Isolation and identification of temperature tolerant phosphate solubilizing bacteria as a potential microbial fertilizer

  • Mohammad Reza SarikhaniEmail author
  • Bahman Khoshru
  • Ralf Greiner
Original Paper


Isolation and identification of temperature tolerant phosphate solubilizing bacteria (TTPSB) and their use as microbial fertilizers was the main goal of the study. In this study, TTPSB were isolated from soil samples treated for 16 h at 55 °C. Their phosphate solubilizing activity was either evaluated in solid media by forming a clear zone (halo) or in liquid media by quantification of the soluble phosphate in the growth medium. Five colonies (RPS4, RPS6, RPS7, RPS8 and RPS9) were identified to be able to form a halo and two of the isolates (RPS9 and RPS7) tolerated a temperature of 55 °C. With tricalcium phosphate (TCP) as the sole P-source, the phosphate solubilizing capacity of RPS9 and RPS7 was determined to be 563.8 and 324.1 mg P L−1 in liquid Sperber medium, respectively. Both bacterial isolates were identified as Pantoea agglomerans by molecular and biochemical characterization. To be used as a microbial fertilizer a carrier system for the temperature tolerant bacteria consisting of rock phosphate, sulfur and bagasse was used. It could be established that the bacterial cell counts of the microbial fertilizers were acceptable for application after storage for 4 months at 28 °C. In a greenhouse experiment using pot cultures, inoculation of maize (S.C.704) with the microbial fertilizers in an autoclaved soil resulted in a significant effect on total fresh and dry weight of the plant root and shoot as well as on the P content of the root and shoot. The effects observed with RPS9 as a component of the microbial fertilizer on plant growth and P nutrition was comparable with the addition of 50% of recommended triple superphosphate (TSP) dose. Using temperature tolerant bacteria in microbial fertilizers will overcome limitations in production and storage of the microbial fertilizers and contribute to a environmentally-friendly agriculture. The temperature tolerant P. agglomerans strain RPS9 was shown to be effective as part of a microbial fertilizer in supporting the growth and P uptake in maize.


Phosphate solubilizing bacteria Temperature tolerance Phosphatic microbial fertilizer P Solubility P Nutrition 



This paper is published as a research project supported by the University of Tabriz Research Affairs Office.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.


  1. Bagyaraj DJ (2004) Quality control and constraints in biofertilizer production technology. In: Kannaiyan S, Kumar K, Govindarajan K (eds) Biofertilizer technology. Scientific Publishers, DelhiGoogle Scholar
  2. Bashan Y, Kamnev AA, de-Bashan LE (2013) A proposal for isolating and testing phosphate-solubilizing bacteria that enhance plant growth. Biol Fertil Soils 49:1–2CrossRefGoogle Scholar
  3. Boratyn GM, Camacho C, Cooper PS, Coulouris G, Fong A, Ma N, Madden TL, Matten WT, McGinnis SD, Merezhuk Y, Raytselis Y (2013) BLAST: a more efficient report with usability improvements. Nucleic Acids Res 41:29–33CrossRefGoogle Scholar
  4. Chen YP, Rekha PD, Arun AB, Shen FT, Lai WA, Young CC (2006) Phosphate solubilizing bacteria from subtropical soil and their tricalcium phosphate solubilizing abilities. Appl Soil Ecol 34:33–41CrossRefGoogle Scholar
  5. Gaind S, Gaur AC (1991) Thermotolerant phosphate solubilizing microorganisms and their interaction with mung bean. Plant Soil 133:141–149CrossRefGoogle Scholar
  6. Gao L, Kong F, Feng C, Wang J, Gao J, Shen G, Zhang C (2016) Isolation, Characterization, and growth promotion of phosphate solubilizing bacteria associated with Nicotiana Tabacum (Tobacco). Pol J Environ Stud 25:993–1003CrossRefGoogle Scholar
  7. Garrity GM, Bell JA, Lilburn TG (2004) Taxonomic outline of the prokaryotes. Bergey’s manual of systematic bacteriology, Springer, New YorkGoogle Scholar
  8. Germida JJ, Janzen HH (1993) Factors affecting the oxidation of elemental sulfur in soils. Fertilizer Res 35(1–2):101–114CrossRefGoogle Scholar
  9. Glick BR (2012) Plant growth-promoting bacteria: mechanisms and applications. Hindawi Publishing Corporation Scientifica, London, pp 1–15Google Scholar
  10. Goldstein AH (1986) Bacterial solubilization of mineral phosphates: historical prospective and future prospects. Am J Altern Agric 1:51–57CrossRefGoogle Scholar
  11. Liu M, Liu X, Cheng BS, Ma XL, Lyu XT, Zhao XF, Ju YL, Min Z, Fang YL (2017) Selection and evaluation of phosphate-solubilizing bacteria from grapevine rhizospheres for use as biofertilizers. Span J Agric Res 14:1106–1116CrossRefGoogle Scholar
  12. Malboobi MA, Zamani K, Lohrasebi T, Sarikhani MR, Samaian A, Sabet MS (2014) Phosphate: the Silent Challenge. Progress Biological Sci 4:1–32Google Scholar
  13. Matthews S, Adzahar MS (2016) Application of phosphate solubilizing microorganisms to increase the solubilisation of rock phosphates in soil. J Trop Agric Food Sci 44:9–18Google Scholar
  14. Motsara MR, Roy RN (2008) Guide to laboratory establishment for plant nutrient analysis. FAO, RomeGoogle Scholar
  15. Nandimath AP, Karad DD, Gupta SG, Kharat AS (2017) Consortium inoculum of five thermo-tolerant phosphate solubilizing Actinomycetes for multipurpose biofertilizer preparation. Iran J Microbiol 9:295–304PubMedPubMedCentralGoogle Scholar
  16. Nautiyal CS, Bhadauria S, Kumar P, Lal H, Mondal R, Verma D (2000) Stress induced phosphate solubilization in bacteria isolated from alkaline soils. FEMS Microbiol Lett 182:291–296CrossRefGoogle Scholar
  17. Nobahar A, Sarikhani MR, Chalabianlou N (2017) Buffering capacity affects phosphorous solubilization assays in rhizobacteria. Rhizosphere 4:119–125CrossRefGoogle Scholar
  18. Pande A, Pandey P, Mehra S, Singh M, Kaushik S (2017) Phenotypic and genotypic characterization of phosphate solubilizing bacteria and their efficiency on the growth of maize (Zea mays L.). Int J Agric Innov Res 5:929–938Google Scholar
  19. Paul EA (2007) Soil microbiology, ecology and biochemistry, 3rd edition. Academic Press is an imprint of Elsevier, AmsterdamCrossRefGoogle Scholar
  20. Rfaki A, Nassiri L, Ibijbijen J (2015) Isolation and characterization of phosphate solubilizing bacteria from the rhizosphere of faba bean (Vicia faba L.) in Meknes Region. Morocco. Brit Microbiol Res J 6:247–254CrossRefGoogle Scholar
  21. Saikrithika S, Krishnaswamy VG, Sujatha B (2016) A Study on isolation of phosphate solubilizing bacterial (PSB) strain from vermicomposted soil and their phosphate solubilizing abilities. Int J Adv Biotechnol Res 7:526–535Google Scholar
  22. Salimpur S, Khvazi K, Nadian H, Miransari M (2010) Enhancing phosphorous availability to canola (Brassica napus L.) Using P solubilizing and sulfur oxidizing bacteria. Aust J Crop Sci 4:330–334Google Scholar
  23. Sarikhani MR, Khoshru B, Oustan S (2016) Efficiency of some bacterial strains in potassium release from mica and phosphate solubilization under in-vitro conditions. Geomicrobiol J 33:832–838CrossRefGoogle Scholar
  24. Sarikhani MR, Oustan S, Ebrahimi M, Aliasgharzad N (2018) Isolation and identification of potassium-releasing bacteria in soil and assessment of their ability to release potassium for plants. Eur J Soil Sci 69:1078–1086CrossRefGoogle Scholar
  25. Sarker A, Talukder NM, Islam MT (2014) Phosphate solubilizing bacteria promote growth and enhance nutrient uptake by wheat. Plant Sci Today 1:86–93CrossRefGoogle Scholar
  26. Selvi KB, Paul JJA, Vijaya V, Saraswathi K (2017) Analyzing the efficacy of phosphate solubilizing microorganisms by enrichment culture techniques. Biochem Mol Biol J 3:1–7Google Scholar
  27. Sperber JI (1958) Solution of apatite by soil microorganisms producing organic acids. Aust J Agric Res 9:782–787CrossRefGoogle Scholar
  28. Vaid SK, Kumar B, Sharma A, Shukla AK, Srivastava PC (2014) Effect of Zn solubilizing bacteria on growth promotion and Zn nutrition of rice. J Soil Sci Plant Nutr 14:889–910Google Scholar
  29. Viruel E, Erazzú LE, Martínez Calsina L, Ferrero MA, Lucca ME, Siñeriz F (2014) Inoculation of maize with phosphate solubilizing bacteria: effect on plant growth and yield. J Soil Sci Plant Nutr 14:819–831Google Scholar
  30. Wang HY, Liu S, Zhai LM, Zhang JZ, Ren TZ, Fan BQ, Liu HB (2015) Preparation and utilization of phosphate biofertilizers using agricultural waste. J Integr Agric 14:158–167CrossRefGoogle Scholar
  31. Walinga I, van Vark W, Houba VJG, van der Lee JJ (1989) Soil and plant analysis, a series of syllabi. Part 7. Plant analysis procedures. Wageningen Agriculture University, WageningenGoogle Scholar
  32. Yadav H, Gothwa RK, Solanki PS, Sinha-Roy Nehra SS, S, and Ghosh P, (2015) Isolation and characterization of thermo-tolerant phosphate-solubilizing bacteria from a phosphate mine and their rock phosphate solubilizing abilities. Geomicrobiol J 32:475–481CrossRefGoogle Scholar
  33. Yulianti E, Rakhmawati A (2017) Screening and characterization of phosphate solubilizing bacteria from isolate of thermophilic bacteria. In: AIP Conference Proceedings. AIP Publishing 1868(1):090015Google Scholar
  34. Zhang J, Wang P, Fang L, Zhang QA, Yan C, Chen J (2017) Isolation and characterization of phosphate-solubilizing bacteria from mushroom residues and their effect on tomato plant growth promotion. Pol J Microbiol 66:57–66CrossRefGoogle Scholar

Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  • Mohammad Reza Sarikhani
    • 1
    Email author
  • Bahman Khoshru
    • 1
  • Ralf Greiner
    • 2
  1. 1.Department of Soil Science, Faculty of AgricultureUniversity of TabrizTabrizIran
  2. 2.Department of Food Technology and Bioprocess EngineeringMax Rubner-Institut, Federal Research Institute of Nutrition and FoodKarlsruheGermany

Personalised recommendations