Asian Journal of Bio Science

Open Access Open Access  Restricted Access Subscription Access
Open Access Open Access Open Access  Restricted Access Restricted Access Subscription Access

Deciphering Soil Diazotrophic Diversity in the Wheat-Maize Cropping System of Punjab Using Morphological, Biochemical and Molecular Techniques


Affiliations
1 Department of Microbiology, Punjab Agricultural University, Ludhiana (Punjab), India
2 Department of Soil Sciences, Punjab Agricultural University, Ludhiana (Punjab), India
3 School of Agricultural Biotechnology, Punjab Agricultural University, Ludhiana (Punjab), India
     

   Subscribe/Renew Journal


Soil is the critical resource as well as a basic medium for the growth of natural micro flora present in it. The soil fertility depends on its physicochemical properties and microbial population diversity. Diazotrophs are the nitrogen fixing bacteria which possess nifH gene that is responsible for coding the nitrogenase enzyme involved in the reduction of atmospheric nitrogen to ammonia. These nitrogen fixing bacteria play an imperative role, function and significance in the soil. Soil samples were collected from the different wheat-maize cropping system of Punjab and analyzed for physicochemical properties as pH, electrical conductivity, organic carbon, soil texture, ammoniacal as well as nitrate nitrogen. Eighty diazotrophic bacteria were isolated on eight different nitrogen free media and characterized culturally, morphologically, biochemically, functionally and using molecular techniques. The diazotrophic nature of the isolates was confirmed by the amplification of nifH gene using two nifH primers viz. nifH1 and nifH2. The nifH positive isolates were further used for 16S rDNA restriction analysis using different enzymes such as Taq1, Rsa I and Hae III. Based on UPGMA clustering, the representative strains were sequenced and identified as Bacillus amyloliquifaciens, Pseudomonas aeruginosa, Bacillus subtilis, Paenibacillus sp., Azotobacter vinelendii, Stenotrophomonas maltophilia, Rhizobium larrymorrei, Flavobacterium anhuiense, Sphingomonas paucimobilis, Paenibacillus panacisoli, Azospirillum sp., Pseudomonas putida, Paenibacillus amyloliticus, Bacillus circulans, Paenibacillus polymyxa and Xanthomonas oryzae.

Keywords

Diazotrophs, nif H, 16S rDNA Sequencing, Physico-Chemical Properties, Soil
Subscription Login to verify subscription
User
Notifications
Font Size


  • Altschul, S.F., Gish, W., Miller, W., Myers, E.W. and Lipman, D.J. (1990). Basic local alignment search tool. J. Mol. Bio., 215 (3) : 403-410.

  • Anand, R.J. (1974). Morphology and genesis of some major soils of the Malva alluvial plain (Punjab). M.Sc. Thesis, Punjab AgriculturalUniversity, Ludhina, Punjab (INDIA).

  • Baldani, J.I., Caruso, L., Baldani, V.L.D., Goi, S.R. and Dobereiner, J. (1997). Recent advances in BNF with non-legume plants. Soil Biol. Biochem., 29 (5-6) : 911-922.

  • Burris, R.H. and Roberts, G.P. (1993). Biological nitrogen fixation. Annu. Rev. Nutr., 13 : 317-335.

  • Chowdhury, S.P., Schmid, M., Hartmann, A. and Tripathi, K. (2007). Identification of diazotrophs in the culturable bacterial community associated with ischolar_mains of Lasiurus sindicus, a perential grass of Thar desert, India. Microb. Ecol., 54 : 82-90.

  • Dhaliwal, G.S. (1991). Comparative studies on methods of determination of carbon and nitrogen in plant and soil samples. M.Sc. Thesis, Punjab Agricultural University, Ludhina, Punjab (INDIA).

  • Edi-Premono, M., Moawad, A.M. and Vlek, P.L.G. (1996). Effect of phosphate solubilizing Pseudomonas putida on the growth of maize and its survival in the rhizosphere. Indones. J. Crop Sci., 11 : 13-23.

  • Garland, J.L. (1996). Analytical approaches to the characterization of samples of microbial communities using patterns of potential (source utilization). Soil Biol. Biochem., 28 (2) : 213-221.

  • Gordon A.S. and Weber, R.P. (1951). Colorimetric estimation of indoleacetic acid. Pl. Physiol., 26 (1) : 192-195.

  • Holt, J.G., Kreig, N.R., Sneath, P.H.A., Staley, J.T. and Williams, S.T. (1994). Bergey’s manual of determinative bacteriology. Williams and Wilkins, Baltmore (U.S.A.).

  • Islam, R., Trivedi, P., Madhaiyan, M., Seshadri, S., Lee, G., Yang, J., Kim, Y., Kim, M., Han, G., Chauhan, P.S. and Sa, T. (2010).

  • Isolation, enumeration and characterization of diazotrophic bacteria from paddy soil sample under long-term fertilizer management experiment. Biol. Fert. Soils, 46 (3) : 261-269.

  • Jackson, M.L. (1973). Soil chemical analysis. Prentice Hall of India, Ltd, New Delhi. 60 pp. 7-33.

  • Jha, B., Thakur, M.C., Grntia, I., Albrecht, V., Stoffels, M., Schmid, M. and Hartmann, A. (2008). Isolation, partial identification and application of diazotrophic rhizobacteria from traditional Indian rice cultivars. Eur. J. Soil Biol., 45 (1) : 62-72.

  • Kuklinshy-Sobral, J., Arauju, W.L., Mendes, R., Geraldi, I.O., Pizzirani-kleiner, A.A. and Azevedo, J.L. (2004). Isolation and characterization of soybean associated bacteria and their potential for plant growth promotion. Environ. Microbiol., 6 (12) : 1244-1251.

  • Lata and Saxena, A.K. (2003). Characterization of plant growth promoting rhizobacteria. In : Training manual on biofertilizer technology (eds.) A.K. Saxena IARI, New Delhi, pp. 24-25.

  • Ozawa, T., Ohwaki, A. and Okumura, K. (2003). Isolation and characterization of diazotrophic bacteria from the surface sterilized ischolar_mains of some legumes. Sci. Rep. Grad. Agric. Biol. Sci., Ozaba Pef Uni., 55 : 29-36.

  • Page, A.L., Miller, R.H. and Koeney, D.R. (1982). Methods of soil analysis. Part 2: Chemical and microbiological properties. Agronomy Monograph No. 9. (American Society of Agronomy Inc., Soil Science Society of America Inc.: Madison, WI).

  • Richard, L.A. (1954). Diagnosis and improvement of saline and alkali soils. Agriculture Hand Book No. 60. (USDA: Washington, DC).

  • Rohalf, F.J. (1998). ‘NTSYS-pc numerical taxonomy and multivariate analysis system. Version 2.02’ (Exeter Publications: Setauket, NY).

  • Selvakumar, G., Joshi, P., Nazim, S., Mishra, P.K. and Gupta, H.S.(2009). Phospahte solubilization and growth promotion by phospahte solubilization and growth promotion by Pseudomonas fragi CSIIRH1 (MTCC 8984) a psychrotolerant bacterium isolate from a high altitude HImalayan rhizosphere. Biologia., 64: 239-245.

  • Somani, L.L. (2005). Handbook of biofertilizers. Agrotech Publishing Academy, Udaipur, Rajasthan (INDIA).

  • Tank, N. and Saraf, M. (2003). Phosphate solubilization, exopolysaccharide production and indole acetic acid secretion by rhizobacteria isolated from Trigonella foenumgraecum. Internat. J Microbiol., 43 (1) : 37-40.

  • Vessey, J.K. (2003). Plant growth promoting rhizobacteria as biofertilizers. Pl. Soil, 255 (2) : 571-86.

  • Walkley, W. and Black, C.A. (1934). An examination of the digtiareff method for determination soil organic matter and a proposed modification of the chromic acid titration method. Soil Sci., 37: 29-38.

  • Wolters, V. (2001). Biodiversity of soil animals and its function. Eur. J. Soil Biol., 37 (4) : 221-227.


Abstract Views: 420

PDF Views: 0




  • Deciphering Soil Diazotrophic Diversity in the Wheat-Maize Cropping System of Punjab Using Morphological, Biochemical and Molecular Techniques

Abstract Views: 420  |  PDF Views: 0

Authors

S.K. Gosal
Department of Microbiology, Punjab Agricultural University, Ludhiana (Punjab), India
G.S. Saroa
Department of Soil Sciences, Punjab Agricultural University, Ludhiana (Punjab), India
Y. Vikal
School of Agricultural Biotechnology, Punjab Agricultural University, Ludhiana (Punjab), India
A. Mehta
Department of Microbiology, Punjab Agricultural University, Ludhiana (Punjab), India

Abstract


Soil is the critical resource as well as a basic medium for the growth of natural micro flora present in it. The soil fertility depends on its physicochemical properties and microbial population diversity. Diazotrophs are the nitrogen fixing bacteria which possess nifH gene that is responsible for coding the nitrogenase enzyme involved in the reduction of atmospheric nitrogen to ammonia. These nitrogen fixing bacteria play an imperative role, function and significance in the soil. Soil samples were collected from the different wheat-maize cropping system of Punjab and analyzed for physicochemical properties as pH, electrical conductivity, organic carbon, soil texture, ammoniacal as well as nitrate nitrogen. Eighty diazotrophic bacteria were isolated on eight different nitrogen free media and characterized culturally, morphologically, biochemically, functionally and using molecular techniques. The diazotrophic nature of the isolates was confirmed by the amplification of nifH gene using two nifH primers viz. nifH1 and nifH2. The nifH positive isolates were further used for 16S rDNA restriction analysis using different enzymes such as Taq1, Rsa I and Hae III. Based on UPGMA clustering, the representative strains were sequenced and identified as Bacillus amyloliquifaciens, Pseudomonas aeruginosa, Bacillus subtilis, Paenibacillus sp., Azotobacter vinelendii, Stenotrophomonas maltophilia, Rhizobium larrymorrei, Flavobacterium anhuiense, Sphingomonas paucimobilis, Paenibacillus panacisoli, Azospirillum sp., Pseudomonas putida, Paenibacillus amyloliticus, Bacillus circulans, Paenibacillus polymyxa and Xanthomonas oryzae.

Keywords


Diazotrophs, nif H, 16S rDNA Sequencing, Physico-Chemical Properties, Soil

References