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Research Article

volume 41 special issue (april 2021) : 195-198,   Doi: 10.18805/ag.D-260

In vitro Inorganic Phosphate Solubilization Tests of Cowpea Root Nodule Bacteria from Ethiopia

G
Girmaye Kenasa
B
B.C. Nandeshwar
F
Fassil Assefa
1Department of Biology, Wollega University, Nekemte, P.O.X – 395, Ethiopia.
Cite article:- Kenasa Girmaye, Nandeshwar B.C., Assefa Fassil (2020). In vitro Inorganic Phosphate Solubilization Tests of Cowpea Root Nodule Bacteria from Ethiopia. Agricultural Science Digest. 41(2021): 195-198. doi: 10.18805/ag.D-260.

ABSTRACT

Background: Phosphorus is one of the limiting plant nutrients in most soil due to its fixation with metal ions both in acidic and alkaline soil. However, some soil bacteria have potential to solubilize the cation-fixed phosphorus and make it available to plants.
Methods: Sixty-six cowpea root nodule bacterial species were tested for inorganic phosphate solubilization (PS). PS potential of the isolates from Ca3 (PO4)2, AlPO4 and FePO4 were tested on four different kinds of agar media. Solublization efficiency was determined as Solubilization Index (SI) on agar media and quantity of bioavailable phosphorous on broth media. 
Result: About 30% of the bacterial isolates, out of which 60% endophytes were Ca3 (PO4)2 solubilizers on Pikovskaya medium but did not grow on media containing AlPO4 and FePO4 as sole P sources. Isolates showed significant variation (p<0.05) in Ca3 (PO4)2 solubilization efficiency on the different culture media and ECE-21 (Pseudomonas putida) was versatile in solubilizing Ca3(PO4)2 from the four test media. However, ECE-26A (Bacillus subtilis) showed the highest PS in broth media (140.8 µg ml-1) indicating the moderate correlation (r=0.5578; p<0.05) of PS between solid and liquid media. Amounts of P solubilized and pH change of the culture filtrate was inversely correlated (r=-0.731; p<0.01). In general, ECE-21 was the most efficient P solubilizer (SI=2.67±0.1; P=87.74±7.4µg ml-1) solublizers and hence can be an ideal candidate as inoculants for the crop production in Ethiopia after field evaluation.

REFERENCES

  1. Abbaszadeh, L.P., Savaghebi, R., Asadi-rahmani, H., Rejali, F., Farahbakhsh, M., Moteshareh-zadeh, B., Omidvari, M. and Lindstrom, K. (2012). Symbiotic effectiveness and plant growth promoting traits in some Rhizobium strains isolated from Phaseolus vulgaris. Plant Growth Regul. 68: 361-370.
  2. Girmaye, K., Fassil, A. and Mussie, H.Y. (2019). Phenotypic and genotypic characteristics of cowpea rhizobia from soils of Ethiopia. African Journal of Biotechnology. 17(42): 1299-1312. 
  3. Gupta, M., Chyi, Y.S., Romero-Severson, J. and Owen, J.L. (1994). Amplification of DNA markers from evolutionarily diverse genomes using single primers of simple-sequence repeats. Theor. Appl. Genet. 89: 998-1006.
  4. Islam, M.T., Deora, A., Hashidoko, Y., Rahman, A., Ito, T. and Tahara, S. (2007). Isolation and identification of potential phosphate solubilizing bacteria from the rhizoplane of Oryza sativa L. cv. BR29 of Bangladesh. Z. Naturforsch. 62: 103-110.
  5. Jadhav, R.N. (2013). Isolation of rhizobia from soybean cultivated in Latur area and study of its phosphate solubilization activity. Biosci. Discovery. 4: 100-103.
  6. Tyagi, M.K., Singh, C.P. and Sharma N.L. (2002). Interactive effect of Rhizobium and Phosphate Solubilizing Micro-organisms on the Solubilization of Insoluble Phosphate. Agric. Sci. Digest. 22: 71-74.
  7. Marra L.M., Soares, C.R., Oliveira, S.M., Ferreira, P.M., Soares, B.L., Carvalho, R.F., Lima, J.M. and Moreira, F.M. (2012). Biological nitrogen fixation and phosphate solubilization by bacteria isolated from tropical soils. Plant and Soil. 35(7): 289-307.
  8. Marra, L.M., de Oliveira, S.M., Soares, C.R. and Moreira, F.M. (2011). Solubilisation of inorganic phosphates by inoculant strains from tropical legumes. Scientia Agricola. 68: 603-609.
  9. Mihalache, G., Zamfirache, M., Mihasan, M., Ivanov, I., Stefan, M. and Raus, L. (2015). Phosphate-solubilizing bacteria associated with runner bean rhizosphere. Arch. Biol. Sci. Belgrade. 67: 793-800.
  10. Nautiyal, C.S. (1999). An efficient microbiological growth medium for screening phosphate solubilizing microorganisms. FEMS Microbiol. Lett. 170: 265-270.
  11. Park, K.H., Lee, C.Y. and Son, H.J. (2009). Mechanism of insoluble phosphate solubilization by Pseudomonas fluorescens RAF15 isolated from ginseng rhizosphere and its plant growth-promoting activities. Lett. Appl. Microbiol. 49: 222-228.
  12. Thakur, D., Kaushal, R. and Shyam, V. (2014). Phosphate solubilising microorganisms: Role in phosphorus nutrition of crop plants. Agri. Review. 35(3): 159-171.
  13. Pikovskaya, R.I. (1948). Mobilization of phosphorus in soil in connection with the vital activity of some microbial species. Mikrobiologiya. 17: 362-370.
  14. Rogers, N.J., Carson, K.C., Glenn, A.R., Dilworth, M.J., Hughes, M.N. and Poole, R.K. (2001). Alleviation of aluminum toxicity to Rhizobium leguminosarum bv. viciae by the hydroxamate siderophore vicibactin. BioMetals. 14: 59-66.
  15. Saida, A., Francisco J.O., Manuel, M., Amin, L., Mohammed, B. and Abdelhay, A. (2015). Isolation and screening of bacteria from rhizospheric soils of rice fields in Northwestern Morocco for different plant growth promotion (PGP) activities: An in vitro study. Int. J. Curr. Microbiol. App. Sci. 4(1): 260-269.
  16. Sengupta, A., Gunri, S.K. Biswas, T. and Saha, J. (2018). Efficacies of freshly isolated phosphate solubilising bacteria (PSB) on growth promotion in groundnut (Arachis hypogaea L.) upon commonly used PSB biofertilizers in eastern India. Legume Research-An International Journal. DOI: 10.18805/LR-4020
  17. Stajkovic, O., Meyer, S.D., Milièiæ, B., Willems, A. and Deliæ, D. (2009). Isolation and characterization of endophytic non-rhizobial bacteria from root nodules of alfalfa (Medicago sativa). Botanica Serbica. 33: 107-114.
  18. Zhu, F., Qu, L., Hong, X. and Sun, X. (2011). Isolation and characterization of a phosphate solubilizing halophilic bacterium Kushneria sp. YCWA18 from Daqiao Saltern on the coast of Yellow Sea of China. Evid. Based Complement. Alternat. Med. 61: 32-50.
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    Research Article

    volume 41 special issue (april 2021) : 195-198,   Doi: 10.18805/ag.D-260

    In vitro Inorganic Phosphate Solubilization Tests of Cowpea Root Nodule Bacteria from Ethiopia

    G
    Girmaye Kenasa
    B
    B.C. Nandeshwar
    F
    Fassil Assefa
    1Department of Biology, Wollega University, Nekemte, P.O.X &ndash; 395, Ethiopia.
    Cite article:- Kenasa Girmaye, Nandeshwar B.C., Assefa Fassil (2020). In vitro Inorganic Phosphate Solubilization Tests of Cowpea Root Nodule Bacteria from Ethiopia. Agricultural Science Digest. 41(2021): 195-198. doi: 10.18805/ag.D-260.

    ABSTRACT

    Background: Phosphorus is one of the limiting plant nutrients in most soil due to its fixation with metal ions both in acidic and alkaline soil. However, some soil bacteria have potential to solubilize the cation-fixed phosphorus and make it available to plants.
    Methods: Sixty-six cowpea root nodule bacterial species were tested for inorganic phosphate solubilization (PS). PS potential of the isolates from Ca3 (PO4)2, AlPO4 and FePO4 were tested on four different kinds of agar media. Solublization efficiency was determined as Solubilization Index (SI) on agar media and quantity of bioavailable phosphorous on broth media. 
    Result: About 30% of the bacterial isolates, out of which 60% endophytes were Ca3 (PO4)2 solubilizers on Pikovskaya medium but did not grow on media containing AlPO4 and FePO4 as sole P sources. Isolates showed significant variation (p<0.05) in Ca3 (PO4)2 solubilization efficiency on the different culture media and ECE-21 (Pseudomonas putida) was versatile in solubilizing Ca3(PO4)2 from the four test media. However, ECE-26A (Bacillus subtilis) showed the highest PS in broth media (140.8 µg ml-1) indicating the moderate correlation (r=0.5578; p<0.05) of PS between solid and liquid media. Amounts of P solubilized and pH change of the culture filtrate was inversely correlated (r=-0.731; p<0.01). In general, ECE-21 was the most efficient P solubilizer (SI=2.67±0.1; P=87.74±7.4µg ml-1) solublizers and hence can be an ideal candidate as inoculants for the crop production in Ethiopia after field evaluation.

    REFERENCES

    1. Abbaszadeh, L.P., Savaghebi, R., Asadi-rahmani, H., Rejali, F., Farahbakhsh, M., Moteshareh-zadeh, B., Omidvari, M. and Lindstrom, K. (2012). Symbiotic effectiveness and plant growth promoting traits in some Rhizobium strains isolated from Phaseolus vulgaris. Plant Growth Regul. 68: 361-370.
    2. Girmaye, K., Fassil, A. and Mussie, H.Y. (2019). Phenotypic and genotypic characteristics of cowpea rhizobia from soils of Ethiopia. African Journal of Biotechnology. 17(42): 1299-1312. 
    3. Gupta, M., Chyi, Y.S., Romero-Severson, J. and Owen, J.L. (1994). Amplification of DNA markers from evolutionarily diverse genomes using single primers of simple-sequence repeats. Theor. Appl. Genet. 89: 998-1006.
    4. Islam, M.T., Deora, A., Hashidoko, Y., Rahman, A., Ito, T. and Tahara, S. (2007). Isolation and identification of potential phosphate solubilizing bacteria from the rhizoplane of Oryza sativa L. cv. BR29 of Bangladesh. Z. Naturforsch. 62: 103-110.
    5. Jadhav, R.N. (2013). Isolation of rhizobia from soybean cultivated in Latur area and study of its phosphate solubilization activity. Biosci. Discovery. 4: 100-103.
    6. Tyagi, M.K., Singh, C.P. and Sharma N.L. (2002). Interactive effect of Rhizobium and Phosphate Solubilizing Micro-organisms on the Solubilization of Insoluble Phosphate. Agric. Sci. Digest. 22: 71-74.
    7. Marra L.M., Soares, C.R., Oliveira, S.M., Ferreira, P.M., Soares, B.L., Carvalho, R.F., Lima, J.M. and Moreira, F.M. (2012). Biological nitrogen fixation and phosphate solubilization by bacteria isolated from tropical soils. Plant and Soil. 35(7): 289-307.
    8. Marra, L.M., de Oliveira, S.M., Soares, C.R. and Moreira, F.M. (2011). Solubilisation of inorganic phosphates by inoculant strains from tropical legumes. Scientia Agricola. 68: 603-609.
    9. Mihalache, G., Zamfirache, M., Mihasan, M., Ivanov, I., Stefan, M. and Raus, L. (2015). Phosphate-solubilizing bacteria associated with runner bean rhizosphere. Arch. Biol. Sci. Belgrade. 67: 793-800.
    10. Nautiyal, C.S. (1999). An efficient microbiological growth medium for screening phosphate solubilizing microorganisms. FEMS Microbiol. Lett. 170: 265-270.
    11. Park, K.H., Lee, C.Y. and Son, H.J. (2009). Mechanism of insoluble phosphate solubilization by Pseudomonas fluorescens RAF15 isolated from ginseng rhizosphere and its plant growth-promoting activities. Lett. Appl. Microbiol. 49: 222-228.
    12. Thakur, D., Kaushal, R. and Shyam, V. (2014). Phosphate solubilising microorganisms: Role in phosphorus nutrition of crop plants. Agri. Review. 35(3): 159-171.
    13. Pikovskaya, R.I. (1948). Mobilization of phosphorus in soil in connection with the vital activity of some microbial species. Mikrobiologiya. 17: 362-370.
    14. Rogers, N.J., Carson, K.C., Glenn, A.R., Dilworth, M.J., Hughes, M.N. and Poole, R.K. (2001). Alleviation of aluminum toxicity to Rhizobium leguminosarum bv. viciae by the hydroxamate siderophore vicibactin. BioMetals. 14: 59-66.
    15. Saida, A., Francisco J.O., Manuel, M., Amin, L., Mohammed, B. and Abdelhay, A. (2015). Isolation and screening of bacteria from rhizospheric soils of rice fields in Northwestern Morocco for different plant growth promotion (PGP) activities: An in vitro study. Int. J. Curr. Microbiol. App. Sci. 4(1): 260-269.
    16. Sengupta, A., Gunri, S.K. Biswas, T. and Saha, J. (2018). Efficacies of freshly isolated phosphate solubilising bacteria (PSB) on growth promotion in groundnut (Arachis hypogaea L.) upon commonly used PSB biofertilizers in eastern India. Legume Research-An International Journal. DOI: 10.18805/LR-4020
    17. Stajkovic, O., Meyer, S.D., Milièiæ, B., Willems, A. and Deliæ, D. (2009). Isolation and characterization of endophytic non-rhizobial bacteria from root nodules of alfalfa (Medicago sativa). Botanica Serbica. 33: 107-114.
    18. Zhu, F., Qu, L., Hong, X. and Sun, X. (2011). Isolation and characterization of a phosphate solubilizing halophilic bacterium Kushneria sp. YCWA18 from Daqiao Saltern on the coast of Yellow Sea of China. Evid. Based Complement. Alternat. Med. 61: 32-50.
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