Ecological Research

, Volume 33, Issue 4, pp 799–810 | Cite as

Crop rotation associating a legume and the nickel hyperaccumulator Alyssum murale improves the structure and biofunctioning of an ultramafic soil

  • Ramez Farajallah Saad
  • Ahmad Kobaissi
  • Gaylord Machinet
  • Geneviève Villemin
  • Guillaume Echevarria
  • Emile BenizriEmail author
Special Feature Ultramafic Ecosystems: Proceedings of the 9th International Conference on Serpentine Ecology


Nickel (Ni) agromining aims to phytoextract heavy metals using hyperaccumulators whilst at the same time rehabilitating ultramafic soils. After removing the bioavailable metal, ultramafic soils are improved in terms of their agronomic properties with the aim of future agricultural uses. The low fertility of ultramafic soils can be compensated by integrating legumes already used in traditional agro-systems because of their importance in soil nitrogen enrichment. However, few studies have evaluated the potential profits of legumes on Ni agromining and their potential benefits on soil biological fertility. Here, we characterized the effect of a crop rotation with two plants, a legume (Vicia sativa) and a hyperaccumulator (Alyssum murale), on the phytoextraction efficiency and on soil structure and biofunctioning. A pot experiment was set up in controlled conditions to grow A. murale and four treatments were tested: rotation with V. sativa (Ro), fertilized mono-culture (FMo), non-fertilized mono-culture (NFMo) and bare soil without plants (BS). No significant difference was found between the Ro and NFMo treatments for the dry biomass yield. However, the Ro treatment showed the highest Ni concentrations ([Ni]) in A. murale shoots compared to FMo and NFMo treatments. The Ro treatment plants had more than twice as many leaves [Ni] compared to FMo. Soil physico-chemical analyses showed that the Ro treatment was better structured and showed the highest presence of bacterial micro-aggregates, as well as less non-aggregated particles. Legumes integration in Ni-agromining systems could be a pioneering strategy to reduce chemical inputs and to improve soil biofunctioning and thus fertility.


Nickel Alyssum murale Legume Soil fertility Soil fractionation Ultrastructural characterization Agromining Crop rotation 



We would like to acknowledge the support of the technical team in the Laboratoire Sols et Environnement, Université de Lorraine. In particular, we appreciate the technical assistance of the joint Microhumus research unit and especially Maxime Maire. We would also like to thank Dr Petra Kidd and her research unit in the Consejo Superior de Investigaciones Científicas (CSIC, Santiago de Compostella, Spain) and especially the Soil Microbiology group. This work was supported by the French National Research Agency through the national ‘Investissements d’avenir’ program, ANR-10-LABX-21-LABEX RESSOURCES21, through the ANR-14-CE04-0005 project ‘Agromine’ and by the European ERA-net FACCE_SURPLUS project ‘AGRONICKEL: Developing Ni agromining on ultramafic land in Europe’, ANR-15-SUSF-0003-05. We acknowledge the Association of Specialization and Scientific Guidance (ASSG, Lebanon) for funding the PhD scholarship for Ramez Farajallah Saad and are grateful to Prof Alan Baker for his advices throughout the submission of this paper.


  1. Abiven S, Menasseri S, Claire Chenu C (2009) The effects of organic inputs over time on soil aggregate stability—a literature analysis. Soil Biol Biochem 41:1–12CrossRefGoogle Scholar
  2. Amezketa E (1999) Soil aggregate stability: a review. J Sustain Agric 14:83–151. CrossRefGoogle Scholar
  3. Andrade G, Mihara KL, Linderman RG, Bethlenfalvay GJ (1998) Soil aggregation status and rhizobacteria in the mycorrhizosphere. Plant Soil 202:89–96. CrossRefGoogle Scholar
  4. Aspiras RB, Allen ON, Harris RF, Chesters G (1971) The role of microorganisms in the stabilization of soil aggregates. Soil Biol Biochem 3:347–353. CrossRefGoogle Scholar
  5. Bani A, Echevarria G, Sulçe S et al (2007) In-situ phytoextraction of Ni by a native population of Alyssum murale on an ultramafic site (Albania). Plant Soil 293:79–89. CrossRefGoogle Scholar
  6. Bani A, Echevarria G, Sulçe S, Morel JL (2015) Improving the agronomy of Alyssum murale for extensive phytomining: a five-year field study. Int J Phytorem 17:117–127. CrossRefGoogle Scholar
  7. Barbaroux R, Mercier G, Blais JF et al (2011) A new method for obtaining nickel metal from the hyperaccumulator plant Alyssum murale. Sep Purif Technol 83:57–65. CrossRefGoogle Scholar
  8. Bernal MR, McGrath SP, Miller AJ, Baker AJM (1994) Comparison of the chemical changes in the rhizosphere of the nickel hyperaccumulator Alyssum murale with the non-accumulator Raphanus sativus. Plant Soil 164:251–259CrossRefGoogle Scholar
  9. Bot A, Benites J (2005) The importance of soil organic matter: key to drought-resistant soil and sustained food production. FAO Soils Bull. CrossRefGoogle Scholar
  10. Caballero R, García C (1996) Cultivo y utilización de la asociación veza-cereal en Castilla-La Mancha. Junta de Comunidades de Castilla-La Mancha, Castilla-La ManchaGoogle Scholar
  11. Centofanti T, Siebecker MG, Chaney RL, Davis AP, Sparks DL (2012) Hyperaccumulation of nickel by Alyssum corsicum is related to solubility of Ni mineral species. Plant Soil 359:71–83. CrossRefGoogle Scholar
  12. Chaney RL, Angle JS, Broadhurst CL et al (2007) Improved understanding of hyperaccumulation yields commercial phytoextraction and phytomining technologies. J Environ Qual 36:1429–1443. CrossRefPubMedGoogle Scholar
  13. Chaney RL, Chen KY, Li YM et al (2008) Effects of calcium on nickel tolerance and accumulation in Alyssum species and cabbage grown in nutrient solution. Plant Soil 311:131–140. CrossRefGoogle Scholar
  14. Chaney RL, Baklanov IA, Ryan TC, Davis AP (2016) Effect of soil volume on Ni hyperaccumulation from serpentine soil by Alyssum corsicum. Int J Phytoremed (submitted) Google Scholar
  15. Chardot V, Massoura ST, Echevarria G, Reeves RD, Morel JL (2005) Phytoextraction potential of the nickel hyperaccumulators Leptoplax emarginata and Bornmuellera tymphaea. Int J Phytoremed 7:323–335CrossRefGoogle Scholar
  16. Cherr CM, Scholberg JMS, McSorley R (2006) Green manure approaches to crop production: a synthesis. Agron J 98:302–319CrossRefGoogle Scholar
  17. Christensen BT (2001) Physical fractionation of soil and structural and functional complexity in organic matter turnover. Eur J Soil Sci 52:345–353. CrossRefGoogle Scholar
  18. Chu J, Zhang T, Chang W, Zhang D, Zulfiqar S, Fu A, Hao Y (2016) Impacts of cropping systems on aggregates associated organic carbon and nitrogen in a semiarid highland agroecosystem. PLoS ONE 11:1–14. CrossRefGoogle Scholar
  19. Coleman DC, Crossley DA, Hendrix PF (2004) Future developments in soil ecology. In: Coleman DC, Crossley DA Jr, Hendrix PF (eds) Fundamentals of soil ecology, 2nd edn. Elsevier Academic Press, Amsterdam, pp 408Google Scholar
  20. Coppens F, Garnier P, Findeling A et al (2007) Decomposition of mulched versus incorporated crop residues: modelling with PASTIS clarifies interactions between residue quality and location. Soil Biol Biochem 39:2339–2350. CrossRefGoogle Scholar
  21. Czarnes S, Dexter AR, Bartoli F (2000) Wetting and drying cycles in the maize rhizosphere under controlled conditions. Mechanics of the root-adhering soil. Plant Soil 221:253–271. CrossRefGoogle Scholar
  22. Dieng A, Baudoin E, Thioulouse J et al (2015) Soil organic matter quality, structure and activity of the denitrifiers community as influenced by decaying mulched crop residues. Appl Ecol Environ Res 13:655–675. CrossRefGoogle Scholar
  23. Echevarria G, Morel J-L, Fardeau JC, Leclerc-Cessac E (1998) Assessment of phytoavailability of nickel in soils. J Environ Qual 27:1064–1070. CrossRefGoogle Scholar
  24. Echevarria G, Massoura S, Sterckeman T, Becquer T, Schwartz C, Morel JL (2006) Assessment and control of the bioavailability of Ni in soils. Environ Toxicol Chem 25:643–651CrossRefPubMedGoogle Scholar
  25. Gao Y, Zhou P, Mao L et al (2010) Effects of plant species coexistence on soil enzyme activities and soil microbial community structure under Cd and Pb combined pollution. J Environ Sci 22:1040–1048. CrossRefGoogle Scholar
  26. Gao Y, Miao CY, Xia J et al (2012) Plant diversity reduces the effect of multiple heavy metal pollution on soil enzyme activities and microbial community structure. Front Environ Sci Eng 6:213–223. CrossRefGoogle Scholar
  27. Gregorich EG, Drury CF, Baldock JA (2001) Changes in soil carbon under long-term maize in monoculture and legume-based rotation. Can J Soil Sci 81:21–31. CrossRefGoogle Scholar
  28. Hargrove WL (1986) Winter legumes as a nitrogen source for no-till grain sorghum. Agron J 78:70–74CrossRefGoogle Scholar
  29. Hattori T (1988) Soil aggregates as microhabitats of microorganisms. Rep Inst Agr Res Tohoku Univ 37:23–36Google Scholar
  30. Haynes RJ (1999) Labile organic matter fractions and aggregate stability under short-term, grass-based leys. Soil Biol Biochem 31:1821–1830. CrossRefGoogle Scholar
  31. Haynes RJ, Francis GS (1993) Changes in microbial biomass C, soil carbohydrate composition and aggregate stability induced by growth of selected crop and forage species under field conditions. Eur J Soil Sci 44(4):665–675CrossRefGoogle Scholar
  32. Hinsinger P, Betencourt E, Bernard L et al (2011) P for two, sharing a scarce resource: soil phosphorus acquisition in the rhizosphere of intercropped species. Plant Physiol 156:1078–1086. CrossRefPubMedPubMedCentralGoogle Scholar
  33. Hycka M (1973) Veza común, su cultivo y utilización. CSIC. Estación Experimental de Aula Dei, ZaragozaGoogle Scholar
  34. Justes E, Bedoussac L, Corre-Hellou G et al (2014) Les processus de complémentarité de niche et de facilitation déterminent le fonctionnement des associations végétales et leur efficacité pour l’acquisition des ressources abiotiques. Innov Agron 40:1–24Google Scholar
  35. Karpenstein-Machan M, Stuelpnagel R (2000) Biomass yield and nitrogen fixation of legumes monocropped and intercropped with rye and rotation effects on a subsequent maize crop. Plant Soil 218:215–232. CrossRefGoogle Scholar
  36. Kidd PS, Álvarez-López V, Becerra-Castro C et al (2017) Potential role of plant-associated bacteria in plant metal uptake and implications in phytotechnologies. Adv Bot Res 83:87–126. CrossRefGoogle Scholar
  37. Kinsbursky RS, Levanon D, Yaron B (1989) Role of fungi in stabilizing aggregates of sewage sludge Amended soils. Soil Sci Soc Am J 53:1086–1091CrossRefGoogle Scholar
  38. Körschens M (2002) Importance of soil organic matter (SOM) for biomass production and environment (a review). Arch Agron Soil Sci 48:89–94. CrossRefGoogle Scholar
  39. Kuzyakov Y (2010) Priming effects: interactions between living and dead organic matter. Soil Biol Biochem 42:1363–1371. CrossRefGoogle Scholar
  40. Li YM, Chaney RL, Brewer EP et al (2003) Phytoextraction of nickel and cobalt by hyperaccumulator Alyssum species grown on nickel-contaminated soils. Environ Sci Technol 37:1463–1468. CrossRefGoogle Scholar
  41. Lindsay WL, Norvell WA (1978) Development of a DTPA soil test for zinc, iron, manganese and copper. Soil Sci Soc Am J 42:421–428CrossRefGoogle Scholar
  42. Liu W, Zhang C, Hu P et al (2016) Influence of nitrogen form on the phytoextraction of cadmium by a newly discovered hyperaccumulator Carpobrotus rossii. Environ Sci Pollut Res 23:1246–1253. CrossRefGoogle Scholar
  43. Lizarazo CI, Yli-Halla M, Stoddard FL (2015) Pre-crop effects on the nutrient composition and utilization efficiency of faba bean (Vicia faba L.) and narrow-leafed lupin (Lupinus angustifolius L.). Nutr Cycl Agroecosystems 103:311–327. CrossRefGoogle Scholar
  44. Lloveras J, Santiveri P, Vendrell A, Torrent D, Ballesta A (2004) Varieties of vetch (Vicia sativa L.) for forage and grain production in Mediterranean areas. In: Ferchichi A, Ferchichi A (eds) Réhabilitation des pâturages et des parcours en milieux méditerranéens. CIHEAM 62, Zaragoza, pp 103–106Google Scholar
  45. Ma Y, Rajkumar M, Freitas H (2009) Isolation and characterization of Ni mobilizing PGPB from serpentine soils and their potential in promoting plant growth and Ni accumulation by Brassica spp. Chemosphere 75:719–725. CrossRefPubMedGoogle Scholar
  46. McNear DH, Chaney RL, Sparks DL (2010) The hyperaccumulator Alyssum murale uses complexation with nitrogen and oxygen donor ligands for Ni transport and storage. Phytochemistry 71:188–200. CrossRefPubMedGoogle Scholar
  47. Metzger L, Levanon D, Mingelgrin U (1987) The effect of sewage sludge on soil structural stability: microbial aspects. Soil Sci Soc Am J 51:346–351CrossRefGoogle Scholar
  48. Moebius BN, Van Es HM, Schindelbeck RR et al (2007) Evaluation of laboratory-measured soil properties as indicators of soil physical quality. Soil Sci 172:895–912. CrossRefGoogle Scholar
  49. Morel JL (2013) Using plants to “micro-mine” metals. Accessed 25 July 2017
  50. Morel JL, Habib L, Plantureux S, Guckert A (1991) Influence of maize root mucilage on soil aggregate stability. Plant Soil 136:111–119. CrossRefGoogle Scholar
  51. Reynolds ES (1963) The use of lead citrate at high pH as an electron-opaque stain in electron microscopy. J Cell Biol 17:208–212. CrossRefPubMedPubMedCentralGoogle Scholar
  52. Rochester IJ, Peoples MB, Hulugalle NR et al (2001) Using legumes to enhance nitrogen fertility and improve soil condition in cotton cropping systems. F Crop Res 70:27–41. CrossRefGoogle Scholar
  53. Rodrigues MÂ, Dimande P, Pereira EL et al (2015) Early-maturing annual legumes: an option for cover cropping in rainfed olive orchards. Nutr Cycl Agroecosyst 103:153–166. CrossRefGoogle Scholar
  54. Rodrigues J, Houzelot V, Ferrari F et al (2016) Life cycle assessment of agromining chain highlights role of erosion control and bioenergy. J Clean Prod 139:770–778. CrossRefGoogle Scholar
  55. Saad R, Kobaissi A, Robin C, Echevarria G, Benizri E (2016) Nitrogen fixation and growth of Lens culinaris as affected by nickel availability: a pre-requisite for optimization of agromining. Environ Exp Bot 131:1–9. CrossRefGoogle Scholar
  56. Saad RF, Kobaissi A, Amiaud B, Ruelle J, Benizri E (2017a) Changes in physicochemical characteristics of a serpentine soil and in root architecture of a hyperaccumulator plant cropped with a legume. J Soils Sediments (submitted) Google Scholar
  57. Saad RF, Kobaissi A, Goux X, Calusinska M, Echevarria G, Kidd P, Benizri E (2017b) Developing Ni-agromining by associating Alyssum murale with a leguminous plant: an in situ experiment on an ultramafic site in Spain. Soil Biol Biochem (submitted) Google Scholar
  58. Sainju UM, Whitehead WF, Singh BP (2003) Cover crops and nitrogen fertilization effects on soil aggregation and carbon and nitrogen pools. Can J Soil Sci 83:155–165. CrossRefGoogle Scholar
  59. Scalise A, Tortorella D, Pristeri A et al (2015) Legume-barley intercropping stimulates soil N supply and crop yield in the succeeding durum wheat in a rotation under rainfed conditions. Soil Biol Biochem 89:150–161. CrossRefGoogle Scholar
  60. Scherer-Lorenzen M, Palmborg C, Prinz A, Schulze ED (2003) The role of plant diversity and composition for nitrate leaching in grasslands. Ecology 84–6:1539–1552CrossRefGoogle Scholar
  61. Shallari S, Echevarria G, Schwartz C, Morel J-L (2001) Availability of nickel in soils for the hyperaccumulator Alyssum murale (Waldst. & Kit.). S Afr J Sci 97:568–570Google Scholar
  62. Sharma AR, Mittra BN (1988) Effect of green manuring and mineral fertilizer on growth and yield of crops in rice-based cropping system on acid lateritic soil. J Agric Sci 110:605–608CrossRefGoogle Scholar
  63. Sieverding E, Leihner DE (1984) Influence of crop rotation and intercropping of cassava with legumes on VA mycorrhizal symbiosis of cassava. Plant Soil 80:143–146. CrossRefGoogle Scholar
  64. Simonin P (2017) Observation des pratiques de fertilisation azotée par un outil d’aide à la décision pour le colza. Oilseeds fats Crop Lipids 24:1–7. CrossRefGoogle Scholar
  65. Singer S, Ewing M (2000) Soil quality. Handbook of soil science. CRC Press, Boca Raton, pp 271–278Google Scholar
  66. Six J, Bossuyt H, Degryze S, Denef K (2004) A history of research on the link between (micro)aggregates, soil biota, and soil organic matter dynamics. Soil Tillage Res 79:7–31CrossRefGoogle Scholar
  67. Snapp SS, Mafongoya PL, Waddington S (1998) Organic matter technologies for integrated nutrient management in smallholder cropping systems of southern Africa. Agric Ecosyst Environ 71:185–200. CrossRefGoogle Scholar
  68. Stevenson FC, Van Kessel C (1996) The nitrogen and non-nitrogen rotation benefits of pea to succeeding crops. Can J Plant Sci 76:735–745. CrossRefGoogle Scholar
  69. Tiemann LK, Grandy AS, Atkinson EE et al (2015) Crop rotational diversity enhances belowground communities and functions in an agroecosystem. Ecol Lett 18:761–771. CrossRefPubMedGoogle Scholar
  70. Tisdall JM, Oades JM (1982) Organic matter and water-stable aggregates in soils. J Soil Sci 33:141–163. CrossRefGoogle Scholar
  71. Valentine RC (1961) Contrast enhancement in the electron microscopy of viruses. Adv Virus Res 8:287–318CrossRefPubMedGoogle Scholar
  72. van der Ent A, Baker AJM, Reeves RD et al (2013) Hyperaccumulators of metal and metalloid trace elements: facts and fiction. Plant Soil 362:319–334. CrossRefGoogle Scholar
  73. van der Ent A, Baker AJM, Reeves RD et al (2015) Agromining: farming for metals in the future? Environ Sci Technol 49:4773–4780. CrossRefPubMedGoogle Scholar
  74. Vergani C, Graf F (2016) Soil permeability, aggregate stability and root growth: a pot experiment from a soil bioengineering perspective. Ecohydrology 9:830–842. CrossRefGoogle Scholar
  75. Vertès F, Hatch D, Velthof G, Taube F, Laurent F, Loiseau P, Recous S (2007) Short-term and cumulative effects of grassland cultivation and carbon cycling in ley-arable rotations. Grassl Sci Eur 12:227–246Google Scholar
  76. Walker TS (2003) Root exudation and rhizosphere biology. Plant Physiol 132:44–51. CrossRefPubMedPubMedCentralGoogle Scholar
  77. Watteau F, Villemin G, Bartoli F et al (2012) 0–20 μm aggregate typology based on the nature of aggregative organic materials in a cultivated silty topsoil. Soil Biol Biochem 46:103–114. CrossRefGoogle Scholar
  78. Whipps JM (2001) Microbial interactions and biocontrol in the rhizosphere. J Exp Bot 52:487–511. CrossRefPubMedGoogle Scholar
  79. Yan F, Schubert S, Mengel K (1996) Soil pH increase due to biological decarboxylation of organic anions. Soil Biol Biochem 28:617–624. CrossRefGoogle Scholar
  80. Yang H, Niu J, Tao J, Gu Y, Zhang Siyuan She C, Chen W, Yang H, Yin H (2016) The Impacts of Different green manure on soil microbial communities and crop health. Preprints. CrossRefGoogle Scholar
  81. Zahran HH (2001) Rhizobia from wild legumes: diversity, taxonomy, ecology, nitrogen fixation and biotechnology. J Biotechnol 9(12):143–153CrossRefGoogle Scholar
  82. Zhang X, Houzelot V, Bani A et al (2014) Selection and combustion of Ni-hyperaccumulators for the phytomining process. Int J Phytorem 16:1058–1072. CrossRefGoogle Scholar
  83. Zhao J, Zeng Z, He X et al (2015) Effects of monoculture and mixed culture of grass and legume forage species on soil microbial community structure under different levels of nitrogen fertilization. Eur J Soil Biol 68:61–68. CrossRefGoogle Scholar

Copyright information

© The Ecological Society of Japan 2017

Authors and Affiliations

  1. 1.Laboratoire « Sols et Environnement »Université de Lorraine, UMR 1120Vandœuvre-lès-NancyFrance
  2. 2.Laboratoire « Sols et Environnement »INRA, UMR 1120Vandœuvre-lès-NancyFrance
  3. 3.Laboratoire « Applied Plant Biotechnology », Faculté des Sciences 1Université LibanaiseBeirutLebanon
  4. 4.Laboratoire « Sols et Environnement », MicrohumusUniversité de Lorraine, UMR 1120Vandœuvre-lès-NancyFrance

Personalised recommendations