Isolation and characterization of PGPRs from organic preparations
Keywords:
Microbial population, Panchagavya, Amritpani, Bacillus pumilus, Alcaligenes faecalisAbstract
Rhizobacteria have been described as plant growth promoting as they offer their plant hosts an additional fortification against pathogen by expression of multiple activities that directly and indirectly inhibit pathogens. Most microbial diversity of the soil ecosystem is confined to the rhizosphere. These rhizospheric microbes have the potential to stimulate plant growth and manage soil and plant health. Nowadays, management of the rhizosphere bacterial population has advanced toward the concept of plant growth promoting rhizobacteria (PGPR) owing to emergence of fungicide-resistant pathogens and health concerns for the producers and consumers. Using PGPR as bio-fertilizer is an efficient approach to replace chemical fertilizers and pesticides for sustainable cultivation. In the present study bacteria isolated from Amritpani and Panchagavya were analysed for their biochemical characters and molecular confirmation using molecular tool. The substrate utilization pattern showed that the characteristics of the majority of the tested strains share a similar pattern. However, few isolates displayed varied patterns. Identification of these isolates showed that most of them belong to genus Bacillus and Alcaligenes spp. This was further confirmed with sequencing of the 16s rDNA gene of the bacteria. The present study has resulted in the isolation of PGPRs Bacillus pumilus (CISH-PGPR96), Bacillus subtilis (CISH-PGPR102) and Alcaligenes faecalis (CISH-PGPR104) isolates that can be utilised for sustainable agriculture production.
Downloads
References
Amalraj, E.L.D., Kumar, P. K., Ahmed, S.K.M.H., Abdul, R. and
Bacteria 108 5.49 ± 0.21 0.86 Fungi 106 0.046 ± 0.001 0.08 Actinomycetes 107 1.31 ± 0.19 0.40 Gram positive bacteria 108 0.30 ± 0.12 0.53 Gram negative bacteria 108 1.35 ± 0.50 0.55 Pseudomonas 107 1.53 ± 1.45 1.07 Rhizobium 106 3.03 ± 0.01 0.36 P-solubilizing microbes 106 4.80 ± 0.001 0.20
Table 2: Different microbial population in Amritpani
Kishore, N. 2013. Microbiological analysis of Panchagavya, vermicompost, and FYM and their effect on plant growth promotion of pigeon pea (Cajanus cajan L.) in India. Organic Agriculture, 3:23-29.
Castro, R.O., Cornejo, H.A.C., Rodriguez, L.M. and Bucio, J.L. 2009.The role of microbial signals in plant growth and development. Plant Signal Behaviour, 4:701-712.
Chadha, S., Rameshwar, Ashlesha, Saini, J.P. and Paul, Y.S. 2012. Vedic Krishi: Sustainable livelihood option for small and
Type of microbe Multiplic ation factor
Microbial population (cfu ml-1) after days of preparation (Mean ± sd)
CD (at
%)
marginal farmers, Indian Journal of Traditional. Knowledge, 11 (3):480-86.
Crielly, E.M., Logan, N.A. and Anderton, A. 1994. Studies on the
Bacteria 108 5.49 ± 0.21 0.86 Fungi 106 0.046 ± 0.001 0.08 Actinomycetes 107 1.31 ± 0.19 0.40 Gram positive bacteria 108 0.30 ± 0.12 0.53 Gram negative bacteria 108 1.35 ± 0.50 0.55 Pseudomonas 107 1.53 ± 1.45 1.07 Rhizobium 106 3.03 ± 0.01 0.36 P-solubilizing microbes 106 4.80 ± 0.001 0.20
Table 3: Identification of bacteria based on maximum similarity with 16sRNA sequences
Organism Name Isolate code NCBI Acc. No. Bacillus subtilis CISH-PGPR 102 KY427114 Alcaligenes faecalis CISH-PGPR 104 KY427117 Bacillus pumilus CISH-PGPR 96 KY523254
Figure 1: Amplification of 1.5kb PCR product of 16sRNA gene in all the isolates and its sequencing
Bacillus flora of milk and milk products. Journal of Applied Microbiology, 77:256-63.
Fred, E.B., Baldwin, I.L., and McCoy, F.1932. Root nodule bacteria and leguminous plants, University of Wisconsin Press, Madison, Wisconsin, U.S.A.
Girija, D., Deepa, K., Xavier, F., Antony, I. and Shidhi, P.R. 2013. Analysis of cow dung microbiota-a metagenomic approach. Indian Journal of Biotechnology, 12:372-78.
Goud, W.D., Hagedorn, C, Bardinelli, T.R. and Zablotowiez, R.M. 1985. New selective medium for enumeration and recovery for fluorescent pseudomonads from various habitats, Applied Environmental Microbiolology, 49:28-32.
Hagedorn, C. and Holt, J.G. 1954. Ecology of soil arthrobactrers in Clarion-Websters toposequence of Iowa, Applied Microbiology, 29:211-18.
Hameeda, B., Harini, G, Rupela, O.P., Wani, S.P. and Reddy, G. 2008. Growth promotion of maize by phosphate solubilizing bacteria isolated from composts and macrofauna. Microbiol Research, 163:234-242.
Israr Ahmad and M. Hamida Nasrin. 2014. Genetic diversity and plant growth promoting activity of salt tolerant rhizobacteria from rice rhizosphere. African Journal of Microbial Research, 8:3204-3215.
Israr Ahmad, Someshwar Bhagat and R.C. Srivastava. 2012. Assessment of rhizobacterial diversity of tomato rhizosphere from South Andaman. Vegetos 25: 102-108.
Journal of Eco-friendly Agriculture 14(2) 2019 21
Isolation and characterization of PGPRs from organic preparations
Jensen, H.L.1954.The Azotobacteriaceae, Bacterial. Review, 18:195- 214.
Khalid, A., Arshad, M. and Zahir, Z.A. 2004. Screening plant growth promoting rhizobacteria for improving growth and yield of wheat. Journal of Applied Microbiology, 96:473- 480.
King, E.O., Ward, M.K. and Rancy, D.E.1954.Two simple media for the demonstration of phycocyanin and fluorescein. Journal of Laboratoroy and Clinical Medicine, 44:301-07.
Mayak, S., Tirosh, T. and Glick, B.R. 2004. Plant growth-promoting bacteria confer resistance in tomato plants to salt stress. Plant Physiology and Biochemistry, 42:565-572.
Nene, Y.L. 2007. Utilizing traditional knowledge in agriculture. National Seminar on Organic Agriculture: Hope of Posterity, July, 13-14, 2007, organized by UPCAR and NCOF, pp. 6-10.
Norris, J.R. and Chapman, H.M.1968. Classification of Azotobacter. In: Identification methods for microbiologists (eds. B.M. Gibbs and D.A. Shapton), Academic Press, London and New York, pp. 19-27.
Okon, Y., Alberecht, S.L. and Burris, R.H. 1977. Methods for growing Spirillum lipoferum and for counting it in pure culture and in association with plants. Applied Environmental Microbiology, 33:85-87.
Pikovskaya, R.E.1948. Mobilization of phosphorous in soil in connection with vital activity of some microbial species. Mikrobiology, 17:363-70.
Radha, T.K. and Rao, D.L.N. 2014. Plant growth promoting bacteria from cow dung based biodynamic preparations. Indian Journal of Microbiology, 54:413-418.
Ram, R. A., Singha, Atul and Kumar, A. 2018. Microbial characterization of cow pat pit and other biodynamic preparations used in biodynamic agriculture. Indian Journal of Agricultural Science, 89: 42-46.
Ram, R.A., Singha Atul and Vaish, S. 2018. Microbial characterization of on-farm produced bio-enhancers used in organic farming, Indian Journal of Agricultural Science, 88: 35-40.
Richardson, A.E., Barea, J.M., McNeill, A.M. and Prigent Combaret, C. 2009. Acquisition of phosphorus and nitrogen in the rhizosphere and plant growth promotion by microorganisms. Plant Soil, 321:305-339.
Ross, I. L., Alami, Y., Harvey, P. R., Achouak, W. and Ryder, M. H. 2000. Genetic diversity and biological control activity of novel species of closely related pseudomonads isolated from wheat field soils in South Australia. Applied Journal of Environmental Microbiology, 66:1609-1616.
Sangeetha, V. and Thevanathan, R. 2010. Biofertilizer potential of traditional and Panchagavya amended with seaweed extract. The Journal of American Science, 6:61-67.
Stutz, E. W., De´fago, G. and Kern, H. 1986. Naturally occurring fluorescent pseudomonads involved in suppression of black root rot of tobacco. Phytopathology, 76: 181-185.
Swaminathan, M. S. and Jana, S. 1997. Biodiversity and Global Food Security. Chennai, India, MacMillan India Ltd. Van Loon, L.C. 2007. Plant responses to plant growth-promoting
rhizobacteria. European Journal of Plant Pathology, 119:243- 254.
Verma, S.C., Ladha, J.K. and Tripathi, A.K. 2001. Evaluation of plant growth promoting and colonization ability of endophytic diazotrophs from deep water rice. Journal of Biotechnology, 91:127-141.
