Growth, accumulation, and antioxidative responses of two Salix genotypes exposed to cadmium and lead in hydroponic culture

  • Xiaohan Xu
  • Baoshan Yang
  • Guanghua QinEmail author
  • Hui WangEmail author
  • Yidan Zhu
  • Kaizhen Zhang
  • Hanqi Yang
Research Article


Cd and Pb are a toxic environmental pollutant, and their elevated concentrations in the waters and soils could exert detriment effects on human health by food chain. In order to evaluate the capacity to heavy metal accumulation and the physiochemical responses of two Salix genotypes, a 35-day hydroponic seedling experiment was implemented with Salix matsudana Koidz. ‘Shidi1’ (A42) and Salix psammophila C. ‘Huangpi1’ (A94) under different concentrations of Cd (15 and 30 μM) or Pb (250 and 300 μM). The results showed that the biomass of A94 severely reduced more than that of A42. The accumulation ability of Cd in different plant organs followed the sequence of leaves > roots > stems. Pb primarily accumulated in the roots for both Salix genotypes (54.27 mg g−1 for A42 and 54.52 mg g−1 for A94). Translocation factors based on accumulation (TF′) for Cd were more than 8.0, while TF′s for Pb were less than 1.0 in both A42 and A94, implying they could be applied in the phytoremediation of Cd-contaminated sites due to their stronger ability to Cd phytoextraction. The stress of Cd or Pb significantly increased malondialdehyde (MDA) contents and increased photosynthetic rates in leaves of two Salix genotypes. Transpiration rates of willow were positively correlated with its Cd translocation. Both catalase (CAT) and peroxidase (POD) activities were suppressed, while the superoxide dismutase (SOD) was boosted with increasing Cd and Pb levels in the leaves and roots of the two willow genotypes, suggesting SOD plays an important role in the removal of ROS. The inconsistency of the changes in enzyme activity suggests that the integrated antioxidative mechanisms regulate the tolerance to Cd and Pb stress.


Salix genotype Cd Pb Phytoremediation Antioxidation mechanism 



Thanks to Dr. Edward C. Mignot, whose mother language is English, served in Shandong University, for linguistic advice.

Funding information

The study was supported by projects from National Natural Science Foundation of China (31870606 and 41877424), Natural Science Foundation of Shandong Provincial, China (ZR2017MD022 and ZR2018MD002), and Special Funds for Forest Science Research in the Public Welfare (201404107).


  1. Aebi H (1984) Catalase in vitro. Methods Enzymol 105:121–126CrossRefGoogle Scholar
  2. Ali B, Mwamba TM, Gill RA, Yang C, Ali S, Daud MK, Wu YY, Zhou WJ (2014) Improvement of element uptake and antioxidative defense in Brassica napus under lead stress by application of hydrogen sulfide. Plant Growth Regul 74:261–273CrossRefGoogle Scholar
  3. Ali A, Guo D, Mahar A, Wang P, Ma F, Shen F, Li R, Zhang Z (2017) Phytoextraction of toxic trace elements by Sorghum bicolor inoculated with Streptomyces pactum (Act12) in contaminated soils. Ecotoxicol Environ Saf 139:202–209CrossRefGoogle Scholar
  4. Amna AN, Masood S, Mukhtar T, Kamran MA, Rafique M, Munis MF, Chaudhary HJ (2015) Differential effects of cadmium and chromium on growth, photosynthetic activity, and metal uptake of Linum usitatissimum in association with Glomus intraradices. Environ Monit Assess 187:1–11CrossRefGoogle Scholar
  5. Arias JA, Peralta-Videa JR, Ellzey JT, Ren M, Viveros MN, Gardea-Torresdey JL (2010) Effects of Glomus deserticola inoculation on Prosopis: enhancing chromium and lead uptake and translocation as confirmed by X-ray mapping, ICP-OES and TEM techniques. Environ Exp Bot 68:139–148CrossRefGoogle Scholar
  6. Arsenov D, Zupunski M, Borisev M, Nikolic N, Orlovic S, Pilipovic A, Pajevic S (2017) Exogenously applied citric acid enhances antioxidant defense and phytoextraction of cadmium by willows (Salix spp.). Water Air Soil Pollut 228:211CrossRefGoogle Scholar
  7. Bauddh K, Kumar A, Srivastava S, Singh RP, Tripathi RD (2016) A study on the effect of cadmium on the antioxidative defense system and alteration in different functional groups in castor bean and Indian mustard. Arch Agron Soil Sci 62:877–891CrossRefGoogle Scholar
  8. Benavides MP, Gallego SM, Tomaro ML (2005) Cadmium toxicity in plants. Braz J Plant Physiol 17:21–34CrossRefGoogle Scholar
  9. Bhatti SS, Kumar V, Sambyal V, Singh J, Nagpal AK (2018) Comparative analysis of tissue compartmentalized heavy metal uptake by common forage crop: a field experiment. Catena 160:185–193CrossRefGoogle Scholar
  10. Brekken A, Steinnes E (2004) Seasonal concentrations of cadmium and zinc in native pasture plants: consequences for grazing animals. Sci Total Environ 326:181–195CrossRefGoogle Scholar
  11. Cao YN, Ma CX, Chen GC, Zhang JF, Xing BS (2017) Physiological and biochemical responses of Salix integra Thunb. under copper stress as affected by soil flooding. Environ Pollut 225:644–653CrossRefGoogle Scholar
  12. Chmielowskabąk J, Gzyl J, Rucińskasobkowiak R, Arasimowiczjelonek M, Deckert J (2014) The new insights into cadmium sensing. Front Plant Sci 5:245Google Scholar
  13. Dogan M, Karatas M, Aasim M (2018) Cadmium and lead bioaccumulation potentials of an aquatic macrophyte Ceratophyllum demersum L.: a laboratory study. Ecotoxicol Environ Saf 148:431–440CrossRefGoogle Scholar
  14. Doumett S, Lamperi L, Checchini L, Azzarello E, Mugnai S, Mancuso S, Petruzzelli G, Del BM (2008) Heavy metal distribution between contaminated soil and Paulownia tomentosa, in a pilot-scale assisted phytoremediation study: influence of different complexing agents. Chemosphere 72:1481–1490CrossRefGoogle Scholar
  15. Farid M, Shakoor MB, Ehsan S, Ali S, Zubair M, Hanif MA (2013) Morphological, physiological and biochemical responses of different plant species to Cd stress. Int J Chem Biochem Sci 3:53–60Google Scholar
  16. Fernández R, Bertrand A, Reis R, Mourato MP, Martins LL, González A (2013) Growth and physiological responses to cadmium stress of two populations of Dittrichia viscosa (L.) Greuter. J Hazard Mater 244–245:555–562CrossRefGoogle Scholar
  17. Fernandez-Fuego D, Keunen E, Cuypers A, Bertrand A, Gonzalez A (2017) Mycorrhization protects Betula pubescens Ehr. from metal-induced oxidative stress increasing its tolerance to grow in an industrial polluted soil. J Hazard Mater 336:119–127CrossRefGoogle Scholar
  18. Guo B, Dai S, Wang R, Guo J, Ding Y, Xu Y (2015) Combined effects of elevated CO2 and Cd-contaminated soil on the growth, gas exchange, antioxidant defense, and Cd accumulation of poplars and willows. Environ Exp Bot 115:1–10CrossRefGoogle Scholar
  19. Han SH, Kim DH, Lee JC (2010) Cadmium and zinc interaction and phytoremediation potential of seven Salix caprea clones. J Ecol Environ 33:245–251CrossRefGoogle Scholar
  20. Hattab S, Hattab S, Flores-Casseres ML, Boussetta H, Doumas P, Hernandez LE, Banni M (2016) Characterisation of lead-induced stress molecular biomarkers in Medicago sativa plants. Environ Exp Bot 123:1–12CrossRefGoogle Scholar
  21. He JL, Qin JJ, Long LY, Ma YL, Li H, Li K, Jiang XN, Liu TX, Polle A, Liang ZS, Lou ZB (2011) Net cadmium flux and accumulation reveal tissue-specific oxidative stress and detoxification in Populus × canescens. Physiol Plant 143:50–63CrossRefGoogle Scholar
  22. Iori V, Pietrini F, Massacci A, Zacchini M (2015) Morphophysiological responses, heavy metal accumulation and phytoremoval ability in four willow clones exposed to cadmium under hydroponics phytoremediation. Springer International Publishing, New York, pp 87–98Google Scholar
  23. Iori V, Pietrini F, Bianconi D, Mughini G, Massacci A, Zacchini M (2017) Analysis of biometric, physiological, and biochemical traits to evaluate the cadmium phytoremediation ability of eucalypt plants under hydroponics. IForest 10:416–421CrossRefGoogle Scholar
  24. Islam E, Liu D, Li T, Yang X, Jin X, Mahmood Q, Tian S, Li J (2007) Effect of Pb toxicity on leaf growth, physiology and ultrastructure in the two ecotypes of Elsholtzia argyi. J Hazard Mater 147:806–816CrossRefGoogle Scholar
  25. Karene G, Huang XD, Bernardr G, Brucem G (2009) Phytoremediation and rhizoremediation of organic soil contaminants: potential and challenges. Plant Sci 176:20–30CrossRefGoogle Scholar
  26. Kaur G, Singh HP, Batish DR, Kohli RK (2013) Lead (Pb)-induced biochemical and ultrastructural changes in wheat (Triticum aestivum) roots. Protoplasma 250:53–62CrossRefGoogle Scholar
  27. Kersten G, Majestic B, Quigley M (2017) Phytoremediation of cadmium and lead-polluted watersheds. Ecotoxicol Environ Saf 137:225–232CrossRefGoogle Scholar
  28. Khan MM, Islam E, Irem S, Akhtar K, Ashraf MY, Iqbal J, Liu D (2018) Pb-induced phytotoxicity in para grass (Brachiaria mutica) and Castor bean (Ricinus communis L.): antioxidant and ultrastructural studies. Chemosphere 200:257–265CrossRefGoogle Scholar
  29. Kim D, Kim HS, You HK, Park HS, Chung HC, Han GD (2008) Responses of hybrid poplar to cadmium stress: photosynthetic characteristics, cadmium and proline accumulation and antioxidant enzyme activity. Acta Biol Cracov 50:95–103Google Scholar
  30. Klang-Westin E, Perttu K (2002) Effects of nutrient supply and soil cadmium concentration on cadmium removal by willow. Biomass Bioenergy 23:415–426CrossRefGoogle Scholar
  31. Kumar A, Prasad M (2018) Plant-lead interactions: transport, toxicity, tolerance, and detoxification mechanisms. Ecotoxicol Environ Saf 166:401–418CrossRefGoogle Scholar
  32. Kumar A, Prasad MNV, Sytar O (2012) Lead toxicity, defense strategies and associated indicative biomarkers in Talinum triangulare grown hydroponically. Chemosphere 89:1056–1065CrossRefGoogle Scholar
  33. Labidi S, Firmin S, Verdin A, Bidar G, Laruelle F, Douay F, Shirali P, Fontaine J, Lounes-Hadj Sahraoui A (2017) Nature of fly ash amendments differently influences oxidative stress alleviation in four forest tree species and metal trace element phytostabilization in aged contaminated soil: a long-term field experiment. Ecotoxicol Environ Saf 138:190–198CrossRefGoogle Scholar
  34. Lamhamdi M, El Galiou O, Bakrim A, Nóvoa-Muñoz JC, Arias-Estévez M, Aarab A, Lafont R (2013) Effect of lead stress on mineral content and growth of wheat (Triticum aestivum) and spinach (Spinacia oleracea) seedlings. Saudi J Biol Sci 20:29–36CrossRefGoogle Scholar
  35. Landberg T, Greger M (2002) Interclonal variation of heavy metal interactions in Salix viminalis. Environ Toxicol Chem 21:2669–2674CrossRefGoogle Scholar
  36. Lei Y, Korpelainen H, Li C (2007) Physiological and biochemical responses to high Mn concentrations in two contrasting Populus cathayana populations. Chemosphere 68:686–694CrossRefGoogle Scholar
  37. Li X, Li N, Yang J, Ye F, Chen F, Chen F (2011) Morphological and photosynthetic responses of riparian plant Distylium chinense seedlings to simulated autumn and winter flooding in Three Gorges Reservoir Region of the Yangtze River, China. Acta Ecol Sin 31:31–39CrossRefGoogle Scholar
  38. Li Y, Zhou C, Huang M, Luo J, Hou X, Wu P, Ma X (2016) Lead tolerance mechanism in Conyza canadensis: subcellular distribution, ultrastructure, antioxidative defense system, and phytochelatins. J Plant Res 129:251–262CrossRefGoogle Scholar
  39. Lunáčková L, Masarovičová E, Král’Ová K, Streško V (2003) Response of fast growing woody plants from family Salicaceae to cadmium treatment. B Environ Contam Tox 70:576–585CrossRefGoogle Scholar
  40. Malá J, Cvrčková H, Máchová P, Dostál J, Šíma P (2010) Heavy metal accumulation by willow clones in short-time hydroponics. J For Sci 56:28–34CrossRefGoogle Scholar
  41. Malecka A, Piechalak A, Tomaszewska B (2009) Reactive oxygen species production and antioxidative defense system in pea root tissues treated with lead ions: the whole roots level. Acta Physiol Plant 31:1053–1063CrossRefGoogle Scholar
  42. Manivasagaperumal R, Balamurugan S, Thiyagarajan G, Sekar J (2011) Effect of zinc on germination, seedling growth and biochemical content of cluster bean (Cyamopsis tetragonoloba (L.) Taub.). Curr Bot 2:11–15Google Scholar
  43. Marques APGC, Rangel AOSS, Castro PML (2009) Remediation of heavy metal contaminated soils: phytoremediation as a potentially promising clean-up technology. Crit Rev Environ Sci Technol 39:622–654CrossRefGoogle Scholar
  44. Mouna F, Laurent L, Najib B, Valerie H, El MM, Didier B, Abdelaziz S (2013) Effect of lead on root growth. Front Plant Sci 4:175Google Scholar
  45. Najeeb U, Jilani G, Ali S, Sarwar M, Xu L, Zhou W (2011) Insights into cadmium induced physiological and ultra-structural disorders in Juncus effusus L. and its remediation through exogenous citric acid. J Hazard Mater 186:565–574CrossRefGoogle Scholar
  46. Nikolić N, Kojić D, Pilipović A, Pajević S, Krstić B, Borišev M, Orlović S (2008) Responses of hybrid poplar to cadmium stress: photosynthetic characteristics, cadmium and proline accumulation and antioxidant enzyme activity. Acta Biol Cracov Ser Bot 2:95–103Google Scholar
  47. Nikolić N, Borišev M, Pajević S, Arsenov D, Zupunski M, Orlovic S, Pilipovic A (2015) Photosynthetic response and tolerance of three willow species to cadmium exposure in hydroponic culture. Arch Biol Sci 67:1411–1420CrossRefGoogle Scholar
  48. Nissim WG, Voicu A, Labrecque M (2014) Willow short-rotation coppice for treatment of polluted groundwater. Ecol Eng 62:102–114CrossRefGoogle Scholar
  49. Pandey N, Sharma CP (2002) Effect of heavy metals Co2+, Ni2+ and Cd2+ on growth and metabolism of cabbage. Plant Sci 163:753–758CrossRefGoogle Scholar
  50. Pandey N, Singh GK (2012) Studies on antioxidative enzymes induced by cadmium in pea plants (Pisum sativum). J Environ Biol 33:201–206Google Scholar
  51. Pandhair V, Sekhon BS (2006) Reactive oxygen species and antioxidants in plants: an overview. J Plant Biochem Biotechnol 15:71–78CrossRefGoogle Scholar
  52. Pellegrini E, Hoshika Y, Dusart N, Cotrozzi L, GéRard J, Nali C, Paoletti E (2019) Antioxidative responses of three oak species under ozone and water stress conditions. Sci Total Environ 647:390–399CrossRefGoogle Scholar
  53. Pietrini F, Zacchini M, Iori V, Pietrosanti L, Bianconi D, Massacci A (2010a) Screening of poplar clones for cadmium phytoremediation using photosynthesis, biomass and cadmium content analyses. Int J Phytoremediat 12:105–120CrossRefGoogle Scholar
  54. Pietrini F, Zacchini M, Iori V, Pietrosanti L, Ferretti M, Massacci A (2010b) Spatial distribution of cadmium in leaves and its impact on photosynthesis: examples of different strategies in willow and poplar clones. Plant Biol 12:355–363CrossRefGoogle Scholar
  55. Pourrut B, Jean S, Silvestre J, Pinelli E (2011) Lead-induced DNA damage in Vicia faba root cells: potential involvement of oxidative stress. Mutat Res/Gene Toxicol Environ Mutagen 726:123–128CrossRefGoogle Scholar
  56. Qin G, Meng X, Wang Q, Tian S (2009) Oxidative damage of mitochondrial proteins contributes to fruit senescence: a redox proteomics analysis. J Proteome Res 8:2449–2462CrossRefGoogle Scholar
  57. Rai VK (2002) Role of amino acids in plant responses to stresses. Biol Plant 45:481–487CrossRefGoogle Scholar
  58. Redovniković IR, De Marco A, Proietti C, Hanousek K, Sedak M, Bilandžić N, Jakovljević T (2017) Poplar response to cadmium and lead soil contamination. Ecotoxicol Environ Saf 144:482–489CrossRefGoogle Scholar
  59. Rinalducci S, Murgiano L, Zolla L (2008) Redox proteomics: basic principles and future perspectives for the detection of protein oxidation in plants. J Exp Bot 59:3781–3801CrossRefGoogle Scholar
  60. Sawidis T (2008) Effect of cadmium on pollen germination and tube growth in Lilium longiflorum and Nicotiana tabacum. Protoplasma 233:95–106CrossRefGoogle Scholar
  61. Schützendübel A, Polle A (2002) Plant responses to abiotic stresses: heavy metal-induced oxidative stress and protection by mycorrhization. J Exp Bot 53:1351–1365Google Scholar
  62. Shahid M, Pourrut B, Dumat C, Nadeem M, Aslam M, Pinel E (2014) Heavy-metal-induced reactive oxygen species: phytotoxicity and physicochemical changes in plants. Rev Environ Contam Toxicol 232:1–44Google Scholar
  63. Sharma SS, Schat H, Vooijs R (1998) In vitro alleviation of heavy metal-induced enzyme inhibition by proline. Phytochemistry 49:1531–1535CrossRefGoogle Scholar
  64. Sharma P, Jha AB, Dubey RS, Pessarakli M (2012) Reactive oxygen species, oxidative damage, and antioxidative defense mechanism in plants under stressful conditions. Aust J Bot 2012:1–26Google Scholar
  65. Shi GR, Cai QS, Liu QQ, Wu L (2009) Salicylic acid-mediated alleviation of cadmium toxicity in hemp plants in relation to cadmium uptake, photosynthesis, and antioxidant enzymes. Acta Physiol Plant 31:969–977CrossRefGoogle Scholar
  66. Sidhu GPS, Singh HP, Batish DR, Kohli RK (2017a) Tolerance and hyperaccumulation of cadmium by a wild, unpalatable herb Coronopus didymus (L.) Sm (Brassicaceae). Ecotoxicol Environ Saf 135:209–215CrossRefGoogle Scholar
  67. Sidhu GPS, Singh HP, Batish DR, Kohli RK (2017b) Effect of lead on oxidative status, antioxidative response and metal accumulation in Coronopus didymus. Plant Physiol Biochem 105:290–296CrossRefGoogle Scholar
  68. Silva JRR, Fernandes AR, Silva Junior ML, Santos CRC, Lobato AKS (2018) Tolerance mechanisms in Cassia alata exposed to cadmium toxicity – potential use for phytoremediation. Photosynthetica 56:495–504CrossRefGoogle Scholar
  69. Stoláriková M, Vaculík M, Lux A, Di Baccio D, Minnocci A, Andreucci A, Sebastiani L (2012) Anatomical differences of poplar (Populus × euramericana clone I-214) roots exposed to zinc excess. Biologia 67:483–489CrossRefGoogle Scholar
  70. Stomp AM, Han KH, Wilbert S, Gordon MP (1993) Genetic improvement of tree species for remediation of hazardous wastes. In Vitro Cell Dev Biol Plant 29:227–232CrossRefGoogle Scholar
  71. Sylvain B, Mikael MH, Florie M, Emmanuel J, Marilyne S, Sylvain B, Domenico M (2016) Phytostabilization of As, Sb and Pb by two willow species (S. viminalis and S. purpurea) on former mine technosols. Catena 136:44–52CrossRefGoogle Scholar
  72. Szabados L, Savouré A (2010) Proline: a multifunctional amino acid. Trends Plant Sci 15:89–97CrossRefGoogle Scholar
  73. Tamás L, Dudíková J, Ďurčeková K, Halušková L, Huttová J, Mistrík I, Ollé M (2008) Alterations of the gene expression, lipid peroxidation, proline and thiol content along the barley root exposed to cadmium. J Plant Physiol 165:1193–1203CrossRefGoogle Scholar
  74. Tanyolaç D, Ekmekçi Y, Unalan S (2007) Changes in photochemical and antioxidant enzyme activities in maize (Zea mays L.) leaves exposed to excess copper. Chemosphere 67:89–98CrossRefGoogle Scholar
  75. Tőzsér D, Magura T, Simon E (2017) Heavy metal uptake by plant parts of willow species: a meta-analysis. J Hazard Mater 336:101–109CrossRefGoogle Scholar
  76. Utmazian MNDS, Wieshammer G, Vega R, Wenzel WW (2007) Hydroponic screening for metal resistance and accumulation of cadmium and zinc in twenty clones of willows and poplars. Environ Pollut 148:155–165CrossRefGoogle Scholar
  77. Vandecasteele B, Meers E, Vervaeke P, De VB, Quataert P, Tack F (2005) Growth and trace metal accumulation of two Salix clones on sediment-derived soils with increasing contamination levels. Chemosphere 58:995–1002CrossRefGoogle Scholar
  78. Vassilev A, Perez-Sanz A, Semane B, Carleer R, Vangronsveld J (2005) Cadmium accumulation and tolerance of two Salix genotypes hydroponically grown in presence of cadmium. J Plant Nutr 28:2159–2177CrossRefGoogle Scholar
  79. Wang SF, Shi X, Sun HJ, Chen YT, Yang XE (2013) Metal uptake and root morphological changes for two varieties of Salix integra under cadmium stress. Acta Ecol Sin 33:6065–6073CrossRefGoogle Scholar
  80. Wu F, Yang W, Zhang J, Zhou L (2010) Cadmium accumulation and growth responses of a poplar (Populus deltoides × Populus nigra) in cadmium contaminated purple soil and alluvial soil. J Hazard Mater 177:268–273CrossRefGoogle Scholar
  81. Yang LP, Zhu J, Wang P, Zeng J, Tan R, Yang YZ, Liu ZM (2018) Effect of Cd on growth, physiological response, Cd subcellular distribution and chemical forms of Koelreuteria paniculata. Ecotoxicol Environ Saf 160:10–18CrossRefGoogle Scholar
  82. Zacchini M, Pietrini F, Mugnozza GS, Iori V, Pietrosanti L, Massacci A (2009) Metal tolerance, accumulation and translocation in poplar and willow clones treated with cadmium in hydroponics. Water Air Soil Pollut 197:23–34CrossRefGoogle Scholar
  83. Zhivotovsky OP, Kuzovkina JA, Schulthess CP, Morris T, Pettinelli D, Ge M (2011) Hydroponic screening of willows (Salix L.) for lead tolerance and accumulation. Inter J Phytoreme 13:75–94CrossRefGoogle Scholar

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© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.School of Water Conservancy and EnvironmentUniversity of JinanJinanChina
  2. 2.Shandong Academy of ForestryJinanChina
  3. 3.The Xiuwen International Academy at JinanJinanChina

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