Skip to main content
Log in

Endophytic nitrogen fixation in sugarcane: present knowledge and future applications

  • Published:
Plant and Soil Aims and scope Submit manuscript

Abstract

In Brazil the long-term continuous cultivation of sugarcane with low N fertiliser inputs, without apparent depletion of soil-N reserves, led to the suggestion that N2-fixing bacteria associated with the plants may be the source of agronomically significant N inputs to this crop. From the 1950s to 1970s, considerable numbers of N2-fixing bacteria were found to be associated with the crop, but it was not until the late 1980s that evidence from N balance and 15N dilution experiments showed that some Brazilian varieties of sugarcane were able to obtain significant contributions from this source. The results of these studies renewed the efforts to search for N2-fixing bacteria, but this time the emphasis was on those diazotrophs that infected the interior of the plants. Within a few years several species of such `endophytic diazotrophs' were discovered including Gluconacetobacter diazotrophicus, Herbaspirillum seropedicae, H. rubrisubalbicansand Burkholderia sp. Work has continued on these endophytes within sugarcane plants, but to date little success has been attained in elucidating which endophyte is responsible for the observed BNF and in what site, or sites, within the cane plants the N2 fixation mainly occurs. Until such important questions are answered further developments or extension of this novel N2-fixing system to other economically important non-legumes (e.g. cereals) will be seriously hindered. As far as application of present knowledge to maximise BNF with sugarcane is concerned, molybdenum is an essential micronutrient. An abundant water supply favours high BNF inputs, and the best medium term strategy to increase BNF would appear to be based on cultivar selection on irrigated N deficient soils fertilised with Mo.

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

Access this article

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

Instant access to the full article PDF.

Similar content being viewed by others

References

  • App A, Santiago T, Daez C, Menguito C, Ventura W, Tirol A, Po J, Watanabe I, Datta S K de 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 

  • Arias O E, Gatti I M, Silva D M, Ruschel A P and Vose P B 1978 Primeiras observaciones al microscópio eletrónico de bacterias fijadoras de N2 en la raiz de la canã de azúcar (Saccharum officinarum L.) Turrialba 28, 203–207.

    Google Scholar 

  • Baldani V L D, Alvarez M A de B, Baldani J I and Döbereiner J 1986a Establishment of inoculated Azospirillum spp. in the rhizosphere and in roots of field grown wheat and sorghum. Plant Soil 90, 35–46

    Google Scholar 

  • Baldani J I, Baldani V L D, Seldin L and Döbereiner J 1986b Characterization of Herbaspirillum seropedicae gen. nov., sp. nov., a root-associated nitrogen-fixing bacterium. Int. J. Syst. Bacteriol. 36, 86–93.

    Google Scholar 

  • Baldani J I, Pot B, Kirchhof G, Falsen E, Baldani V L D, Olivares F L, Hoste B, Kersters K, Hartmann A, Gillis M and Döbereiner J 1996 Emended description of Herbaspirillum; inclusion of (Pseudomonas) rubrisubalbicans, a mild plant pathogen, as Herbaspirillum rubrisubalbicans comb. nov. and classification of a group of clinical isolates (EF group 1) as Herbaspirillum species 3. Int. J. Syst. Bacteriol. 46, 802–810.

    PubMed  Google Scholar 

  • Baldani J I, Caruso L, Baldani V L D, Goi S R and Döbereiner J 1997 Recent advances in BNF with non-legume plants. Soil Biol. Biochem. 29, 911–922.

    Google Scholar 

  • Baldani V L D, Baldani J I and Döbereiner J 1983 Effects of Azospirillum inoculation on root infection and nitrogen incorporation in wheat. Can. J. Microbiol. 29, 924–929.

    Google Scholar 

  • Baldani V L D, Baldani J I and Döbereiner J 1987 Inoculation of field-grown wheat (Triticum aestivum) with Azospirillum spp. in Brazil. Biol. Fertil. Soils. 4(1), 37–40.

    Google Scholar 

  • Baldani V L D, Reis V M, Stoffels M, Kirchhof G, Schmid M, Boddey L H, Baldani J I, Oliveira E de, Dobereiner J and Hartmann A 2002 Description of two new nitrogen-fixing, plant-associated bacterial species in the genus Burkholderia, Burkholderia brasilensis sp. nov. and Burkholderia tropicalis sp. nov. Int. J. Syst. Evol. Microb. (in press).

  • Boddey L H, Dart P J, Goi S R and Baldani J I 1998 Ocorrência de bactérias diazotróficas endofíticas no cultivar Q151 de cana-deaçúcar cultivada na Austrália. In Proc. V Simpósio Brasileiro de Microbiologia do Solo, FERTBIO 98, Caxambu, MG, 11 a 16 de Outubro, 1998, Abstract No 772.

  • Boddey R M 1993 'Green' Energy from Sugar Cane. Chemistry and Industry (London) 17 May, No. 10, pp. 355–358.

    Google Scholar 

  • Boddey R M, Peoples M B, Palmer B and Dart P J 2000 Use of the 15N natural abundance technique to quantify biological nitrogen fixation by woody perennials Nutrient Cycl. Agroecosyst. 57, 235–270.

    Google Scholar 

  • Boddey R M, Polidoro J C, Resende A S, Alves B J R and Urquiaga S 2001 Use of the 15N natural abundance technique for the quantification of the contribution of N2 fixation to grasses and cereals. Aust. J. Plant Physiol. 28, 889–895.

    Google Scholar 

  • Christopher W N and Edgerton C W 1932 Bacterial stripe diseases of sugarcane in Louisiana. J. Agric. Res. 41, 259.

    Google Scholar 

  • Döbereiner J 1961 Nitrogen-fixing bacteria of the genus Beijerinckia Derx in the rhizosphere of sugar cane. Plant Soil 15, 211–216.

    Google Scholar 

  • Döbereiner J 1988 Isolation and identification of root associated diazotrophs. Plant Soil 110, 207–212.

    Google Scholar 

  • Döbereiner J 1992 History and new perspectives of diazotrophs in association with non-leguminous plants. Symbiosis 13, 1–13.

    Google Scholar 

  • Döbereiner J and Ruschel A P 1958 Uma nova espécie de Beijerinkia. Rev. Biol. 1, 261–272.

    Google Scholar 

  • Franke I H, Fegan M, Hayward C, Leonard G, Stackebrandt E and Sly L I 1999 Description of Gluconacetobacter sacchari sp. nov., a new species of acetic acid bacterium isolated from the leaf sheath of sugar cane and from the pink sugar-cane mealy bug. Int. J. Syst. Bacteriol. 49, 1681–1693.

    PubMed  Google Scholar 

  • Fuentes-Ramirez L E, Caballero-Mellado J, Sepúlveda J and Martínez-Romero E 1999. Colonization of sugarcane by Gluconacetobacter diazotrophicus is inhibited by high N-fertilization. FEMS Microbiol. Ecol. 29, 117–128.

    Google Scholar 

  • Fuentes-Ramirez L E, Bustillos-Cristales R, Tapia-Hernández A, Jiménez-Salgado T, Wang E T, Martinez-Romero E and Caballero-Mellado J 2001 Novel nitrogen-fixing acetic acid bacteria Gluconacetobacter johannae sp. nov. and Gluconacetobacter azotocaptans sp. nov., associated with coffee plants. Int. J. Syst. Evol. Microb. 51, 1305–1314.

    Google Scholar 

  • Galloway J H 1989 The Sugar Cane Industry: An Historical Geography from its Origins to 1914. Cambridge Univ. Press, UK.

    Google Scholar 

  • Gillis M, Kerters K, Hoste B, Janssens D, Kroppenstedt R M, Stephan M P, Teixeira K R S, Döbereiner J and Deley J 1989 Gluconacetobacter diazotrophicus sp. nov. a nitrogen fixing acetic acid bacterium associated with sugar cane. Int. J. Syst. Bacteriol. 39, 361–364.

    Google Scholar 

  • Gillis M, Döbereiner J, Pot B, Goor M, Falsen E, Hoste B, Reinhold B and Kersters K 1991 Taxonomy relationship between 'Pseudomonas' rubrisubalbicans, some clinical isolates (EF group 1), Herbaspirillum seropedicae and 'Aquaspirillum' autotrophicuim. In Nitrogen Fixation. Eds. M Polsinelli, R Materassi, M Vicenzini. pp. 292–294. Kluwer Acad. Publ. Dordrecht, Netherlands.

    Google Scholar 

  • Gonzalez M S and Barraquio W L 2000 Isolation and characterization of Gluconacetobacter diazotrophicus (Gillis) in Saccharum officinarum L.,S. spontaneum L., and Erianthus sp. The Philippine Agricultural Scientist 83, 173–181.

    Google Scholar 

  • Graciolli L A, Freitas J R de and Ruschel A P 1983 Bactérias fixadoras de nitrogênio nas raízes, caules e folhas de cana-de-açúcar (Saccharum sp.). Rev. Microbiol. 14, 191–196.

    Google Scholar 

  • Handley L L and Scrimgeour C M. Terrestrial plant ecology and 15N natural abundance: The present limits to interpretation for uncultivated systems with original data from a Scottish old field. Adv. Ecol. Res. 27, 133–212.

  • Hegazi N A, Eid M, Farag R S and Monib M 1979 Asymbiotic N2-fixation in the rhizosphere of sugar cane planted under semi-arid conditions of Egypt. Rev. Ecol. Biol. Sol. 16(1), 23–37.

    Google Scholar 

  • Hendre R R, Iyor R S, Kotwalm M, Kluspe S S and Mascarenhas A F 1983 Rapid multiplication of sugar cane by tissue culture. Sugar Cane 1, 5–8.

    Google Scholar 

  • Högberg P 1997 Tansley review no 95. 15N natural abundance in soil-plant systems. New Phytol. 137, 179–203.

    Google Scholar 

  • James E K and Olivares F L 1998 Infection and colonization of sugar cane and other graminaceous plants by endophytic diazotrophs. Crit. Rev. Plant Sci. 17, 77–119.

    Google Scholar 

  • James E K, Olivares F L, Baldani J I and Döbereiner J 1997 Herbaspirillum, an endophytic diazotroph colonising vascular tissue in leaves of Sorghum bicolor L. Moench. J. Exp. Bot. 48, 785–797.

    Google Scholar 

  • 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 Gluconacetobacter diazotrophicus. J. Exp. Bot. 45, 757–766.

    Google Scholar 

  • Jimenez-Salgado T, Fuentes-Ramirez L E, Tapia-Hernandez A, Mascarua-Esparza M A, Martinez-Romero E and Caballero-Mellado J 1997 Coffea arabica L., a new host plant for Gluconacetobacter diazotrophicus and isolation of other nitrogenfixing Gluconacetobacteria. Appl. Environ. Microbiol. 63, 3676–3683.

    PubMed  Google Scholar 

  • 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 

  • Macedo I de C 1998 Greenhouse gas emissions and energy balances in bio-ethanol production and utilization in Brazil (1996). Biomass Bioenergy 14, 77–81.

    Google Scholar 

  • Machado G R, da Silva W M and Irvine J E 1987 Sugar cane breeding in Brazil: the Copersucar Program. In Copersucar International Sugarcane Breeding Workshop, Cooperativa de Produtores de Cana, Açúcar e Álcool do Estado de São Paulo Ltda, São Paulo, SP, Brazil. pp. 215–232.

  • Matsui E, Vose P B, Rodrigues N S 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. Eds. P B Vose and A P Ruschel. Vol II, pp. 153–161. CRC press, Boca Raton, Florida, USA.

    Google Scholar 

  • Muthukumarasamy R, Revathi G and Lakshminarasimhan C 1999 Influence of N fertilisation on the isolation of Gluconacetobacter diazotrophicus and Herbaspirillum spp. from Indian sugarcane varieties. Biol. Fertil. Soil, 29, 157–164.

    Google Scholar 

  • Olivares F L, James E K, Baldani J I and Döbereiner J 1997 Infection of mottled stripe disease susceptible and resistant varieties of sugar cane by the endophytic diazotroph Herbaspirillum. New Phytol. 135, 723–737.

    Google Scholar 

  • Oliveira A L M, Urquiaga S, Dobereiner J and Baldani J I 2002 The effect of inoculating endophytic N2-fixing bacteria on micropropagated sugarcane plants. Plant Soil. 242, 205–215.

    Google Scholar 

  • Oliveira O C, Urquiaga S and Boddey R M 1994 Burning cane: the long term effects. Int. Sug. J. 96, 272–275.

    Google Scholar 

  • Paula M A de, Döbereiner J and Siqueira J O 1989 Nutrição e produção de batata-doce micropropagada e inoculada com fungo micorrízico VA e bactérias diazotróficas</del>. In Reuniao Brasileira Sobre Micorrizas. 3, 1989. Anais. Piracicaba, USP/CENA, 1989. p 26.

    Google Scholar 

  • Peoples M B, Palmer B, Lilley D M, Duc L M and Herridge D F 1996 Application of N-15 and xylem ureide methods for assessing N2 fixation of three shrub legumes periodically pruned for forage. Plant Soil 182, 125–137.

    Google Scholar 

  • Peoples M B, Turner G L, Shah Z, Shah S H, Aslam M, Ali S, Markey S L, Afandi F, Schwenke G D & Herridge D F 1997 Evaluation of the 15N natural abundance technique for measuring N2 fixation in experimental plots and farmer's fields. In Proceedings of an International Workshop on Managing Legume Nitrogen Fixation in Cropping Systems of Asia. Eds. O P Rupela, C Johansen and D F Herridge. pp. 57–75. ICRISAT, Hyderabad, India.

    Google Scholar 

  • Purchase B S 1980 Nitrogen fixation associated with sugarcane. In Proceedings of the South African Sugar Technologists Association: 173–176.

  • 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 

  • Reis V M, Olivares F L, Oliveira A L M, Reis Junior F B, Baldani J I and Dobereiner J 1999 Technical approaches to inoculate micropropagated sugarcane plants with Acetobacter diazotrophicus. Plant Soil 206, 205–211.

    Google Scholar 

  • Reis V M, Baldani J I, Baldani V L D and Döbereiner J 2000 Biological dinitrogen fixation in Gramineae and palm trees. Crit. Rev. Plant Sci. 19, 227–247.

    Google Scholar 

  • Reis Junior F B, Silva L G, Reis V M and Dobereiner J 2000a Ocorrência de bactérias diazotróficas em diferentes genótipos de cana-de-açúcar. Pesquisa Agropecuária Brasileira. 35, 985–994.

    Google Scholar 

  • Reis Junior F B dos, Reis V M, Urquiaga S and Döbereiner J 2000b Influence of nitrogen fertilisation on the population of diazotrophic Herbaspirillum spp. and Gluconacetobacter diazotrophicus in sugar cane (Saccharum spp.) Plant Soil 219, 153–159.

    Google Scholar 

  • Rennie R J, Freitas J R d, Ruschel A P and Vose P B 1982 Isolation and identification of N2-fixing bacteria associated with sugar cane (Saccharum sp.). Can. J. Microbiol. 28, 462–467.

    Google Scholar 

  • Robertson S, Kingston G, Dart P J and Brown S 2000 Occurrence of Endophytic Bacteria in Australian Sugarcane. Poster Presented at 8th Int. Symp Nitrogen Fixation with Non-Legumes, 3–7 Dec, Sydney, Australia. Abstract book. p. 140.

  • 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 

  • Seldin L, Van Elsas J D and Penido E G C 1984 Bacillus azotofixans sp. nov., a nitrogen-fixing species from Brazilian soils and grass roots. Int. J. Syst. Bacteriol. 34, 451–456.

    Google Scholar 

  • Sevilla M, Burris R H, Gunapala N and Kennedy C 2001 Comparison of benefit to sugarcane plant growth and 15N2 incorporation following inoculation of sterile plants with Acetobacter diazotrophicus wild-type and mutant strains. Molec. Plant-Microbe Interact. 14, 358–366.

    Google Scholar 

  • Shearer G and Kohl D H 1986 N2-fixation in field settings: estimations based on natural 15N abundance. Aust. J. Plant Physiol. 13, 699–756.

    Google Scholar 

  • Stephan M P, Oliveira M, Teixeira K R S, Martinez Drets G and Döbereiner J 1991 Physiology and dinitrogen fixation of Acetobacter diazotrophicus. FEMS. Microbiol. Lett. 77, 67–72.

    Google Scholar 

  • Tapia-Hernandez A, Bustillos-Cristales M R, Jimenez-Salgado T, Caballero-Mellado J and Fuentes-Ramirez L E 2000 Natural endophytic occurrence of Gluconacetobacter diazotrophicus in pineapple plants. Microb. Ecol. 39, 49–55.

    PubMed  Google Scholar 

  • Unkovich M J, Pate J S, Sanford P & Armstrong E L 1994 Potential precision of the 15N natural abundance method in field estimates of nitrogen fixation by crop and pasture legumes in south-west Australia. Aust. J. Agric. Res. 45, 119–132.

    Google Scholar 

  • 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 

  • Ventura T S, Bravo M, Daez C, Ventura V, Watanabe I and App A A 1986 Effects of N fertilizers, straw, and dry fallow on the nitrogen balance of a flooded soil planted with rice. Plant Soil 93, 405–411.

    Google Scholar 

  • 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. Experimental Agriculture 13, 305–316.

    Google Scholar 

  • Yoneyama T, Muraoka T, Kim T H, Dacanay E V and Nakanishi Y 1997 The natural 15N abundance of sugarcane nad neighbouring plants in Brazil, the Philippines and Miyako (Japan). Plant Soil 189, 239–244.

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Robert M. Boddey.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Boddey, R.M., Urquiaga, S., Alves, B.J. et al. Endophytic nitrogen fixation in sugarcane: present knowledge and future applications. Plant and Soil 252, 139–149 (2003). https://doi.org/10.1023/A:1024152126541

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1024152126541

Navigation