Genotoxicity response of Vicia faba seedlings to cadmium in soils as characterized by direct soil exposure and micronucleus test

  • Lang Chen
  • Shankui Yuan
  • Xingang Liu
  • Xinxin Zhou
  • Yanming Zhou
  • Yufang SongEmail author


To overcome the drawbacks of the Vicia faba root tip micronucleus test in soil using the solution extract method, we conducted a potting experiment by direct soil exposure. Cadmium was spiked into 3 typical soils (brown soil, red soil, and black soil) to simulate environmental concentrations (0.625, 1.25, 2.5, 5, and 10 mg kg−1). Multiple Vicia faba tissues (primary root tips, secondary root tips, and leaf tips) were sampled, and mitotic index (MI), chromosome aberration frequency (CA), and micronucleus frequency (MN) were used as endpoints after a seedling period of 5 days. The results showed a response between Cd concentrations and multiple sampling tissues of Vicia faba, and the secondary root tips responded to Cd stress the most, followed by primary root tips and leaf tips. Soil physicochemical properties (e.g., pH, total phosphorus, total organic carbon, etc.) influenced the genotoxicity of Cd, and pH was the dominant factor, which resulted in the genetic toxicity response of Cd in soils in the order: red soil > brown soil > black soil. The lowest observable effect concentration (LOEC) of Cd was 1.25 mg kg−1 for both brown soil and red soil and 2.5 mg kg−1 for black soil. In view of this, we suggested that soil properties should be considered in evaluating genotoxicity risk of Cd in soil, especially with soil pH range, and the secondary root tips should be taken as suitable test tissues in the MN test due to its more sensible response feature to Cd stress in soil.


  1. 1.

    The best sampling tissue for micronucleus test of Vicia faba seedlings was secondary root tips, followed by primary root and leaf tips, and the later may be used for in situ monitoring.

  2. 2.

    Soil pH did have influence on the genotoxicity effects of cadmium, and the NOECs of Cd on Vicia faba were 0.625 (red soil), 0.625 (brown soil) and 1.25 (black soil) mg kg−1, respectively.

  3. 3.

    The existing Cd limits in soil may not be protective enough for higher plants and even human beings.



Direct soil exposure Genotoxicity Vicia faba Micronucleus test Secondary root tip Cadmium 



This research was financially supported by the National Key R&D Program of China, 2018YFD0200100 and the Natural Science Foundation of China (Grant No. 31801768 and 31861133021).


This research was financially supported by the National Key R&D Program of China, 2018YFD0200100 and the Natural Science Foundation of China (Grant No. 31801768 and 31861133021).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

This manuscript does not contain any studies with human participants or animals performed by any of the authors. All authors have read and approved this version of the article, and due care has been taken to ensure the integrity of the work. Neither the entire paper nor any part of its content has been published or has been accepted elsewhere. And it is not being submitted to any other journal.

Informed consent

This manuscript does not contain any studies with human participants or animals performed by any of the authors.


  1. An YJ (2004) Soil ecotoxicity assessment using cadmium sensitive plants. Environ Pollut 127:21–26CrossRefGoogle Scholar
  2. Béraud E, Cotelle S, Leroy P, Férard JF (2007) Genotoxic effects and induction of phytochelatins in the presence of cadmium in Vicia faba roots. Mutat Res 633:112–116CrossRefGoogle Scholar
  3. Bao SD (2000) The method of the soil and agriculture chemical analysis. China Agriculture Press, BeijingGoogle Scholar
  4. Bilodeau-Gauthier S, Houle D, Gagnon C, Côté B, Messier C (2011) Assessment of sugar maple tree growth in relation to the partitioning of elements in xylem along a soil acidity gradient. For Ecol Manag 261:95–104CrossRefGoogle Scholar
  5. Bray RH, Kurtz LT (1945) Determination of total, organic, and available forms of phosphorus in soils. Soil Sci 59:39–45CrossRefGoogle Scholar
  6. Bremner JM, Mulvaney CS (1982) Nitrogen total. In: Method of soil analysis—part 2: chemical and microbiological methods (2nd edn). 595–624 Springer-Verlag, BerlinGoogle Scholar
  7. Chang XX (1999) Cytogenetic toxic effects of Cd2 + and Al3+ on root tip of Vicia faba. Agro-Environ Prot 18:1–3Google Scholar
  8. Chen L, Song YF, Zhang W, Xiu ying LI, Wang L, Pu hui JI, Yang XX (2008) Assessment of toxicity effects for cadmium contamination in soils by means of multi-indexes. Huan Jing Ke Xue 29:2606–2612Google Scholar
  9. Claire-Emmanuelle MR, Maritxu G, Marie C, Sylvie C, Eric P (2009) New direct contact approach to evaluate soil genotoxicity using the Vicia faba micronucleus test. Chemosphere 77:345–350CrossRefGoogle Scholar
  10. Cotelle S, Masfaraud JFO, Férard JF (1999) Assessment of the genotoxicity of contaminated soil with the Allium/Vicia-micronucleus and the Tradescantia-micronucleus assays. Mut Res-FUND MOL 426:167–171CrossRefGoogle Scholar
  11. Cui LT, Geng SG, Li ZW (2006) Status and control of cadmium pollution in farmland soil in China. Mod Agric Sci Technol 125:184–185. 2006Google Scholar
  12. Degrassi F, Rizzoni M (1982) Micronucleus test in Vicia faba root tips to detect mutagen damage in fresh-water pollution. Mutat Res 97:19–33CrossRefGoogle Scholar
  13. Dehn PF, White CM, Conners DE, Shipkey G, Cumbo TA (2004) Characterization of the human hepatocellular carcinoma (HEPG2) cell line as an in vitro model for cadmium toxicity studies. In Vitro Cell Dev Biol Anim 40:172–182CrossRefGoogle Scholar
  14. Demarco A, Desimone CM, Lorenzoni P (1995) Influence of soil characteristics on the clastogenic activity of maleic hydrazide in root tips of vicia faba. Mutat Res Genet Toxicol 344:5–12CrossRefGoogle Scholar
  15. Deng SP, Zhao YT, Zhu CH, Zeng MS, Shu-Fang FU, Guang-Li, Li (2012) Effect of cadmium on the antioxidant enzyme activity and lipid peroxidation in sanguinolaria acuta. Acta Hydrobiol Sinica 36:689–695Google Scholar
  16. Depledge MH (1990) New approaches in ecotoxicology: can interindividual physiological variability be used as a tool to investigate pollution effects? Ambio 19:251–252Google Scholar
  17. Duan CQ, Wang HX (1995) Cytogenetical toxical effects of heavy metals on Vicia faba and inquires into the Vicia-micronucleus. Acta Bot Sinica 37:14–24Google Scholar
  18. Feng SL, Wang XM, Wei GJ, Peng PG, Yang Y, Gao ZH (2007) Leachates of municipal solid waste incineration bottom ash from Macao: heavy metal concentrations and genotoxicity. Chemosphere 67:1133–1137CrossRefGoogle Scholar
  19. Flores-Maya S, Gómez-Arroyo S, Calderón-Segura ME, Villalobos-Pietrini R, Waliszewski SM, Cruz LGDL (2005) Promutagen activation of triazine herbicides metribuzin and ametryn through Vicia faba metabolism inducing sister chromatid exchanges in human lymphocytes in vitro and in V. faba root tip meristems. Toxicol In Vitro 19:243–251CrossRefGoogle Scholar
  20. Foltête AS, Masfaraud JF, Férard JF, Cotelle S (2012) Is there a relationship between early genotoxicity and life-history traits in Vicia faba exposed to cadmium-spiked soils? Mutat Res 747:159–163CrossRefGoogle Scholar
  21. He ZL, Yang XE, Stoffella PJ (2005) Trace elements in agroecosystems and impacts on the environment. J Trace Elem Med Biol 19:125–140CrossRefGoogle Scholar
  22. Iqbal M (2016) Vicia faba bioassay for environmental toxicity monitoring: a review. Chemosphere 144:785–802CrossRefGoogle Scholar
  23. ISO (2013) 29200-Soil quality e assessment of genotoxic effects on higher plants -Vicia faba micronucleus test. ISO, SwitzerlandGoogle Scholar
  24. Ma J, Shen JL, Liu QX, Fang F, Cai HS, Guo CH (2014) Risk assessment of petroleum-contaminated soil using soil enzyme activities and genotoxicity to Vicia faba. Ecotoxicology 23:665–673CrossRefGoogle Scholar
  25. Kristen U (1997) Use of higher plants as screens for toxicity assessment. Toxicol In Vitro 11:181–191CrossRefGoogle Scholar
  26. Lagriffoul A, Mocquot B, Mench M, Vangronsveld J (1998) Cadmium toxicity effects on growth, mineral and chlorophyll contents, and activities of stress related enzymes in young maize plants (Zea mays L.). Plant Soil 200:241–250CrossRefGoogle Scholar
  27. Leme DM, Marin-Morales MA (2009) Allium cepa test in environmental monitoring: a review on its application. Mutat Res 682:71–81CrossRefGoogle Scholar
  28. Liu Y, Min G, Wei H, Zhe L (2017) Evaluation and quantification of genotoxicity of urban waters by using Vicia faba bioassays. Chem Ecol 33:1–15CrossRefGoogle Scholar
  29. Lucia G, Hakima T, Mohammed M, Leonardo C, Chiara G, Stefania F (2011) Genotoxicity evaluation of effluents from textile industries of the region Fez-Boulmane, Morocco: a case study. Ecotoxicol Environ Saf 74:2275–2283CrossRefGoogle Scholar
  30. Luo HF (2017) Study on BMD of cadmium in urine, rice and soil based on human renal function. Jiaotong University, BeijingGoogle Scholar
  31. Ma LJ, Zhang Y, Bu N, Wang SH (2010) Alleviation effect of alginate-derived oligosaccharides on Vicia faba root tip cells damaged by cadmium. Bull Environ Contamination Toxicol 84:161–164CrossRefGoogle Scholar
  32. Ma TH (1982) Vicia cytogenetic tests for environmental mutagens. A report of the U.S. Environmental Protection Agency Gene-Tox Program. Mutat Res 99:257–271CrossRefGoogle Scholar
  33. Manier N, Deram A, Curieux FL, Marzin D (2009) Comparison between new wild plant trifolium repens and Vicia faba on their sensitivity in detecting the genotoxic potential of heavy metal solutions and heavy metal-contaminated soils. Water Air Soil Pollut 202:343–352CrossRefGoogle Scholar
  34. Manzo S, Picione FDL, Rocco A, Carotenuto R, Maisto G (2010) Urban and agricultural soil ecotoxicity and heavy metal contamination. Fresenius Environ Bull 09:1749–1755Google Scholar
  35. MOA (2006) Soil testing Part 6: method for determination of organic matter. China Agricultural Press, BeijingGoogle Scholar
  36. Ohe T, Watanabe T, Wakabayashi K (2004) Mutagens in surface waters: a review. Mutat Res 567:109–149CrossRefGoogle Scholar
  37. Pansu DM, Gautheyrou J (2006) Handbook of soil analysis. Springer-Verlag, BerlinCrossRefGoogle Scholar
  38. Patlolla AK, Berry A, May LB, Tchounwou PB (2012) Genotoxicity of silver nanoparticles in Vicia faba: a pilot study on the environmental monitoring of nanoparticles. Int J Environ Res Public Health 9:1649–1662CrossRefGoogle Scholar
  39. Richardson SD, Demarini DM, Manolis K, Pilar F, Esther M, Carolina L, Clara B, Dick H, Kees M, A Bruce M (2010) What’s in the pool? A comprehensive identification of disinfection by-products and assessment of mutagenicity of chlorinated and brominated swimming pool water. Environ Health Perspect 118:1523–1530CrossRefGoogle Scholar
  40. Serpil U, Ayla C, F Ozlem C, Aysin GZ (2006) Cadmium-induced genotoxicity, cytotoxicity and lipid peroxidation in Allium sativum and Vicia faba. Mutagenesis 21:77–81CrossRefGoogle Scholar
  41. Shahid M, Dumat C, Khalid S, Niazi NK, Antunes PMC (2016) Cadmium bioavailability, uptake, toxicity and detoxification in soil-plant system. Rev Environ Contam Toxicol 241:73–137Google Scholar
  42. Shahid M, Dumat C, Pourrut B, Abbas G, Shahid N, Pinelli E (2015) Role of metal speciation in lead-induced oxidative stress to Vicia faba roots. Russ J Plant Physiol 62:448–454CrossRefGoogle Scholar
  43. Shahid M, Pinelli E, Pourrut B, Silvestre J, Dumat C (2011) Lead-induced genotoxicity to Vicia faba L. roots in relation with metal cell uptake and initial speciation. Ecotoxicol Environ Saf 74:78–84CrossRefGoogle Scholar
  44. Silva AJD, Gouveia-Neto ADS, Silva-Jr EAD (2012) LED-Induced chlorophyll fluorescence spectral analysis for the early detection and monitoring of cadmium toxicity in maize plants. Water Air Soil Pollut 223:3527–3533CrossRefGoogle Scholar
  45. Song YF, Gong P, Wilke BM, Zhang W, Song XY, Sun TH, Ackland ML (2007) Genotoxicity assessment of soils from wastewater irrigation areas and bioremediation sites using the Vicia faba root tip micronucleus assay. J Environ Monit 9:182–186CrossRefGoogle Scholar
  46. Song YF, Kai JR, Song XY, Zhang W, Li LL (2015) Long-term toxic effects of deltamethrin and fenvalerante in soil. J Hazard Mater 289:158–164CrossRefGoogle Scholar
  47. Strickland RC (1979) Cadmium uptake by pinus resinosa ait. pollen and the effect on cation release and membrane permeability. Plant Physiol 64:366–370CrossRefGoogle Scholar
  48. Tang X, Gu X, Wen Y (2018) Study on teratogenic effect of nitrobenzene on Vicia faba root tip cells. Laboratory Cell Biological Technic 23–31Google Scholar
  49. Turner L, Choplin F, Dugard P, Hermens J, Jaeckh R, Marsmann M, Roberts D (1987) Structure-activity relationships in toxicology and ecotoxicology: an assessment. Toxicol In Vitro 1:143–171CrossRefGoogle Scholar
  50. Wagner GJ (1993) Accumulation of cadmium in crop plants and its consequences to human health. Adv Agron 51:173–212CrossRefGoogle Scholar
  51. Wang H (1999) Clastogenicity of chromium contaminated soil samples evaluated by Vicia root-micronucleus assay. Mutat Res 426:147–149CrossRefGoogle Scholar
  52. Wang GQ, Deng SP, Feng YH, Zheng LP, Zhang Y, Lin YS (2015) Comparative study on soil environmental standards for heavy metals in China and other countries: cadmium. J Ecol Rural Environ 31:808–821Google Scholar
  53. Wu J, Geilfus CM, Pitann B, Mühling KH (2016) Silicon-enhanced oxalate exudation contributes to alleviation of cadmium toxicity in wheat. Environ Exp Bot 131:10–18CrossRefGoogle Scholar
  54. Yi M, Yi H, Li H, Wu L (2010) Aluminum induces chromosome aberrations, micronuclei, and cell cycle dysfunction in root cells of Vicia faba. Environ Toxicol 25:124–129Google Scholar
  55. Zhang X, Rui H, Zhang F, Hu Z, Yan X, Shen Z (2018) Overexpression of a functionalVicia sativaPCS1 homolog increases cadmium tolerance and phytochelatins synthesis in arabidopsis. Front Plant Sci 9:107CrossRefGoogle Scholar
  56. Zhan FD, Qin L, Guo Xh, Tan JB, Liu NN, Zu YQ, Li Y (2016) Cadmium and lead accumulation and low-molecular-weight organic acids secreted by roots in an intercropping of a cadmium accumulator Sonchus asper L. with Vicia faba L. Rsc Adv 6:33240–33248CrossRefGoogle Scholar
  57. Zhang XX, Rui HY, Zhang FQ, Hu ZB, Xia Y, Shen ZG (2018) Overexpression of a functional Vicia sativa PCS1 homolog increases cadmium tolerance and phytochelatins synthesis in arabidopsis. Front Plant Sci 9:107CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Lang Chen
    • 1
  • Shankui Yuan
    • 2
  • Xingang Liu
    • 1
  • Xinxin Zhou
    • 2
  • Yanming Zhou
    • 2
  • Yufang Song
    • 3
    Email author
  1. 1.State Key Laboratory for Biology of Plant Disease and Insect Pests, Institute of Plant ProtectionChinese Academy of Agricultural SciencesBeijingChina
  2. 2.Institute for the Control of AgrochemicalsMinistry of Agriculture and Rural AffairsBeijingChina
  3. 3.Key Laboratory of Pollution Ecology and Environmental Engineering, Institute of Applied EcologyChinese Academy of SciencesShenyangP.R. China

Personalised recommendations