 1585–1596. https://doi.org/10.1016/j.bbapap.2011.06.018Mukherjee T., Hanes J., Tews I., Ealick S.E., Begley T.P. Pyridoxal phosphate: Biosynthesis and catabolism. Biochimica et Biophysica Acta (BBA) — Proteins and Proteomics. 2011; 1814(11): 1585–1596. https://doi.org/10.1016/j.bbapap.2011.06.018

The authors declare that there are no conflicts of interest present.