Plant and Soil

, Volume 315, Issue 1–2, pp 285–296 | Cite as

Facilitated legume nodulation, phosphate uptake and nitrogen transfer by arbuscular inoculation in an upland rice and mung bean intercropping system

  • Yuefeng Li
  • Wei Ran
  • Ruiping Zhang
  • Shubin Sun
  • Guohua XuEmail author
Regular Article


Intercropping of upland rice with short-duration grain legumes has major advantages in increasing crop yields and soil productivity. However, the contribution of arbuscular mycorrhizas, the common mutualistic symbiosis between most crops and mycorrhizal fungi, is not fully understood in intercropping systems. We assayed the contribution of inoculation of the arbuscular mycorrhizal fungus (AMF) Glomus caledonium on nutrient acquisition and biomass yield. Using the method of plastic film and nylon net partition and tracing 15N transferred between the intercropped upland rice (Oryza sativa ssp. Japonica Nipponbare) and mung bean (Vigna radiata L. Chuanyuan), we compared the intercropping, with separation of the whole root systems by a plastic film, with and without a barrier of nylon net to allow penetration of the fungal hyphae. Intercropping significantly improved the formation of arbuscular mycorrhizas, particularly in the upland rice roots. The improved formation of mycorrhizas by the intercropping increased total P uptake by 57% in rice, total P and N acquisition by 65% and 64% respectively in mung bean, and nodulation by 54% in mung bean. The percentage of total 15N transfer from mung bean to rice leaves was increased from 5.4% to 15.7% by inoculation with AMF. In contrast, there was only 2.7% of 15N transfer from rice to mung bean and no AMF effect on N transfer. It is concluded that cereal and legume crop intercropping increase mycorrhiza formation, which in turn improves nodulation, N and P acquisition and N transfer in the legumes.


Arbuscular mycorrhizal fungi Nutrient transfer Hyphal links 15Rice Mung bean Intercropping 



This work was supported by China 863 program (2006AA10Z134), National Natural Science Foundation of China. We thank Ms. Juan Zhu and Mr. Guiyun Zhang for technical support, Dr. Xiaolin Li from China Agricultural University and Dr. Xiangui Lin from Nanjing Soil Science Institute, CAS for providing mycorrhizal fungal inoculum, Dr. Uzi Kafkafi for revising the manuscript.


  1. Af Geijersstam L, Mårtensson A (2006) Nitrogen fixation and residual effects of field pea intercropped with oats. Acta Agricult Scand B 56:186–196Google Scholar
  2. Aggarwal PK, Garrity DP, Liboon SP, Morris RA (1992) Resource use and plant interactions in a rice-mungbean intercrop. Agron J 84:71–78Google Scholar
  3. Allen LH Jr, Albrecht SL, Colon-Guasp W, Covell SA, Baker JT, Pan D et al (2003) Methane emissions of rice increased by elevated carbon dioxide and temperature. J Environ Qual 32:1978–1991PubMedGoogle Scholar
  4. Ames RN, Reid CPP, Porter LK, Cambardella C (1983) Hyphal uptake and transport of nitrogen from two 15N-labelled sources by Glomus mosseae, a vesicular–arbuscular mycorrhizal fungus. New Phytol 95:381–396 doi: 10.1111/j.1469-8137.1983.tb03506.x CrossRefGoogle Scholar
  5. Barker SJ, Tagu D, Delp G (1998) Regulation of root and fungal morphogenesis in mycorrhizal symbioses. Plant Physiol 116:1201–1207 doi: 10.1104/pp.116.4.1201 CrossRefGoogle Scholar
  6. Bethlenfalvay GJ, Reyes-Solis MG, Camel SB, Ferrera-Cerrato R (1991) Nutrient transfer between the root zones of soybean and maize plants connected by a common mycorrhizal mycelium. Physiol Plant 82:423–432 doi: 10.1111/j.1399-3054.1991.tb02928.x CrossRefGoogle Scholar
  7. Bouman BAM, Humphreys E, Tuong TP, Baker RB (2007) Rice and water. Adv Agron 92:187–237 doi: 10.1016/S0065-2113(04)92004-4 CrossRefGoogle Scholar
  8. Bremner JM (1965) Chapter 83: total nitrogen. In: Black CA (ed) Methods of soil analysis: part 2. chemical and microbiological properties. American Society of Agronomy, Madison, WI, pp 1149–1178Google Scholar
  9. Chakraborty N, Sarkar GM, Lahiri SC (2000) Methane emission from rice paddy soils, aerotolerance of methanogens and global thermal warming. Environmentalist 20:343–350 doi: 10.1023/A:1006734101607 CrossRefGoogle Scholar
  10. Chalk PM (1998) Dynamics of biologically fixed N in legume–cereal rotations: a review. Aust J Agric Res 49:303–316 doi: 10.1071/A97013 CrossRefGoogle Scholar
  11. Clark RB, Zeto SK (2000) Mineral acquisition by arbuscular mycorrhizal plants. J Plant Nutr 23:867–902CrossRefGoogle Scholar
  12. Dhillion SS (1992) Dual inoculation of pretransplant stage Oryza sativa L. plants with indigenous vesicular–arbuscular mycorrhizal fungi and fluorescent Pseudomonas spp. Biol Fertil Soils 13:147–151 doi: 10.1007/BF00337340 CrossRefGoogle Scholar
  13. Fumoto T, Kobayashi K, Li C, Yagi K, Hasegawa T (2008) Revising a process-based biogeochemistry model (DNDC) to simulate methane emission from rice paddy fields under various residue management and fertilizer regimes. Glob Change Biol 14:382–402Google Scholar
  14. Gao X, Kuyper TW, Zou C, Zhang FS, Hoffland E (2007) Mycorrhizal responsiveness of aerobic rice genotypes is negatively correlated with their zinc uptake when nonmycorrhizal. Plant Soil 290:283–291 doi: 10.1007/s11104-006-9160-x CrossRefGoogle Scholar
  15. Gavito ME, Curtis PS, Mikkelsen TN, Jakobsen I (2000) Atmospheric CO2 and mycorrhiza effects on biomass allocation and nutrient uptake of nodulated pea (Pisum sativum L.) plants. J Exp Bot 51:1931–1938 doi: 10.1093/jexbot/51.352.1931 PubMedCrossRefGoogle Scholar
  16. Ghosh PK, Bandyopadhyay KK, Wanjari RH, Manna MC, Misra AK, Mohanty M et al (2007) Legume effect for enhancing productivity and nutrient use-efficiency in major cropping systems—an Indian perspective: a review. J Sustain Agric 30:61–86 doi: 10.1300/J064v30n01_07 Google Scholar
  17. Govindarajulu M, Pfeffer PE, Jin H, Abubaker J, Douds DD, Allen JW et al (2005) Nitrogen transfer in the arbuscular mycorrhizal symbiosis. Nature 435:819–823 doi: 10.1038/nature03610 PubMedCrossRefGoogle Scholar
  18. Güimil S, Chang HS, Zhu T, Sesma A, Osbourn A, Roux C, Ioannidis V, Oakeley EJ, Docquier M, Descombes P, Briggs SP, Paszkowski U (2005) Comparative transcriptomics of rice reveals an ancient pattern of response to microbial colonization. Proc Natl Acad Sci USA 102:8066–8070PubMedCrossRefGoogle Scholar
  19. Hamel C, Furlan V, Smith DL (1991) N2-fixation and transfer in a field grown mycorrhizal corn and soybean intercrop. Plant Soil 133:177–185CrossRefGoogle Scholar
  20. Hamel C, Furlan V, Smith DL (1992) Mycorrhizal effects on interspecific plant competition and nitrogen transfer in legume-grass mixtures. Crop Sci 32:991–996Google Scholar
  21. Hardarson G, Danso SKA, Zapata F (1988) Dinitrogen fixation measurements in alfalfa-ryegrass swards using nitrogen-15 and influence of the reference crop. Crop Sci 28:101–105Google Scholar
  22. Harrison MJ (1999) Molecular and cellular aspects of the arbuscular mycorrhizal symbiosis. Annu Rev Plant Bio 50:361–389CrossRefGoogle Scholar
  23. Haugen LM, Smith SE (1992) The effect of high temperature and fallow period on infection of mung bean and cashew roots by the vesicular–arbuscular mycorrhizal fungus Glomus intraradices. Plant Soil 145:71–80CrossRefGoogle Scholar
  24. Hauggaard-Nielsen H, Jensen ES (2005) Facilitative root interactions in intercrops. Plant Soil 274:237–250CrossRefGoogle Scholar
  25. Hause B, Fester T (2005) Molecular and cell biology of arbuscular mycorrhizal symbiosis. Planta 221:184–196PubMedCrossRefGoogle Scholar
  26. He XH, Critchley C, Bledsoe C (2003) Nitrogen transfer within and between plants through common mycorrhizal networks (CMNs). Crit Rev Plant Sci 22:531–567CrossRefGoogle Scholar
  27. Herdler S, Kreuzer K, Scheu S, Bonkowski M (2008) Interactions between arbuscular mycorrhizal fungi (Glomus intraradices, Glomeromycota) and amoebae (Acanthamoeba castellanii, Protozoa) in the rhizosphere of rice (Oryza sativa). Soil Biol Biochem 40:660–668CrossRefGoogle Scholar
  28. Hoagland DL, Arnon DI (1950) The water culture method of growing plants without soil. Cali Agri Experi Station Cir 347Google Scholar
  29. Ikram A, Jensen ES, Jakobsen I (1994) No significant transfer of N and P from Pueraria phaseoloides to Hevea brasiliensis via hyphal links of arbuscular mycorrhiza. Soil Biol Biochem 26:1541–1547CrossRefGoogle Scholar
  30. Israel DW (1987) Investigation of the role of phosphorus in symbiotic dinitrogen fixation. Plant Physiol 84:835–840PubMedCrossRefGoogle Scholar
  31. Israel DW (1993) Symbiotic dinitrogen fixation and host-plant growth during development of and recovery from phosphorus deficiency. Physiol Plant 88:294–300CrossRefGoogle Scholar
  32. Jensen ES (1996) Grain yield, symbiotic N2 fixation and interspecific competition for inorganic N in pea-barley intercrops. Plant Soil 182:25–38CrossRefGoogle Scholar
  33. Johansen A, Jensen ES (1996) Transfer of N and P from intact or decomposing roots of pea to barley interconnected by an arbuscular mycorrhizal fungus. Soil Biol Biochem 28:73–81CrossRefGoogle Scholar
  34. Johansen A, Jakobsen I, Jensen ES (1993) Hyphal transport by a vesicular–arbuscular mycorrhizal fungus of N applied to the soil as ammonium or nitrate. Biol Fertil Soils 16:66–70CrossRefGoogle Scholar
  35. John MK (1970) Colorimetric determination of phosphorus in soil and plant material with ascorbic acid. Soil Sci 11:214–220CrossRefGoogle Scholar
  36. Johnson NC, Graham JH, Smith FA (1997) Functioning of mycorrhizal associations along the mutualism-parasitism continuum. New Phytol 135:575–585CrossRefGoogle Scholar
  37. Johnson D, Leake JR, Ostle N, Ineson P, Read DJ (2002a) In situ 13CO2 pulse-labelling of upland grassland demonstrates a rapid pathway of carbon flux from arbuscular mycorrhizal mycelia to the soil. New Phytol 153:327–334CrossRefGoogle Scholar
  38. Johnson D, Leake JR, Read DJ (2002b) Transfer of recent photosynthate into mycorrhizal mycelium of an upland grassland: Short-term respiratory losses and accumulation of 14C. Soil Biol Biochem 34:1521–1524CrossRefGoogle Scholar
  39. Kasiamdari RS, Smith SE, Smith FA, Scott ES (2002) Influence of the mycorrhizal fungus, Glomus coronatum, and soil phosphorus on infection and disease caused by binucleate Rhizoctonia and Rhizoctonia solani on mung bean (Vigna radiata). Plant Soil 238:235–244CrossRefGoogle Scholar
  40. Kawai Y, Yamamoto Y (1986) Increase in the formation and nitrogen fixation of soybean nodules by vesicular–arbuscular mycorrhiza. Plant Cell Physiol 27:399–405Google Scholar
  41. Khasa P, Furlan V, Fortin JA (1992) Response of some tropical plant species to endomycorrhizal fungi under field conditions. Trop Agri 69:279–283Google Scholar
  42. Kwabiah AB (2005) Biological efficiency and economic benefits of pea–barley and pea–oat intercrops. J Sustain Agri 25:117–128CrossRefGoogle Scholar
  43. Lauk R, Lauk E (2008) Pea-oat intercrops are superior to pea–wheat and pea–barley intercrops. Acta Agricult Scand B 58:139–144Google Scholar
  44. Laxminarayana K, Munda GC (2004) Performance of rice (Oryza sativa) and maize (Zea mays)-based cropping systems under mid-hills of Mizoram. Indian J Agro 49:230–232Google Scholar
  45. Li XL, George E, Marschner H (1991) Extension of the phosphorus depletion zone in VA-mycorrhizal white clover in a calcareous soil. Plant Soil 136:41–48CrossRefGoogle Scholar
  46. Li H, Smith SE, Holloway RE, Zhu Y, Smith FA (2006) Arbuscular mycorrhizal fungi contribute to phosphorus uptake by wheat grown in a phosphorus-fixing soil even in the absence of positive growth responses. New Phytol 172:536–543PubMedCrossRefGoogle Scholar
  47. Lin XG, Shuguang W, Yaqin S (2001) Tolerance of VA mycorrhizal fungi to soil acidity. Pedosphere 11:105–113Google Scholar
  48. Lithourgidis AS, Dhima KV, Vasilakoglou IB, Dordas CA, Yiakoulaki MD (2007) Sustainable production of barley and wheat by intercropping common vetch. Agron Sustain Dev 27:95–99CrossRefGoogle Scholar
  49. Marschner H, Dell B (1994) Nutrient uptake in mycorrhizal symbiosis. Plant Soil 159:89–102Google Scholar
  50. Mårtensson AM, Rydberg I, Vestberg M (1998) Potential to improve transfer of N in intercropped systems by optimising host-endophyte combinations. Plant Soil 205:57–66CrossRefGoogle Scholar
  51. Martin RC, Astatkie T, Cooper JM (1998) The effect of soybean variety on corn-soybean intercrop biomass and protein yields. Can J Plant Sci 78:289–294Google Scholar
  52. Martins MA, Cruz AF (1998) The role of the external mycelial network of arbuscular mycorrhizal fungi: III. A study of nitrogen transfer between plants interconnected by a common mycelium. Rev Microbiol 29:289–294CrossRefGoogle Scholar
  53. Mikkelsen DS, DeDatta SK (1991) Rice culture. In: Luh BS (ed) In rice production. Van Nostrand Reinhold, New York, pp 103–186Google Scholar
  54. Newbould P, Rangeley A (1984) Effect of lime, phosphorus and mycorrhizal fungi on growth, nodulation and nitrogen fixation by white clover (Trifolium repens) grown in UK hill soils. Plant Soil 76:105–114CrossRefGoogle Scholar
  55. Ofosu-Budu KG, Noumura K, Fujita K (1995) N2 fixation, N transfer and biomass production of soybean cv. bragg or its supernodulating nts1007 and sorghum mixing-cropping at two rates of N fertilizer. Soil Biol Biochem 27:311–317CrossRefGoogle Scholar
  56. Olivera M, Tejera N, Iribarne C, Ocana A, Lluch C (2004) Growth, nitrogen fixation and ammonium assimilation in common bean (Phaseolus vulgaris): effect of phosphorus. Physiol Plant 121:498–505CrossRefGoogle Scholar
  57. Oroka FO, Omoregie AU (2007) Competition in a rice–cowpea intercrop as affected by nitrogen fertilizer and plant population. Scientia Agricola 64:621–629CrossRefGoogle Scholar
  58. Paszkowski U, Kroken S, Roux C, Briggs SP (2002) Rice phosphate transporters include an evolutionarily divergent gene specifically activated in arbuscular mycorrhizal symbiosis. Proc Natl Acad Sci USA 99:13324–11329PubMedCrossRefGoogle Scholar
  59. Phillip JM, Hayman DS (1970) Improved procedures for clearing roots and staining parasitic and vesicular arbuscular mycorrhizal fungi for rapid assessment of infection. T Brit Myco Soc 55:158–160Google Scholar
  60. Purakayastha TJ, Chhonkar PK (2001) Influence of vesicular–arbuscular mycorrhizal fungi (Glomus etunicatum L.) on mobilization of zinc in wetland rice (Oryza sativa L.). Biol Fertil Soils 33:323–327CrossRefGoogle Scholar
  61. Raimam MP, Albino U, Cruz MF, Lovato GM, Spago F, Ferracin TP, Lima DS, Goulart T, Bernardi CM, Miyauchi M, Nogueira MA, Andrade G (2007) Interaction among free-living N-fixing bacteria isolated from Drosera villosa var. villosa and AM fungi (Glomus clarum) in rice (Oryza sativa). Appl Soil Ecol 35:25–34CrossRefGoogle Scholar
  62. Reeves M (1992) The role of VAM fungi in nitrogen dynamics in maize–bean intercrops. Plant Soil 144:85–92CrossRefGoogle Scholar
  63. Ryan MH, McCully ME, Huang CX (2007) Relative amounts of soluble and insoluble forms of phosphorus and other elements in intraradical hyphae and arbuscules of arbuscular mycorrhizas. Funct Plant Biol 34:457–464CrossRefGoogle Scholar
  64. Sarkar RK, Sanyal SR (2000) Production potential and economic feasibility of sesame (Sesamum indicum)-based intercropping system with pulse and oilseed crops on rice fallow land. Indian J Agro 45:545–550Google Scholar
  65. Sawers RJH, Gutjahr C, Paszkowski U (2008) Cereal mycorrhiza: an ancient symbiosis in modern agriculture. Trends Plant Sci 13:93–97PubMedGoogle Scholar
  66. Sarr PS, Khouma M, Sene M, Guisse A, Badiane AN, Yamakawa T (2008) Effect of pearl millet-cowpea cropping systems on nitrogen recovery, nitrogen use efficiency and biological fixation using the 15N tracer technique. Soil Sci Plant Nutr 54:142–147Google Scholar
  67. Shen QR, Chu GX (2004) Bi-directional nitrogen transfer in an intercropping system of peanut with rice cultivated in aerobic soil. Biol Fertil Soils 40:81–87CrossRefGoogle Scholar
  68. Sieverding E, Leihner DE (1984) Influence of crop rotation and intercropping of cassava with legumes on VA mycorrhizal symbiosis of cassava. Plant Soil 80:143–146CrossRefGoogle Scholar
  69. Smith SE, Read DJ (1997) The symbionts forming VA mycorrhizas. In Mycorrhizal symbiosis, 2nd edn. Academic, London, pp 11–32Google Scholar
  70. Smith SE, Dickson S, Smith FA (2001) Nutrient transfer in arbuscular mycorrhizas: How are fungal and plant processes integrated? Funct Plant Biol 28:683–694Google Scholar
  71. Smith SE, Smith FA, Jakobsen I (2003) Mycorrhizal fungi can dominate phosphate supply to plants irrespective of growth responses. Plant Physiol 133:16–20PubMedCrossRefGoogle Scholar
  72. Smith SE, Smith FA, Jakobsen I (2004) Functional diversity in arbuscular mycorrhizal (AM) symbioses: The contribution of the mycorrhizal P uptake pathway is not correlated with mycorrhizal responses in growth or total P uptake. New Phytol 162:511–524CrossRefGoogle Scholar
  73. Solaiman MZ, Hirata H (1997) Responses of directly seeded wetland rice to arbuscular mycorrhizal fungi inoculation. J Plant Nutr 20:1479–1487CrossRefGoogle Scholar
  74. Sprent JI, James EK (2007) Legume evolution: where do nodules and mycorrhizas fit in? Plant Physiol 144:575–581PubMedCrossRefGoogle Scholar
  75. Toomsan B, Cadisch G, Srichantawong M, Thongsodsaeng C, Giller KE, Limpinuntana V (2000) Biological N2 fixation and residual N benefit of pre-rice leguminous crops and green manures. Neth J Agr Sci 48:19–29Google Scholar
  76. Trouvelot A, Kough JL, Gianinazzi-Pearson V (1986) Mesure de taux de mycorhization VA d’un systeme radiculaire. Recherche de méthodes d’estimation ayant une signification fonctionnelle. In: Gianinazzi-Pearson V, Gianinazzi S (eds) Physiological and genetical aspects of mycorrhizae. INRA, Paris, pp 217–221Google Scholar
  77. Zhang XH, Zhu YG, Chen BD, Lin AJ, Smith SE, Smith FA (2005) Arbuscular mycorrhizal fungi contribute to resistance of upland rice to combined metal contamination of soil. J Plant Nutr 28:2065–2077CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2008

Authors and Affiliations

  • Yuefeng Li
    • 1
  • Wei Ran
    • 1
  • Ruiping Zhang
    • 1
  • Shubin Sun
    • 1
  • Guohua Xu
    • 1
    Email author
  1. 1.State Key Laboratory of Crop Genetics and Germplasm Enhancement, College of Resources and Environmental SciencesNanjing Agricultural UniversityNanjingChina

Personalised recommendations