Abstract
This study aimed to investigate the mode of action of cadmium (Cd) toxicity at cell level, especially at early stages of plant exposure. Tomato seedlings were cultivated in growth media containing from 0.1 to 70 µM CdCl2 for 24 h. Mitotic index, chromosome abnormality, DNA integrity and organization of tubulin-based structures were assessed in root cells. As higher the Cd concentration in the growth media, higher was the DNA damage intensity and the occurrence of chromosomal abnormalities that included chromosome lost, bridges, stickiness, C-metaphase and polyploidy. The profile of chromosomal aberrations also varied with elevated Cd concentration, being observed increases in the frequency of chromosome stickiness. The mitotic index was reduced at the lowest Cd concentration, but such reduction was statistically similar to that detected at the highest concentration, suggesting that mitotic depression is a rapid outcome and, at same time, a Cd-induced effect that is limited at the first 24 h of direct root exposure to this metal. Under exposure to 20 µM CdCl2, heterogenous distribution of the spindle fibers, formation of two spindle complexes in both of the cell poles, absence of centrosome center, polarization of the spindle fibers during cell division, and non-uniform tubulin deposition in microtubule and phragmoplast were noticed. The results indicate that the tubulin-dependent components of cytoskeleton are Cd targets, and the sensitivity of tubulin-based structures to Cd exposure depends on cell cycle phase. Moreover, DNA damage intensity and chromosomal abnormality profile can be employed as markers of Cd toxicity level.
References
Abdalla KO, Thomson JA, Rafudeen MS (2009) Protocols for nuclei isolation and nuclear protein extraction from the resurrection plant Xerophyta viscosa for proteomic studies. Anal Biochem 384:365–367
Alves LA, Monteiro CC, Carvalho RF, Ribeiro PC, Tezotto T, Azevedo RA, Gratão PL (2017) Cadmium stress related to root-to-shoot communication depends on ethylene and auxin in tomato plants. Environ Exp Bot 134:102–115
Araújo RP, Almeida A-AF, Pereira LS, Mangabeira PAO, Souza JO, Pirovani CP, Ahnert D, Baligar VC (2017) Photosynthetic, antioxidative, molecular and ultrastructural responses of young cacao plants to Cd toxicity in the soil. Ecotoxicol Environ Saf 144:148–157
Arya SK, Mukherjee A (2014) Sensitivity of Allium cepa and Vicia faba towards cadmium toxicity. J Soil Sci Plant Nutr 14:447–458
Avila-Ospina L, Moison M, Yoshimoto K, Daubresse CM (2014) Autophagy, plant senescence, and nutrient recycling. J Exp Bot 65:3799–811
Bandyopadhyay A, Mukherjee A (2011) Sensitivity of Allium and Nicotiana in cellular and acellular comet assays to assess differential genotoxicity of direct and indirect acting mutagens. Ecotoxicol Environ Saf 74:860–865
Bayçu G, Gevrek-Kürüm N, Moustaka J, Csatári I, Rognes SE, Moustakas M (2017) Cadmium-zinc accumulation and photosystem II responses of Noccaea caerulescens to Cd and Zn exposure. Environ Sci Pollut Res Int 24:2840–2850
Borges KLR, Hippler FWR, Carvalho MEA, Nalin RS, Matias FI, Azevedo RA (2019) Nutritional status and root morphology of tomato under Cd-induced stress: comparing contrasting genotypes for metal-tolerance. Sci Hortic 246:518–527
Borges KLR, Salvato F, Alcântara BK, Nalin RS, Piotto FA, Azevedo RA (2018) Temporal dynamic responses of roots in contrasting tomato genotypes to cadmium tolerance. Ecotoxicology 27:245–258
Cabral Pinto MMS, Marinho-Reis AP, Almeida A, Ordens CM, Silva MMVG, Freitas S, Simões MR, Moreira PI, Dinis PA, Diniz ML, Ferreira da Silva EA, Condesso de Melo MT (2017) Human predisposition to cognitive impairment and its relation with environmental exposure to potentially toxic elements. Environ Geochem Health 40:1767–1784
Carvalho MEA, Piotto FA, Franco MR, Borges KLR, Gaziola SA, Castro PRC, Azevedo RA (2018c) Cadmium toxicity degree on tomato development is associated with disbalances in B and Mn status at early stages of plant exposure. Ecotoxicology 27:1293–1302
Carvalho MEA, Piotto FA, Franco MR, Martinelli AP, Cuypers A, Azevedo RA (2019) Relationship between Mg, B and Mn status and tomato tolerance against Cd toxicity. J Environ Manag 240:84–92
Carvalho MEA, Piotto FA, Gaziola SA, Jacomino AP, Jozefczak M, Cuypers A, Azevedo RA (2018a) New insights about cadmium impacts on tomato: plant acclimation, nutritional changes, fruit quality and yield. Food Energy Secur 7:e00131
Carvalho MEA, Piotto FA, Nogueira ML, Gomes-Junior FG, Chamma HMCP, Pizzaia D, Azevedo RA (2018b) Cadmium exposure triggers genotype-dependent changes in seed vigor and germination of tomato offspring. Protoplasma 255:989–999
Cerda H, Delincée H, Haine H, Rupp H (1997) The DNA ‘comet assay’ as a rapid screening technique to control irradiated food. Mutat Res 375:167–81
Fan JL, Wei XG, Wan LC, Zhang LY, Zhao XQ, Liu WZ, Hao HQ, Zhang HY (2011) Disarrangement of actin filaments and Ca2+ gradient by CdCl2 alters cell wall construction in Arabidopsis thaliana root hairs by inhibiting vesicular trafficking. J Plant Physiol 168:1157–1167
Fidalgo F, Freitas R, Ferreira R, Pessoa AM, Teixeira J (2011) Solanum nigrum L. antioxidant defence system isozymes are regulated transcriptionally and postranslationally in Cd-induced stress. Environ Exp Bot 72:312–319
Gallego SM, Pena LB, Barcia RA, Azpilicueta CE, Iannone MF, Rosales EP, Zawoznik MS, Groppa MD, Benavides MP (2012) Unravelling cadmium toxicity and tolerance in plants: insight into regulatory mechanisms. Environ Exp Bot 83:33–46
Gichner T, Patková Z, Kim J (2003) DNA damage measured by the comet assay in eight agronomic plants. Biol Plant 47:185–188
Gratão PL, Monteiro CC, Tezotto T, Carvalho RF, Alves LR, Peters LP, Azevedo RA (2015) Cadmium stress antioxidant responses and root-to-shoot communication in grafted tomato plants. Biometals 28:803–816
Gratão PL, Polle A, Lea PJ, Azevedo RA (2005) Making the life of heavy metal stressed plants a little easier. Funct Plant Biol 32:481–494
Gzyl J, Chmielowska-Bak J, Przymusinski R, Gwóźdź EA (2015) Cadmium affects microtubule organization and post-translational modifications of tubulin in seedlings of soybean (Glycine max L.). Front Plant Sci 6:937
Hamada T (2007) Microtubule-associated proteins in higher plants. J Plant Res 120:79–98
Hendrix S, Keunen E, Mertens AIG, Beemster GTS, Vangronsveld J, Cuypers A (2018) Cell cycle regulation in different leaves of Arabidopsis thaliana plants grown under control and cadmium-exposed conditions. Environ Exp Bot 155:441–452
Horio T, Murata T (2014) The role of dynamic instability in microtubule organization. Front Plant Sci 5:511
Hossain Z, Huq F (2002) Studies on the interaction between Cd2+ ions and DNA. J Inorg Biochem 90:85–96
Iakimova ET, Woltering EJ, Kapchina-Toteva VM, Harren FJM, Cristescu SM (2008) Cadmium toxicity in cultured tomato cells—role of ethylene, proteases and oxidative stress in cell death signaling. Cell Biol Int 32:1521–1529
Kato FH, Carvalho MEA, Gaziola SA, Piotto FA, Azevedo RA (2020) Lysine metabolism and amino acid profile in maize grains from plants subjected to cadmium exposure. Sci Agric 77:e20180095
Leme DM, Marin-Morales MA (2009) Allium cepa test in environmental monitoring: a review on its application. Mutat Res 682:71–81
Liu D, Xue P, Meng Q, Zou J, Gu J, Jiang W (2009) Pb/Cu effects on the organization of microtubule cytoskeleton in interphase and mitotic cells of Allium sativum L. Plant Cell Rep 28:695–702
Marques DN, Carvalho MEA, Piotto FA, Batagin-Piotto KD, Nogueira ML, Gaziola SA, Azevedo RA (2019) Antioxidant defense response in plants to cadmium stress. In: Hasanuzzaman M, Prasad MNV, Nahar M (eds) Cadmium tolerance in plants: agronomic, molecular, signaling, and omic approaches, 1st edn. Academic Press, Cambridge, p 423–461
Marschner P (2012) Marschner’s mineral nutrition of higher plants. Academic Press, San Diego
Michaeli S, Galili G (2014) Degradation of organelles or specific organelle components via selective autophagy in plant cells. Int J Mol Sci 15:7624–7638
Mondin M, Santos-Serejo JA, Aguiar-Perecin MLR (2007) Karyotype characterization of Crotalaria juncea (L.) by chromosome banding and physical mapping of 18S-5.8S-26S and 5S rRNA gene sites. Genet Mol Biol 30:65–72
Nogueirol RC, Monteiro FA, Gratão PL, Silva BKA, Azevedo RA (2016) Cadmium application in tomato: nutritional imbalance and oxidative stress. Water Air Soil Pollut 227:210
Nogueirol RC, Monteiro FA, Souza Jr JC, Azevedo RA (2018) NO3 −/NH4 + proportions affect cadmium bioaccumulation and tolerance of tomato. Environ Sci Pollut Res 25:13916–13928
O’Lexy R, Kasai K, Clark N, Fujiwara T, Sozzani R, Gallaghe KL (2018) Exposure to heavy metal stress triggers changes in plasmodesmatal permeability via deposition and breakdown of callose. J Exp Bot 69:3715–3728
Oldenburg DJ, Bendich AJ (2015) DNA maintenance in plastids and mitochondria of plants. Front Plant Sci 6:883
Parrotta L, Cresti M, Cai G (2014) Accumulation and post-translational modifications of plant tubulins. Plant Biol 16:521–527
Piotto FA, Carvalho MEA, Souza LA, Rabêlo FHS, Franco MR, Batagin-Piotto KB, Azevedo RA (2018) Estimating tomato tolerance to heavy metal toxicity: cadmium as study case. Environ Sci Pollut Res 25:27535–27544
Pompeu GB, Vilhena MB, Gratão PL, Carvalho RF, Rossi ML, Martinelli AP, Azevedo RA (2017) Abscisic acid-deficient sit tomato mutant responses to cadmium-induced stress. Protoplasma 254:771–783
R Development Core Team (2014) R: a language and environment for statistical computing. R Foundation for Statistical Computing, Vienna
Sebastian A, Prasad MNV (2016) Modulatory role of mineral nutrients on cadmium accumulation and stress tolerance in Oryza sativa L. seedlings. Environ Sci Pollut Res 23:1224–1233
Seth CS, Misraa V, Chauhan LKS, Singh RR (2008) Genotoxicity of cadmium on root meristem cells of Allium cepa: cytogenetic and comet assay approach. Ecotoxicol Environ Safe 71:711–716
Shi HP, Feng Y, Wang YL, Tsang PKE (2014) Effect of cadmium on cytogenetic toxicity in hairy roots of Wedelia trilobata L. and their alleviation by exogenous CaCl2. Environ Sci Pollut Res 21:1436–1443
Shi Q, Wang J, Zou J, Jiang Z, Wu H, Wang J, Jiang W, Liu D (2016) Cadmium localization and its toxic effects on root tips of barley. Zemdirbyste-Agricultue 103:151–158
Siegel S, Castellan NJ (1988) Non parametric statistics for the behavioural sciences. MacGraw Hill Int., New York, NY, p 213–214
Singh S, Parihar P, Singh R, Singh VP, Prasad SM (2016) Heavy metal tolerance in plants: role of transcriptomics, proteomics, metabolomics, and ionomics. Front Plant Sci 6:1143
Soares C, Carvalho MEA, Azevedo RA, Fidalgo F (2019) Plants facing oxidative challenges—a little help from the antioxidant networks. Environ Exp Bot 161:4–25
Souza LA, Camargos LS, Carvalho MEA (2018) Toxic metal phytoremediation using high biomass non-hyperaccumulator crops: new possibilities for bioenergy resources. In: Matichenkov V (ed) Phytoremediation: methods, management, assessment. Nova Science, New York, NY, p 1–25
Wang QL, Liu DH, Yue JY (2016) The uptake of cadmium by Allium cepa var. agrogarum L. and its effects on chromosome and nucleolar behavior in root tip cells. Phyton 85:155–161
Wang Y, Ji Y, Fu Y, Guo H (2018) Ethylene-induced microtubule reorientation is essential for fast inhibition of root elongation in Arabidopsis. J Integr Plant Biol 60:864–877
Yakimova ET, Kapchina-Toteva VM, Laarhoven L-J, Harren FM, Woltering EJ (2006) Involvement of ethylene and lipid signalling in cadmium-induced programmed cell death in tomato suspension cells. Plant Physiol Biochem 44:581–589
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
Zhou J, Yao J, Joshi HC (2012) Attachment and tension in the spindle assembly checkpoint. J Cell Sci 115:3547–3555
Acknowledgements
This work was funded in part by Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP—Grant number 2009/54676-0 and scholarship number 2013/15217-5) and the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior—Brasil (CAPES)—Finance Code 001. We also thank the National Council for Scientific and Technological Development—CNPq, Brazil and FAPESP, Brazil, for the fellowship and scholarships, respectively.
Author contributions
DP and FAP designed the experiment. DP and MLN carried out the experiments. MFR performed the statistical analysis of the data. DP, FAP, MLN, MM, MEAC and RAA interpreted the results and wrote the manuscript. FAP and RAA assisted during the research. All authors (except MFR, in memorium) read and approved the final manuscript.
Author information
Authors and Affiliations
Corresponding author
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.
In memoriam: Millor Fernandes Rosario
Rights and permissions
About this article
Cite this article
Pizzaia, D., Nogueira, M.L., Mondin, M. et al. Cadmium toxicity and its relationship with disturbances in the cytoskeleton, cell cycle and chromosome stability. Ecotoxicology 28, 1046–1055 (2019). https://doi.org/10.1007/s10646-019-02096-0
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10646-019-02096-0