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Plant and Soil

, Volume 174, Issue 1–2, pp 195–209 | Cite as

Biological nitrogen fixation associated with sugar cane and rice: Contributions and prospects for improvement

  • R. M. Boddey
  • O. C. de Oliveira
  • S. Urquiaga
  • V. M. Reis
  • F. L. de Olivares
  • V. L. D. Baldani
  • J. Döbereiner
Article

Abstract

15N isotope and N balance studies performed over the last few years have shown that several Brazilian varieties of sugarcane are capable of obtaining over 60% of their nitrogen (<150 kg N ha-1 year-1) from biological nitrogen fixation (BNF). This may be due to the fact that this crop in Brazil has been systematically bred for high yields with low fertilizer N inputs. In the case of wetland rice, N balance experiments performed both in the field and in pots suggest that 30 to 60 N ha-1 crop-1 may be obtained from plant-associated BNF and that different varieties have different capacities to obtain N from this source. 15N2 incorporation studies have proved that wetland rice can obtain at least some N from BNF and acetylene reduction (AR) assays also indicate differences in N2-fixing ability between different rice varieties. However in situ AR field estimates suggest plant-associated BNF inputs to be less than 8 kg N ha-1 crop-1. The problems associated with the use of the 15N dilution technique for BNF quantification are discussed and illustrated with data from a recent study performed at EMBRAPA-CNPAB. Although many species of diazotrophs have been isolated from the rhizosphere of both sugarcane and wetland rice, the recent discovery of endophytic N2-fixing bacteria within roots, shoots and leaves of both crops suggests, at least in the case of sugarcane, that these bacteria may be the most important contributors to the observed BNF contributions. In sugarcane both Acetobacter diazotrophicus and Herbaspirillum spp. have been found within roots and aerial tissues and these microorganisms, unlike Azospirillum spp. and other rhizospheric diazotrophs, have been shown to survive poorly in soil. Herbaspirillum spp. are found in many graminaceous crops, including rice (in roots and aerial tissue), and are able to survive and pass from crop to crop in the seeds. The physiology, ecology and infection of plants by these endophytes are fully discussed in this paper. The sugarcane/endophytic diazotroph association is the first efficient N2-fixing system to be discovered associated with any member of the gramineae. As yet the individual roles of the different diazotrophs in this system have not been elucidated and far more work on the physiology and anatomy of this system is required. However, the understanding gained in these studies should serve as a foundation for the improvement/development of similar N2-fixing systems in wetland rice and other cereal crops.

Key words

Acetobacter diazotrophicus biological nitrogen fixation endophytic bacteria Herbaspirillum spp. sugar cane wetland rice 

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References

  1. Al-Mallah M K, Davey M R and Cocking E C 1989 Formation of nodular structures on rice seedlings by rhizobia. J. Exp. Bot. 40, 473–478.Google Scholar
  2. App A A, Watanabe I, Alexander M, Ventura W, Daez C, Santiago T and De Datta S K 1980 Nonsymbiotic nitrogen fixation associated with the rice plant in flooded soils. Soil Sci. 130, 283–289.Google Scholar
  3. App A A, Watanabe I, Ventura T S, Bravo M and Jurey C D 1986 The effect of cultivated and wild rice varieties on the nitrogen balance of flooded soil. Soil Sci. 141, 448–452.Google Scholar
  4. App A A, Santiago T, Daez C, Menguito C, Ventura W, Tirol A, Po J, Watanabe I, De Datta S K and Roger P 1984 Estimation of the nitrogen balance for irrigated rice and the contribution of phototrophic nitrogen fixation. Field Crops Res. 9, 17–27.Google Scholar
  5. Azeredo D F, Bolsanello J, Weber M and Vieira J R 1986 Nitrogênio em cana-planta, doses e fracionamento. STAB 4, 26–32.Google Scholar
  6. Baldani J I, Baldani V L D, Seldin L and Döbereiner J 1986 Characterization of Herbaspirillum seropedicae gen. nov., sp. nov., a root-associated nitrogen-fixing bacterium. Int. J. Syst. Bacteriol. 36, 86–93.Google Scholar
  7. Baldani J I, Pereira P A A, Rocha R E M and Döbereiner J 1981 Especificidade na infecçÁo de raizes por Azospirillum spp. em plantas com via fotossintetica C3 e C4. Pesq. Agropec. Bras. 16, 325–330.Google Scholar
  8. Baldani V L D and Döbereiner J 1980 Host-plant specificity in the infection of cereals with Azospirillum spp. Soil Biol. Biochem. 12, 433–439.Google Scholar
  9. Baldani V L D, Baldani J I, Olivares F L and Döbereiner J 1992a Identification and ecology of Herbaspirillum seropedicae and the closely related Pseudomonas rubrisubalbicans. Symbiosis 13, 65–73.Google Scholar
  10. Baldani V L D, James E, Baldani J I and Döbereiner J 1992b Localization of the N2-fixing bacteria Herbaspirillum seropedicae within root cells of rice. An. Acad. Bras. Cienc. 64–431.Google Scholar
  11. Bally R, Thomas-Bauzon D, Heulin T and Balandreau J 1983 Determination of the most frequent N2-fixing bacteria in a rice rhizosphere. Can. J. Microbiol. 29, 881–887.Google Scholar
  12. Barber D A and Lynch J M 1977 Microbial growth in the rhizosphere. Soil Biol. Biochem. 9, 305–308.Google Scholar
  13. Barber L E, Tjepkema J D, Russell S A and Evans H J 1976 Acetylene reduction (nitrogen fixation) associated with corn inoculated with Spirillum. Appl. Environ. Microbiol. 32, 108–113.Google Scholar
  14. Barraquio W L, Daroy M L G, Tirol A C, Ladha J K and Watanabe I 1986 Laboratory acetylene reduction assay for relative measurement of N2-fixing activities associated with field-grown wetland rice plants. Plant and Soil 90, 359–372.Google Scholar
  15. Barraquio W L, Guzman M Rde, Barrion M and Watanabe I 1982 Population of aerobic heterotrophic nitrogen fixing bacteria associated with wetland and dryland rice. Appl. Environ. Microbiol. 43, 124–128.Google Scholar
  16. Barraquio W L, Ladha J K and Watanabe I 1983 Isolation and identification of N2-fixing Pseudomonas associated with wetland rice. Can. J. Microbiol. 29, 867–873.Google Scholar
  17. Bennett J and Ladha J K 1992 Introduction: Feasibility of nodulation and nitrogen fixation in rice. In Nodulation and Nitrogen Fixation in Rice. Eds. G S Khush and B Bennett. pp 1–14. IRRI, Manila.Google Scholar
  18. Berkum Pvan and Bohlool B B 1980 Evaluation of nitrogen fixation by bacteria in association with roots of tropical grasses. Microbiol. Rev. 44, 491–517.Google Scholar
  19. Boddey R M 1981 Biological nitrogen fixation in the rhizosphere of lowland rice. PhD thesis, University of the West Indies, St Augustine, Trinidad. 275p.Google Scholar
  20. Boddey R M 1987 Methods for quantification of nitrogen fixation associated with gramineae. CRC Crit. Rev. Plant Sci. 6, 209–266.Google Scholar
  21. Boddey R M 1995 Biological nitrogen fixation in sugar cane: A key to energetically viable bio-fuel production. CRC Crit. Rev. Plant Sci. (In press).Google Scholar
  22. Boddey R M and Ahmad N 1981 Seasonal variations in nitrogenase activity of various rice varieties measured with an in situ acetylene reduction technique in the field. In Associative N2 Fixation. Eds. P B Vose and A P Ruschel. pp 219–229. CRC press. Boca Raton, Florida.Google Scholar
  23. Boddey R M and Döbereiner J 1988 Nitrogen fixation associated with grasses and cereals: recent results and perspectives for future and research. Plant and Soil 108, 53–65.Google Scholar
  24. Boddey R M, Quilt P and Ahmad N 1978 Acetylene reduction in the rhizosphere of rice: methods of assay. Plant and Soil 50, 567–574.Google Scholar
  25. Boddey R M, Baldani V L D, Baldani J I and Döbereiner J 1986 Effect of inoculation of Azospirillum spp. on the nitrogen assimilation of field grown wheat. Plant and Soil 95, 109–121.Google Scholar
  26. Boddey R M, Urquiaga S, Reis V and Döbereiner J 1991 Biological nitrogen fixation associated with sugar cane. Plant and Soil 137, 111–117.Google Scholar
  27. Boreau M 1977 Application de la cromatographie en phase gazeuse á l'etude de l'exudation racinaire du riz. Cah. ORSTOM Sér. Biol. 12, 75–81.Google Scholar
  28. Cavalcante V A and Döbereiner J 1988 A new acid-tolerant nitrogen-fixing bacterium associated with sugarcane. Plant and Soil 108, 23–31.Google Scholar
  29. Chalk P M 1985 Estimation of N2 fixation by isotope dilution: An appraisal of techniques involving 15N enrichment and their application. Soil Biol. Biochem. 17, 389–410.Google Scholar
  30. Cojho E H, Reis V M, Schenberg A C G and Döbereiner J 1993 Interactions of Acetobacter diazotrophicus with an amylolytic yeast in nitrogen-free batch culture. FEMS Microbiol. Lett. 106, 341–346.Google Scholar
  31. Da Silva J G, Serra G E, Moreira J R, Gonçalves J C, Goldemberg J 1978 Energy balance for ethyl alcohol production from crops. Science 210, 903–906.Google Scholar
  32. De P K and Sulaiman M 1950 Influence of algal growth in the rice fields on the yield of crop. Indian J. Agric. Sci. 20, 327–342.Google Scholar
  33. Dénarié J F and Roche P 1991 Rhizobium nodulation signals. In Molecular signals in plant-microbe communications. Ed. D P S Verma. pp. 295–324. CRC Press, Boca Raton, Florida.Google Scholar
  34. Desomer J, Holsters B, Reinhold-Hurek B and Van Monatgu M 1992 Exploring new soil bacteria. In Nodulation and Nitrogen Fixation in Rice. Eds. G S Khush and B Bennett. pp 77–82. IRRI, Manila.Google Scholar
  35. Döbereiner J 1961 Nitrogen-fixing bacteria of the genus Beijerinckia Derx in the rhizosphere of sugar cane. Plant and Soil 15, 211–216.Google Scholar
  36. Döbereiner J and Day J M 1976 Associative symbiosis in tropical grasses: Characterization of microorganisms and dinitrogen fixing sites. In Nitrogen Fixation. Eds. W E Newton and C J N Nyman. pp 518–538. Washington State Univ. Press, Pullman, Washington.Google Scholar
  37. Döbereiner J and Ruschel A P 1958 Uma nova espécie de Beijerinckia. R. Biol. 1, 261–272.Google Scholar
  38. Döbereiner J and Ruschel A P 1961 Inoculação do arroz com bactérias fixadoras de nitrogênio do gênero Beijerinckia Derx. R. Bras. Biol. 21, 397–407.Google Scholar
  39. Döbereiner J, Baldani V L D, Olivares F and de Reis V M 1994 Endophytic diazotrophs: The key to BNF in gramineous plants. In Nitrogen Fixation with Non-Legumes. Eds. N A Hegazi, M Fayez and M Monib. pp 395–408. Am. Univ. in Cairo Press, Cairo, Egypt.Google Scholar
  40. Döbereiner J, Reis V and Lazarine A C 1988 A new N2 fixing bacteria in association with cereals and sugarcane. In Nitrogen Fixation: Hundred years after. Eds. H Bothe, F JDe Bruijn and W E Newton. pp 717–722. Gustav Fischer, Stuttgart.Google Scholar
  41. Dowson-Day M, Ashurst J L, Watts J, Dixon R A and Merrick M J 1991 Studies of the potential for expression of nitrogenase Fe-protein in cells of higher plants. In Nitrogen Fixation. Eds. M Polsinelli, R Materassi and M Vincenzini. pp 659–669. Kluwer Academic Publ., Dordrecht.Google Scholar
  42. Eskew D L, Eaglesham A R J and App A A 1981 Heterotrophic N2 fixation and distribution of newly fixed nitrogen in a rice-flooded soil system. Plant Physiol. 68, 48–52.Google Scholar
  43. Firth P, Thitipoca H, Suthipradit S, Wetselaar R and Beech D F 1973 Nitrogen balance studies in the Central Plain of Thailand. Soil Biol. Biochem. 5, 41–46.Google Scholar
  44. Fuentes-Ramirez L E, Jiminez-Salgado T, Abarca-Ocampo I R and Caballero Mellado J 1993 Acetobacter diazotrophicus, an indolacetic acid-producing bacterium isolated from sugarcane cultivars in Mexico. Plant and Soil 154, 145–150.Google Scholar
  45. Giller K E 1987 Use and abuse of the acetylene reduction assay for measurement of ‘associative’ nitrogen fixation. Soil Biol. Biochem. 19, 783–784.Google Scholar
  46. Gillis M, Döbereiner J, Pot B, Goor M, Falsen E, Hoste B, Reinhold B and Kersters K 1991 Taxonomic relationships between [Pseudomonas] rubrisubalbicans, sorhe clinical isolates (EF group 1), Herbaspirillum seropedicae and [Aquaspirillum] autotrophicum. In Nitrogen Fixation. Eds. M Polsinelli, R Materassi and M Vincenzini. pp 292–294. Kluwer Academic Publ., Dordrecht.Google Scholar
  47. Gillis M, Kerters B, Hoste D J, Kroppenstedt R M, Stephan M P, Teixeira K R S, Döbereiner J and De Ley J 1989 Acetobacter diazotrophicus sp. nov. a nitrogen fixing acetic acid bacterium associated with sugar cane. Int. J. Syst. Bacteriol. 39, 361–364.Google Scholar
  48. Graciolli L A, Freitas J Rde and Ruschel A P 1983 Bactérias fixadoras de nitrogênio nas raizes, caules e folhas de cana-de-açúcar (Saccharum sp.). R. Microbiol. (São Paulo) 14, 191–196.Google Scholar
  49. Hurek T, Reinhold-Hurek B, van Montagu M and Kellenberger E 1991 Infection of intact roots of Kallar grass and rice seedlings by ‘Azoarcus’. Dev. Plant Soil Sci. 48, 235–242.Google Scholar
  50. Ito O, Cabrera D and Watanabe I 1980 Fixation of dinitrogen-15 associated with rice plants. Appl. Environ. Microbiol. 39, 554–558.Google Scholar
  51. James E K, Reis V M, Olivares F L, Baldani J I and Döbereiner J 1994 Infection of sugar cane by the nitrogen-fixing bacterium Acetobacter diazotrophicus. J. Exp. Bot. 45, 757–766.Google Scholar
  52. Kennedy I R and Tchan Y T 1992 Biological nitrogen fixation in non-leguminous field crops: Recent advances. Plant and Soil 141, 93–118.Google Scholar
  53. Koch B L 1977 Associative nitrogenase activity by some Hawaiian grass roots. Plant and Soil 47, 703–706.Google Scholar
  54. Koyama T and App A A 1979 Nitrogen balance in flooded rice soils. In Nitrogen and Rice. pp 95–104, IRRI, Manila.Google Scholar
  55. Ladha J K, Barraquio W L and Watanabe I 1982 Immunological techniques to identify Azospirillum associated with wetland rice. Can. J. Microbiol. 28, 478–485.Google Scholar
  56. Ladha J K, Barraquio W L and Watanabe I 1983 Isolation and identification of nitrogen-fixing Enterobacter clocae and Klebsiella panticola associated with rice plants. Can. J. Microbiol. 29, 1301–1308.Google Scholar
  57. Ladha J K, Tirol A C, Daroy M L G, Caldo G and Watanabe I 1987a Plant-associated N2 fixation (C2H2-reduction) by five rice varieties, and relationship with plant growth characters as affected by straw incorporation. Soil Sci. Plant Nutr. 32, 91–96.Google Scholar
  58. Ladha J K, Tirol Padre A, Punzalan G and Watanabe I 1987b Nitrogen fixing (C2H2-reducing) activity and plant growth characters of 16 wetland rice varieties. Soil Sci. Plant Nutr. 33, 187–200.Google Scholar
  59. Lee K K, Castro T and Yoshida T 1977 Nitrogen fixation throughout growth, and varietal differences in nitrogen fixation by the rhizosphere of rice planted in pots. Plant and Soil 48, 613–619.Google Scholar
  60. Li R and MacRae I C 1992 Specific identification and enumeration of Acetobacter diazotrophicus in sugarcane. Soil Biol. Biochem. 24, 413–419.Google Scholar
  61. Lima E, Boddey R M and Döbereiner J 1987 Quantification of biological nitrogen fixation associated with sugar cane using a 15N aided nitrogen balance. Soil Biol. Biochem. 19, 165–170.Google Scholar
  62. Matsui E, Vose P B, Rodrigues N S R and Ruschel A P 1981 Use of 15N enriched gas to determine N2-fixation by undisturbed sugar cane plant in the field. In Associative N2 Fixation. Vol. 2. Eds. P B Vose and A P Ruschel. pp 153–161. CRC press, Boca Raton, Florida.Google Scholar
  63. Mc Auliffe C, Chamblee D S, Uribe Arango H and Woodhouse W W 1958 Influence of inorganic nitrogen on nitrogen fixation by legumes as revealed by N15. Agron. J. 50, 334–337.Google Scholar
  64. Nayak D N, Ladha J K and Watanabe I 1986 The fate of marker Azospirillum lipoferum inoculated into rice and its effect on growth, yield and N2 fixation of plants studies by acetylene reduction, 15N2 feeding and 15N dilution techniques. Biol. Fertil. Soils 2, 7–14.Google Scholar
  65. Ohta H and Hattori T 1983 Agromonas oligotrophica gen. nov., sp. nov., a nitrogen-fixing oligotrophic bacterium. Antonie Leeuwenhoek 49, 429–446.Google Scholar
  66. Olivares F L, Baldani V L D, Baldani J I and Döbereiner J 1993 Ecology of Herbaspirillum spp. and ways of infection and colonization of cereals with these endophytic diazotrophs. In Poster at 6th Int. Symp. Nitrogen Fixation with Non-legumes. Ismailia, Egypt. 6–10 Sept. Program and Abstracts, 118p.Google Scholar
  67. Oliveira E 1992 Estudo da associação entre bactérias diazotróficas e arroz. MSc Thesis, Univ. Fed. Rural Rio de Janeiro, Itaguai, RJ.Google Scholar
  68. Oliveira O C de 1994 Quantificação da fixaçao biológica de nitrogênio em arroz (Oryza sativa, L.) inundado. MSc Thesis, Univ. Fed. Rural Rio de Janeiro, Itaguaí, RJ.Google Scholar
  69. Oliveira O Cde, Urquiaga S and Boddey R M 1994 Burning cane: the long term effects. Int. Sugar J. 96, 272–275.Google Scholar
  70. Omar A M N, Richard C, Weinhard P and Balandreau J 1989 Using the spermosphere model technique to describe the dominant nitrogen-fixing microflora associated with wetland rice in two Egyptian soils. Biol. Fertil. Soils 7, 158–163.Google Scholar
  71. Orlando Filho J, Haag H P and Zambello EJr 1980 Crescimento e absorção de macronutrientes pela cana-de-açúcar, variedade CB41–76, em função da idade, em solos do Estado de São Paulo. Bol. Tec. Planalsucar, Piracicaba, São Paulo 2, 1–128.Google Scholar
  72. Paula M A de, Döbereiner J and Siqueira J O 1989 Efeito da inoculaçao com fungo micorrizico VA e bactérias diazotróficas no crescimento e produçao de batata-doce. In 22nd Congresso Brasileiro de Ciência do Solo, Recife, SBCS. pp 109. Programa e resumos.Google Scholar
  73. Paula M Ade, Reis V M and Döbereiner J 1991 Interactions of Glomus clarum with Acetobacter diazotrophicus in infection of sweet potato (Ipomoea batatas), sugarcane (Sacchrum spp.) and sweet sorghum (Sorghum vulgare). Biol. Fertil. Soils 11, 111–115.Google Scholar
  74. Pimentel J P, Olivares F, Pitard R M, Urquiaga S, Akiba F and Döbereiner J 1991 Dinitrogen fixation and infection of grass leaves by Pseudomonas rubrisubalbicans and Herbaspirillum seropedicae. Plant and Soil 137, 61–65.Google Scholar
  75. Purchase B S 1980 Nitrogen fixation associated with sugarcane, Proc. S. Afr. Sugar Technol. Assoc., June. pp 173–176.Google Scholar
  76. Qui Y S, Zhou S P and Mo X Z 1981 Study of nitrogen fixing bacteria associated with rice root. 1. Isolation and identification of organisms. Acta Microbiol. Sinica 21, 468–472.Google Scholar
  77. Reinhold-Hurek B, Hurek T, Gillis M, Hoste B, Vancanneyt M, Kersters K and De Ley J 1993 Azoarcus gen. nov., a nitrogen fixing Proteobacteria associated with roots of Kallar grass (Leptochloa fusca (L.) Kunth), and description of two species Azoarcus indigens sp. nov. and Azoarcus communis sp. nov. Int. J. Syst. Bacteriol. 43, 574–588.Google Scholar
  78. Reis V M, Olivares F L and Döbereiner J 1994 Improved methodology for isolation of Acetobacter diazotrophicus and confirmation of its endophytic habitat. World J. Microbiol. Biotechnol. 10, 101–104.Google Scholar
  79. Reis V M, Zang Y and Burris R H 1990 Regulation of nitrogenase activity by ammonium and oxygen in Acetobacter diazotrophicus, An. Acad. Bras. Cienc. 62, 317.Google Scholar
  80. Rinaudo G and Dommergues Y 1971 Validité de l'estimation de la fixation biologique de l'azote dans la rhizosphere par la methode de reduction de l'acetylene. Ann. Inst. Pasteur 121, 93–99.Google Scholar
  81. Roger P A and Watanabe I 1986 Technologies for utilizing biological nitrogen fixation in wetland rice: potentialities, current usage, and limiting factors. Fert. Res. 9, 39–77.Google Scholar
  82. Rolfe B G and Bender G L 1990 Evolving a Rhizobium for non-legume nodulation. In Nitrogen Fixation: Achievements and Objectives. Eds. P M Gresshof, L E Roth, G Stacey and W L Newton. pp 779–780. Chapman and Hall, New York.Google Scholar
  83. Ruschel A P, Henis Y and Salati E 1975 Nitrogen-15 tracing of N-fixation with soil-grown sugar cane seedlings. Soil Biol. Biochem. 7, 181–182.Google Scholar
  84. Ruschel A P, Victoria R L, Salati E and Henis Y 1978 Nitrogen fixation in sugar cane (Saccharum officinarum). In Environmental Role of Nitrogen-fixing Blue-green Algae and Asymbiotic Bacteria. Ed. U Granhall. pp 297–303. Ecological Bulletins, Swedish Natural, Stockholm. (Ecological Bulletins/NFR 26).Google Scholar
  85. Sampaio E V S B, Salgado I H and Bettany J 1984 Dinâmica de nutrientes em cana-de-açúcar. I. Eficiência da utilizaçao de uréia (15N) em aplicaçao única ou parcelada. Pesq. Agropec. Bras. 19, 943–949.Google Scholar
  86. Sen J 1929 Is bacterial association a factor in nitrogen assimilation by rice plants? Agric. J. India 24, 229–231.Google Scholar
  87. Stanford G and Ayres A S 1964 The internal nitrogen requirement of sugarcane. Soil Sci. 98, 338–344.Google Scholar
  88. Stephan M P, Oliveira M, Teixeira K R S, Martinez Dretz G and Döbereiner J 1991 Physiology and dinitrogen fixation of Acetobacter diazotrophicus. FEMS Microbiol. Lett. 77, 67–72.Google Scholar
  89. Teixeira K R S, Stephan M P and Döbereiner J 1987 Physiological studies of Saccharobacter nitrocaptans a new acid tolerant N2-fixing bacterium. In Poster presented at 4th International Symposium on Nitrogen Fixation with Non-legumes, Rio de Janeiro. Final program abstracts.Google Scholar
  90. Tjepkema J and Van Berkum P 1977 Acetylene reduction by soil cores of maize and sorghum in Brazil. Appl. Environ. Microbiol. 33, 626–629.Google Scholar
  91. Urquiaga S, Cruz K H S and Boddey R M 1992 Contribution of nitrogen fixation to sugar cane: Nitrogen-15 and nitrogen balance estimates. Soil Sci. Soc. Am. J. 56, 105–114.Google Scholar
  92. Ventura T S, Bravo M, Daez C, Ventura V, Watanabe I and App A 1986 Effects of N-fertilizers, straw, and dry fallow on the nitrogen balance of a flooded soil planted with rice. Plant and Soil 93, 405–411.Google Scholar
  93. Ventura W and Watanabe I 1983 15N dilution technique of assessing the contribution of nitrogen fixation to rice plant. Soil Sci. Plant Nutr. 29, 123–130.Google Scholar
  94. Walcott J J, Chauviroj M, Chinchest A, Choticheuy P, Ferraris R and Norman B W 1977 Long-term productivity of intensive rice cropping systems on the central plain of Thailand. Exp. Agric. 13, 305–316.Google Scholar
  95. Watanabe I, Deguzman M R and Cabrera D A 1981 The effect of nitrogen fertilizer on N2 fixation in the paddy field measured by in situ acetylene reduction assay. Plant and Soil 59, 135–140.Google Scholar
  96. Watanabe I, Lee K and Guzman Mde 1978a Seasonal change of N2 fixing rate in rice field assayed by in situ acetylene reduction technique. II. Estimate of nitrogen fixation associated with rice plants. Soil Sci. Plant Nutr. 24, 465–471.Google Scholar
  97. Watanabe I, Lee K K and Alimagno B V 1978b Seasonal change of N2-fixing rate in rice field assayed by in situ acetylene reduction technique. I. Experiments in long-term fertility plots. Soil Sci. Plant Nutr. 24, 1–13.Google Scholar
  98. Watanabe I, Yoneyama T, Padre B and Ladha J K 1987a Difference in natural abundance of 15N in several rice (Oryza sativa L.) varieties: Applications for evaluating N2 fixation. Soil Sci. Plant Nutr. 33, 407–415.Google Scholar
  99. Watanabe I, So R, Ladha J K, Fujimura Y, Katayama and Kuraishi H 1987b A new nitrogen-fixing species of pseudomonad: Pseudomonas diazotrophicus sp. nov. isolated from the root of wetland rice. Can. J. Microbiol. 33, 670–678.Google Scholar
  100. Willis W H and Green V EJr 1948 Movement of nitrogen in flooded soil planted to rice. Soil Sci. Soc. Am. Proc. 13, 229–237.Google Scholar
  101. Witty J F 1983 Estimating N2 fixation in the field using 15N-labelled fertilizer: some problems and solutions. Soil Biol. Biochem. 15, 631–639.Google Scholar
  102. Yoshida T and Ancajas R R 1970 Application of the acetylene reduction method in nitrogen fixation studies. Soil Sci. Plant Nutr. 16, 234–237.Google Scholar
  103. Yoshida T and Ancajas R R 1973 Nitrogen-fixing activity in upland and flooded rice fields. Soil Sci. Soc. Am. Proc. 37, 42–46.Google Scholar
  104. Yoshida T and Yoneyama T 1980 Atmospheric dinitrogen fixation in the flooded rice rhizosphere as determined by the N-15 isotope technique. Soil Sci. Plant Nutr. 26, 551–560.Google Scholar

Copyright information

© Kluwer Academic Publishers 1995

Authors and Affiliations

  • R. M. Boddey
    • 1
  • O. C. de Oliveira
    • 1
  • S. Urquiaga
    • 1
  • V. M. Reis
    • 1
  • F. L. de Olivares
    • 1
  • V. L. D. Baldani
    • 1
  • J. Döbereiner
    • 1
  1. 1.EMBRAPA-Centro Nacional de Pesquisa de AgrobiologiaRio de JaneiroBrazil

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