Plant and Soil

, Volume 351, Issue 1–2, pp 389–403 | Cite as

Arbuscular mycorrhizal fungal diversity in perennial pastures; responses to long-term lime application

  • Y. J. Guo
  • Y. Ni
  • H. Raman
  • B. A. L. Wilson
  • G. J. Ash
  • A. S. Wang
  • G. D. Li
Regular Article


Background and aims

We investigated the genetic diversity of arbuscular mycorrhizal fungi (AMF) in soils and the roots of Phalaris aquatica L., Trifolium subterraneum L., and Hordeum leporinum Link growing in limed and unlimed soil, the influence of lime application on AMF colonization and the relationship between AMF diversity and soil chemical properties.


The sampling was conducted on a long-term liming experimental site, established in 1992, in which lime was applied every 6 years to maintain soil pH (in CaCl2) at 5.5 in the 0–10 cm soil depth. Polymerase chain reaction, cloning and sequencing techniques were used to investigate the diversity of AMF.


Altogether, 438 AMF sequences from a total of 480 clones were obtained. Sequences of phylotypes Aca/Scu were detected exclusively in soil, while Glomus sp. (GlGr Ab) and an uncultured Glomus (UnGlGr A) were detected only in plant roots. Glomus mosseae (GlGr Aa) was the dominant AMF in the pastures examined; however, the proportion of G. mosseae was negatively correlated with soil pH, exchangeable Ca and available P. Generally, diversity of the AMF phylotypes was greater in the bulk unlimed soil and plants from this treatment when compared to the limed treatments.


Long-term lime application changed soil nutrient availability and increased AMF colonization, but decreased AMF phylotype diversity, implying that soil chemistry may determine the distribution of AMF in acid soils. Future studies are required to explore the functions of these AMF groups and select the most efficient AMF for sustainable farming in acid soils.


Diversity Glomeromycota Hordeum leporinum Phalaris aquatica Soil acidity Trifolium subterraneum 


  1. Abbott LK, Robson AD (1977) Infectivity and effectiveness of vesicular-carbuncular mycorrhizal fungi: effect of inoculums type. Aust J Agr Res 32:631–639CrossRefGoogle Scholar
  2. Akinrinde EA (2008) Lime and phosphorus effects in maize (Zea mays L.) production. Res Crops 9:547–553Google Scholar
  3. Alguacil MM, Lozano Z, Campoy MJ, Rold A (2010) Phosphorus fertilisation management modifies the biodiversity of AM fungi in a tropical savanna forage system. Soil Biol Biochem 42:1114–1122CrossRefGoogle Scholar
  4. Alguacil MM, Torres MP, Torrecillas E, Diaz G, Roldan A (2011) Plant type differently promote the arbuscular mycorrhizal fungi biodiversity in the rhizosphere after revegetation of a degraded, semiarid land. Soil Biol Biochem 43:167–173CrossRefGoogle Scholar
  5. Alkan N, Gadkar V, Yarden O, Kapulnik Y (2006) Analysis of quantitative interactions between two species of arbuscular mycorrhizal fungi, Glomus mosseae and G. intraradices, by real-time PCR. Appl Environ Microb 72:4192–4199CrossRefGoogle Scholar
  6. Altschul SF, Madden TL, Schäffer AA, Zhang JH, Zhang Z, Miller W, Lipman DJ (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 25:3389–3402PubMedCrossRefGoogle Scholar
  7. Bardgett RD, Leemans DK, Cook R, Hobbs PJ (1997) Seasonality of the soil biota of grazed and ungrazed hill grasslands. Soil Biol Biochem 29:1285–1294CrossRefGoogle Scholar
  8. Bartolomeesteban H, Schenck N (1994) Spore germination and hyphal growth of arbuscular mycorrhizal fungi in relation to soil aluminum saturation. Mycologia 86:217–226CrossRefGoogle Scholar
  9. Bhadalung NN, Suwanarit A, Dell B, Nopamornbodi O, Thamchaipenet A, Rungchuang J (2005) Effects of long-term NP-fertilization on abundance and diversity of arbuscular mycorrhizal fungi under a maize cropping system. Plant Soil 270:371–382CrossRefGoogle Scholar
  10. Callaway RM, Mahall BE, Wicks C, Pankey J, Zabinski C (2003) Soil fungi and the effects of an invasive forb on grasses: neighbor identity matters. Ecology 84:129–135CrossRefGoogle Scholar
  11. Chang CS, Sung JM (2004) Nutrient uptake and yield responses of peanuts and rice to lime and fused magnesium phosphate in an acid soil. Field Crops Res 89:319–325CrossRefGoogle Scholar
  12. Clapp JP, Young JPW, Merryweather JW, Fitter AH (1995) Diversity of fungal symbionts in arbuscular mycorrhizas from a natural community. New Phytol 130:259–265CrossRefGoogle Scholar
  13. Clapp JP, Rodriguez A, Dodd JC (2001) Inter- and intra-isolate rRNA large subunit variation in Glomus coronatum spores. New Phytol 149:539–554CrossRefGoogle Scholar
  14. Clark RB (1997) Arbuscular mycorrhizal adaptation, spore germination, root colonization, and host plant growth and mineral acquisition at low pH. Plant Soil 192:15–22CrossRefGoogle Scholar
  15. Colwell J (1963) The estimation of the phosphorus fertilizer requirements of wheat in southern New South Wales by soil analysis. Aust J Exp Agric Anim Husb 3:190–198CrossRefGoogle Scholar
  16. Cuenca G, Lovera M (2010) Seasonal variation and distribution at different soil depths of arbuscular mycorrhizal fungi spores in a tropical sclerophyllous shrubland. Botany-Botanique 88:54–64CrossRefGoogle Scholar
  17. Cui MY, Caldwell MM (1996) Facilitation of plant phosphate acquisition by arbuscular mycorrhizas from enriched soil patches.2. Hyphae exploiting root-free soil. New Phytol 133:461–467CrossRefGoogle Scholar
  18. Da Silva GA, Lumini E, Costa Maia L, Bonfante P, Bianciotto V (2006) Phylogenetic analysis of Glomeromycota by partial LSU rDNA sequences. Mycorrhiza 16:183–189PubMedCrossRefGoogle Scholar
  19. Dhillion SS, Gardsjord TL (2004) Arbuscular mycorrhizas influence plant diversity, productivity, and nutrients in boreal grasslands. Can J Bo 82:104–114CrossRefGoogle Scholar
  20. Foy CD (1988) Plant adaptation to acid, aluminum-toxic soils. Comm Soil Sci Plant Anal 19:959–987CrossRefGoogle Scholar
  21. Gillman G, Sumpter E (1986) Modification to the compulsive exchange method for measuring exchange characteristics of soils. Aust J Soil Res 24:61–66CrossRefGoogle Scholar
  22. Gotelli N, Colwell RK (2001) Quantifying biodiversity: Procedures and pitfalls in the measurement and comparison of species richness. Ecol Lett 4:379–391CrossRefGoogle Scholar
  23. Grayston SJ, Wang SQ, Campbell CD, Edwards AC (1998) Selective influence of plant species on microbial diversity in the rhizosphere. Soil Biol Biochem 30:369–378CrossRefGoogle Scholar
  24. Guo Y, Ni Y, Huang J (2010) Effects of rhizobium, arbuscular mycorrhiza and lime on nodulation, growth and nutrient uptake of lucerne in acid purplish soil in China. Trop Grassl 44:109–114Google Scholar
  25. Haling R, Simpson R, Delhaize E, Hocking P, Richardson A (2010) Effect of lime on root growth, morphology and the rhizosheath of cereal seedlings growing in an acid soil. Plant Soil 327:199–212CrossRefGoogle Scholar
  26. Hamel C, Dalpe Y, Lapierre C, Simard RR, Smith DL (1996) Endomycorrhizae in a newly cultivated acidic meadow: Effects of three years of barley cropping, tillage, lime, and phosphorus on root colonization and soil infectivity. Biol Fertil Soils 21:160–165CrossRefGoogle Scholar
  27. Hijri I, Sykorova Z, Oehl F, Ineichen K, Mader P, Wiemken A, Redecker D (2006) Communities of arbuscular mycorrhizal fungi in arable soils are not necessarily low in diversity. Mol Ecol 15:2277–2289PubMedCrossRefGoogle Scholar
  28. Husband R, Herre EA, Turner SL, Gallery R, Young JPW (2002) Molecular diversity of arbuscular mycorrhizal fungi and patterns of host association over time and space in a tropical forest. Mol Ecol 11:2669–2678PubMedCrossRefGoogle Scholar
  29. Jansa J, Smith F, Smith S (2008) Are there benefits of simultaneous root colonization by different arbuscular mycorrhizal fungi? New Phytol 177:779–789PubMedCrossRefGoogle Scholar
  30. Johnson NC (1993) Can fertilization of soil select less mutualistic mycorrhizae. Ecol Appl 3:749–757CrossRefGoogle Scholar
  31. Johnson D, Ijdo M, Anderson IC, Alexander IJ (2005a) Do plant communities influence microbial diversity and function? Comp Biochem Physiol Mol Integr Physiol 141:S220–S220Google Scholar
  32. Johnson D, Leake JR, Read DJ (2005b) Liming and nitrogen fertilization affects phosphatase activities, microbial biomass and mycorrhizal colonisation in upland grassland. Plant Soil 271:157–164CrossRefGoogle Scholar
  33. Johnson D, Anderson IC, Williams A, Whitlock R, Grime JP (2010) Plant genotypic diversity does not beget root-fungal species diversity. Plant Soil 336:107–111CrossRefGoogle Scholar
  34. Krebs C (1989) Ecological methodology. HarperCollins, New YorkGoogle Scholar
  35. Li GD, Singh RP, Brennan JP, Helyar KR (2010a) A financial analysis of lime application in a long-term agronomic experiment on the south-western slopes of New South Wales. Crop Past Sci 61:12–23CrossRefGoogle Scholar
  36. Li L-F, Li T, Zhang Y, Zhao Z-W (2010b) Molecular diversity of arbuscular mycorrhizal fungi and their distribution patterns related to host-plants and habitats in a hot and arid ecosystem, southwest China. FEMS Microbiol Ecol 71:418–427PubMedCrossRefGoogle Scholar
  37. Lux H, Cumming J (2001) Mycorrhizae confer aluminium resistance to tulip-poplar seedlings. Can J Forest Res 31:694–702Google Scholar
  38. Martinez-Garcia LB, Pugnaire FI (2011) Arbuscular mycorrhizal fungi host preference and site effects in two plant species in a semiarid environment. Appl Soil Ecol 48:313–317CrossRefGoogle Scholar
  39. Martini JA, Mutters RG (1985) Effect of lime rates on nutrient availability, mobility, and uptake during the soybean-growing season.1. aluminum, manganese, and phosphorus. Soil Sci 139:219–226CrossRefGoogle Scholar
  40. Morton JB (1995) Taxonomic and phylogenetic divergence among 5 scutellospora species based on comparative developmental sequences. Mycologia 87:127–137CrossRefGoogle Scholar
  41. Nei M, Kumar S (2000) Molecular evolution and phylogenetics. Oxford University Press, New YorkGoogle Scholar
  42. Oehl F, Laczko E, Bogenrieder A, Stahr K, Bosch R, van der Heijden M, Sieverding E (2010) Soil type and land use intensity determine the composition of arbuscular mycorrhizal fungal communities. Soil Biol Biochem 42:724–738CrossRefGoogle Scholar
  43. Olsson P, Hansson M, Burleigh S (2006) Effect of P availability on temporal dynamics of carbon allocation and glomus intraradices high-affinity P transporter gene induction in arbuscular mycorrhiza. Appl Environ Microb 72:4115–4120CrossRefGoogle Scholar
  44. Payne R, Harding S, Murray D, Soutar D, Baird D, Glaser A, Channing I, Welham S, Gilmour A, Thompson R, Webster R (2010) The Guide to GenStat Release 13, Part 2: statistics. VSN International, Hemel HempsteadGoogle Scholar
  45. Porter W, Robson A, Abbott L (1987) Factors controlling the distribution of vesicular arbuscular mycorrhizal fungi in relation to soil-pH. J Appl Ecol 24:663–672CrossRefGoogle Scholar
  46. Raznikiewicz H, Carlgren K, Martensson A (1994) Impact of phosphorus fertilization and liming on the presence of arbuscular mycorrhizal spores in a swedish long-term field experiment. Swed J Agr Res 24:157–164Google Scholar
  47. Rodriguez A, Dougall T, Dodd JC, Clapp JP (2001) The large subunit ribosomal RNA genes of Entrophospora infrequens comprise sequences related to two different glomalean families. New Phytol 152:159–167CrossRefGoogle Scholar
  48. Rosendahl S, McGee P, Morton JB (2009) Lack of global population genetic differentiation in the arbuscular mycorrhizal fungus Glomus mosseae suggests a recent range expansion which may have coincided with the spread of agriculture. Mol Ecol 18:4316–4329PubMedCrossRefGoogle Scholar
  49. Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, New YorkGoogle Scholar
  50. Schwarzott D, Walker C, Schüssler A (2001) Glomus, the largest genus of the arbuscular mycorrhizal fungi (Glomales), is nonmonophyletic. Mol Phylogenet Evolut 21:190–197Google Scholar
  51. Sen R, Hepper CM, Azcon-Aguilar C, Rosendahl S (1990) Competition between introduced and indigenous mycorrhizal fungi (Glomus spp.) for root colonization of leek. Agr Eco Envir 29:355–359CrossRefGoogle Scholar
  52. Smith S, Read D (1997) Mycorrhizal symbiosis. Academic, LondonGoogle Scholar
  53. Soil Survey Staff (2006) Keys to soil taxonomy. In USDA-Natural resources conservation service, Washington, DCGoogle Scholar
  54. Sonjak S, Udovic M, Wraber T, Likar M, Regvar M (2009) Diversity of halophytes and identification of arbuscular mycorrhizal fungi colonising their roots in an abandoned and sustained part of Secovlje salterns. Soil Biol Biochem 41:1847–1856CrossRefGoogle Scholar
  55. Stackebrandt E, Goebel BM (1994) Taxonomic note: a place for DNA-DNA reassociation and 16S rRNA sequence analysis in the present species definition in bacteriology. Int J Syst Bacteriol 44:846–849CrossRefGoogle Scholar
  56. Tamura K, Dudley J, Nei M, Kumar S (2007) MEGA4: Molecular Evolutionary Genetics Analysis (MEGA) software version 4.0. Mol Biol Evol 24:1596–1599PubMedCrossRefGoogle Scholar
  57. Troeh ZI, Loynachan TE (2009) Diversity of arbuscular mycorrhizal fungal species in soils of cultivated soybean fields. Agro J 101:1453–1462CrossRefGoogle Scholar
  58. Trouvelot A, Kough J, Gianinazzi-Pearson V (1986) Mesure du taux de mycorhization VA d’un système radiculaire. Recherche de méthodes d’estimation ayant une signification fonctionnelle. In: Gianinazzi-Pearson V, Gianinazzi S (eds) Physiological and genetical aspects of mycorrhizae. INRA Press, Paris, pp 217–221Google Scholar
  59. Trouvelot S, van Tuinen D, Hijri M, Gianinazzi-Pearson V (1999) Visualization of ribosomal DNA loci in spore interphasic nuclei of glomalean fungi by fluorescence in situ hyhridization. Mycorrhiza 8:203–206CrossRefGoogle Scholar
  60. Unkovich MJ, Sanford P, Pate JS (1996) Nodulation and nitrogen fixation by subterranean clover in acid soils as influenced by lime application, toxic aluminium, soil mineral N, and competition from annual ryegrass. Soil Biol Biochem 28:639–648CrossRefGoogle Scholar
  61. Vandenkoornhuyse P, Leyval C, Bonnin I (2001) High genetic diversity in arbuscular mycorrhizal fungi: evidence for recombination events. Heredity 87:243–253PubMedCrossRefGoogle Scholar
  62. Vandenkoornhuyse P, Ridgway KP, Watson IJ, Fitter AH, Young JPW (2003) Co-existing grass species have distinctive arbuscular mycorrhizal communities. Mol Ecol 12:3085–3095PubMedCrossRefGoogle Scholar
  63. van der Heijden MGA, Klironomos JN, Ursic M, Moutoglis P, Streitwolf-Engel R, Boller T, Wiemken A, Sanders IR (1998) Mycorrhizal fungal diversity determines plant biodiversity, ecosystem variability and productivity. Nature 396:69–72CrossRefGoogle Scholar
  64. Wakelin SA, Gregg AL, Simpson RJ, Li GD, Riley IT, McKay AC (2009) Pasture management clearly affects soil microbial community structure and N-cycling bacteria. Pedobiologia 52:237–251CrossRefGoogle Scholar
  65. Wenke L (2008) N, P contribution and soil adaptability of four arbuscular mycorrhizal fungi. Acta Agr Scand B-S P 58:285–288Google Scholar
  66. Whelan A, Alexander M (1986) Effects of low pH and high Al, Mn and Fe levels on the survival of Rhizobium trifolii and the nodulation of subterranean clover. Plant Soil 92:363–371CrossRefGoogle Scholar
  67. Yano K, Takaki M (2005) Mycorrhizal alleviation of acid soil stress in the sweet potato (Ipomoea batatas). Soil Biol Biochem 37:1569–1572CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2011

Authors and Affiliations

  • Y. J. Guo
    • 1
    • 2
  • Y. Ni
    • 3
  • H. Raman
    • 2
  • B. A. L. Wilson
    • 2
  • G. J. Ash
    • 2
  • A. S. Wang
    • 2
  • G. D. Li
    • 2
  1. 1.Faculty of Animal Science and TechnologySouthwest UniversityChongqingChina
  2. 2.EH Graham Centre for Agricultural Innovation (an alliance between Charles Sturt University and NSW Department of Primary Industries)Wagga WaggaAustralia
  3. 3.Faculty of Agronomy and Bio-technologySouthwest UniversityChongqingChina

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