Skip to main content
SpringerLink
Log in

Cultivable heterotrophic N2-fixing bacterial diversity in rice fields in the Yangtze River Plain

  • Original Paper
  • Published:
Biology and Fertility of Soils Aims and scope Submit manuscript

Abstract

Heterotrophic N2-fixing bacteria are a potentially important source of N2 fixation in rice fields due to the moist soil conditions. This study was conducted at eight sites along a geographic gradient of the Yangtze River Plain in central China. A nitrogen-free solid malate-sucrose medium was used to isolate heterotrophic N2-fixing bacteria. Numbers of the culturable N2-fixing bacteria expressed as CFU (colony forming units) ranged between 1.41±0.42×106 and 1.24±0.23×108 in the sampled paddy field sites along the plain. Thirty strains with high ARA (acetylene reduction activity) were isolated and purified; ARA of the strains varied from 0.9 to 537.8 nmol C2H4 culture-1 h-1, and amounts of 15N fixed ranged between 0.008 and 0.4866 mg·culture-1·day-1. According to morphological and biochemical characteristics, 14 strains were identified as the genus Bacillus, 2 as Burkholderia, 1 as Agrobacterium, 4 as Pseudomonas, 2 as Derxia, 1 as Alcaligenes, 1 as Aeromonas, 2 as Citrobacter, and 3 strains belonged to the corynebacter-form group.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1.

Similar content being viewed by others

References

  • Anderson GR (1955) Nitrogen fixation by Pseudomonas-like soil bacteria. J Bacteriol 70:129–133

    CAS  Google Scholar 

  • Asanuma S, Tanaka H, Yatazawa M (1980) Pseudomonas cepacia: a characteristic rhizoplane microorganism in rice plant. Soil Plant Nutr 26:71–78

    Google Scholar 

  • Balandreau J (1983) Microbiology of the association. Can J Microbiol 29:851–859

    Google Scholar 

  • Balandreau J (1986) Ecological factors and adaptive processes in N2-fixing bacterial populations of the plant environment. Plant Soil 90:73–92

    Google Scholar 

  • Balandreau J, Roger P (1996) Some comments about a better use of biological nitrogen fixation in rice cultivation. In: Rahman M, Podder AK, van Hove C, Begum ZNT, Heulin T, Hartmann A (eds) Biological nitrogen fixation associated with rice production. Kluwer, Dordrecht, pp 1–12

  • Barber LE, Evans HJ (1976) Characterization of a nitrogen-fixing bacterial strain from the roots of Digitaria sanguinalis. Can J Microbiol 22:254–260

    CAS  PubMed  Google Scholar 

  • Barraquio WL, Guzman MR, Barrion M, Watanbe I (1982) Population of aerobic heterotrophic nitrogen fixing bacteria associated with wetland and dryland rice. Appl Environ Microbiol 43:124–128

    CAS  Google Scholar 

  • Barraquio WL, Ladha JK, Watanabe J (1983) Isolation and identification of N2-fixing Pseudomonas associated with wetland rice. Can J Microbiol 29:867–873

    CAS  PubMed  Google Scholar 

  • Berge O, Heulin T, Balandreau J (1991) Diversity of diazotroph populations in the rhizosphere of maize (Zea mays L.) growing on different French soil. Biol Fertil Soils 11:210–215

    Google Scholar 

  • Boddey RM, de Oliveira OC, Urquiaga S, Reis VM, Olivares FL, Baldani VLD, Döbereiner J (1995) Biological nitrogen fixation associated with sugar cane and rice: contributions and prospects for improvement. Plant Soil 174:195–209

    CAS  Google Scholar 

  • Boddey RM, Alves B, Urquiaga S (1998) Evaluation of biological nitrogen fixation associated with non-legumes. In: Malik KA, Mirza MS, Ladha JK (eds) Nitrogen fixation with non-legumes. Kluwer, Dordrecht, pp 287–305

  • Bohlool BB, Ladha JK, Garrity DP, George T (1992) Biological nitrogen fixation for sustainable agriculture: a perspective. Plant Soil 141:1–11

    CAS  Google Scholar 

  • Brejda JJ, Kremer RJ, Brown JR (1994) Indications of associative nitrogen fixation in eastern gamagrasses. J Range Manage 47:192–195

    Google Scholar 

  • Burris RH (1975) The acetylene-reduction technique. In: Stewart WDP (ed) Nitrogen fixation by free-living micro-organisms. Cambridge University Press, London, pp 249–258

  • Chalk PM (1991) The contribution of associative and symbiotic nitrogen fixation to the nitrogen nutrition of non-legumes. Plant Soil 132:29–39

    CAS  Google Scholar 

  • Cowan ST (1974) Cowan and Steel's manual for the identification of medical bacteria, 2nd edn. Cambridge University Press, Cambridge pp 137–180

  • Cui Z, Su B, Cai M, Hai W, Xie G (1996) Studies on the resource of N2-fixing bacteria in North-China paddy (in Chinese). J China Agric Univ 1:63

    Google Scholar 

  • Döbereiner J (1989) Isolation and identification of root associated diazotrophs. In: Skinner FA (ed) Nitrogen fixation with non-legumes. Kluwer, Dordrecht, pp 103–108

  • Döbereiner J, Pedrosa FO (1987) Nitrogen-fixing bacteria in non-leguminous crop plants. Science Tech, Madison, Wis., pp 63–77

  • Estrada SP de los, Bustillos CR, Caballero MJ (2001) Burkholderia, a genus rich in plant-associated nitrogen fixers with wide environmental and geographic distribution. Appl Environ Microbiol 67:2790–2798

    Article  PubMed  Google Scholar 

  • Garcia JL, Roussos S, Gauthier D, Rinaudo G, Mandel M (1983) Taxonomical study of free-living N2-fixing bacteria isolated from the endorhizosphere of rice. Annal Microbiol Inst Pasteur 134B:329–346

    CAS  Google Scholar 

  • Giller KE, Cadisch G (1995) Future benefits from biological nitrogen fixation: an ecological approach to agriculture. Plant Soil 174:255–277

    Google Scholar 

  • Gogotov JN, Schlegel HG (1974) N2-fixation by chemoautotrophic hydrogen bacteria. Arch Microbiol 97:359–362

    CAS  PubMed  Google Scholar 

  • Hardarson G, Danso SKA (1993) Methods for measuring biological nitrogen fixation in grain legumes. Plant Soil 152:19–23

    Google Scholar 

  • Hardy RWF, Holston RD (1977) Methods for measurement of dinitrogen fixation. In: Hardy RWF, Gibson AH (eds) Treatise on dinitrogen fixation. Wiley, New York, pp 451–485

  • Hayward AC (1964) Characteristics of Preudomonas solanacearum. J Appl Bacteriol 27:265–277

    Google Scholar 

  • Herman RP, Provencio K, Torrez R, Seager GM (1994) Seasonal and spatial population dynamics of the nitrogen efficient guild in a desert Bajada grassland. Appl Environ Microbiol 60:1160–1165

    CAS  PubMed  Google Scholar 

  • Holt JG, Krieg NR, Sneath PHA (eds) (1994) Bergey's manual of determinative bacteriology, 9th edn. Williams and Wilkins, Baltimore, Md.

  • Kammen A van (1997) Biological nitrogen fixation for sustainable agriculture. In: Legocki A, Bothe H, Pühler A (eds) Biological fixation of nitrogen for ecology and sustainable agriculture. Springer, Berlin Heidelberg New York, pp 177–178

  • Krieg NR, Holt JG (eds) (1984) Bergey's manual of systematic bacteriology, 9th edn, vol. 1. Williams and Wilkins, Baltimore, Md.

  • Line MA (1990) Identification of nitrogen-fixing enterobacteria from living sassafras ( Atherospermum moschastum Labill) trees. Plant Soil 125:149–152

    CAS  Google Scholar 

  • Mahadevappa M, Shenoy VV (2000). Towards nitrogen fixing rice (Oryza sativa). Adv Agric Res India 13:131–139

    Google Scholar 

  • Malik KA, Bilal P, Rasul G, Mahwood K, Sajjad MI (1991) Associative N2-fixation in plants growing in saline sodic soils and its relative quantification based on 15N natural abundance. Plant Soil 137:67–74

    Google Scholar 

  • Mark BP, Craswell ET (1992) Biological nitrogen fixation: investments, expectations and actual contributions to agriculture. Plant Soil 141:13–39

    CAS  Google Scholar 

  • Omar AMN, Richard C, Weinhard P, Balandreau J (1989) Using the spermosphere mogel technique to describe the dominant nitrogen-fixing microflora associated with wetland rice in two Egyptian soils. Biol Fertil Soils 7:158–163

    CAS  Google Scholar 

  • Ostle AG, Holt G (1982) Nile blue A as a fluorescent stain for poly-β-hydrooxybutrate. Appl Environ Microbiol 44:238–241

    CAS  PubMed  Google Scholar 

  • Oyaizu-Masuchi Y, Komagata K (1988) Isolation of free-living nitrogen-fixing bacteria from the rhizosphere of rice. J Gen Appl Microbiol 34:127–164

    CAS  Google Scholar 

  • Parkinson D, Gray TRG, Williams J (1971) Methods for studying the ecology of soil micro-organisms. Blackwell, Oxford, pp 64–66

  • Paul EA, Newton JD (1961) Studies of aerobic non-symbiotic nitrogen-fixing bacteria. Can J Microbiol 7:7–13

    CAS  Google Scholar 

  • Peoples MB, Herridge DF, Ladha JK (1995) Biological nitrogen fixation: an efficient source of nitrogen for sustainable agricultural production. Plant Soil 174:3–28

    CAS  Google Scholar 

  • Postgate J (1998) Nitrogen fixation, 3rd edn. Cambridge University Press, Cambridge, pp 28–33

  • Proctor MH, Wilson PW (1958) Nitrogen fixation by gram-negative bacteria. Nature 182:891

    CAS  Google Scholar 

  • Puente ME, Bashan Y (1994) The desert epiphyte Tillandsia recurvata harbours the nitrogen-fixing bacterium Pseudomonas stutzeri. Can J Bot 72:406–408

    Google Scholar 

  • Rao SRV, Dart PJ (1979) Nitrogen fixation associated with sorghum and millet. In: Vose PB, Alaides P (eds) Associative N2-fixation, vol 1. CRC, Boca Raton, Fla., pp 119–129

  • Rao VR, Charyulu PBBN, Nayak DN, Ramakrishna C (1979) 15N2 incorporation in rice soils and by Azospirillum lipoferum from rice root association. In: Vose PB, Ruschel AP (eds) Associated N2-fixaton. CRC, Boca Raton, Fla., pp 197–204

  • Seldin L, van Elsas JD, Penido EGCR (1984) Bacillus azotofixans sp. nov. a nitrogen species from Brazilian soil and grass roots. Int J Syst Bacteriol 34:451–456

    CAS  Google Scholar 

  • Skerman VBD (1967) A guide to the identification of the genera bacteria, 2nd edn. Williams and Wilkins, Baltimore, Md., pp 219–220, 276–280

  • Sokal RR, Rohlf FJ (1969) Biometry: principles, practices and statistics in biological research. Freeman, San Francisco, Calif.

    Google Scholar 

  • Sprent JI (1979a) The biology of nitrogen-fixing organisms. McGraw-Hill, London, pp 103–105

  • Sprent JI (1979b) The biology of nitrogen-fixing organisms. McGraw-Hill, London, pp 6–8

  • Tarrand JJ, Kreig NR, Döbereiner J (1978) A taxonomic study of the Spirillum lipoferum group, with descriptions of a new genus, Azopirillum gen. nov. and two species, Azospirillum lipoferum (Boijerinck) comb. nov. and Azospirillum brasilens sp. nov. Can J Microbiol 24:967–980

    Google Scholar 

  • Trolldenier G (1977) Influence of some environmental factors on nitrogen fixation in the rhizosphere of rice. Plant Soil 47:203–217

    CAS  Google Scholar 

  • Vance CP (1997) Enhanced agricultural sustainability through biological nitrogen fixation. In: Legocki A, Bothe H, Pühler A (eds) Biological fixation of nitrogen for ecology and sustainable agriculture. Springer, Berlin, Heidelberg, pp 179–186

  • Watanabe I, Lee KK, Alimagne BV (1978) Seasonal change of N2-fixing rate in rice field assayed by in situ acetylene reduction technique. Experiments in long-term fertility plots. Soil Sci Plant Nutr 24:1–13

    CAS  Google Scholar 

  • Xie G, Steinberger Y (2002) Dynamics of nitrogen-efficient guild and its relationship to nitrogen and carbon pattern in desert loessial and sandy soil ecosystems. Arid Land Res Manage 16:69–81

    Article  Google Scholar 

  • Yabuuchi E, Kosako Y, Oyaizu H, Yano I, Hotta H, Hashimoto Y, Ezaki T, Arakawa M (1992) Proposal of Burkholderia gen. nov. and transfer of seven species of the genus Pseudomonas homology group II to the new genus, with the type species Burkholderia cepacia comb. nov. Microbiol Immunol 36:1251–1275

    PubMed  Google Scholar 

  • You CB, Wang HX (1990) Rhizoshpere nitrogen fixation in wetland rice. Acta Phytophysiol Sin 16:209–214

    Google Scholar 

  • Zhang J, Wang L, Li N (1992) Identification of Agrobacterium sp.. with nitrogenase activity (in Chinese). Chin Sci Bull 18:1725–1726

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. College of Crop Science of China Agricultural University, , 100094, Beijing, China

    Guang Hui Xie & Guang Can Tao

  2. Institute of Microbiology, Academia Scinica, 100080, Beijing, China

    Miao Ying Cai

  3. Laboratory of Soil Ecology, Faculty of Life Sciences of Bar-Ilan University, 52900, Ramat-Gan, Israel

    Yosef Steinberger

Authors
  1. Guang Hui Xie
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Miao Ying Cai
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Guang Can Tao
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Yosef Steinberger
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Guang Hui Xie.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Xie, G.H., Cai, M.Y., Tao, G.C. et al. Cultivable heterotrophic N2-fixing bacterial diversity in rice fields in the Yangtze River Plain. Biol Fertil Soils 37, 29–38 (2003). https://doi.org/10.1007/s00374-002-0565-2

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00374-002-0565-2

Keywords

Search

Navigation