Selection and identification of Pseudomonas and Bacillus Rhizobacteria with Bioinoculant potential for Sorghum
Befekadu Teshome, Eleni Belay, Barkelegne Mengesha, Zerihun Tsegaye, Edwin K. Akley & Awuku Frederick
Abstract
This study aimed to select and identify elite strains of plant growth-promoting rhizobacteria (PGPR) among phosphate-solubilizing bacteria (PSB) conserved at the Ethiopian Biodiversity Institute. In-vitro screening for plant growth-promoting traits and abiotic stress tolerance identified 12 promising PGPR isolates. These strains exhibited phosphate solubilization (index 1.8–3.2), auxin production, nitrogen fixation, and ammonia production. All isolates showed drought tolerance, and some exhibited salt tolerance (up to 10% NaCl). 16S rDNA sequencing identified Pseudomonas frederiksbergensis, Pseudomonas fluorescens, Pseudomonas rhodesiae, Pseudomonas azotoformans, and Bacillus altitudinis. These strains have strong potential as bioinoculants for sustainable sorghum cultivation. Future greenhouse and field trials are recommended to validate their effectiveness.
Keywords
identification, Sorghum, plant growth, rhizobacteria
Author information & Contribution
Befekadu Teshome. Correspondent author. Ethiopian Biodiversity Institute, Microbial Biodiversity Research, Addis Ababa, Ethiopia. E-mail: Beftesh@gmail.com
Eleni Belay. Ethiopian Biodiversity Institute, Microbial Biodiversity Research, Addis Ababa, Ethiopia.
Barkelegne Mengesha. Ethiopian Biodiversity Institute, Microbial Biodiversity Research, Addis Ababa, Ethiopia.
Zerihun Tsegaye. Ethiopian Biodiversity Institute, Microbial Biodiversity Research, Addis Ababa, Ethiopia.
Edwin K. Akley. Savana Agricultural Research Institute, Soil Microbiology Department, Tamale, Ghana.
Awuku Frederick. Savana Agricultural Research Institute, Soil Microbiology Department, Tamale, Ghana.
"Author 1 conceptualized and finalized the research framework. Author 1 drafted the manuscript, and Author 1, 4 and 5 revised it. Author 1, 2, 3, 4 gathered the data, and Author 1 and 6 analyzed and interpreted the data. All authors contributed to the writing of the final version of the manuscript."
Disclosure statement
No potential conflict of interest was reported by the author(s).
Funding
This work is funded by the Bio-Bridge Initiative of the Secretariat of the Convention on the Biological Diversity.
Institutional Review Board Statement
Not Applicable
AI Declaration
AI tools were not used in writing this paper.
Notes
Acknowledgement
References
Adesemoye, A.O., & Kloepper, J.W. (2009). Plant-microbe’s interactions in enhanced fertilizer use efficiency. Appl Microbiol Biotechnol, 85(1), 1–12. https://doi.org/10.1007/s00253-009-2196-0
Ahmad, H. M., Fiaz, S., Hafeez, S., Zahra, S., Shah, A. N., Gul, B., Aziz, O., Mahmood-Ur- Rahman, Fakhar, A., Rafique, M., Chen, Y., Yang, S. H., & Wang, X. (2022). Plant growth-promoting rhizobacteria eliminate the effect of drought stress in plants: A review. Frontiers in Plant Science, 13, 875774. https://doi.org/10.3389/fpls.2022.875774
Akhtar A, Hisamuddin, Robab, M.I., Sharf, A. & Sharf, R. (2012). Plant growth promoting rhizobacteria: An overview. J. Nat. Prod. Plant Resour, 2(1), 19-31. https://doi.org/10.3390/microorganisms11041088
Ansari, F., Jabeen, M. & Ahmad, I. (2021). Pseudomonas azotoformans FAP5, a novel biofilm-forming PGPR strain, alleviates drought stress in wheat plant. International Journal of Environmental Science and Technology, 18. https://doi.org/10.1007/s13762-020-03045-9
Babalola, O.O. (2010). Beneficial bacteria of agricultural importance. Biotechnol Lett, 32, 1559–1570. https://doi.org/10.1007/s10529-010-0347-0
Barriuso J., Solano, B.R., José, A.L., Lobo, A.P., García-Villaraco, A., & Gutiérrez Mañero, F.J. (2008). Ecology, genetic diversity and screening strategies of plant growth promoting rhizobacteria (PGPR). In: I. Ahmad, J. Pichtel, & S. Hayat, Plant-Bacteria Interactions. Strategies and Techniques to Promote Plant Growth. Wiley Online Library. https://doi.org/10.1002/9783527621989.ch1
Cappuccino, J.C., & Sherman, N. (1992). Microbiology: A Laboratory Manual, 3rd edn. Benjamin/cummings Pub.
Costa, R., Götz, M., Mrotzek, N., Lottmann, J., Berg, G., & Smalla, K. (2005). Effects of site and plant species on rhizosphere community structure as revealed by molecular analysis of microbial guilds. FEMS Microbiology and Ecology, 56, 236–249. https://doi.org/10.1111/j.1574-6941.2005.00026.x
Chiu-Chung, Y., & Bhagwath, A. (2003). Exploring the microbial potentiality to augment soil fertility in Taiwan. 6th ESAFS International Conference Soil Management Technology on Low productivity and degraded soils. https://doi.org/10.13140/RG.2.1.1055.9205
Chatterjee, P., Samaddar, S., Anandham, R., Kang, Y., Kim, K., Selvakumar, G., & Sa, T. (2017). Beneficial soil bacterium Pseudomonas frederiksbergensis Os261 augments salt tolerance and promotes red pepper plant growth. Frontiers in Plant Science, 8. https://pubmed.ncbi.nlm.nih.gov/28523010/
Dardanelli, M. S., Carletti, S. M., Paulucci, N. S., Medeot, D. B., Rodriguez Ca´ceres, E. A., Vita, & Garcia, M. B., (2010). Benefits of plant growth-promoting rhizobacteria and rhizobia in agriculture. In D. K. Maheshwari, (Ed.), Plant Growth and Health Promoting Bacteria, Microbiology Monographs, 18, 1-20. Berlin, Germany: Springer Berlin Heidelberg.
Datta M., Palit R., Sengupta, C., Kumar, M., & Banerjee, S. (2011). Plant growth promoting rhizobacteria enhance growth and yield of Chili (Capsicum annuum L.) under field conditions. Australian Journal of Crop Science, 5,531–536.
Egamberdiyeva, D, & Islam, K.R. (2008). Salt tolerant rhizobacteria: plant growth promoting traits and physiological characterization within ecologically stressed environment. In: Ahmad, I, Pichtel, J, Hayat, S. (eds) Plant-bacteria interactions: strategies and techniques to promote plant growth. Wiley-VCH, Weinheim, 257–281
Elfira, Y., Kusmiyati, F. & Budiharjo, A. (2020). The Effect of Bacillus altitudinis P-10
combination treatments on the plant growth and seed quality of Corn (Zea mays L). Bioma, 22(2), 80-187. https://doi.org/10.14710/bioma.22.2.180-187
Felsenstein, J. (1985). Confidence limits on phylogenies: An approach using the bootstrap. Evolution, 39(4), 783-791. https://doi.10.1111/j.1558-5646.1985.tb00420.x
Gise`, G., Laguerre, G., Mavingui, P., Allard, M.R., Charnay, M.P., Louvrier, P., Mazurier, S.I., Rigottier-Gois, L., Noe¨, N., & Amarger, N. (1996). Typing of Rhizobia by PCR DNA fingerprinting and PCR-Restriction Fragment Length Polymorphism Analysis of Chromosomal and Symbiotic Gene Regions: Application to Rhizobium leguminosarum and its different biovars. Applied and Environmental Microbiology, 62(6). https://doi.org/10.1128/aem.62.6.2029-2036.1996
Gupta, S., Meena, M.K. & Datta, S. (2014). Isolation, characterization of plant growth promoting bacteria from the plant Chlorophytum borivilianum and in-vitro screening for activity of nitrogen fixation, phosphate solubilization and IAA production. Int.J.Curr.Microbiol.App.Sci, (7), 1082-1090.
Gusain, P., & Bhandari, B.S. (2019). Rhizosphere associated PGPR functioning. J Pharmacognosy and Phytochem, 8(5), 1181-1191.
Hart, M.M. & Trevors, J.T. (2005). Microbe management: Application of mycorrhyzal fungi in sustainable agriculture. Frontiers in Ecology & the Environment, 3(10), 533-539.
Idris, A., Labuschagne, N. & Korsten, L. (2009). Efficacy of rhizobacteria for growth promotion in sorghum under greenhouse conditions and selected modes of action studies. Journal of Agricultural Science, 147, 17–30. https://doi.10.1017/S0021859608008174
Jime´nez-Salgado T, Fuentes-Ramrez LE, Tapia-Herna´ndez A, Mascarua-Esparza MA, Martnez-Romero E, & Caballero-Mellado J. (1997). Coffea arabica L., a new host plant for Acetobacter diazotrophicus and isolation of nitrogen-fixing acetobacteria. Applied Environmental Microbiology, 63(9), 3676–3683. https://doi.10.1128/aem.63.9.3676-3683.1997
Kimura, M. (1980). A simple method for estimating evolutionary rate of base substitutions through comparative studies of nucleotide sequences. Journal of Molecular Evolution, 16(2),111-120. https://doi.10.1007/BF01731581
Kloepper, J.W., Reddy, S.M., Rodreguez-Kabana, R., Kenney, D.S., Kokalis- Burelle, N., & Ochoa, N.M. (2004.) Application for rhizobacteria in transplant production and yield enhancement. Acta Horticulturae, 631(28), 217-229. https://doi.org/10.17660/ActaHortic.2004.631.28
Krieg, N.R., Staley, J.T., Brown, D.R., Hedlund, B.P., Paster, B.J., Ward, N.L., Ludwig, W., & Whitman, W.B. (2010). Bergey’s manual of systematic bacteriology (Second Edition). Springer. https://doi.org/10.1007/978-0-387-68572-4
Kumar, G.P., MirHassanAhmed, S.K., Desai, S., Amalraj, E.L.l, & Rasul, A. (2014). In- vitro screening for abiotic stress tolerance in potent biocontrol and plant growth promoting strains of Pseudomonas and Bacillus spp. Int. J. of Bact, 195946. http://dx.doi.org/10.1155/2014/195946
Kumar, P., Desai, S., Amalraj, L.D., Ahmed, M.H. & Reddy, G. (2012). Plant growth promoting Pseudomonas spp. from diverse agro-ecosystems of India for Sorghum bicolor. Journal of Biofertilizers & Biopesticides, 7. https://doi.org/10.4172/2155-6202.s7-001
Kumar, S., Stecher, G., Li, M., Knyaz, C., & Tamura, K. (2018). MEGA X: Molecular evolutionary genetics analysis across computing platforms. Molecular Biology and Evolution, 35(6),1547-1549. https://doi.org/10.1093/molbev/msy096
Kumar, T., Bibhuti, A., Mishra, B., Annapurna, K., Kumar, D., Upendra, V., & Editors, K. (2017). Advances in soil microbiology: Recent trends and future prospects 2. Springer Nature Singapore Pte Ltd. https://doi.org/10.1007/978-981-10-7380-9
Lone, R., Hassan, N., Bashir, B, Rohela, G.K., & Malla, N.A. (2023). Role of growth elicitors and microbes in stress management and sustainable production of Sorghum. Plant Stress, 9 (2023), 100179. https://doi.org/10.1016/j.stress.2023.100179
Lucy, M., Reed, E., & Glick, B.R. (2004). Applications of free-living plant growth-promoting rhizobacteria. Antonie van Leeuwenhoek, 86(1), 1-25. https://doi.org/10.1023/B:ANTO.0000024903.10757.6e
Lugtenberg, B.J., Dekkers, L, & Bloemberg, G.V. (2001). Molecular determinants of rhizosphere colonization by Pseudomonas. Ann Rev Phytopathol, 39, 461–490. https://doi.org/10.1146/annurev.phyto.39.1.461
Maheshwari, D.K., Dubey, R.C., Aeron, A., Kumar, B., Kumar, S., Tewari, S., & Arora, N.K. (2012). Integrated approach for disease management and growth enhancement of Sesamum indicum L. utilizing Azotobacter chroococcum TRA2 and chemical fertilizer. World J Microbiol Biotechnol, 28 (10), 3015–3024. https://doi.org/10.1007/s11274-012-1112-4
Mampallil, L.J., Faizal, M.H., & Anith, K.N. (2017). Bacterial bioagents for insect pest management. J. Entomol Zool Stud, 5(6), 2237-2244.
Manasa, T.K, Reddy, R., & Triveni, S. (2017). Isolation and characterization of Pseudomonas fluorescens isolates from different rhizosphere soils of Telangana. Journal of Pharmacognosy and Phytochemistry, 6(3), 224-229.
Matson, P.A., Parton, W.J., Power, A.G., & Swift, M.J. (1997). Agricultural intensification and ecosystem properties. Science, 277(5325),504-509. https://doi.org/10.1126/science.277.5325.504
Monk, J., Gerard, E., Young, S., Widdup, K., & O’Callaghan, M. (2009). Isolation and identification of plant growth-promoting bacteria. Proceedings of the New Zealand Grassland Association, 71, 211-216.
Muleta, D., Assefa, F., Borjesson, E., & Granhall, U. (2013) Phosphate-solubilising rhizobacteria associated with Coffea arabica L. in natural coffee forests of southwestern Ethiopia. Journal of the Saudi Society of Agricultural Sciences, 12(1),73-84. https://doi.org/10.1016/j.jssas.2012.07.002
Oo, K.T., Win, T.T., Khai, A.A. & Fu, P.C. (2020) Isolation, screening and molecular characterization of multifunctional plant growth promoting rhizobacteria for a sustainable agriculture. American Journal of Plant Sciences, 11, 773-792. https://doi.org/10.4236/ajps.2020.116055
Patten, C., & Glick, B. (2002). Role of Pseudomonas putida indoleacetic acid in development of the host plant root system. Appl. Environ. Microb, 68(8), 3795–3801. https://doi.org/10.1128/AEM.68.8.3795-3801.2002
Pikovskaya’s, R.I. (1948). Mobilization of phosphorus in soil connection with the vital activity of some microbial species. Microbiologiya, 17, 362–370.
Raj, D.P. & Cherian, N. (2013). Isolation and characterization of rhizobacteria from forest soils for in vitro study on crop growth enhancement. Int J Curr Sci E, 3(4), 19-23.
Rashid, M., Khalil, S., Ayub, N., Alam, S., & Latif, F. (2004). Organic acid production and phosphate solubilization by phosphate solubilizing microorganisms (PSM) under in vitro conditions. Pakistan Journal of Biological Sciences, 7(2), 187-196. https://doi.org/10.3923/pjbs.2004.187.196
Rodríguez, H., Fraga, R., Gonzalez, T. & Bashan, Y. (2006). Genetics of phosphate solubilization and its potential applications for improving plant growth-promoting bacteria. Plant Soil, 287, 15-21. https://doi.org/10.1007/s11104-006-9056-9
Saharan, B.S., & Nehra, V. (2011). Plant Growth Promoting Rhizobacteria: A critical review. Life Sciences and Medicine Research, 1-20.
Sheng, X.F. (2005). Growth promotion and increased potassium uptake of cotton and rape by a potassium releasing strain of Bacillus edaphicus. Soil Biol Biochem, 37(2005), 1918–1922. https://doi.org/10.1016/j.soilbio.2005.02.026
Singh, D., Ghosh, P., Kumar, J., & Kumar, A. (2019). Plant Growth-Promoting Rhizobacteria (PGPRs): Functions and Benefits. In: Singh, D., Gupta, V., Prabha, R. (eds) Microbial Interventions in Agriculture and Environment. Springer, Singapore. https://doi.org/10.1007/978-981-13-8383-0_7
Thanh, D.T.N., & Diep, C.N. (2014). Isolation and identification of rhizospheric bacteria in acrisols of maize (Zea mays L.) in the eastern of South Vietnam. American Journal of Life Sciences, 2(2), 82-89.
Verma, T. & Pal, P. (2020). Isolation and screening of rhizobacteria for various plant growth promoting attributes. Journal of Pharmacognosy and Phytochemistry, 9(1),1514-1517. https://doi.org/10.22271/phyto.2020.v9.i1z.10678
Vessey, J.K. (2003). Plant growth promoting rhizobacteria as biofertilizers. Plant & Soil, 255, 571-586. https://doi.org/10.1023/A:1026037216893
Weisburg, W.G., Barns, S.M., Pelletier, D. A., & Lane, D.J. (1991). 16S Ribosomal DNA Amplification for Phylogenetic Study. Journal of Bacteriology,173(2), 697-703. https://doi.org/10.1128/jb.173.2.697-703.1991
Wen, Y., Jin, Y., Wang, J., & Cai, L. (2015). MiSeq sequencing analysis of bacterial community structures in wastewater treatment plants. Pol. J. Environ. Stud, 24 (4), 1809-1815. https://doi.org/10.15244/pjoes/38456
Widawati, S., & Suliasih. (2018). The effect of plant growth promoting rhizobacteria (pgpr) on germination and seedling growth of Sorghum bicolor L. Moench. IOP Conference Series: Earth and Environmental Science, 166(2018), 012022. https://doi.org/10.1088/1755-1315/166/1/012022
Yamaoka-Yano, D.M., Amorim, E., Valarini, P.J., Melo, I.S., & Kosako, Y. (1988). Characterization of rhizobacteria from Citrus rhizosphere to control Phytophthora nicotinae var. parasitica and P. citrophthora. Nature Biotechnology, 6(3), 282-286.
Yang, J., Kloepper, J. W., & Ryu, C. M. (2009b). Rhizosphere bacteria help plants tolerate abiotic stress. Trends in Plant Science, 14(1), 1–4. https://doi.org/10.1016/j.tplants.2008.10.004
Ye, S., Yan, R., Li, X., Lin, Y., Yang, Z., Ma, Y. & Ding, Z. (2022). Biocontrol potential of Pseudomonas rhodesiae GC-7 against the root-knot nematode Meloidogyne graminicola through both antagonistic effects and induced plant resistance. Front. Microbiol, 13 – 2022. https://doi.org/10.3389/fmicb.2022.1025727
Zahid, M., Abbasi, M.K., Hameed, S. & Rahim, N. (2015) Isolation and identification of indigenous plant growth promoting rhizobacteria from Himalayan region of Kashmir and their effect on improving growth and nutrient contents of maize (Zeamays L.) Frontiers in Microbiology, 6(207). https://doi.org/10.3389/fmicb.2015.00207
Zahir, Z.A., Munir, A., Asghar, H.N., Shaharoona, B., & Arshad, M. (2008). Effectiveness of rhizobacteria containing ACC deaminase for growth promotion of peas (Pisum sativum) under drought conditions. J Microbiol Biotechnol, 18(5), 958–963.
Cite this article:
Teshome, B., Belay, E., Mengesha, B., Tsegaye, Z., Akley, E.K. & Frederick, A. (2025). Selection and identification of Pseudomonas and Bacillus Rhizobacteria with Bioinoculant potential for Sorghum cultivation. International Journal of Science, Technology, Engineering and Mathematics, 5(2), 38-59. https://doi.org/10.53378/ijstem.353191
License:
This work is licensed under a Creative Commons Attribution (CC BY 4.0) International License.
Find this article in:
Most read articles
- Social media usage and the academic performance of Filipino junior high school students
- Exploring the factors influencing commuters’ satisfaction and the use of public utility buses in Quezon City, Philippines
- A narrative exploration of romantic experiences and ideal relationship standards among Filipino Gen Z
- Students’ exposure to social media and their radical involvement on the societal issues in the Philippines
- ChatGPT integration significantly boosts personalized learning outcomes: A Philippine study
- Tiktok made me book it: The impact of Tiktok on tourism destination selection of generation Z and millennials in Manila
- Senior high school students’ awareness and literacy on computer software applications
- Self-perception of ABM students towards their academic, social and emotional college preparedness
- Enhancing financial literacy among Pantawid Pamilyang Pilipino Program beneficiaries
- Emotional intelligence and leadership efficacy of university student leaders