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Photosynthesis light-independent reactions are sensitive biomarkers to monitor lead phytotoxicity in a Pb-tolerant Pisum sativum cultivar

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Abstract

Lead (Pb) environmental contamination remains prevalent. Pisum sativum L. plants have been used in ecotoxicological studies, but some cultivars showed to tolerate and accumulate some levels of Pb, opening new perspectives to their use in phytoremediation approaches. However, the putative use of pea plants in phytoremediation requires reliable toxicity endpoints. Here, we evaluated the sensitivity of a large number of photosynthesis-related biomarkers in Pb-exposed pea plants. Plants (cv. “Corne de Bélier”) were exposed to Pb concentrations up to 1,000 mg kg−1 soil during 28 days. The photosynthetic potential biomarkers that were analyzed included pigments, chlorophyll (Chl) a fluorescence, gas exchange, ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) activity, and carbohydrates. Flow cytometry (FCM) was also used to assess the morpho-functional status of chloroplasts. Finally, Pb-induced nutrient disorders were also evaluated. Net CO2 assimilation rate (A) and RuBisCO activity decreased strongly in Pb-exposed plants. Plant dry mass (DM) accumulation, however, was only reduced in the higher Pb concentrations tested (500 and 1,000 mg kg−1 soil). Pigment contents increased solely in plants exposed to the largest Pb concentration, and in addition, the parameters related to the light-dependent reactions of photosynthesis, F v/F m and ΦPSII, were not affected by Pb exposure. In contrast to this, carbohydrates showed an overall tendency to increase in Pb-exposed plants. The morphological status of chloroplasts was affected by Pb exposure, with a general trend of volume decrease and granularity increase. These results point the endpoints related to the light-independent reactions of photosynthesis as more sensitive predictors of Pb-toxicity than the light-dependent reactions ones. Among the endpoints related to the light-independent photosynthesis reactions, RuBisCO activity and A were found to be the most sensitive. We discuss here the advantages of using these parameters as biomarkers for Pb toxicity in plants. Finally, we report that, despite showing physiological disorders, these cultivar plants survived and accumulated high doses of Pb, and their use in environmental/decontamination studies is open to debate.

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References

  • Alzandi AR (2012) The use of Pisum sativum L as bioindicator for lead ions detoxification. Int J Sci Technol Res 1(2):42–45

    Google Scholar 

  • An YJ (2004) Soil ecotoxicity assessment using cadmium sensitive plants. Environ Pollut 127:21–6

    Article  CAS  Google Scholar 

  • Aravind P, Prasad MN, Malec P, Waloszek A, Strzalka K (2009) Zinc protects Ceratophyllum demersum L. (free-floating hydrophyte) against reactive oxygen species induced by cadmium. J Trace Elem Med Biol 23:50–60

    Article  CAS  Google Scholar 

  • Bah A, Sun H, Chen F, Zhou J, Dai H, Zhang G, Wu F (2010) Comparative proteomic analysis of Typha angustifolia leaf under chromium, cadmium and lead stress. J Hazard Mater 184:191–203

    Article  CAS  Google Scholar 

  • Berti W, Cunningham S (1997) In-place inactivation of Pb in Pb-contaminated soils. Environ Sci Technol 31:1359–1364

    Article  CAS  Google Scholar 

  • Bertrand M, Poirier I (2005) Photosynthetic organisms and excess of metals. Photosynthetica 43:345–353

    Article  CAS  Google Scholar 

  • Bibi M, Hussain M (2005) Effect of copper and lead on photosynthesis and plant pigments in black gram [Vigna mungo (L.) Hepper]. Bull Environ Contam Toxicol 74:1126–33

    Article  CAS  Google Scholar 

  • Breckle SW, Kahle H (1992) Effects of toxic heavy-metals (Cd, Pb) on growth and mineral-nutrition of beech (Fagus sylvatica L). Vegetatio 101:43–53

    Article  Google Scholar 

  • Burzynski M, Klobus G (2004) Changes of photosynthetic parameters in cucumber leaves under Cu, Cd, and Pb stress. Photosynthetica 42:505–510

    Article  CAS  Google Scholar 

  • Capelo A, Santos C, Loureiro S, Pedrosa M (2012) Phytotoxicity of lead on Lactuca sativa: effects on growth, mineral nutrition, photosynthetic activity and oxidant metabolism. Fresenius Envir Bull 21:450–459

    CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  • Choudhury NK, Behera RK (2001) Photoinhibition of photosynthesis: role of carotenoids in photoprotection of chloroplast constituents. Photosynthetica 39:481–488

    Article  CAS  Google Scholar 

  • Clijsters H, Van Assche F (1985) Inhibition of photosynthesis by heavy metals. Photosynth Res 7:31–40

    Article  CAS  Google Scholar 

  • Devi R, Mundiral N, Gupta A, Kaur N (2013) Effect of exogenous lead on growth and carbon metabolism of pea (Pisum sativum L) seedlings. Physiol Mol Biol Plants 19(1):81–89

    Article  CAS  Google Scholar 

  • Dias MC, Brüggemann W (2010) Limitations of photosynthesis in Phaseolus vulgaris under drought stress: gas exchange, chlorophyll fluorescence and Calvin cycle enzymes. Photosynthetica 48:96–102

    Article  CAS  Google Scholar 

  • Dias MC, Monteiro C, Moutinho-Pereira J, Correia C, Gonçalves B, Santos C (2013) Cadmium toxicity affects photosynthesis and plant growth at different levels. Acta Physiol Plant 35:1281–1289

    Article  CAS  Google Scholar 

  • Gupta K, Huang H, Corpas F (2013) Lead tolerance in plants: strategies for phytoremediation. Environ Sci Pollut Res 20:2150–2161

    Article  CAS  Google Scholar 

  • Hall JL (2002) Cellular mechanism of heavy metal detoxification and tolerance. J Exp Bot 53:1–11

    Article  CAS  Google Scholar 

  • Hardison DW Jr, Ma LQ, Luongo T, Harris WG (2004) Lead contamination in shooting range soils from abrasion of lead bullets and subsequent weathering. Sci Total Envir 328:175–183

    Article  CAS  Google Scholar 

  • ISO (1993) Soil quality—determination of the effects of pollutants on soil flora—Part 1: method for the measurement of inhibition of root growth. ISO 11269-1. Genève, Switzerland

  • ISO (1995) Soil quality—determination of the effects of pollutants on soil flora—Part 2: effects of pollutants on the emergence and growth of higher plants. ISO 11269-2:1995. Genève, Switzerland

  • John R, Ahmad P, Gadgil K, Sharma S (2009) Heavy metal toxicity: effect on plant growth, biochemical parameters and metal accumulation by Brassica juncea L. Int J Plant Prod 3:65–76

    CAS  Google Scholar 

  • Jones MGK, Outlaw WH, Lowry OH (1977) Enzymic assay of 10-7 to 10-14 moles of sucrose in plant-tissues. Plant Physiol 60:379–383

    Article  CAS  Google Scholar 

  • Kaznina NM, Laidinen GF, Titov AF, Talanov AV (2005) Effect of lead on the photosynthetic apparatus of annual grasses. Biol Bull 32:147–150

    Article  CAS  Google Scholar 

  • Kosobrukhov A, Knyazeva I, Mudrik V (2004) Plantago major plants responses to increase content of lead in soil: growth and photosynthesis. Plant Growth Regul 42:145–151

    Article  CAS  Google Scholar 

  • Krishna R, Dan T, Saxena P (2000) A fragrant solution to soil remediation. Int J Phytorem 2:117–132

    Article  Google Scholar 

  • Krupa Z, Baszynski T (1985) Requirement of galactolipids for photosystem I activity in lyophilized spinach chloroplasts. Biochim Biophys Acta 408:26–34

    Article  Google Scholar 

  • Kučera T, Horáková H, Šonská A (2008) Toxic metal ions in photoautotrophic organisms. Photosynthetica 46:481–489

    Article  Google Scholar 

  • Lee KR, Roh KS (2003) Influence of cadmium on RuBisCO activation in Canavalia ensiformis L. leaves. Biotechnol Bioproc E 8:94–100

    Article  CAS  Google Scholar 

  • Li J, Xie ZM, Zhu YG, Naidu R (2005) Risk assessment of heavy metal contaminated soil in the vicinity of a lead/zinc mine. J Environ Sci (China) 17:881–5

    CAS  Google Scholar 

  • Lichtenhaler HK (1987) Chlorophylls and carotenoids: pigments of photosynthetic biomembranes. Meth Enzymol 148:350–382

    Article  Google Scholar 

  • Lilley RM, Walker DA (1974) Improved spectrophotometric assay for ribulose-bis-phosphate carboxylase. Biochim Biophys Acta 358:226–229

    Article  CAS  Google Scholar 

  • Loureiro S, Santos C, Pinto G, Costa A, Monteiro A, Soares A, Nogueira A (2006). Toxicity Assessment of Two Soils from Jales Mine (Portugal) Using Plants: Growth and Biochemical Parameters. Arch. Environ Contam Toxicol 50:182–190

  • Malecka A, Piechalak A, Mensinger A, Hannc A, Balalkiewicz D, Tomaszewska B (2012) Antioxidant defense system in Pisum sativum roots exposed to heavy metals (Pb, Cu, Cd, Zn). Pol J Environ Stud 2:1721–1730

    Google Scholar 

  • Monteiro M, Santos C, Mann M, Soares A, Lopes T (2007) Evaluation of cadmium genotoxicity in Lactuca sativa L. using nuclear microsatellites. Environ Exp Bot 60(3):421–427

  • Monteiro MS, Santos C, Soares AMVM, Mann RM (2009) Assessment of biomarkers of cadmium stress in lettuce. Ecotox Environ Safe 72:811–818

    Article  CAS  Google Scholar 

  • Monteiro C, Santos C, Pinho S, Dias C (2012) Cadmium-induced cyto- and genotoxicity are organ-dependent in lettuce. Chem Res Tox 25:1423–1434

    Article  CAS  Google Scholar 

  • Moustakas M, Lanaras T, Symeonidis L, Karataglis S (1994) Growth and photosynthetic characteristics of field-grown Avena sativa under copper and lead stress. Photosynthetica 30:389–396

    CAS  Google Scholar 

  • OECD (2006) Terrestrial plant test: seedling emergence and seedling growth test. OECD-208. France

  • OEHHA (2009) Integrated Risk Assessment Branch, office of environmental health hazard assessment. California Environmental Protection Agency, USA, 153pp

    Google Scholar 

  • Päivöke AEA, Simola LK (2001) Arsenate toxicity to Pisum sativum: mineral nutrients, chlorophyll content, and phytase activity. Ecotoxicol Environ Saf 49:111–121

    Article  Google Scholar 

  • Parys E, Romanowska E, Siedlecka M, Poskuta JW (1998) The effect of lead on photosynthesis and respiration in detached leaves and in mesophyll protoplasts of Pisum sativum. Acta Physiol Plant 20:313–322

    Article  CAS  Google Scholar 

  • Peralta-Videa JR, Lopez ML, Narayan M, Saupe G, Gardea-Torresdey J (2009) The biochemistry of environmental heavy metal uptake by plants: implications for the food chain. Int J Biochem Cell Biol 41:1665–1677

    Article  CAS  Google Scholar 

  • Piechalak A, Tomaszewskaa B, Barakiewicz D (2003) Enhancing phytoremediative ability of Pisum sativum by EDTA application. Phytochemistry 64:1239–1251

    Article  CAS  Google Scholar 

  • Podazza G, Rosa M, Gonzalez JA, Hilal M, Prado FE (2006) Cadmium induces changes in sucrose partitioning, invertase activities, and membrane functionality in roots of Rangpur lime (Citrus limonia L. Osbeck). Plant Biol 8:706–714

    Article  CAS  Google Scholar 

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

    CAS  Google Scholar 

  • Prasad M, Strzalka S (2002) Physiology and biochemistry of metal toxicity and tolerance in plants. Kluwer Academic, Dordrecht

    Book  Google Scholar 

  • Prüss-Üstün A, Campbell-Lendrum D, Corvalán C, Woodward A (2003) Assessing the environmental burden of disease at national and local levels. In: Fewtrell L, Kaufmann R, Prüss-Üstün A (eds), World Health Organization, Protection of the Human Environment, Geneva

  • Rodriguez E, Azevedo R, Fernandes P, Santos C (2011) Cr(VI) induces DNA damage, cell cycle arrest and polyploidization. Chemical Res Toxicol 24:1040–1047

    Article  CAS  Google Scholar 

  • Rodriguez E, Santos C, Azevedo R, Moutinho-Pereira J, Correia C, Dias MC (2012) Chromium (VI) induces toxicity at different photosynthetic levels in pea. Plant Physiol Biochem 53:94–100

    Article  CAS  Google Scholar 

  • Rodriguez E, Azevedo R, Moreira H, Souto L, Santos C (2013a) Pb2+ exposure induced microsatellite instability in Pisum sativum in a locus related with glutamine metabolism. Plant Physiol Biochem 62:19–22

    Article  CAS  Google Scholar 

  • Rodriguez E, Azevedo R, Moreira H, Souto L, Santos C (2013b) Pb2+ exposure induced microsatellite instability in Pisum sativum in a locus related with glutamine metabolism. Plant Physiol Biochem 62:19–22

    Article  CAS  Google Scholar 

  • Romanowska E, Wasilewska W, Fristedt R, Vener A, Zienkiewicz M (2012) Phosphorylation of PSII proteins in maize thylakoids in the presence of Pb ions. J Plant Physiol 169(4):345–352

    Article  CAS  Google Scholar 

  • Ruley A, Sharma N, Sahi S, Sajwan K (2006) Effects of lead and chelators on growth, photosynthetic activity and Pb uptake in Sesbania drummondii grown in soil. Environ Pollut 144:11–18

    Article  CAS  Google Scholar 

  • Silva S, Pinto-Carnide O, Martins-Lopes P, Guedes-Pinto H, Santos C (2010) Differential aluminium changes on nutrient accumulation and root differentiation in an Al sensitive vs. tolerant wheat. Environ Exp Bot 68:91–98

    Article  CAS  Google Scholar 

  • Sims DA, Gamon JA (2002) Relationships between leaf pigment content and spectral reflectance across a wide range of species, leaf structures and developmental stages. Remote Sens Environ 81:337–354

    Article  Google Scholar 

  • Stitt M, Bulpin PV, Rees TA (1978) Pathway of starch breakdown in photosynthetic tissues of Pisum sativum. Biochim Biophys Acta 544:200–214

    Article  CAS  Google Scholar 

  • Tawinteung N, Parkpian P, DeLaune RD, Jugsujinda A (2005) Evaluation of extraction procedures for removing lead from contaminated soil. J Environ Sci Health 40(2):385–407

    Article  Google Scholar 

  • Tripathy BC, Bhatia B, Mohanty P (1981) Inactivation of chloroplast photosynthetic electron-transport activity by Ni2+. Biochim Biophys Acta 638:217–224

    Article  CAS  Google Scholar 

  • Visioli G, Marmiroli N (2013) The proteomics of heavy metal hyperaccumulation by plants. J Proteomics 79:133–45

  • Vojtĕchová M, Leblová S (1991) Uptake of lead and cadmium by maize seedlings and the effect of heavy-metals on the activity of phosphoenolpyruvate carboxylase isolated from maize. Biol Plantarum 33:386–394

    Article  Google Scholar 

  • Von Caemmerer S, Farquhar GD (1981) Some relationships between the biochemistry of photosynthesis and the gas-exchange of leaves. Planta 153:376–387

    Article  Google Scholar 

  • Wang C, Wang X, Tian Y, Xue Y, Xu X, Sui Y, Yu H (2008) Oxidative stress and potential biomarkers in tomato seedlings subjected to soil lead contamination. Ecotoxicol Environ Saf 71:685–691

    Article  Google Scholar 

  • WHO (2010) Childhood lead poisoning 2010. ISBN 978 92 4 150033 3

  • Yanagiya T, Imura N, Enomoto S, Kondo Y, Himeno S (2000) Suppression of a high affinity transport system for manganese in cadmium-resistant metallothionein-null cells. J Pharmacol Exp Therap 292:1080–1086

  • Zeng LS, Liao M, Chen CL, Huang CY (2006) Effects of lead contamination on soil microbial activity and rice physiological indices in soil-Pb-rice (Oryza sativa L.) system. Chemosphere 65(4):567–574

    Article  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported by the Portuguese Foundation for Science and Technology (FCT) FCT/PTDC/AAC-AMB/112804/2009, BioRem: integration of multiple BIOmarkers of toxicity in an assay of phytoremediation in contaminated sites. FCT also supported a post-doctoral fellowship of M. C. Dias (SFRH/BPD/41700/2007), E. Rodriguez (SFRH/BPD/75696/2011), and J.M.P. Ferreira de Oliveira (SFRH/BPD/74868/2010).

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Authors and Affiliations

  1. Department of Biology, Laboratory of Biotechnology and Cytometry, Campus Universitário de Santiago, University of Aveiro, CESAM, 3810, Aveiro, Portugal

    Eleazar Rodriguez, Raquel Azevedo, José Miguel Pimenta Ferreira de Oliveira & Maria Celeste Dias

  2. Laboratory of Biotechnology and Cytometry, University Aveiro & CESAM, Campus Universitário de Santiago, 3810, Aveiro, Portugal

    Maria da Conceição Santos

  3. Department of Biology and Environment, University of Trás-os-Montes e Alto Douro, CITAB, Apartado 1013, 5001-801, Vila Real, Portugal

    Carlos Correia & José Moutinho-Pereira

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  1. Eleazar Rodriguez
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  2. Maria da Conceição Santos
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  3. Raquel Azevedo
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  5. José Moutinho-Pereira
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Correspondence to Maria da Conceição Santos.

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Rodriguez, E., da Conceição Santos, M., Azevedo, R. et al. Photosynthesis light-independent reactions are sensitive biomarkers to monitor lead phytotoxicity in a Pb-tolerant Pisum sativum cultivar. Environ Sci Pollut Res 22, 574–585 (2015). https://doi.org/10.1007/s11356-014-3375-9

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