Effects of inoculation with a commercial microbial inoculant Bacillus subtilis C-3102 mixture on rice and barley growth and its possible mechanism in the plant growth stimulatory effect
Koki Toyota 1, C-D,F
More details
Hide details
Graduate School of Bio-Applications and Systems Engineering, Tokyo University of Agriculture and Technology, Japan
Process Development Laboratories, Asahi Group Holdings, Japan
A - Research concept and design; B - Collection and/or assembly of data; C - Data analysis and interpretation; D - Writing the article; E - Critical revision of the article; F - Final approval of article
Submission date: 2018-12-25
Acceptance date: 2019-06-04
Online publication date: 2019-06-27
Corresponding author
Koki Toyota   

Graduate School of Bio-Applications and Systems Engineering, Tokyo University of Agriculture and Technology, Naka, 184-8588, Koganei, Japan
Journal of Plant Protection Research 2019;59(2):193-205
The effects of a microbial inoculant (Thervelics®: a mixture of cells of Bacillus subtilis C-3102 and carrier materials) on rice (Oryza sativa cv. Milkyprincess) and barley (Hordeum vulgare cv. Sachiho Golden) were evaluated in four pot experiments. In the first and second experiments, the dry matter production of rice and barley increased significantly by 10–20% with the inoculation of the mixture at a rate of 107 cfu ⋅ g–1 soil compared with the non-inoculated control. In the third experiment, the growth promoting effects of the mixture, the autoclaved mixture and the carrier materials were compared. The dry mater production of rice grains was the highest in the mixture, and it was significantly higher in the three treatments than in the control, suggesting that the carrier materials may also have a plant growth promoting effect and the living cells might have an additional stimulatory effect. To confirm the efficacy of the living cells in the mixture, only B. subtilis C-3102 cells were used in the fourth experiment. In addition, to estimate the mechanisms in growth promotion by B. subtilis C-3102, three B. subtilis strains with similar or different properties in the production of indole-3-acetic acid (IAA), protease and siderophore and phosphatesolubilizing ability were used as reference strains. Only B. subtilis C-3102 significantly increased the dry matter production of rice grains and the soil protease activity was consistently higher in the soil inoculated with B. subtilis C-3102 throughout the growing period. These results indicate that the microbial inoculant including live B. subtilis C-3102 may have growth promoting effects on rice and barley.
The authors have declared that no conflict of interests exist.
Al-Taweil H.I., Osman M., Aidi A.H., Wan M.W.Y. 2009. Development of microbial inoculants and the impact of soil application on rice seedlings growth. American Journal of Agricultural and Biological Sciences 4 (4): 79–82.
Arkhipova T.N., Prinsen E., Veselov S.U., Martinenko E.V., Melentiev A.I., Kudoyarova G.R. 2007. Cytokine producing bacteria enhance plant growth in drying soil. Plant and Soil 292 (1): 305–315. DOI: 10.1007/s11104-007-9233-5.
Cabangon R.J., Castillo E.G., Tuong T.P. 2011. Chlorophyll meter-based nitrogen management of rice grown under alternate wetting and drying irrigation. Field Crops Research 121 (1): 136–146. DOI: ttps://doi.org/10.1016/j.fcr.2010.12.002.
Chandler S., Hese N.V., Coutte F., Jacques P., Hofte M., Vleesschauwer D.D. 2015. Role of cyclic lipopeptides produced by Bacillus subtilis in mounting induced immunity in rice (Oryza sativa L.). Physiological and Molecular Plant Pathology 91: 20–30. DOI: 10.1016/j.pmpp.2015.05.010.
Chauhan H., Bagyaraj D.J., Selvakumar G., Sundaram S.P. 2015. Novel plant growth promoting rhizobacteria-prospects and potential. Applied Soil Ecology 95: 38–53. DOI: 10.1016/j.apsoil.2015.05.011.
Chen F., Wang M., Zheng Y., Lue J., Yang X., Wang X. 2010. Quantitative changes of plant defense enzymes and phytohormone in biocontrol of cucumber Fusarium wilt by Bacillus subtilis B759. World Journal of Microbiology and Biotechnology 26 (4): 675–684. DOI: https://doi.org/10.1007/s11274....
Cherif-Silini H., Silini A., Yahiaoui B., Ouzari I., Boudabous A. 2016. Phylogenetic and plant-growth promoting characteristics of Bacillus isolates from the wheat rhizosphere. Annals of Microbiology 66 (3): 1087–1097. DOI: https://doi.org/10.1007/s13213....
Compant S., Clement C., Sessitsch A. 2010. Plant growth-promoting bacteria in the rhizo- and endosphere of plants: their role, colonization, mechanisms involved and prospects for utilization. Soil Biology and Biochemistry 42 (5): 669–678. DOI: https://doi.org/10.1016/j.soil....
Dias D.C., Leonardo A.F.G., Tachibana L., Correa C.F., Bordon I.C.A.C., Romagosa E., Ranzani-Paiva M.J.T. 2012. Effect of incorporating probiotics into the diet of matrinxa (Brycon amazonicus) breeders. Journal of Applied Ichthyology 28 (1): 40–45. DOI: https://doi.org/10.1111/j.1439....
El-Bendary M.A., Hamed H.A., Moharam M.E. 2016. Potential of Bacillus isolates as bio-control agent against some fungal phytopathogens. Biocatalysis and Agricultural Biotechnology 5: 173–178. DOI: https://doi.org/10.1016/j.bcab....
Fritts C.A., Kersey J.H., Motl M.A., Kroger E.C., Yan F., Si J., Jiang Q., Campos M.M., Waldroup A.L., Waldroup P.W. 2000. Bacillus subtilis C-3102 (Calsporin) improves live performance and microbiological status of broiler chickens. The Journal of Applied Poultry Research 9 (2): 149–155. DOI: 10.1093/japr/9.2.149}.
Garcia-Marengoni N., Menezes-Albuquerque D. 2015. Quantification of intestinal bacteria, operating cost and performance of fingerlings Nile tilapia subjected to probiotics. Latin American Journal of Aquatic Research 43 (3): 367–373. DOI: 10.3856/vol43-issue3-fulltext-23.
Gardan L., David C., Morel M., Glickmann E., Abughorrah M., Petit A., Dessaux Y. 1992. Evidence for a correlation between auxin production and host plant-species among strains of Pseudomonas syringae subsp. savastanoi. Applied and Environmental Microbiology 58 (5): 1780–1783.
Ge B., Liu B., Nwet T.T., Zhao W., Shi L., Zhang K. 2016 Bacillus methylotrophicus strains NKG-1, isolated from Changbai Mountain, China, has potential applications as a biofertilizer or biocontrol agents. PLoS One 11 (11): 1–13. DOI: 10.1371/journal.pone.0166079.
Gupta M., Kiran S., Gultai A., Singh B., Tewari R. 2012. Isolation and identification of phosphate solubilizing bacteria able to enhance the growth and aloin-A biosynthesis of Aloe barbadensis Miller. Microbiological Research 167 (4): 358–363. DOI: 10.1016/j.micres.2012.02.004.
Habibi S., Djedid S., Prongjunthuek K., Mortuza M.F., Ohkama-Ohtsu N., Sekimoto H., Yokoyoma T. 2014. Physiological and genetic characterization of rice nitrogen fixer PGPR isolated from rhizosphere soils of different crops. Plant and Soil 379 (1–2): 51–66. DOI: 10.1007/s11104-014-2035-7.
Hanif M.K., Hameed S., Imran A., Naqqash T., Shahid M., Van Elsas J.D. 2015. Isolation and characterization of a β-propeller gene containing phosphobacterium Bacillus subtilis strain KPS-11 for growth promotion of potato (Solanum tubersum L.). Frontiers in Microbiology 6: 583. DOI: 10.3389/fmicb.2015.00583.
Hatanaka M., Nakamura Y., Maathuis A.J.H., Venema K., Murota I., Yamamoto N. 2012. Influence of Bacillus subtilis C-3102 on microbiota in a dynamic in vitro model of the gastrointestinal tract simulating human conditions. Beneficial Microbes 3 (3): 229–236. DOI: https://doi.org/10.3920/BM2012....
He S.X., Zhang Y., Xu L., Yang Y.L., Marubashi T., Zhou Z.G., Yao B. 2013. Effects of dietary Bacillus subtilis C-3102 on the production, intestinal cytokine expression and autochthonous bacteria of hybrid tilapia Oreochromis niloticus female × Oreochromis aureus male. Aquaculture 412: 125–130. DOI: 10.1016/j.aquaculture.2013.06.028.
Islam S., Akanda A.B., Prova A., Islam M.T., Hossain M. 2016. Isolation and identification of plant growth promoting rhizobacteria and their effect on plant growth promotion and disease suppression. Frontiers in Microbiology 6: 1360. DOI: 10.3389/fmicb.2015.01360.
Jeong J.S., Kim I.H. 2014. Effect of Bacillus subtilis C-3102 spores as a probiotic feed supplement on growth performance, noxious gas emission, and intestinal microflora in broilers. Poultry Science 93 (12): 3097–3103. DOI: https://doi.org/10.3382/ps.201....
Karnwal A. 2017. Isolation and identification of plant growth promoting rhizobacteria from maize (Zea mays L.) rhizosphere and their plant growth promoting effect on rice (Oryza sativa L.). Journal of Plant Protection Research 57 (2): 144–151. DOI: 10.1515/jppr-2017-0020.
Kembhavi A.A., Kulkarni A., Pant A. 1993. Salt-tolerant and thermostable alkaline protease from Bacillus subtilis NCIM No. 64. Applied Biochemistry and Biotechnology 38 (1–2): 83–92. DOI: https://doi.org/10.1007/BF0291....
Khedher S.B., Kilani-Feki O., Dammak M., Jabnoun-Khiareddine H., Daami-Remadi M., Tounsi S. 2015. Efficacy of Bacillus subtilis V26 as a biological control agent against Rhizoctonia solani on potato. Comptes Rendus Biologies 338 (12): 784–792. DOI: https://doi.org/10.1016/j.crvi....
Khochamit N., Siripornadulsil S., Sukon P., Siripornadulsil W. 2015. Antibacterial activity and genotypic-phenotypic characteristics of bacteriocin-producing Bacillus subtilis KKU213: potential as a probiotic strain. Microbiological Research 170: 36–50. DOI: https://doi.org/10.1016/j.micr....
Kumar K.V.K., Yellareddygari S.K.R., Reddy M.S., Kloepper J.W., Lawrence K.S., Zhou X.G., Sudini H., Groth D.E., Raju S.K., Miller M.E. 2012a. Efficacy of Bacillus subtilis MBI 600 against sheath blight caused by Rhizoctonia solani and on growth and yield of rice. Rice Science 19 (1): 55–63. DOI: https://doi.org/10.1016/S1672-....
Kumar P., Dubey R.C., Maheshwari D.K. 2012b. Bacillus strains isolated from rhizosphere showed plant growth promoting and antagonistic activity against phytopathogens. Microbiological Research 167 (8): 493–499. DOI: https://doi.org/10.1016/j.micr....
Ladd J.N., Butler J.H.A. 1972. Short-term assays of soil proteolytic enzyme activities using proteins and dipeptide derivatives as substrates. Soil Biology and Biochemistry 4 (1): 19–30. DOI: https://doi.org/10.1016/0038-0....
Lee Y.S., Kim K.Y. 2015. Antagonistic potential of Bacillus pumilus L1 against root-knot nematode, Meloidogyne arenaria. Journal of Phytopathology 164: 29–39. doi.org/10.1111/jph.12421.
Lin Y., Du D., Si C., Zhao Q., Li Z., Li P. 2014. Potential biocontrol Bacillus sp. strains isolated by an improved method from vinegar waste compost exhibited antibiosis against fungal pathogens and promote growth of cucumber. Biological Control 71: 7–15. DOI: 10.1016/j.biocontrol.2013.12.010.
Marubashi T., Imabayashi T., Maruta, K. 2009. Bacterial translocation inhibitor and method of inhibiting bacterial translocation. United Sates Patent Application Publication. US 2009/0022690 A1.
Meng Q., Jiang H., Hao J. 2016 Effects of Bacillus velezensis strain BAC03 in promoting plant growth. Biological Control 98: 18–26. DOI: 10.1016/j.biocontrol.2016.03.010.
Meurer R.F.P., Leal P.C., da Rocha C., Bueno I.J.M., Maiorka A., Dahlke F. 2010 Evaluation of the use of probiotics in diets with or without growth promoters for broiler chicks. Revista Brasileira de Zootecnia 39 (2): 2687–2690. DOI: http://dx.doi.org/10.1590/S151....
Park K., Park J.W., Lee S.W., Balaraju K. 2013. Disease suppression and growth promotion in cucumber induced by integrating PGPR agent Bacillus subtilis strain B4 and chemical elicitor ASM. Crop Protection 54: 199–205. DOI: 10.1016/j.cropro.2013.08.017.
Perez-Miranda S., Cabirol N., George-Tellez R., Zamudio-Rivera L.S., Fernandez F.J. 2007. O-CAS, a fast and universal method for siderophore detection. Journal of Microbiological Methods 70 (1): 127–131. DOI: 10.1016/j.mimet.2007.03.023.
Qiao J.Q., Wu H.J., Huo R., Gao X.W., Borriss R. 2014. Stimulation of plant growth and biocontrol by Bacillus amyloliquefaciens subsp. plantarum FZB42 engineered for improved action. Chemical and Biological Technologies in Agriculture 1: 12. DOI: doi.org/10.1186/s40538-014-0012-2.
Rainer B. 2011. Use of plant-associated Bacillus strains as biofertilizers and biocontrol agents in agriculture. p. 41–76. In: “Bacteria in Agrobiology: Plant Growth Responses” (D.K. Maheshwari, ed.). Springer-Verlag Berlin, Heidelberg, Germany.
Robin A., Vansuyt G., Hinsinger P., Meyer J.M., Briat J.F., Lemancean P. 2008 Iron dynamics in the rhizosphere: consequences for plant health and nutrition. Advances in Agronomy 99: 183–225. DOI: 10.1016/s0065-2113(08)00404-5.
Sanghavi P., Patel H., Vaishnav D., Oza T., Dave G., Kunjadia P., Sheth N. 2016. A novel alkaline keratinase from Bacillus subtilis DP1with potential utility in cosmetic formulation. International Journal of Biological Macromolecules 87: 256–262. DOI: 10.1016/j.ijbiomac.2016.02.067.
Shakeel M., Rais A., Hassan M.N., Hafeez F.Y. 2015. Root associated Bacillus sp. improves growth, yield and zinc translocation for Basmati rice (Oryza sativa) varieties. Frontiers in Microbiology 6: 1286. DOI: 10.3389/fmicb.2015.01286.
Sharma K.M., Kumar R., Panwar S., Kumar A. 2017. Microbial alkaline proteases: optimization of production parameters and their properties. Journal of Genetic Engineering and Biotechnology 15 (1): 115–126. DOI: doi.org/10.1016/j.jgeb.2017.02.001.
Sotome T., Kawada N., Kato T., Sekiwa T., Nishigawa H., Natsuaki, Kimura K., Maeoka Y., Nagamine T., Kobayashi S., Wada Y., Yoshida T. 2010. The current and new strains of barley yellow mosaic virus (BaYMV) in Tochigi Prefecture. Japanese Journal of Crop Science 79 (1): 29–36. DOI: https://doi.org/10.1626/jcs.79....
Suarez D.E.C., Gigon A., Freitas R.P., Lavelle P., Velasquez E., Blouin M. 2014. Combined effects of earthworms and IAA-producing rhizobacteria on plant growth and development. Applied Soil Ecology 80: 100–107. DOI: 10.1016/j.apsoil.2014.04.004.
Suslow T.V., Schroth M.N. 1982. Role of deleterious rhizobacteria as minor pathogens in reducing crop growth. Phytopathology 72 (1): 111–115. DOI: 10.1094/Phyto-72-111.
Toyota K. 2015. Bacillus-related spore formers: attractive agents for plant growth promotion. Microbes and Environments 30 (3): 205–207. DOI: 10.1264/jsme2.ME3003rh.
Uttatree S., Charoenpanich J. 2016. Isolation and characterization of a broad pH- and temperature-active, solvent and surfactant stable protease from a new strain of Bacillus subtilis. Biocatalysis and Agricultural Biotechnology 8 (C): 32–38. DOI: 10.1016/j.bcab.2016.08.003.
Uyeno Y., Shigemori S., Shimosato T. 2015. Effect of probiotics/prebiotics on cattle health and productivity. Microbes and Environments 30 (2): 126–132. DOI: 10.1264/jsme2.ME14176.
Wang H.L., Wen K., Zhao X.Y., Wang X.D., Li A.Y., Hong H.Z. 2009. The inhibitory activity of endophytic Bacillus sp. strain CHM1 against plant pathogenic fungi and its plant growth-promoting effect. Crop Protection 28 (8): 634–639. DOI: 10.1016/j.cropro.2009.03.017.
Yu X., Ai C., Xin L., Zhou G. 2011. The siderophore-producing bacterium, Bacillus subtilis CAS15, has a biocontrol effect on Fusarium wilt and promotes the growth of pepper. European Journal of Soil Biology 47 (2): 138–145.
Journals System - logo
Scroll to top