Free Standard AU & NZ Shipping For All Book Orders Over $80!
Register      Login
Crop and Pasture Science Crop and Pasture Science Society
Plant sciences, sustainable farming systems and food quality
RESEARCH ARTICLE

Nitrogen cycling in summer active perennial grass systems in South Australia: non-symbiotic nitrogen fixation

V. V. S. R. Gupta A C , S. J. Kroker A , M. Hicks A , C. W. Davoren A , K. Descheemaeker A B and R. Llewellyn A
+ Author Affiliations
- Author Affiliations

A CSIRO Agriculture Flagship, PMB No. 2, Glen Osmond, SA 5064, Australia.

B Land Production Systems, Wageningen University, PO Box 430, 6700 AK, Wageningen, The Netherlands.

C Corresponding author. Email: Gupta.Vadakattu@csiro.au

Crop and Pasture Science 65(10) 1044-1056 https://doi.org/10.1071/CP14109
Submitted: 8 April 2014  Accepted: 30 May 2014   Published: 7 October 2014

Abstract

Non-symbiotic nitrogen (N2) fixation by diazotrophic bacteria is a potential source for biological N inputs in non-leguminous crops and pastures. Perennial grasses generally add larger quantities of above- and belowground plant residues to soil, and so can support higher levels of soil biological activity than annual crops. In this study, the hypothesis is tested that summer-active perennial grasses can provide suitable microsites with the required carbon supply for N2 fixation by diazotrophs, in particular during summer, through their rhizosphere contribution. In a field experiment on a Calcarosol at Karoonda, South Australia, during summer 2011, we measured populations of N2-fixing bacteria by nifH-PCR quantification and the amount of 15N2 fixed in the rhizosphere and roots of summer-active perennial grasses. Diazotrophic N2 fixation estimates for the grass roots ranged between 0.92 and 2.35 mg 15N kg–1 root day–1. Potential rates of N2 fixation for the rhizosphere soils were 0.84–1.4 mg 15N kg–1 soil day–1 whereas the amount of N2 fixation in the bulk soil was 0.1–0.58 mg 15N kg–1 soil day–1. Populations of diazotrophic bacteria in the grass rhizosphere soils (2.45 × 106 nifH gene copies g–1 soil) were similar to populations in the roots (2.20 × 106 nifH gene copies g–1 roots) but the diversity of diazotrophic bacteria was significantly higher in the rhizosphere than the roots. Different grass species promoted the abundance of specific members of the nifH community, suggesting a plant-based selection from the rhizosphere microbial community. The results show that rhizosphere and root environments of summer-active perennial grasses support significant amounts of non-symbiotic N2 fixation during summer compared with cropping soils, thus contributing to biological N inputs into the soil N cycle. Some pasture species also maintained N2 fixation in October (spring), when the grasses were dormant, similar to that found in soils under a cereal crop. Surface soils in the rainfed cropping regions of southern Australia are generally low in soil organic matter and thus have lower N-supply capacity. The greater volume of rhizosphere soil under perennial grasses and carbon inputs belowground can potentially change the balance between N immobilisation and mineralisation processes in the surface soils in favour of immobilisation, which in turn contributes to reduced N losses from leaching.

Additional keywords: free-living bacteria, nifH, nitrogen fixation, non-symbiotic, microbial activity, perennial grasses.


References

Angus JF, Gault RR, Peoples MB, Stapper M, van Herwaarden AF (2001) Soil water extraction by dryland crops, annual pastures and lucerne in south-eastern Australia. Australian Journal of Agricultural Research 52, 183–192.
Soil water extraction by dryland crops, annual pastures and lucerne in south-eastern Australia.Crossref | GoogleScholarGoogle Scholar |

Berendsen RL, Pieterse CM, Bakker PAHM (2012) The rhizosphere microbiome and plant health. Trends in Plant Science 17, 478–486.
The rhizosphere microbiome and plant health.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC38Xms12rs70%3D&md5=9cb38aa8779505b244b39de094e00748CAS | 22564542PubMed |

Bowen GD, Rovira AD (1999) The rhizosphere and its management to improve plant growth. Advances in Agronomy 66, 1–102.
The rhizosphere and its management to improve plant growth.Crossref | GoogleScholarGoogle Scholar |

Buckley DH, Huangyutitham V, Hsu S-F, Nelson TA (2007) Stable isotope probing with 15N2 reveals novel noncultivated diazotrophs in soil. Applied and Environmental Microbiology 73, 3196–3204.
Stable isotope probing with 15N2 reveals novel noncultivated diazotrophs in soil.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2sXlslSnuro%3D&md5=7a88bb95b6e348e363f20978f13a7546CAS | 17369332PubMed |

Bürgmann H, Widmer F, Sigler WV, Zeyer J (2004) New molecular screening tools for analysis of free-living diazotrophs in soil. Applied and Environmental Microbiology 70, 240–247.
New molecular screening tools for analysis of free-living diazotrophs in soil.Crossref | GoogleScholarGoogle Scholar | 14711647PubMed |

Chowdhury SP, Schmid M, Hartmann A, Tripathi AK (2009) Diversity of 16S-rRNA and nifH genes derived from rhizosphere soil and roots of an endemic drought tolerant grass, Lasiurus sindicus. European Journal of Soil Biology 45, 114–122.
Diversity of 16S-rRNA and nifH genes derived from rhizosphere soil and roots of an endemic drought tolerant grass, Lasiurus sindicus.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD1cXhsFahurjO&md5=3881419738062e145e89fba6a163a504CAS |

Clarke KR, Ainsworth M (1993) A method of linking multivariate community structure to environmental variables. Marine Ecology Progress Series 92, 205–219.
A method of linking multivariate community structure to environmental variables.Crossref | GoogleScholarGoogle Scholar |

Cleveland CC, Townsend AR, Schimel DS, Fisher H, Howarth RW, Hedin LO, Perakis SS, Latty EF, Von Fischer JC, Elseroad A, Wasson MF (1999) Global patterns of terrestrial biological nitrogen (N2) fixation in natural ecosystems. Global Biogeochemical Cycles 13, 623–645.
Global patterns of terrestrial biological nitrogen (N2) fixation in natural ecosystems.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK1MXkslOru7s%3D&md5=4ca182c640928a8c9689dac042268b5bCAS |

Coelho MRR, Marriel IE, Jenkins SN, Lanyon SC, Seldin L, O’Donnel AG (2009) Molecular detection and quantification of nifH gene sequences in the rhizosphere of sorghum (Sorghum bicolor) sown with two levels of nitrogen fertilizer. Applied Soil Ecology 42, 48–53.
Molecular detection and quantification of nifH gene sequences in the rhizosphere of sorghum (Sorghum bicolor) sown with two levels of nitrogen fertilizer.Crossref | GoogleScholarGoogle Scholar |

Coleman DC, Crossley DA, Jr, Hendrix PF (2004) ‘Fundamentals of soil ecology.’ 2nd edn (Elsevier: San Diego, CA, USA)

Dear BS, Ewing MA (2008) The search for new pasture plants to achieve more sustainable production systems in southern Australia. Australian Journal of Experimental Agriculture 48, 387–396.
The search for new pasture plants to achieve more sustainable production systems in southern Australia.Crossref | GoogleScholarGoogle Scholar |

Descheemaeker K, Llewellyn R, Moore A, Whitbread A (2014) Summer-growing perennial grasses are a potential new feed source in the low rainfall environment of southern Australia. Crop & Pasture Science 65, 1033–1043.

Ellis SL, Ryan MH, Angus JF, Pratley JE (2008) Soil nitrogen and water dynamics in crops following perennial pastures under drought conditions. In ‘Global Issues Paddock Action. Proceedings 14th Australian Agronomy Conference’. September 2008, Adelaide, S. Aust. (Ed. MJ Unkovich) (Australian Society of Agronomy/The Regional Institute Ltd: Gosford, NSW) Available at: www.regional.org.au/au/asa/2008/concurrent/assessing-yield-potential/5928_ellissl.htm

Gaby JC, Buckley DH (2011) A global census of nitrogenase diversity. Environmental Microbiology 13, 1790–1799.
A global census of nitrogenase diversity.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3MXhtVajur3K&md5=8de31b6f211583138f17fc5522776988CAS | 21535343PubMed |

Grace PR, Ladd JN, Skjemstad JO (1994) The effect of management practices on soil organic matter dynamics. In ‘Soil biota: management in sustainable farming systems’. (Eds CE Pankhurst, BM Doube, VVSR Gupta) pp. 162–171. (CSIRO Publishing: Melbourne)

Gregory PJ, Atwell BJ (1991) The fate of carbon in pulse-labelled crops of barley and wheat. Plant and Soil 136, 205–213.

Gupta VVSR (2012) Diversity and functional capability of free-living N fixing bacteria in southern Australian soils. In ‘The power of the small. 14th International Symposium of Microbial Ecology’. 19–24 August, Copenhagen, Denmark. (ISME: Wageningen, The Netherlands)

Gupta VVSR, Hicks M (2011) Diversity and activity of free-living bacteria in South Australian soils. In ‘Rhizosphere 3 International Conference’. 25–30 September, Perth, W. Aust. (University of Western Australia: Perth) Available at: http://rhizosphere3.com/conference-program/Rhizosphere/pdf/RHIZO_Program.pdf

Gupta VVSR, Roget DK (2004) Understanding soil biota and biological functions: management of soil biota for improved benefits to crop production and environmental health. In ‘Soil Biology in Agriculture. Proceedings of Workshop on Current Research into Soil Biology in Agriculture’. Tamworth Sustainable Farming Training Centre, 11–12 August 2004. (Ed. R Lines-Kelly) pp. 1–7. (NSW Dept of Primary Industries: Orange, NSW)

Gupta VVSR, Roper MM (2010) Protection of free-living nitrogen fixing bacteria within the soil matrix. Soil & Tillage Research 109, 50–54.
Protection of free-living nitrogen fixing bacteria within the soil matrix.Crossref | GoogleScholarGoogle Scholar |

Gupta VVSR, Roper MM, Roget DK (2006) Potential for non-symbiotic N2-fixation in different agroecological zones of southern Australia. Australian Journal of Soil Research 44, 343–354.
Potential for non-symbiotic N2-fixation in different agroecological zones of southern Australia.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD28XmtFKgs7k%3D&md5=b9e34e21de95681bb839444e2573a34eCAS |

Gupta VVSR, Penton CR, Lardner R, Tiedje J (2010) Catabolic and genetic diversity of microbial communities in Australian soils are influenced by soil type and stubble management. In ‘Soil solutions for a changing world. Proceedings of the 19th World Congress of Soil Science’. 1–10 August 2010, Brisbane, Qld. pp. 1–4. (IUSS) Available at: www.iuss.org

Gupta VVSR, Rovira AD, Roget DK (2011) Principles and management of soil biological factors for sustainable rainfed farming systems. In ‘Rainfed farming systems’. (Eds P Tow, I Cooper, I Partridge, C Birch) pp. 149–184. (Springer Science and Business Media: Berlin, Heidelberg)

Hamelin J, Fromin N, Tarnawski S, Teyssier-Cuvelle S, Aragno M (2002) nifH gene diversity in the bacterial community associated with the rhizosphere of Molinia coerulea, an oligonitrophilic perennial grass. Environmental Microbiology 4, 477–481.
nifH gene diversity in the bacterial community associated with the rhizosphere of Molinia coerulea, an oligonitrophilic perennial grass.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD38XntFSltLc%3D&md5=ece2eccc52b59945f707ae4129567466CAS | 12153588PubMed |

Hirsch PR, Miller AJ, Dennis PG (2013) Do root exudates exert more influence on rhizosphere bacterial community structure than other rhizodeposits. Molecular Microbial Ecology of the Rhizosphere 1, 229–242.
Do root exudates exert more influence on rhizosphere bacterial community structure than other rhizodeposits.Crossref | GoogleScholarGoogle Scholar |

Hoyle FC, Murphy DV (2006) Seasonal changes in microbial function and diversity associated with stubble retention versus burning. Australian Journal of Soil Research 44, 407–424.
Seasonal changes in microbial function and diversity associated with stubble retention versus burning.Crossref | GoogleScholarGoogle Scholar |

Hsu S-F, Buckley DH (2009) Evidence for the functional significance of diazotroph community structure in soil. The ISME Journal 3, 124–136.
Evidence for the functional significance of diazotroph community structure in soil.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD1MXms12gsbk%3D&md5=62aa2e094c7291e9e7d009ab7e8aec84CAS | 18769458PubMed |

Hurek T, Handley LL, Reinhold-Hurek B, Piche Y (2002) Azoarcus grass endophytes contribute fixed nitrogen to the plant in an unculturable state. Molecular Plant-Microbe Interactions 15, 233–242.
Azoarcus grass endophytes contribute fixed nitrogen to the plant in an unculturable state.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD38XitF2gtbw%3D&md5=dcb779886c45d5e22aad601d6418b4b2CAS | 11952126PubMed |

Isbell RF (2002) ‘The Australian Soil Classification.’ (CSIRO Publishing: Melbourne)

Izquierdo JA, Nusslein K (2006) Distribution of extensive nifH gene diversity across physical soil microenvironments. Microbial Ecology 51, 441–452.
Distribution of extensive nifH gene diversity across physical soil microenvironments.Crossref | GoogleScholarGoogle Scholar | 16645928PubMed |

Janik LJ, Forrester ST, Rowson AJ (2009) The prediction of soil chemical and physical properties from mid-infrared spectroscopy and combined partial least-squares regression and neural networks (PLS-NN) analysis. Chemometrics and Intelligent Laboratory Systems 97, 179–188.
The prediction of soil chemical and physical properties from mid-infrared spectroscopy and combined partial least-squares regression and neural networks (PLS-NN) analysis.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD1MXmtlWmtLY%3D&md5=abc47ef20a7445c74ddfe76e63d2ed45CAS |

Jensen BB, Cox RP (1983) Direct measurements of steady-state kinetics of cyanobacterial N2 uptake by membrane-leak mass spectrometry and comparisons between nitrogen fixation and acetylene reduction. Applied and Environmental Microbiology 45, 1331–1337.

Jones DL, Nguyen C, Finlay RD (2009) Carbon flow in the rhizosphere: carbon trading at the soil-root interface. Plant and Soil 321, 5–33.
Carbon flow in the rhizosphere: carbon trading at the soil-root interface.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD1MXos1enu7c%3D&md5=f20c5a931c6f271a402f8c29481e25f9CAS |

Kennedy IR, Islam N (2001) The current and potential contribution of asymbiotic nitrogen fixation to nitrogen requirements on farms: a review. Australian Journal of Experimental Agriculture 41, 447–457.
The current and potential contribution of asymbiotic nitrogen fixation to nitrogen requirements on farms: a review.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD3MXkt1Crtrc%3D&md5=0628a0ebe37eaab896cd10f5001f789eCAS |

Kennedy IR, Choudhury ATMA, Kecskes ML (2004) Non-symbiotic bacterial diazotrophs in crop-farming systems: can their potential for plant growth promotion be better exploited? Soil Biology & Biochemistry 36, 1229–1244.
Non-symbiotic bacterial diazotrophs in crop-farming systems: can their potential for plant growth promotion be better exploited?Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2cXls1WmsbY%3D&md5=3a45eb12a7d1b1e8ba9e073ebe2bfc25CAS |

Knox Ok, Gupta VVSR, Lardner R (2009) Cotton cultivar selection impacts on microbial diversity and function. In ‘Aspects of applied biology. Positive plant microbial interactions in relation to plant performance and ecosystem function’. Vol. 98. pp. 129–136. (The Association of Applied Biologists: Warwick, UK)

Kuzyakov Y, Domanski G (2000) Carbon input by plants into the soil. Review. Journal of Plant Nutrition and Soil Science 163, 421–431.
Carbon input by plants into the soil. Review.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD3cXmtFyrurc%3D&md5=02aa081c4ac094398ecec65f62d82070CAS |

Lambers H (1987) Growth, respiration, exudation and symbiotic associations: the fate of carbon translocated to the roots. In ‘Root development and function—effects of the physical environment’. (Eds PJ Gregory, JV Lake, DA Rose) pp. 125–145. (Cambridge University Press: Cambridge, UK)

Lawes RA, Robertson MJ (2012) Effect of subtropical perennial grass pastures on nutrients and carbon in coarse-textured soils in a Mediterranean climate. Soil Research 50, 551–561.
Effect of subtropical perennial grass pastures on nutrients and carbon in coarse-textured soils in a Mediterranean climate.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC38Xhs12jsrjN&md5=87c45f05c8e0d2c7d1f048d05d2d0e36CAS |

Lawes RA, Ward PR, Ferris D (2014) Pasture cropping with C4 grasses in a barley–lupin rotation can increase production. Crop & Pasture Science 65, 1002–1015.

Lovell CR, Piceno YM, Quattro JM, Bagwell CE (2000) Molecular analysis of diazotroph diversity in the rhizosphere of the smooth cordgrass, Spartina alterniflora. Applied and Environmental Microbiology 66, 3814–3822.
Molecular analysis of diazotroph diversity in the rhizosphere of the smooth cordgrass, Spartina alterniflora.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD3cXmsVWqu7c%3D&md5=e6aa4a8e1d3d3c512eb157a73098bd83CAS | 10966395PubMed |

Margalef R (1958) Information theory in ecology. General Systems 3, 36–71.

Matejovic I (1997) Determination of carbon and nitrogen in samples of various soils by the dry combustion. Communications in Soil Science and Plant Analysis 28, 1499–1511.
Determination of carbon and nitrogen in samples of various soils by the dry combustion.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK2sXnsVCltbg%3D&md5=35834d869ed53a2ec1d3319c7af6185eCAS |

Mirza BS, Potisap C, Nusslein K, Bohannan BJM, Rodrigues JLM (2014) Response of free-living nitrogen-fixing microorganisms to land use change in the Amazon rainforest. Applied and Environmental Microbiology 80, 281–288.
Response of free-living nitrogen-fixing microorganisms to land use change in the Amazon rainforest.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC2cXltFGnsQ%3D%3D&md5=754bc13a823162b259e5422ce68d67f1CAS | 24162570PubMed |

Nelson DR, Mele PM (2006) The impact of crop residue amendments and lime on microbial community structure and nitrogen-fixing bacteria in the wheat rhizosphere. Australian Journal of Soil Research 44, 319–329.
The impact of crop residue amendments and lime on microbial community structure and nitrogen-fixing bacteria in the wheat rhizosphere.Crossref | GoogleScholarGoogle Scholar |

Peoples MB, Boddey RM, Herridge DF (2002) Quantification of nitrogen fixation. In ‘Nitrogen fixation at the millennium’. (Ed. GJ Leigh) pp. 357–389. (Elsevier Science: Amsterdam)

Pereira e Silva MC, Semenov AV, van Elsas JD, Salles JF (2011) Seasonal variations in the diversity and abundance of diazotrophic communities across soils. FEMS Microbiology Ecology 77, 57–68.
Seasonal variations in the diversity and abundance of diazotrophic communities across soils.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3MXosFKjsb4%3D&md5=9655062de01c0f20531874d2994a3f64CAS | 21385188PubMed |

Pielou EC (1975) ‘Ecological diversity.’ (Wiley Press: New York)

Reed HE, Martiny JBH (2007) Testing the functional significance of microbial composition in natural communities. FEMS Microbiology Ecology 62, 161–170.
Testing the functional significance of microbial composition in natural communities.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2sXht1yns73K&md5=d29b4f7f3f41badc4e87722c08987c2eCAS | 17937673PubMed |

Reis VM, dos Reis FB, Quesada DM, de Oliveira OCA, Alves BJR, Urquiaga S, Boddey RM (2001) Biological nitrogen fixation associated with tropical pasture grasses. Australian Journal of Plant Physiology 28, 837–844.

Roёsch LFW, Camargo FAO, Bento FM, Triplett EW (2008) Biodiversity of diazotrophic bacteria within the soil, root and stem of field-grown maize. Plant and Soil 302, 91–104.
Biodiversity of diazotrophic bacteria within the soil, root and stem of field-grown maize.Crossref | GoogleScholarGoogle Scholar |

Roёsch LFW, Fulthorpe RR, Jaccques RJS, Bento FM, Camargo FAdeO (2010) Biogeography of diazotrophic bacteria in soils. World Journal of Microbiology and Biotechnology 26, 1503–1508.
Biogeography of diazotrophic bacteria in soils.Crossref | GoogleScholarGoogle Scholar |

Roper MM, Ladha JK (1995) Biological N2 fixation by heterotrophic and phototrophic bacteria in association with straw. Plant and Soil 174, 211–224.
Biological N2 fixation by heterotrophic and phototrophic bacteria in association with straw.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK2MXotFalsL0%3D&md5=ff13207f18d6216b5639183174c8d755CAS |

Roper MM, Smith NA (1991) Straw decomposition and nitrogenase activity (C2H2 reduction) by free-living microorganisms from soil: effects of pH and clay content. Soil Biology & Biochemistry 23, 275–283.
Straw decomposition and nitrogenase activity (C2H2 reduction) by free-living microorganisms from soil: effects of pH and clay content.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK3MXltlWlsrw%3D&md5=95efe36d3202beab1fa2a0921df2d293CAS |

Roper MM, Fillery IRP, Jongepier R, Sanford P, Macdonald LM, Sanderman L, Baldock JA (2013) Allocation into soil organic matter fractions of 14C captured via photosynthesis by two perennial grass pastures. Soil Research 51, 748–759.
Allocation into soil organic matter fractions of 14C captured via photosynthesis by two perennial grass pastures.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3sXhvF2ktbbO&md5=767b1b2e1212eb2a1ca25e873e3d4546CAS |

Rösch C, Mergel A, Bothe H (2002) Biodiversity of denitrifying and dinitrogen-fixing bacteria in an acid forest soil. Applied and Environmental Microbiology 68, 3818–3829.
Biodiversity of denitrifying and dinitrogen-fixing bacteria in an acid forest soil.Crossref | GoogleScholarGoogle Scholar | 12147477PubMed |

Sanderman J, Fillery IRP, Jongepier R, Massalsky A, Roper MM, Macdonald LM, Maddern T, Murphy DV, Wilson BR, Baldock JA (2013) Carbon sequestration under subtropical perennial pastures I: Overall trends. Soil Research 51, 760–770.
Carbon sequestration under subtropical perennial pastures I: Overall trends.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3sXhvF2ktbnN&md5=e43266f09839f51adcd0e44ba8ef1715CAS |

Shannon CE, Weaver W (1949) ‘The mathematical theory of communication.’ (The University of Illinois Press: Urbana, IL, USA)

Smalla K, Oros-Sichler M, Milling A, Heuer H, Baumgarte S, Becker R, Neuber G, Kropf S, Ulrich A, Tebbe C (2007) Bacterial diversity of soils assessed by DGGE, T-RFLP and SSCP fingerprints of PCR-amplified 16S rRNA gene fragments: Do the different methods provide similar results. Journal of Microbiological Methods 69, 470–479.
Bacterial diversity of soils assessed by DGGE, T-RFLP and SSCP fingerprints of PCR-amplified 16S rRNA gene fragments: Do the different methods provide similar results.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2sXkvFGhurw%3D&md5=35ec6422b6b4b4229bc8dd9d587e7d3bCAS | 17407797PubMed |

Soares RA, Roёsch LFW, Zanatta G, Camargo FAdeO, Passaglia LMP (2006) Occurrence and distribution of nitrogen fixing bacterial community associated with oat (Avena sativa) assessed by molecular and microbiological techniques. Applied Soil Ecology 33, 221–234.
Occurrence and distribution of nitrogen fixing bacterial community associated with oat (Avena sativa) assessed by molecular and microbiological techniques.Crossref | GoogleScholarGoogle Scholar |

Swinnen J (1994) Rhizodeposition and turnover of root-derived organic material in barley and wheat under conventional and integrated management. Agriculture, Ecosystems & Environment 51, 115–128.
Rhizodeposition and turnover of root-derived organic material in barley and wheat under conventional and integrated management.Crossref | GoogleScholarGoogle Scholar |

Syme H, Botwright Acuna TL, Abrecht D, Wade LJ (2007) Nitrogen contributions in a windmill grass (Chloris truncate)–wheat (Triticum aestivum L.) system in south-western Australia. Australian Journal of Soil Research 45, 635–642.
Nitrogen contributions in a windmill grass (Chloris truncate)–wheat (Triticum aestivum L.) system in south-western Australia.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2sXhsVamtbnN&md5=448080eadea08cb685fe7cf8c8f0c268CAS |

Tan Z, Hurek T, Reinhold-Hurek B (2003) Effect of N-fertilization, plant genotype and environmental conditions on nifH gene pools in roots of rice. Environmental Microbiology 5, 1009–1015.
Effect of N-fertilization, plant genotype and environmental conditions on nifH gene pools in roots of rice.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD3sXos1CltbY%3D&md5=5585ec1de485eba7e5ce530ddbd2da53CAS | 14510855PubMed |

Wakelin SA, Gupta VVSR, Forrester ST (2010) Regional and local factors affecting diversity, abundance and activity of free-living, N2-fixing bacteria in Australian agricultural soils. Pedobiologia 53, 391–399.
Regional and local factors affecting diversity, abundance and activity of free-living, N2-fixing bacteria in Australian agricultural soils.Crossref | GoogleScholarGoogle Scholar |

Warwick RM, Platt HM, Clarke KR, Agard J, Gobin J (1990) Analysis of macrobenthic and meiobenthic community structure in relationship to pollution and disturbance in Hamilton Harbour, Bermuda. Journal of Experimental Marine Biology and Ecology 138, 119–142.
Analysis of macrobenthic and meiobenthic community structure in relationship to pollution and disturbance in Hamilton Harbour, Bermuda.Crossref | GoogleScholarGoogle Scholar |

Whipps JM (1990) Carbon economy. In ‘The rhizosphere’. (Ed. JM Lynch) pp. 59–97. (Wiley: Chichester, UK)

Wu L, Ma K, Lu Y (2009) Prevalence of betaproteobacterial sequences in nifH gene pools associated with roots of modern rice cultivars. Microbial Ecology 57, 58–68.
Prevalence of betaproteobacterial sequences in nifH gene pools associated with roots of modern rice cultivars.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD1cXhsFWrsrjI&md5=9d51046225ce7f317eb28ab211e735e1CAS | 18548184PubMed |

Zehr JP, Jenkins BD, Short SM, Steward GF (2003) Nitrogenase gene diversity and microbial community structure: a cross-system comparison. Environmental Microbiology 5, 539–554.
Nitrogenase gene diversity and microbial community structure: a cross-system comparison.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD3sXlvFChs7s%3D&md5=32f20a5120584d4763f38406d309f25bCAS | 12823187PubMed |

Zhan J, Sun Q (2012) Diversity of free-living nitrogen-fixing microorganisms in the rhizosphere and non-rhizosphere of pioneer plants growing on wastelands of copper mine tailings. Microbiological Research 167, 157–165.
Diversity of free-living nitrogen-fixing microorganisms in the rhizosphere and non-rhizosphere of pioneer plants growing on wastelands of copper mine tailings.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC38XitVChsrk%3D&md5=73a6f453b1c7e03b43e74407c46b0f0cCAS | 21665448PubMed |

Zou Y, Zhang J, Yang D, Chen X, Zhao J, Xiu W, Lai X, Li G (2011) Effects of different land use patterns on nifH genetic diversity of soil nitrogen-fixing microbial communities in Leymus chinensis steppe. Acta Ecologica Sinica 31, 150–156.
Effects of different land use patterns on nifH genetic diversity of soil nitrogen-fixing microbial communities in Leymus chinensis steppe.Crossref | GoogleScholarGoogle Scholar |