Isolation and molecular characterization of Enterobacter species ENKS2 and its phosphate solubilisation from rhizospheric soil

Santhanalakshmi Krishnamoorthy¹ , Natarajan Ekambaram²

Section:Research Paper, Product Type: Isroset-Journal
Vol.5 , Issue.6 , pp.20-25, Dec-2018

CrossRef-DOI:   https://doi.org/10.26438/ijsrbs/v5i6.2025

Online published on Jan 01, 2018

Copyright © Santhanalakshmi Krishnamoorthy, Natarajan Ekambaram . This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
 

View this paper at   Google Scholar | DPI Digital Library

XML View     PDF Download

How to Cite this Paper

454 Views    269 Downloads    169 Downloads

Abstract :
In this present study, the potential of a phosphate solubilising bacterium (PSB) was isolated from rhizosphere soil. The phosphate solubilisation was grown in Pikovskaya’s agar medium and the isolate was purified in AT salt minimal medium and further it was cultured in NBRIP liquid media for characterization, identification processes. Based on the morphological, biochemical characterization and 16S rRNA gene sequencing the phosphate solubilising bacterium was identified as Enterobacter species ENKS2. The maximum phosphate solubilisation was in 96h at pH 7.3 in NBRIP liquid media. The Enterobacter species ENKS2 optimized phosphate solubilisation was at pH 7 (281µg/cfuX106), temperature at 30°C (268µg/cfuX106) at pH 6.8, salt conditions was 0.5% (169µg/cfuX106) at pH 7.1, carbon source xylose (213µg/cfuX106) at pH 6.3 and nitrogen source Pottassium nitrate (212µg/cfuX106) at pH 6.1. The different time intervals were followed to maximum phosphate solubilisation in 96h (531µg/cfuX106) at pH 5.8. Hence, this study was useful to develop the biofertilizer for improves the soil, plant growth and reduce the chemical fertilizer usages.

Key-Words / Index Term :
Phosphate solubilisation, Rhizospheric soil, Enterobacter sp., 16S rRNA gene sequencing, acid phosphates

References :
[1]. Illmer P, Schinner F. Solubilization of inorganic phosphates by microorganisms isolated from forest soil. Soil Biol Biochem 1992;24: pp389–95.
[2]. Tewari SK, Das B, Mehrotra S. Cultivation of medicinal plants—tool for rural development. J Rural Tech 2004;3: pp147–50.
[3]. Alam, S., Khalil, S., Ayub, N. and Rashid, M. 2002. In vitrosolubilization of inorganic phosphate by phosphate solubilizing microorganism (PSM) from maize rhizosphere. Intl. J. Agric. Biol. 4 : pp 454-458.
[4]. Jeon JS, Lee SS, Kim HY, Ahn TS, Song HG (2003). Plant growth promotion in soil by some inoculated microorganisms. J. Microbiol.,41: pp 271-276.
[5]. Rodríguez H, Fraga R (1999). Phosphate solubilizing bacteria and their role in plant growth promotion. Biotechnol. Advan., 17: pp 319-339.
[6]. Son HJ, Park GT, Cha MS, Heo MS (2006). Solubilization of insoluble inorganic phosphates by a novel salt- and pH-teolerant Pantoea agglomerans R-42 isolated from soybean rhizosphere. Biores. Technol., 97: pp 204-210.
[7]. Omar SA. The role of rock-phosphate-solubilizing fungi and vesiculararbuscular–amycorrhiza (VAM) in growth of wheat plants fertilized with rock phosphate. World J Microb Biot 1998; 14:211–8.
[8]. Narula N, Kumar V, Behl RK, Duebel AA, Gransee A, Merbach W. Effect of P solubilising Azotobacter chroococcum on N, P, K uptake in P responsive wheat genotypes grown under green house conditions. J Plant Nutr Soil Sci 2000;163:393–8.
[9]. Whitelaw MA. Growth promotion of plants inoculated with phosphate solubilizingfungi. Adv Agron 2000; 69:99–151.
[10]. Bunemann, E.K., Bossio, D.A., Smithson, P.C., Frossard, E.and Oberson, A. 2004. Microbial community composition and substrate use in a highly weathered soil as affected by crop rotation and P fertilization.SoilBiol. Biochem. 36 : 889-901.
[11]. Chen, Y.P., Rekha, P.D., Arunshen, A.B., Lai, W.A. and Young, C.C. 2006. Phosphate solubilizing bacteriafrom subtropical soil and their tricalcium phosphate solubilizing abilities. Appl. Soil Ecol. 34 : pp 33-41.
[12]. Gyaneshwar, P., Kumar, G.N., Parekh, L.J. and Poole, P. S. 2002. Role of soil microorganisms in improving P nutrition of plants.Plant Soil. 245 : pp 83-93.
[13]. Krieg, N.R. 1984. Bergey’s manual of systemic bacteriology, Vol. I. Williams and Wilkins. Baltimore, U.S.A.
[14]. Kumar, R., Pandey, S., Pandey, A. 2006. Plant roots and carbon sequesteration. Curr. Sci. 91: 885–890.
[15]. Pradhan, N. and Sukla, L.B. 2005. Solubilization of inorganic phosphate by fungi isolated from agriculture soil. African J. Biotechnol. 5: pp 850-854.
[16]. Pérez E, Sulbarán M, Ball MM, Yarzábal LA (2007). Isolation and characterization of mineral phosphate-solubilizing bacteria naturally colonizing a limonitic crust in the south-eastern Venezuelan region. Soil Biol. Biochem., 39: pp 2905-2914.
[17]. Yi Y, Huang W, Ge Y (2008). Exopolysaccharide: a novel importantfactor in the microbial dissolution of tricalcium phosphate. World J.Microbiol. Biotechnol., 24: pp 1059-1065.
[18]. López-Gutiérrez JC, Toro M, López-Hernández D (2004). Seasonality of organic phosphorus mineralization in the rhizosphere of the native savanna grass, Trachypogon plumosus. Soil Biol. Biochem., 36:1675-1684.
[19]. Olsen SR, Cole CV, Watanabe FS, Dean LA (1954). Estimation of available phosphorus in soils by extraction with sodium bicarbonate, US Government Printing Office, USDA Washington, D.C. Circ.
[20]. American Public Health Association (1995). Standard methods for examination of water and waste water. In Eaton AD, Clesceri LS, Greenberg AE (eds), 19th Ed., Washington, D.C.
[21]. Pikovskaya RI (1948). Mobilization of phosphorus in soil in connection with the vital activity of some microbial species. Microbiology, 17: pp 362-370.
[22]. Johri JK, Surange S, Nautiyal CS (1999). Occurrence of salt, pH, and temperature-tolerant, phosphate solubilizing bacteria in alkaline soils.Curr. Microbiol., 39: pp 89-93.
[23]. Nautiyal CS (1999). An efficient microbiological growth medium for screening phosphate solubilizing microorganisms. FEMS Microbiol. Lett., 170: pp 265-270.
[24]. Brenner JD, Farmer III JJ (2005) Order XIII, Enterobacteriales. In Brenner DJ, Krieg NR, Staley JT (Eds) Bergey’s, Springer, New York.Man. Syst. Bacteriol., 2: pp 587-850.
[25]. Collee JC, Miles RS, Watt B (1996). Tests for the identification of bacteria. In Collee JG, Marmion BP, Fraser AG, Simmons A (Eds), Churchill Livingstone Inc, New York, pp. 131-149.
[26]. López-Gutiérrez JC, Toro M, López-Hernández D (2004). Seasonality of organic phosphorus mineralization in the rhizosphere of the native savanna grass, Trachypogon plumosus. Soil Biol. Biochem., 36: pp 1675-1684.
[27]. Janisiewicz WJ, Bors B (1995). Development of a microbial community of bacterial and yeast antagonist to control wound-invading postharvest pathogens of fruits. Appl. Environ. Microbiol., 61: 3261- 3267.
[28]. Arvind Kumar, Poonam Bhargava and Lal Chand Rai (2010). Isolation and molecular characterization of phosphate solubilizing Enterobacter and Exiguobacterium species from paddy fields of Eastern Uttar Pradesh, India. African Journal of Microbiology Research Vol. 4(9), pp. 820-829.
[29]. Benlloch S, López-López A, Casamayor EO, Øvreås L, Goddard V, Daae FL, Smerdon G, Massana R, Joint I, Thingstad F, Pedrós-Alió C, Rodríguez-Valera F (2002). Prokaryotic genetic diversity throughout the salinity gradient of a coastal solar saltern. Environ. Microbiol., 4: pp 349-360.
[30]. Nautiyal CS (1999). An efficient microbiological growth medium for screening phosphate solubilizing microorganisms. FEMS Microbiol. Lett., 170: pp 265-270.
[31]. J. Murphy, J.P. Riley, Anal. Chim. Act. 27 (1962) pp 31–36.
[32]. M.A. Tabatabi, J.M. Bremner, Soil Biol. Biochem. 1 (1969) pp 301–307.