Skip to main content
Log in

Genotoxic effects and proteomic analysis on Allium cepa var. agrogarum L. root cells under Pb stress

  • Published:
Ecotoxicology Aims and scope Submit manuscript

Abstract

Ionic lead (Pb) in the environment has accumulated due to anthropogenic activities, causing a potential threat to plants and plant consumers. We conducted this study to reveal the molecular mechanism of Pb stress response in plants. The effects of Pb (5.0 and 15.0 μM) on mitosis, DNA replication, gene expression and proteins in root-tip cells of Allium cepa var. agrogarum L. were addressed. The results indicated that root growth was inhibited dramatically in Pb treatment groups. Chromosomal aberrations were observed and the mitotic index decreased during Pb treatments at different concentrations. The accumulation of reactive oxygen species (ROS) in onion roots was induced by Pb stress. Pb increased DNA damage and suppressed cell cycle progression. The above toxic effects got more serious with increasing Pb concentration and prolonging exposure time. A total of 17 proteins were expressed differentially between control and Pb exposure groups. Under Pb treatment, the decreased expression of Anx D1 indicated decreased defensive response; the decreased expression of SHMT1 indicated decreased respiration; the decreased expression of COMT2 indicated decreased response of other funtions; the increased expression of NDPK indicated increased transcription and protein synthesis; the increased expression of PR1 and CHI1 indicated increased pathogen invasion; the increased expression of ORC5 and MPK5 indicated the reduced DNA replicating activity; the decreased expression of POLD1 indicated the reduced DNA repair activity. Our results provide new insights at the proteomic level into the Pb-induced responses, defensive responses and toxic effects, and provide new molecular markers of the early events of plant responses to Pb toxicity.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8

Similar content being viewed by others

References

  • Attanayake CP, Hettiarachchi GM, Harms AMR, Presley DR, Martin S, Pierzynski GM (2014) Field evaluations on soil plant transfer of lead from an urban garden soil. J Environ Qual 43:475–487

    Google Scholar 

  • Bhattacharjee S (2005) Reactive oxygen species and oxidative burst: Roles in stress, senescence and signal transduction in plants. Curr Sci 89:1113–1121

    CAS  Google Scholar 

  • Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254

    CAS  Google Scholar 

  • Brunet J, Varrault G, Zuilyfodil Y, Repellin A (2009) Accumulation of lead in the roots of grass pea (Lathyrus sativus L.) plants triggers systemic variation in gene expression in the shoots. Chemosphere 77:1113–1120

    CAS  Google Scholar 

  • Buell CR, Somerville S (1997) Use of Arabidopsis recombinant inbred lines reveals a monogenic and a novel digenic resistance mechanism to Xanthomonas campestris pv campestris. Plant J 12:21–29

    CAS  Google Scholar 

  • Cenkci S, Cigerci IH, Yildiz M, Ozay C, Bozdag A, Terzi H (2010) Lead contamination reduces chlorophyll biosynthesis and genomic template stability in Brassica rapa L. Environ Exp Bot 67:467–473

    CAS  Google Scholar 

  • Colak N, Torun H, Gruz J, Strnad M, Ayaz FA (2019) Exogenous N-Acetylcysteine alleviates heavy metal stress by promoting phenolic acids to support antioxidant defence systems in wheat roots. Ecotoxicol Environ Saf 181:49–59. https://doi.org/10.1016/j.ecoenv.2019.05.052

    Article  CAS  Google Scholar 

  • Davis JM, Wu H, Cooke JEK, Reed J, Luce KS, Michler CH (2002) Pathogen challenge, salicylic acid, and jasmonic acid regulate expression of chitinase gene homologs in pine. Mol Plant-Microbe Interact 15:380–387

    CAS  Google Scholar 

  • Diaztrivino S, Castellano MADM, Sanchez MADLP, Ramirezparra E, Desvoyes BND, Gutierrez C (2005) The genes encoding Arabidopsis ORC subunits are E2F targets and the two ORC1 genes are differently expressed in proliferating and endoreplicating cells. Nucleic Acids Res 33:5404–5414

    CAS  Google Scholar 

  • Ding C, Li X, Zhang T, Wang X (2015) Transfer model of lead in soil–carrot (Daucus carota L.) system and food safety thresholds in soil. Environ Toxicol Chem 34:2078–2086

    CAS  Google Scholar 

  • Fahr M, Laplaze L, Bendaou N, Hocher V, Mzibri ME, Bogusz D, Smouni A (2013) Effect of lead on root growth. Front Plant Sci 4:175–175

    Google Scholar 

  • Fang Y et al. (2017) Protection mechanism of Se-containing protein hydrolysates from Se-enriched rice on Pb2+-induced apoptosis in PC12 and RAW264.7 cells. Food Chem 219:391–398. https://doi.org/10.1016/j.foodchem.2016.09.131

    Article  CAS  Google Scholar 

  • Gaudet P, Livstone MS, Lewis SE, Thomas PD (2011) Phylogenetic-based propagation of functional annotations within the Gene Ontology consortium. Brief Bioinforma 12:449–462

    Google Scholar 

  • Ghosh M et al. (2016) Effects of ZnO nanoparticles in plants: Cytotoxicity, genotoxicity, deregulation of antioxidant defenses, and cell-cycle arrest. Mutat Res/Genet Toxicol Environ Mutagenesis 807:25–32. https://doi.org/10.1016/j.mrgentox.2016.07.006

    Article  CAS  Google Scholar 

  • Gorg A, Weiss W, Dunn MJ (2004) Current two-dimensional electrophoresis technology for proteomics. Proteomics 4:3665–3685

    Google Scholar 

  • Hattab S, Chouba L, Kheder M, Mahouachi T, Boussetta H (2009) Cadmium- and copper-induced DNA damage in Pisum sativum roots and leaves as determined by the comet assay. Plant Biosyst 143:S6–S11. https://doi.org/10.1080/11263500903187035

    Article  Google Scholar 

  • Hollingworth D, Candel AM, Nicastro G, Martin SR, Briata P, Gherzi R, Ramos A (2012) KH domains with impaired nucleic acid binding as a tool for functional analysis. Nucleic Acids Res 40:6873–6886. https://doi.org/10.1093/nar/gks368

    Article  CAS  Google Scholar 

  • Hossain Z, Komatsu S (2013) Contribution of proteomic studies towards understanding plant heavy metal stress response. Front Plant Sci 3:310–310

    Google Scholar 

  • Hwang J, Oh C, Kang B (2013) Translation elongation factor 1B (eEF1B) is an essential host factor for Tobacco mosaic virus infection in plants. Virology 439:105–114

    CAS  Google Scholar 

  • Indovina P, Marcelli E, Pentimalli F, Tanganelli P, Tarro G, Giordano A (2013) Mass spectrometry-based proteomics: the road to lung cancer biomarker discovery. Mass Spectrom Rev 32:129–142. https://doi.org/10.1002/mas.21355

    Article  CAS  Google Scholar 

  • Islam E et al. (2008) Effect of Pb toxicity on leaf growth, physiology and ultrastructure in the two ecotypes of Elsholtzia argyi. J Hazard Mater 154:914–926

    CAS  Google Scholar 

  • Islam E, Yang X, Li T, Liu D, Jin X, Meng F (2007) Effect of Pb toxicity on root morphology, physiology and ultrastructure in the two ecotypes of Elsholtzia argyi. J Hazard Mater 147:806–816

    CAS  Google Scholar 

  • Jiang L, Wang Y, Li S (2007) Application of the comet assay to measure DNA damage induced by UV radiation in the hydrophyte, Spirodela polyrhiza. Physiologia Plant 129:652–657

    CAS  Google Scholar 

  • Konca K et al. (2003) A cross-platform public domain PC image-analysis program for the comet assay. Mutat Res-Genet Toxicol Environ Mutagenesis 534:15–20

    CAS  Google Scholar 

  • Kopittke PM, Asher CJ, Kopittke RA, Menzies NW (2007) Toxic effects of Pb2+ on growth of cowpea (Vigna unguiculata). Environ Pollut 150:280–287

    CAS  Google Scholar 

  • Kumaravel T, Jha AN (2006) Reliable Comet assay measurements for detecting DNA damage induced by ionising radiation and chemicals. Mutat Res-Genet Toxicol Environ Mutagenesis 605:7–16

    CAS  Google Scholar 

  • Lee S, Lee E, Yang E, Lee J, Park AR, Song W, Park O (2004) Proteomic identification of annexins, calcium-dependent membrane binding proteins that mediate osmotic stress and abscisic acid signal transduction in Arabidopsis. Plant Cell 16:1378–1391. https://doi.org/10.1105/tpc.021683

    Article  CAS  Google Scholar 

  • Li F, Liu C, Yang Y, Bi X, Liu T, Zhao Z (2012) Natural and anthropogenic lead in soils and vegetables around Guiyang city, southwest China: A Pb isotopic approach. Sci Total Environ 431:339–347

    CAS  Google Scholar 

  • Li J, Oulee TM, Raba R, Amundson RG (1993) Arabidopsis flavonoid mutants are hypersensitive to UV-B irradiation. Plant Cell 5:171–179

    CAS  Google Scholar 

  • Liu N, Lin Z-F, Mo H (2012) Metal (Pb, Cd, and Cu)-induced reactive oxygen species accumulations in aerial root cells of the Chinese Banyan (Ficus microcarpa). Ecotoxicology 21:2004–2011. https://doi.org/10.1007/s10646-012-0935-y

    Article  CAS  Google Scholar 

  • Livramento KGD, Borém FM, José AC, Santos AV, Livramento DED, Alves JD, Paiva LV (2017) Proteomic analysis of coffee grains exposed to different drying process. Food Chem 221:1874–1882. https://doi.org/10.1016/j.foodchem.2016.10.069

    Article  CAS  Google Scholar 

  • Lu N et al. (2019) Genome-wide analysis of the Catalpa bungei caffeic acid O-methyltransferase (COMT) gene family: identification and expression profiles in normal, tension, and opposite wood. PeerJ 7:e6520

    Google Scholar 

  • Lyu G et al. (2020) Quantitative proteomic analyses identify STO/BBX24-related proteins induced by UV-B. Int J Mol Sci 21:2496

    Google Scholar 

  • Moon H et al. (2003) NDP kinase 2 interacts with two oxidative stress-activated MAPKs to regulate cellular redox state and enhances multiple stress tolerance in transgenic plants. Proc Natl Acad Sci USA 100:358–363

  • Nasreddine L, Parent-Massin D (2002) Food contamination by metals and pesticides in the European Union. Should we worry? Toxicol Lett 127:29–41

    CAS  Google Scholar 

  • Ning C, Qin R, Chen D, Bjorn LO, Li S (2016) Application of in-house virtual protein database performed in genomic-proteomic combined research on heavy-metal stressed onion roots. Biotechnol Lett 38:1293–1300

    CAS  Google Scholar 

  • Palmer S et al. (2015) The effects of lead sources on oral bioaccessibility in soil and implications for contaminated land risk management. Environ Pollut 198:161–171

    CAS  Google Scholar 

  • Paul N, Chakraborty S, Sengupta M (2014) Lead toxicity on non-specific immune mechanisms of freshwater fish Channa punctatus. Aquat Toxicol 152:105–112

    CAS  Google Scholar 

  • Pourrut B, Perchet G, Silvestre J, Cecchi M, Guiresse M, Pinelli E (2008) Potential role of NADPH-oxidase in early steps of lead-induced oxidative burst in Vicia faba roots. J Plant Physiol 165:571–579. https://doi.org/10.1016/j.jplph.2007.07.016

    Article  CAS  Google Scholar 

  • Pourrut B, Shahid M, Dumat C, Winterton P, Pinelli E (2011) Lead uptake, toxicity, and detoxification in plants. Rev Environ Contamination Toxicol 213:113–136

    CAS  Google Scholar 

  • Qin R, Ning C, Björn LO, Li S (2015a) Proteomic analysis of Allium cepa var. agrogarum L. roots under copper stress. Plant Soil 401:197–212

    Google Scholar 

  • Qin R, Wang C, Chen D, Björn LO, Li S (2015b) Copper‐induced root growth inhibition of Allium cepa var. agrogarum L. involves disturbances in cell division and DNA damage. Environ Toxicol Chem 34:1045–1055

    CAS  Google Scholar 

  • Rodriguez E, Azevedo R, Fernandes P, Santos CAO (2011) Cr(VI) induces DNA damage, cell cycle arrest and polyploidization: a flow cytometric and comet assay study in Pisum sativum. Chem Res Toxicol 24:1040–1047

    CAS  Google Scholar 

  • 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 74:78–84

    CAS  Google Scholar 

  • Sidoli S et al. (2015) SWATH analysis for characterization and quantification of histone post-translational modifications molecular & cellular proteomics. https://doi.org/10.1074/mcp.O1114.046102

  • Soltys D, Rudzinskalangwald A, Kurek W, Gniazdowska A, Sliwinska E, Bogatek R (2011) Cyanamide mode of action during inhibition of onion (Allium cepa L.) root growth involves disturbances in cell division and cytoskeleton formation. Planta 234:609–621

    CAS  Google Scholar 

  • Song Y, Cui J, Zhang H, Wang G, Zhao F, Shen Z (2012) Proteomic analysis of copper stress responses in the roots of two rice (Oryza sativa L.) varieties differing in Cu tolerance. Plant Soil 366:647–658

    Google Scholar 

  • Tak H, Negi S, Rajpurohit YS, Misra HS, Ganapathi TR (2020) MusaMPK5, a mitogen activated protein kinase is involved in regulation of cold tolerance in banana. Plant Physiol Biochem 146:112–123

    CAS  Google Scholar 

  • Voll LM, Jamai A, Renne P, Voll H, Mcclung CR, Weber APM (2005) The photorespiratory Arabidopsis shm1 mutant is deficient in SHM1. Plant Physiol 140:59–66

    Google Scholar 

  • Xue Y, Peijnenburg W, Huang J, Wang D, Jin Y (2018) Trophic transfer of Cd from duckweed (Lemna minor L.) to tilapia (Oreochromis mossambicus). Environ Toxicol Chem 37:1367–1377

    CAS  Google Scholar 

  • Yadav SK (2010) Heavy metals toxicity in plants: An overview on the role of glutathione and phytochelatins in heavy metal stress tolerance of plants. South Afr J Bot 76:167–179. https://doi.org/10.1016/j.sajb.2009.10.007

    Article  CAS  Google Scholar 

  • Yang Y, Jung J, Song W, Suh H, Lee Y (2000) Identification of rice varieties with high tolerance or sensitivity to lead and characterization of the mechanism of tolerance. Plant Physiol 124:1019–1026

    CAS  Google Scholar 

  • Zhang F et al. (2014) SWATH™- and iTRAQ-based quantitative proteomic analyses reveal an overexpression and biological relevance of CD109 in advanced NSCLC. J Proteom 102:125–136

    CAS  Google Scholar 

  • Zhang S, Zhang H, Qin R, Jiang W, Liu D (2009) Cadmium induction of lipid peroxidation and effects on root tip cells and antioxidant enzyme activities in Vicia faba L. Ecotoxicology 18:814–823

    CAS  Google Scholar 

  • Zhao Y, Wu J, Shang D, Ning J, Zhai Y, Sheng X, Ding H (2015) Subcellular distribution and chemical forms of cadmium in the edible seaweed, Porphyra yezoensis. Food Chem 168:48–54

    CAS  Google Scholar 

Download references

Acknowledgements

This work was supported by the National Natural Science Foundation of China (grant 31670266), the Leading Scientists Project of Guangdong Province, the Guangdong Pearl River Scholar Funded Scheme (2012) and the Innovation Project of Graduate School of South China Normal University (2016lkxm10). We thank Prof. Lars Olof Björn (Lund University) and Prof. Paul Giller (University College Cork) for English editing of the paper.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Shaoshan Li.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Additional information

Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

These authors contributed equally: Guizhen Lyu, Dongbing Li

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Lyu, G., Li, D., Li, S. et al. Genotoxic effects and proteomic analysis on Allium cepa var. agrogarum L. root cells under Pb stress. Ecotoxicology 29, 959–972 (2020). https://doi.org/10.1007/s10646-020-02236-x

Download citation

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10646-020-02236-x

Keywords

Navigation