Phenotypic and genotypic characterization of inhabitant PGPR strains of Pseudomonas from apple orchards
Article Main
Abstract
Aim of present research was to isolate and characterize the Pseudomonas strains phenotypically and genotypically from the rhizospheric soil of apple orchard at Maggota (Shimla district) Himachal Pradesh. Phenotypic identification of the test isolates was based on morphological, physiological and biochemical characterization of the bacteria followed by genotypic analysis using rRNA gene sequencing and RAPD-PCR analysis. The fourteen Pseudomonas sp. isolates were screened out for various plant growth promoting activities such as siderophore production, antifungal activity, phosphate solubilisation, HCN and ammonia production, production of plant growth regulators and lytic enzymes. Isolates showed production of plant growth regulators (auxins, gibberellins and cytokinins) in the range of 19.67-83.33?g/ml, 21.00-58.67 ?g/ml and 12.33-43.33 ?g/ml respectively. Pseudomonas strains showed phosphate solubilising activity in the range of 12.33-63.33 Pi ?g/ml, 53.66-93.44 % SU siderophore production and 11.33-96.33mm (diameter) protease activity in plate assay. Five Pseudomonas isolates i.e. An-16-kul, An-1-mag, An-2-mag, An-3-mag and An-6-mag showed maximum antifungal activity against plant pathogenic fungi. Therefore, the aim of present investigation was to study multifarious plant growth promoting qualities of Pseudomonas sp. and to select more efficient PGPR strain of fluorescent Pseudomonas sp. which can be further used as biofertilizer.
Article Details
Article Details
Keywords
Genotype, PGPR, Phenotype, Phosphate solubilization, Pseudomonas sp., Siderophore
References
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Bai, Y., Zhou X. and Smith, D. L. (2003). Enhanced soybean plant growth resulting from co-inoculation of Bacillus strains with Bradyrhizobium japonicum. Crop Science, 43: 1774-1781.
Bakker, and Schippers. (1987). Microbial cyanide production in the rhizosphere in relation to potato yield reduction and Pseudomonas spp. Mediated plant growth stimulation. Soil Biology and Biochemistry, 19: 451-457.
Bray, R. H. and Kurtz, L. T. (1945). Determination of total organic available forms of phosphorus in soil. Soil Science, 23: 343-353.
Chaiharn, M., Chunhaleuchanon, S. and Lumyong, S. (2009). Screening siderophore producing bacteria as potential biological control agent for fungal rice pathogens in Thailand. World Journal of Microbiology and Biotechnology, 25: 1919-1928.
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Glick, B. R. (1995). Enhancement of plant growth by free living bacteria. Canadian Journal of Microbiology, 41: 109-117.
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Pikovsakaya , R. E.(1948). Mobilization of phosphorus in soil in connection with vital activity of some microbial species. Microbiologia, 17: 362-370.
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Ribeiro, C. M. and Cardoso, E. J. (2011). Isolation, selection and characterization of root associated growth promoting bacteria in Brazil Pine (Araucaria angustifolia). Microbial Research, 167: 69-78.
Sajjad, Mirza, M., Ahmad, W., Latif, F., Haurat, J. and Bally, R. (2001). Isolation, partial characterization, and the effect of plant growth promoting bacteria (PGPB) on micro-propagated sugarcane in vitro. Plant and Soil Science, 237: 47-54.
Schwyan, B. and Neilands, J. B. (1987). Universal chemical assay for the detection and determination of siderophores. Analytic Biochemistry, 28(8): 751-759.
Shalini, and Srivastava, R. (2008). Screening for antifungal activity of Pseudomonas fluorescens against phytopathogenic fungi. The International Journal of Microbiology, 5: 2p.
Sharma, S., Prashad, D. and Kaur, M. (2014). Isolation of fluorescent Pseudomonas strain from temperate zone of Himachal Pradesh and their evaluation as plant growth promoting rhizobacteria (PGPR).The Bioscan, 9(2):323-328.
Vandamme, P., Pot, B., Gibbs, M., DeVos, P., Kersters, K. and Swings, J. (1996). Polybasic taxonomy: a consensus approach to bacterial systematic. Microbiological Reviews, 60: 407-438.
Vessey, J. K. (2003). Plant growth promoting rhizobacteria as biofertilizers. Plant and Soil, 255: 571-586.
Vincent, J. M. (1947). Distribution of fungal hyphae in presence of certain inhibitors. Nature, 150-850.
Weller, D. M., Raaijmakers, J. Mcspadden, Gardener, B. and Thomashow, L. S. (2002). Microbial populations responsible for specific soil suppressive to plant pathogens. Annual Review of Phytopathology, 40: 309-348.
Widmer, F., Seidler, R. J., Gillevet, P. M., Watrud, L. S. and DiGiovanni. G. D. (1998). Highly selective PCR protocol for detecting 16S rRNA genes of the genus Pseudomonas (Sensu Stricto) in environmental samples. Applied and Environmental Microbiology 64(7): 2545-2553.
Altschul, S. F., Gish, W., Miller, W., Myers, E. W. and Lipman, D. J. (1990). Basic local alignment search tool. Journal of Molecular Biology, 215: 403-441.
Aneja, K. R. (2003). Experiments in microbiology, plant pathology and biotechnology. 4th rev. ed. New Age International: New Delhi. 607 p.
Bai, Y., Zhou X. and Smith, D. L. (2003). Enhanced soybean plant growth resulting from co-inoculation of Bacillus strains with Bradyrhizobium japonicum. Crop Science, 43: 1774-1781.
Bakker, and Schippers. (1987). Microbial cyanide production in the rhizosphere in relation to potato yield reduction and Pseudomonas spp. Mediated plant growth stimulation. Soil Biology and Biochemistry, 19: 451-457.
Bray, R. H. and Kurtz, L. T. (1945). Determination of total organic available forms of phosphorus in soil. Soil Science, 23: 343-353.
Chaiharn, M., Chunhaleuchanon, S. and Lumyong, S. (2009). Screening siderophore producing bacteria as potential biological control agent for fungal rice pathogens in Thailand. World Journal of Microbiology and Biotechnology, 25: 1919-1928.
Clark, A. G. and Lanigan, C. M. S. (1993). Prospects for estimating nucleotidic divergence with RAPDs. Molecular Biology and Evolution, 10: 1096-1111.
Glick, B. R. (1995). Enhancement of plant growth by free living bacteria. Canadian Journal of Microbiology, 41: 109-117.
Gopalakrishnan, S., Humayun, P., Kiran, B. K., Kannan, I. G. K. and Vidya, M. S. (2011). Evaluation of bacteria isolated from rice rhizosphere for biological control of charcoal rot of sorghum caused by Macrophomina phaseolina (Tassi) Goid. World Journal of Microbial Biotechnology, 27: 1313-1321.
Gordon, S. A. and Weber, R. P. (1951). Colorimetric estimation of indoleacetic acid. Plant Physiology, 26: 192-195.
Gutierrez Mañero, F. J., Ramos Solano, B., Probanza, A., Mehouachi, J., Tadeo, F. R. and Talon, M. (2001). The plant growth-promoting rhizobacteria Bacillus pumilus and Bacillus licheniformis produce high amounts of physiologically active gibberellins. Physiology Plant, 111: 1-7.
Holbrook, A. A., Edge, W. L. W. and Bailey, F. (1961). Spectrophotometric method for determination of gibberellic acid in gibberellins, ACS Washington, D.C., pp. 159-167.
Kaur, M., Gupta, M., Tripatathi, K. A. K. and Gupta, K. G. (1989). Lytic effect of Pseudomonas aeruginosa elastase on gram positive and gram negative bacteria: entrablatt Bakt. Indian Journal Microbiology, 34: 855-859.
King, E. O., Ward, M. K. and Raney, D. E. (1954). Two simple media for the demonstration of pyocyanin and fluorescin. Journal of Laboratory and Chemical Medicine, 44: 301-307.
Lata, and Saxena, A. K. (2003). Characterization of plant growth promoting rhizobacteria. In: Training manual on Biofertilizer Technology (eds.) A K Saxena. IARI Delhi, pp. 24-25.
Letham, A. (1971). Regulator of cell division in plant tissues. XII. A cytokinin bioassay using excised radish cotyledons. Physiology Plant, 25: 391-396.
Lucas, G. J. A., Probanza, A., Ramos, B. and Gutierrez, M. F. J. (2001). Genetic variability of rhizobacteria from wild populations of four Lupinus species based on PCRRAPDs. Journal of Plant Nutrition and Soil Science, 164: 1-7.
Lucy, M., Reed, E. and B. Glick. (2004). Application of free living plant growth-promoting rhizobacteria. Antonie van Leeuwenhoek, 86: 1-25.
Mina, D., Koche, Gade, R. M. and Deshmukh, A. G. (2013). Antifungal activity of secondary metabolites produced by Pseudomonas fluorescens. The Bioscan, 8(2): 723-726.
Nei, M. and Miller, J. C. (1990). A simple method for estimating average number of nucleotide substitutions within and between populations from restriction data. Genetics, 125: 873-879.
Pikovsakaya , R. E.(1948). Mobilization of phosphorus in soil in connection with vital activity of some microbial species. Microbiologia, 17: 362-370.
Ramette, A., Moënne, Loccoz, Y. and Défago, G. (2003). Prevalence of fluorescent pseudomonads producing antifungal phloroglucinols and/or hydrogen cyanide in soils naturally suppressive or conducive to tobacco black root rot. FEMS Microbiology Ecology, 44: 35-43.
Ribeiro, C. M. and Cardoso, E. J. (2011). Isolation, selection and characterization of root associated growth promoting bacteria in Brazil Pine (Araucaria angustifolia). Microbial Research, 167: 69-78.
Sajjad, Mirza, M., Ahmad, W., Latif, F., Haurat, J. and Bally, R. (2001). Isolation, partial characterization, and the effect of plant growth promoting bacteria (PGPB) on micro-propagated sugarcane in vitro. Plant and Soil Science, 237: 47-54.
Schwyan, B. and Neilands, J. B. (1987). Universal chemical assay for the detection and determination of siderophores. Analytic Biochemistry, 28(8): 751-759.
Shalini, and Srivastava, R. (2008). Screening for antifungal activity of Pseudomonas fluorescens against phytopathogenic fungi. The International Journal of Microbiology, 5: 2p.
Sharma, S., Prashad, D. and Kaur, M. (2014). Isolation of fluorescent Pseudomonas strain from temperate zone of Himachal Pradesh and their evaluation as plant growth promoting rhizobacteria (PGPR).The Bioscan, 9(2):323-328.
Vandamme, P., Pot, B., Gibbs, M., DeVos, P., Kersters, K. and Swings, J. (1996). Polybasic taxonomy: a consensus approach to bacterial systematic. Microbiological Reviews, 60: 407-438.
Vessey, J. K. (2003). Plant growth promoting rhizobacteria as biofertilizers. Plant and Soil, 255: 571-586.
Vincent, J. M. (1947). Distribution of fungal hyphae in presence of certain inhibitors. Nature, 150-850.
Weller, D. M., Raaijmakers, J. Mcspadden, Gardener, B. and Thomashow, L. S. (2002). Microbial populations responsible for specific soil suppressive to plant pathogens. Annual Review of Phytopathology, 40: 309-348.
Widmer, F., Seidler, R. J., Gillevet, P. M., Watrud, L. S. and DiGiovanni. G. D. (1998). Highly selective PCR protocol for detecting 16S rRNA genes of the genus Pseudomonas (Sensu Stricto) in environmental samples. Applied and Environmental Microbiology 64(7): 2545-2553.
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Phenotypic and genotypic characterization of inhabitant PGPR strains of Pseudomonas from apple orchards. (2015). Journal of Applied and Natural Science, 7(2), 893-902. https://doi.org/10.31018/jans.v7i2.703
