Abstract
Lead (Pb) persists among the most hazardous contaminant metals. Pb-induced genotoxic effects remain a matter of debate as they are a major cause of plant growth impairment, but assessing Pb genotoxicity requires the selection of Pb-sensitive genotoxic biomarkers. Seedlings of the ecotoxicological model species Pisum sativum L. were exposed to Pb2+ (≤ 2000 mg L−1). Flow cytometry (FCM) revealed that 28 days after, Pb2+ arrested root cell cycle at G2 but no eu/aneuploidies were found. Comet assay and FCM-clastogenicity assays showed that Pb2+ increased DNA breaks in roots at concentrations as low as 20 mg L−1. Leaves showed no variation in DNA-ploidy or cell cycle progression but had increased DNA breaks at the highest Pb2+ dose. We conclude that both Comet assay and the full-peak coefficient of variation (FPCV) were the most relevant endpoints of Pb-phytogenotoxicity. Also, the Pb-induced DNA breaks may be related with the arrest at the G2-checkpoint. Data will be relevant to better define Pb2+ ecogenotoxicological effects and their measuring tools and may contribute to a regulatory debate of this pollutant limits.
This is a preview of subscription content, access via your institution.




References
Azevedo H, Gomes C, Pinto G, Santos C (2005) Cadmium effects in sunflower: nutrient imbalances in leaves and calluses. J Pant Nutr 28(12):2233–2241. https://doi.org/10.1080/01904160500324808
Azevedo R, Rodriguez E, Mendes RJ, Mariz-Ponte N, Sario S, Lopes JC, Ferreira de Oliveira JMP, Santos C (2018) Inorganic Hg toxicity in plants: a comparison of different genotoxic parameters. Plant Physiol Biochem 125:247–254. https://doi.org/10.1016/j.plaphy.2018.02.015
Cao X, Wang H, Zhuang D, Zhu H, Du Y, Cheng Z, Cui W, Rogers HJ, Zhang Q, Jia C, Yang Y, Tai P, Xie F, Liu W (2018) Roles of MSH2 and MSH6 in cadmium-induced G2/M checkpoint arrest in Arabidopsis roots. Chemosphere 201:586–594. https://doi.org/10.1016/j.chemosphere.2018.03.017
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. https://doi.org/10.1016/j.envexpbot.2009.10.001
Collins AR, Oscoz AA, Brunborg G, Gaivao I, Giovannelli L, Kruszewski M, Smith CC, Stetina R (2008) The comet assay: topical issues. Mutagenesis 23:143–151. https://doi.org/10.1093/mutage/gem051
Dias MC, Mariz-Ponte N, Santos C (2019) Lead induces oxidative stress in Pisum sativum plants and changes the levels of phytohormones with antioxidant role. Plant Physiol Biochem 37:121–129. https://doi.org/10.1016/j.plaphy.2019.02.005
Edelstein M, Ben-Hur M (2018) Heavy metals and metalloids: sources, risks and strategies to reduce their accumulation in horticultural crops. Sci Hortic 234:431–444. https://doi.org/10.1016/j.scienta.2017.12.039
Ghani A, Khan I, Ahmed I, Mustafa I, Abd-Ur-Rehman MN (2015) Amelioration of lead toxicity in Pisum sativum (L.) by foliar application of salicylic acid. J\ Environ Anal Toxicol 5(292):2161–0525. https://doi.org/10.4172/2161-0525.1000292
Gichner T, Patkova Z, Szakova J, Znidar I, Mukherjee A (2008a) DNA damage in potato plants induced by cadmium, ethyl methanesulphonate and gamma-rays. Environ Exp Bot 62:113–119. https://doi.org/10.1016/j.envexpbot.2007.07.013
Gichner T, Znidar I, Szakova J (2008b) Evaluation of DNA damage and mutagenicity induced by lead in tobacco plants. Mutat Res Genet Toxicol Environ 652:186–190. https://doi.org/10.1016/j.mrgentox.2008.02.009
Hardison D Jr, Ma LQ, Luongo T, Harris WG (2004) Lead contamination in shooting range soils from abrasion of lead bullets and subsequent weathering. Sci Total Environ 328:175–183. https://doi.org/10.1016/j.scitotenv.2003.12.013
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
Jeong H, Kim H, Jang T (2016) Irrigation water quality standards for indirect wastewater reuse in agriculture: a contribution toward sustainable wastewater reuse in South Korea. Water 8(4):169. https://doi.org/10.3390/w8040169
Kumar A, Prasad M, Achary M, Panda B (2013) Elucidation of lead-induced oxidative stress in Talinum triangulare roots by analysis of antioxidant responses and DNA damage at cellular level. Environ Sci Pollut Res 20:4551–4561. https://doi.org/10.1007/s11356-012-1354-6
Liu D, Li T, Jin X, Yang X, Islam E, Mahmood Q (2008) Lead induced changes in the growth and antioxidant metabolism of the lead accumulating and non-accumulating ecotypes of Sedum alfredii. J Integr Plant Biol 50:129–140. https://doi.org/10.1111/j.1744-7909.2007.00608.x
López-Orenes A, Dias MC, Ferrer MÁ, Calderón A, Moutinho-Pereira J, Correia C, Santos C (2018) Different mechanisms of the metalliferous Zygophyllum fabago shoots and roots to cope with Pb toxicity. Environ Sci Pollut Res 25(2):1319–1330. https://doi.org/10.1007/s11356-017-0505-1
Loureiro J, Capelo A, Brito G, Rodriguez E, Silva S, Santos C (2007) Micropropagation of Juniperus phoenicea L adult plants and analyses of ploidy stability and DNA content in micropragation plants. Biol Plant 51:7–14. https://doi.org/10.1007/s10535-007-0003-2
Malar S, Manikandan R, Favas P, Vikram Sahi S, Venkatachalam P (2014) Effect of lead on phytotoxicity, growth, biochemical alterations and its role on genomic template stability in Sesbania grandiflora: a potential plant for phytoremediation. Ecotoxicol Environ Saf 108:249–257. https://doi.org/10.1016/j.ecoenv.2014.05.018
MAOTDR, Ministério do Ambiente, do Ordenamento do Território e do Desenvolvimento Regional (2009) DL 277/2009, 192-2-10-2009, 7154-7165
Monteiro MS, Rodriguez E, Loureiro J, Mann R, Soares A, Santos C (2010) Flow cytometric assessment of Cd genotoxicity in three plants with different metal accumulation and detoxification capacities. Ecotoxicol Environ Saf 73:1231–1237. https://doi.org/10.1016/j.ecoenv.2010.06.020
Piechalak A, Tomaszewska B, Baralkiewicz D (2003) Enhancing phytoremediative ability of Pisum sativum by EDTA application. Phytochemistry 64:1239–1251. https://doi.org/10.1016/S0031-9422(03)00515-6
Rodriguez E, Azevedo R, Moreira H, Souto L, Santos C (2013) Pb2+ exposure induced microsatellite instability in Pisum sativum in a locus related with glutamine metabolism. Plant Physiol Biochem 62:19–22. https://doi.org/10.1016/j.plaphy.2012.10.006
Santos C, Pourrut B, Oliveira J (2015) The use of comet assay in plant toxicology: recent advances. Front Genet 30(6):216. https://doi.org/10.3389/fgene.2015.00216
Seth C, Misra V, Chauhan L (2012) Accumulation, detoxification, and genotoxicity of heavy metals in Indian mustard (Brassica Juncea L). Int J Phytoremed 14:1):1–1)13. https://doi.org/10.1080/15226514.2011.555799
Shahid M, Dumat C, Pourrut B, Pinelli E (2014) Assessing the effect of metal speciation on lead toxicity to Vicia faba pigment contents. J Geochem Explor 144:290–297. https://doi.org/10.1016/j.gexplo.2014.01.003
Silva S, Santos C, Matos M, Pinto-Carnide O (2012) Al toxicity mechanism in tolerant and sensitive rye genotypes. Environ Exp Bot 75:89–97. https://doi.org/10.1016/j.envexpbot.2011.08.017
Silva S, Pinto G, Santos C (2017) Low doses of Pb affected Lactuca sativa photosynthetic performance. Photosynthetica 55(1):50–57. https://doi.org/10.1007/s11099-016-0220-z
Tariq SR, Ashraf A (2016) Comparative evaluation of phytoremediation of metal contaminated soil of firing range by four different plant species. Arab J Chem 9(6):806–814. https://doi.org/10.1016/j.arabjc.2013.09.024
Tawinteung N, Parkpian P, DeLaune R, Jugsujinda A (2005) Evaluation of extraction procedures for removing lead from contaminated soil. J Environ Sci Health A 40(2):385–407. https://doi.org/10.1081/ESE-200045631
Tóth G, Hermann T, Silva M, Montanarella L (2016) Heavy metals in agricultural soils of the European Union with implications for food safety. Environ Int 88:299–309. https://doi.org/10.1016/j.envint.2015.12.017
Zeng P, Guo Z, Xiao X, Peng C, Feng W, Xin L, Xu Z (2018) Phytoextraction potential of Pteris vittata L. co-planted with woody species for As, Cd, Pb and Zn in contaminated soil. Sci Total Environ 650:594–603. https://doi.org/10.1016/j.scitotenv.2018.09.055
Zhang X, Li M, Yang H, Li X, Cui Z (2018) Physiological responses of Suaeda glauca and Arabidopsis thaliana in phytoremediation of heavy metals. J Environ Manag 223:132–139. https://doi.org/10.1016/j.jenvman.2018.06.025
Funding
This work was funded by FEDER/COMPETE [POCI/01/0145/FEDER/007265; FCT/MEC PT2020 UID/QUI/5006/2019]; Fundação para a Ciência e Tecnologia funded Nuno Mariz-Ponte, SFRH/BD/138187/2018; Sara Sario, SFRH/BD/138186/2018; and Rafael J. Mendes, SFRH/BD/133519/2017.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interest.
Additional information
Responsible editor: Gangrong Shi
Publisher’s note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Rodriguez, E., Sousa, M., Gomes, A. et al. Genotoxic endpoints in a Pb-accumulating pea cultivar: insights into Pb2+ contamination limits. Environ Sci Pollut Res 26, 32368–32373 (2019). https://doi.org/10.1007/s11356-019-06465-4
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11356-019-06465-4
Keywords
- Cytostaticity
- Comet assay
- DNA breaks
- Lead
- Phytogenotoxicity
- Pisum sativum