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Assessment of genetic and functional relationship of antagonistic fluorescent pseudomonads of rice rhizosphere by repetitive sequence, protein coding sequence and functional gene analyses

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Abstract

Antagonistic fluorescent pseudomonads isolated from rice rhizospheric soil were characterized using biochemical, taxonomical and molecular tools. Production of cyclopropane fatty acid (CFA) was correlated with their antagonistic potential. Strains were grouped into 18 different genotypes on the basis of amplified ribosomal DNA restriction analysis (ARDRA) and repetitive (rep)-PCR based genotypic fingerprinting analyses. High phylogenetic resolution among antagonistic fluorescent pseudomonad strains was obtained based on the DNA gyrase B subunit (gyrB) and RNA polymerase sigma factor 70 (rpoD) gene sequence analyses. Combined gyrB and rpoD sequence analysis resulted in the accurate estimation of molecular phylogeny and provided a significant correlation between the genetic distances among strains. Present study demonstrated the genetic and functional relationship of fluorescent pseudomonads. The knowledge on genetic and functional potential of fluorescent pseudomonads associated with rice rhizosphere is useful to understand their ecological role and for their utilization in sustainable agriculture.

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References

  • Anjaiah, V., P. Cornelis, and N. Koedam. 2003. Effect of genotype and root colonization in biological control of Fusarium wilts in pigeonpea and chickpea by Pseudomonas aeruginosa PNA1. Can. J. Microbiol. 149, 85–91.

    Article  Google Scholar 

  • Ayyadurai, N., P. Ravindra Naik, and N. Sakthivel. 2007. Functional characterization of antagonistic fluorescent pseudomonads associated with rhizospheric soil of rice (Oryza sativa L.). J. Microbiol. Biotechnol. 117, 919–927.

    Google Scholar 

  • Ayyadurai, N., P. Ravindra Naik, M. Sreehari Rao, R. Sunish Kumar, S.K. Samrat, M. Manohar, and N. Sakthivel. 2006. Isolation and characterization of a novel banana rhizosphere bacterium as fungal antagonist and microbial adjuvant in micropropagation of banana. J. Appl. Microbiol. 1100, 926–937.

    Article  Google Scholar 

  • Bano, N. and J. Mussarrat. 2003. Characterization of a new Pseudomonas aeruginosa strain NJ-15 as a potential biocontrol agent. Curr. Microbiol. 146, 324–328.

    Article  Google Scholar 

  • Berg, G., A. Krechel, M. Ditz, R. Sikora, A. Ulrich, and J. Hallmann. 2005. Endophytic and ectophytic potato-associated bacterial communities differ in structure and antagonistic function against plant pathogenic fungi. FEMS Microbiol. Ecol. 151, 215–229.

    Google Scholar 

  • Berg, G., K. Opelt, C. Zachow, J. Lottmann, M. Gotz, R. Costa, and K. Smalla. 2006. The rhizosphere effect on bacterial antagonistic towards the pathogenic fungus Verticillium differs depending on plant species and site. FEMS Microbiol. Ecol. 156, 250–261.

    Google Scholar 

  • Berg, G., N. Roskot, A. Steidle, L. Eberl, A. Zock, and K. Smalla. 2002. Plant-dependent genotypic and phenotypic diversity of antagonistic rhizobacteria isolated from different Verticillium host plants. Appl. Environ. Microbiol. 168, 3328–3338.

    Article  Google Scholar 

  • Bhattacharya, D., P.M. Sarma, S. Krishnan, S. Mishra, and B. Lal. 2003. Evaluation of genetic diversity among Pseudomonas citronellolis strains isolated from oily sludge-contaminated sites. Appl. Environ. Microbiol. 169, 1435–1441.

    Article  Google Scholar 

  • Bossis, E., P. Lemanceau, X. Latour, and L. Garden. 2000. The Taxonomy of Pseudomonas fluorescens and Pseudomonas putida: current status and need for revision. Agronomie 20, 51–63.

    Article  Google Scholar 

  • Bric, M., M. John, R. Bostock, and R.E. Silverstone. 1991. Rapid in situ assay for indoleacetic acid production by bacteria immobilized on a nitrocellulose membrane. Appl. Environ. Microbiol. 157, 535–538.

    Google Scholar 

  • Brown, J.L., T. Brown, T.A. Ross, N. McMeekin, and P.D. Nichols. 1997. Acid habituation of Escherichia coli and the potential role of cyclopropane fatty acids in low pH tolerance. Int. J. Food Microbiol. 137, 163–173.

    Article  Google Scholar 

  • Cai, J.J., D.K. Smith, X. Xia, and K. Yuen. 2005. MBEToolbox: a Matlab toolbox for sequence data analysis in molecular biology and evolution. BMC Bioinfor. 16, 63–71.

    Google Scholar 

  • Costa, R., N.C.M. Gomes, E. Krogerrecklenfort, K. Opelt, G. Berg, and K. Smalla. 2007. Pseudomonas community structure and antagonistic potential in the rhizosphere: insights gained by combining phylogenetic and functional gene-based analyses. Environ. Microbiol. 19, 2260–2273.

    Article  Google Scholar 

  • Delorme, S., L. Philippot, V. Edel-Hermann, C. Deulvot, C. Mougel, and P. Lemanceau. 2003. Comparative genetic diversity of the narG, nosZ, and 16S rRNA genes in fluorescent pseudomonads. Appl. Environ. Microbiol. 169, 1004–1012.

    Article  Google Scholar 

  • de Souza, J.T. and J. M. Raaijmakers. 2003. Polymorphisms within the PrnD and PltC genes from pyrrolnitrin and pyoluteorin-producing Pseudomonas and Burkholderia spp. FEMS Microbiol. Ecol. 143, 21–34.

    Google Scholar 

  • Dye, D.W. 1962. The inadequacy of the usual determinative tests for identification of Xanthomonas spp. Newzealand J. Sci. 15, 393–416.

    Google Scholar 

  • Ellis, R.J., T.M. Timms Wilson, and M.J. Bailey. 2000. Identification of conserved traits in fluorescent pseudomonads with antifungal activity. Environ. Microbiol. 12, 274–284.

    Article  Google Scholar 

  • Emmert, E.A.B. and J. Handelsman. 1999. Biocontrol of plant disease: A (Gram) positive perspective. FEMS Microbiol. Lett. 1171, 1–9.

    Article  Google Scholar 

  • Gorden, S.A. and R.P. Weber. 1951. Colorimetric estimation of indoleacetic acid. Plant Physiol. 126, 192–195.

    Article  Google Scholar 

  • Grogan, D.W. and J.E. Cronan. 1997. Cyclopropane ring formation in membrane lipids of bacteria. Microbiol. Mol. Biol. Rev. 161, 429–441.

    Google Scholar 

  • Higgins, D.G., A.T. Bleashy, and R. Fuchs. 1992. Clustal V: Improved software for multiple sequence alignment. Comput. Appl. Biosci. 18, 189–191.

    Google Scholar 

  • Keel, C., U. Schnider, M. Maurhofer, C. Voisard, M. Burger, P. Wirthner, D. Haas, and G. Defago. 1992. Suppression of root disease by Pseudomonas fluorescens CHAO: importance of bacterial secondary metabolite 2,4-diacetylphloroglucinol. Mol. Plant. Microbe Interact. 15, 4–13.

    Google Scholar 

  • Kimura, M. 1980. A simple method for estimating evolutionary rates of base substitutions through comparative studies of nucleotide sequences. J. Mol. Evol. 116, 111–120.

    Article  Google Scholar 

  • King, J.E. 1936. The colorimetric determination of phosphorus. Biochem. J. 126, 292–297.

    Google Scholar 

  • King, E.O., M.K. Ward, and D.E. Raney. 1954. Two simple media for demonstration of pyocyanin and fluorescein. J. Lab. Clin. Med. 144, 301–307.

    Google Scholar 

  • Kumar, B.S.D. and H.S. Dube. 1992. Seed bacterization with a fluorescent Pseudomonas for enhanced plant growth, yield and disease control. Soil Biol. Biochem. 124, 539–542.

    Article  Google Scholar 

  • Kumar, S., K. Tamura, I.B. Jakobsen, and M. Nei. 2001. MEGA2: molecular evolutionary genetics analysis software. Bioinfor. 117, 1244–1245.

    Article  Google Scholar 

  • Laguerre, G., L. Rigottier-Gois, and P. Lemanceau. 1994. Fluorescent Pseudomonas species categorized by using polymerase chain reaction (PCR) / restriction fragment analysis of 16S rDNA. Mol. Ecol. 13, 479–487.

    Article  Google Scholar 

  • Leach, J.E., M.L. Rhoades, C.M.V. Cruz, F.F. White, T.W. Mew, and H. Leung. 1992. Assessment of genetic diversity and population structure of Xanthomonas oryzae with a repetitive DNA element. Appl. Environ. Microbiol. 158, 2188–2195.

    Google Scholar 

  • Lottmann, J. and G. Berg. 2001. Phenotypic and genotypic characterization of antagonistic bacteria associated with roots of transgenic and non transgenic potato plants. Microbiol. Res. 1156, 75–82.

    Article  Google Scholar 

  • Nei, M. and T. Gojobori. 1986. Simple methods for estimating the numbers of synonymous and nonsynonymous nucleotide substitutions. Mol. Biol. Evol. 13, 418–426.

    Google Scholar 

  • Patten, C.L. and B.R. Glick. 2002. Role of Pseudomonas putida indoleacetic acid in development of the host plant root system. Appl. Environ. Microbiol. 168, 3795–3801.

    Article  Google Scholar 

  • Penrose, D.M. and B.R. Glick. 2002. Methods for isolating and characterizing ACC deaminase containing plant growth promoting rhizobacteria. Physiol. Plantarum. 1118, 10–15.

    Google Scholar 

  • Raaijmakers, J., D.M. Weller, and L.S. Thomashow. 1997. Frequency of antibiotic producing Pseudomonas spp. in natural environments. Appl. Environ. Microbiol. 163, 881–887.

    Google Scholar 

  • Rainey, P.B. and E.R. Moxon. 2000. When being hyper keeps you fit. Science 288, 1186–1187.

    Article  CAS  PubMed  Google Scholar 

  • Rangarajan, S., P. Loganathan, L.M. Saleena, and S. Nair. 2001. Diversity of pseudomonads isolated from three different plant rhizospheres. J. Appl. Microbiol. 191, 742–749.

    Article  Google Scholar 

  • Ravindra, N.P., N. Sahoo, D. Goswami, N. Ayyadurai, and N. Sakthivel. 2008. Genetic and functional diversity among fluorescent pseudomonads isolated from the rhizosphere of banana. Microb. Ecol. 156, 492–504.

    Google Scholar 

  • Ravindra, N.P. and N. Sakthivel. 2006. Functional characterization of a novel hydrocarbonoclastic Pseudomonas sp. strain PUP6 with plant-growth-promoting traits and antifungal potential. Res. Microbiol. 1157, 538–546.

    Google Scholar 

  • Saitou, N. and M. Nei. 1987. The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol. Biol. Evol. 14, 406–425.

    Google Scholar 

  • Sakthivel, N. and S.S. Gnanamanickam. 1987. Evaluation of Pseudomonas fluorescens for suppression of sheath rot disease and for enhancement of grain yields in rice (Oryza sativa L.). Appl. Environ. Microbiol. 153, 2056–2059.

    Google Scholar 

  • Schwyn, B. and J.B. Neiland. 1987. Universal chemical assay for the detection and determination of siderophores. Analy. Biochem. 1160, 47–56.

    Article  Google Scholar 

  • Sharp, P.M. 1997. In search of molecular darwinism. Nature 385, 111–112.

    Article  PubMed  Google Scholar 

  • Sunish, K.R., N. Ayyadurai, P. Pandiaraja, A.V. Reddy, Y. Venkateshwarlu, O. Prakash, and N. Sakthivel. 2005. Characterization of antifungal metabolite produced by a new strain Pseudomonas aeruginosa PuPa3 that exhibits broad-spectrum antifungal activity and biofertility traits. J. Appl. Microbiol. 198, 145–154.

    Google Scholar 

  • Thomashow, L.S., D.M. Weller, R.F. Bonsall, and L.S. Pierson. 1990. Production of the antibiotic phenazine-1-carboxylic acid of fluorescent Pseudomonas species in the rhizosphere of wheat. Appl. Environ. Microbiol. 156, 908–912.

    Google Scholar 

  • Weisburg, W.G., S.M. Barns, and D.J. Lane. 1991. 16S ribosomal DNA amplification for phylogenetic study. J. Bacteriol. 1173, 697–703.

    Google Scholar 

  • Yamamoto, S. and S. Harayama. 1998. Phylogenetic relationships of Pseudomonas putida strains deduced from the nucleotide sequences of gyrB, rpoD and 16S rRNA genes. Int. J. Syst. Bacteriol. 148, 813–819.

    Article  Google Scholar 

  • Yamamoto, S., H. Kasai, D.L. Arnold, R.W. Jackson, A. Vivian, and S. Harayama. 2000. Phylogeny of the genus Pseudomonas: intrageneric structure reconstructed from the nucleotide sequences of gyrB and rpoD genes. Microbiology 146, 2385–2394.

    CAS  PubMed  Google Scholar 

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Pathma, J., Ayyadurai, N. & Sakthivel, N. Assessment of genetic and functional relationship of antagonistic fluorescent pseudomonads of rice rhizosphere by repetitive sequence, protein coding sequence and functional gene analyses. J Microbiol. 48, 715–727 (2010). https://doi.org/10.1007/s12275-010-0064-3

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