Skip to main content
SpringerLink
Log in

Overexpression of PtPCS enhances cadmium tolerance and cadmium accumulation in tobacco

  • Original Paper
  • Published:
Plant Cell, Tissue and Organ Culture (PCTOC) Aims and scope Submit manuscript

Abstract

Phytochelatins chelate heavy metal ions to decrease their toxicity. The chelates are then transferred to, and stored in, the vacuole. Phytochelatin synthase (PCS), which is involved in phytochelatin synthesis, is thought to be a key enzyme for phytoremediation. In this study, a PCS gene encoding phytochelatin synthase was cloned from poplar (Populus tomentosa Carr.), a widely grown model woody plant that accumulates high levels of heavy metals, especially cadmium. Poplar is considered to have potential applications in phytoremediation. The full-length PtPCS cDNA (1512-bp) encoded a polypeptide of 503 amino acid residues. The PtPCS cDNA was transferred into tobacco by Agrobacterium-mediated leaf disk transformation. The transgenic and wild-type (WT) lines of tobacco were subjected to a one time Cd treatment (90 μmol Cd2+) for 30 days, and then evaluated to determine their Cd tolerance. We evaluated morphological and physiological indices including leaf relative electrolyte leakage, malondialdehyde content, total superoxide dismutase activity, chlorophyll content and root activity. Compared with WT plants, the transgenic plants expressing PtPCS grew better in the Cd treatment and showed significantly higher Cd tolerance. Compared with WT plants, the transgenic lines accumulated higher concentrations of Cd (1.7 to 3.0-fold higher Cd concentration in roots; 1.24 to 2.28-fold higher Cd concentration in leaves). However, the transfer coefficient was lower in the transgenic lines than in wild type. We concluded that PtPCS encodes a functional PCS that may be involved in Cd tolerance and accumulation, but not in Cd transport.

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

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  • Benavides MP, Gallego SM, Tomaro ML, Braz J (2005) Cadmium toxicity in plants. Braz J Plant Physiol 17:21–34

    Article  CAS  Google Scholar 

  • Bernarda C, Roosensa N, Czernicb P, Lebrunb M, Verbruggen N (2004) A novel CPx-ATPase from the cadmium hyperaccumulator Thlaspi caerulescens. FEBS Lett 569:140–148

    Article  Google Scholar 

  • Bhuiyan MSU, Min SR, Jeong WJ, Sultana S, Choi KS, Lee Y, Liu JR (2011a) Overexpression of a yeast cadmium factor 1 (YCF1) enhances heavy metal tolerance and accumulation in Brassica juncea. Plant Cell Tissue Organ Cult 105:85–91

    Article  CAS  Google Scholar 

  • Bhuiyan MSU, Min SR, Jeong WJ, Sultana S, Choi KS, Song WY, Lee Y, Lim YP, Liu JR (2011b) Overexpression of AtATM3 in Brassica juncea confers enhanced heavy metal tolerance and accumulation. Plant Cell Tissue Organ Cult 107:69–77

    Article  CAS  Google Scholar 

  • Brunetti P, Zanella L, Proia A, De Paolis A, Falasca G, Altamura MM, Sanità di Toppi L, Costantino P, Cardarelli M (2011) Cadmium tolerance and phytochelatin content of Arabidopsis seedlings over-expressing the phytochelatin synthase gene AtPCS1. J Exp Bot 62:5509–5519

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Clemens S, Kim EJ, Neumann D, Schroeder JI (1999) Tolerance to toxic metals by a gene family of phytochelatin synthases from plants and yeast. EMBO J 18:3325–3333

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Clemens S, Bloss T, Vess C, Neumann D, Nies DH, Zur Nieden U (2002) A transporter in the endoplasmic reticulum of Schizosaccharomyces pombe cells mediates zinc storage and differentially affects transition metal tolerance. J Biol Chem 277:18215–18221

    Article  CAS  PubMed  Google Scholar 

  • Cobbett CS (2000) Phytochelatin biosynthesis and function in heavy-metal detoxification. Curr Opin Plant Biol 3:211–216

    Article  CAS  PubMed  Google Scholar 

  • Curie C, Cassin G, Couch D, Divol F, Higuchi K, Jean ML, Misson J, Schikora A, Czernic P, Mari S (2009) Metal movement within the plant: contribution of nicotianamine and yellow stripe 1-like transporters. Ann Bot-Lond 103:1–11

    Article  CAS  Google Scholar 

  • DalCorso G, Farinati S, Maistri S, Furini A (2008) How plants cope with cadmium: staking all on metabolism and gene expression. J Integr Plant Biol 50:1268–1280

    Article  CAS  PubMed  Google Scholar 

  • Franchi N, Piccinni E, Ferro D, Basso G, Spolaore B, Santovito G, Ballarin L (2014) Characterization and transcription studies of a phytochelatin synthase gene from the solitary tunicate Ciona intestinalis exposed to cadmium. Aquat Toxicol 152:47–56

    Article  CAS  PubMed  Google Scholar 

  • Gasic K, Korban SS (2007a) Expression of Arabidopsis phytochelatin synthase in Indian mustard (Brassica juncea) plants enhances tolerance for Cd and Zn. Planta 225:1277–1285

    Article  CAS  PubMed  Google Scholar 

  • Gasic K, Korban SS (2007b) Transgenic Indian mustard (Brassica juncea) plants expressing an Arabidopsis phytochelatin synthase (AtPCS1) exhibit enhanced As and Cd tolerance. Plant Mol Biol 64:361–369

    Article  CAS  PubMed  Google Scholar 

  • Glaeser H, Coblenz A, Kruczek R, Ruttke I, Ebert-Jung A, Wolf K (1991) Glutathione metabolism and heavy metal detoxification in Schizosaccharomyces pombe. Curr Genet 19:207–213

    Article  CAS  Google Scholar 

  • Grill E, Winnacker EL, Zenk MH (1985) Phytochelatins: the principal heavy-metal complexing peptides of higher plants. Science 230:674–676

    Article  CAS  PubMed  Google Scholar 

  • Hasegawa I, Terada E, Sunairi M, Wakita H, Shinmachi F, Noguchi A, Nakajima M, Yazaki J (1997) Genetic improvement of heavy metal tolerance in plants by transfer of the yeast metallothionein gene (CUP1). In: Ando T, Fujita K, Mae T, Matsumoto H, Mori S, Sekiya J (eds) Plant nutrition for sustainable food production and environment. Kluwer Academic Publishers, Dordrecht, pp 391–395

    Chapter  Google Scholar 

  • He J, Li H, Luo J, Ma C, Li S, Qu L, Gai Y, Jiang X, Janz D, Polle A, Tyree M, Luo ZB (2013) A transcriptomic network underlies microstructural and physiological responses to cadmium in Populus × canescens. Plant Physiol 162:424–439

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • He J, Li H, Ma C, Zhang Y, Polle A, Rennenberg H, Cheng X, Luo ZB (2015) Overexpression of bacterial γ-glutamylcysteine synthetase mediates changes in cadmium influx, allocation and detoxification in poplar. New Phytol 205:240–254

    Article  CAS  PubMed  Google Scholar 

  • Heiss S, Wachter A, Bogs J, Cobbett C, Rausch T (2003) Phytochelatin synthase (PCS) protein is induced in Brassica juncea leaves after prolonged Cd exposure. J Exp Bot 54:1833–1839

    Article  CAS  PubMed  Google Scholar 

  • Lee S, Moon JS, Ko TS, Petros D, Goldsbrough PB, Korban SS (2003) Overexpression of Arabidopsis phytochelatin synthase paradoxically leads to hypersensitivity to cadmium stress. Plant Physiol 131:656–663

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Lee S, Kim YY, Lee Y, An G (2007) Rice P1B-type heavy-metal ATPase, OsHMA9, is a metal efflux protein. Plant Physiol 145:831–842

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Li ZS, Lu YP, Zhen RG, Szczypka M, Thiele DJ, Rea PA (1997) A new pathway for vacuolar cadmium sequestration in Saccharomyces cerevisiae: YCF1-catalyzed transport of bis(glutathionato) cadmium. Proc Natl Acad Sci 94:42–47

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Li J, Guo J, Xu W, Ma M (2006) Enhanced cadmium accumulation in transgenic tobacco expressing the phytochelatin synthase gene of Cynodon dactylon L. J Integr Plant Biol 48:928–937

    Article  CAS  Google Scholar 

  • Lin YF, Aarts MG (2012) The molecular mechanism of zinc and cadmium stress response in plants. Cell Mol Life Sci 69:3187–3206

    Article  CAS  PubMed  Google Scholar 

  • Liu GY, Zhang YX, Chai TY (2011) Phytochelatin synthase of Thlaspi caerulescens enhanced tolerance and accumulation of heavy metals when expressed in yeast and tobacco. Plant Cell Rep 30:1067–1076

    Article  CAS  PubMed  Google Scholar 

  • Liu YX, Wang XT, Su XD, Dai L, Zhang ZY, Xu JC (2012) Cloning and expression analysis of a phytochelatin synthase gene (PtPCS) in Populus tomentosa Carr. Mol Plant Breed 10:174–183 (in Chinese)

    CAS  Google Scholar 

  • Martínez M, Bernal P, Almela C, Vélez D, García-Agustín P, Serrano R, Navarro-Aviñó J (2006) An engineered plant that accumulates higher levels of heavy metals than Thlaspi caerulescens, with yields of 100 times more biomass in mine soils. Chemosphere 64:478–485

    Article  PubMed  Google Scholar 

  • Meyer CL, Peisker D, Courbot M, Craciun AR, Cazalé AC, Desgain D, Schat H, Clemens S, Verbruggen N (2011) Isolation and characterization of Arabidopsis halleri and Thlaspi caerulescens phytochelatin synthases. Planta 234:83–95

    Article  CAS  PubMed  Google Scholar 

  • Migocka M, Papierniak A, Kosatka E, Kłobus G (2011) Comparative study of the active cadmium efflux systems operating at the plasma membrane and tonoplast of cucumber root cells. J Exp Bot 62:4903–4916

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Park J, Song WY, Ko D, Eom Y, Hansen TH, Schiller M, Lee TG, Martinoia E, Lee Y (2012) The phytochelatin transporters AtABCC1 and AtABCC2 mediate tolerance to cadmium and mercury. Plant J 69:278–288

    Article  CAS  PubMed  Google Scholar 

  • Pence NS, Larsen PB, Ebbs SD, Letham DLD, Lasat MM, Garvin DF, Eide D, Kochian LV (2000) The molecular physiology of heavy metal transport in the Zn/Cd hyperaccumulator Thlaspi caerulescens. Proc Natl Acad Sci 97:4956–4960

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Polle A, Klein T, Kettner C (2013) Impact of cadmium on young plants of Populus euphratica and P. × canescens, two poplar species that differ in stress tolerance. New For 44:13–22

    Article  Google Scholar 

  • Pomponi M, Censi V, Di Girolamo V, De Paolis A, di Toppi LS, Aromolo R, Costantino P, Cardarelli M (2006) Overexpression of Arabidopsis phytochelatin synthase in tobacco plants enhances Cd2+ tolerance and accumulation but not translocation to the shoot. Planta 223:180–190

    Article  CAS  PubMed  Google Scholar 

  • Rea PA (2007) Plant ATP-binding cassette transporters. Annu Rev Plant Biol 58:347–375

    Article  CAS  PubMed  Google Scholar 

  • Rea PA (2012) Phytochelatin synthase: of a protease a peptide polymerase made. Physiol Plant 145:154–164

    Article  CAS  PubMed  Google Scholar 

  • Robinson BH, Mills TM, Petit D, Fung LE, Green SR, Clothier BE (2000) Natural and induced cadmium-accumulation in poplar and willow: implications for phytoremediation. Plant Soil 227:301–306

    Article  CAS  Google Scholar 

  • Salt DE, Rauser WE (1995) MgATP-dependent transport of phytochelatins across the tonoplast of oat roots. Plant Physiol 107:1293–1301

    PubMed Central  CAS  PubMed  Google Scholar 

  • Shukla D, Kesari R, Mishra S, Dwivedi S, Tripathi RD, Nath P, Trivedi PK (2012) Expression of phytochelatin synthase from aquatic macrophyte Ceratophyllum demersum L. enhances cadmium and arsenic accumulation in tobacco. Plant Cell Rep 31:1687–1699

    Article  CAS  PubMed  Google Scholar 

  • Shukla D, Kesari R, Tiwari M, Dwivedi S, Tripathi RD, Nath P, Trivedi PK (2013) Expression of Ceratophyllum demersum phytochelatin synthase, CdPCS1, in Escherichia coli and Arabidopsis enhances heavy metal (loid)s accumulation. Protoplasma 250:1263–1272

    Article  CAS  PubMed  Google Scholar 

  • Thomine S, Wang R, Ward JM, Crawford NM, Schroeder JI (2000) Cadmium and iron transport by members of a plant metal transporter family in Arabidopsis with homology to Nramp genes. Proc Natl Acad Sci 97:4991–4996

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Vatamaniuk OK, Mari S, Lu YP, Rea PA (1999) AtPCS1, a phytochelatin synthase from Arabidopsis: isolation and in vitro reconstitution. Proc Natl Acad Sci 96:7110–7115

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Vatamaniuk OK, Mari S, Lang A, Chalasani S, Demkiv LO, Rea PA (2004) Phytochelatin synthase, a dipeptidyltransferase that undergoes multisite acylation with γ-glutamylcysteine during catalysis: stoichiometric and site-directed mutagenic analysis of Arabidopsis thaliana PCS1-catalyzed phytochelatin synthesis. J Biol Chem 279:22449–22460

    Article  CAS  PubMed  Google Scholar 

  • Vögeli-Lange R, Wagner GJ (1990) Subcellular localization of cadmium and cadmium-binding peptides in tobacco leaves: implication of a transport function for cadmium-binding peptides. Plant Physiol 92:1086–1093

    Article  PubMed Central  PubMed  Google Scholar 

  • Vrbová M, Kotrba P, Horáček J, Smýkal P, Švábová L, Větrovcová M, Smýkalová I, Griga M (2013) Enhanced accumulation of cadmium in Linum usitatissimum L. plants due to overproduction of metallothionein α-domain as a fusion to β-glucuronidase protein. Plant Cell Tissue Organ Cult 112:321–330

    Article  Google Scholar 

  • Wang F, Wang Z, Zhu C (2012) Heteroexpression of the wheat phytochelatin synthase gene (TaPCS1) in rice enhances cadmium sensitivity. Acta Biochim Biophys Sin 44:886–893

    Article  CAS  PubMed  Google Scholar 

  • Xu X, Lv QM, Shang JJ, Pang ZQ, Zhou ZQ, Wang J, Jiang GH, Tao Y, Xu Q, Li XB, Zhao XF, Li SG, Xu JC, Zhu LH (2014) Excavation of Pid3 orthologs with differential resistance spectra to Magnaporthe oryzae in rice resource. PLoS One 9:e93275

    Article  PubMed Central  PubMed  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. S Afr J Bot 76:167–179

    Article  CAS  Google Scholar 

  • 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 197:23–34

    Article  CAS  Google Scholar 

  • Zhao C, Xu J, Li Q, Li S, Wang P, Xiang F (2014) Cloning and characterization of a Phragmites australis phytochelatin synthase (PaPCS) and achieving Cd tolerance in tall fescue. PLoS One 9:e103771

    Article  PubMed Central  PubMed  Google Scholar 

Download references

Acknowledgments

The authors wish to thank Beijing Natural Science Foundation (#5122019) for funding support.

Author information

Author notes

    Authors and Affiliations

    1. National Engineering Laboratory for Tree Breeding, Beijing Forestry University, Beijing, 100083, China

      Yongkun Chen, Yuxia Liu, Yana Ding, Xiaotong Wang & Jichen Xu

    Authors
    1. Yongkun Chen
      View author publications

      You can also search for this author in PubMed Google Scholar

    2. Yuxia Liu
      View author publications

      You can also search for this author in PubMed Google Scholar

    3. Yana Ding
      View author publications

      You can also search for this author in PubMed Google Scholar

    4. Xiaotong Wang
      View author publications

      You can also search for this author in PubMed Google Scholar

    5. Jichen Xu
      View author publications

      You can also search for this author in PubMed Google Scholar

    Corresponding author

    Correspondence to Jichen Xu.

    Additional information

    Chen Yongkun and Liu Yuxia have contributed equally to this work.

    Rights and permissions

    Reprints and permissions

    About this article

    Check for updates. Verify currency and authenticity via CrossMark

    Cite this article

    Chen, Y., Liu, Y., Ding, Y. et al. Overexpression of PtPCS enhances cadmium tolerance and cadmium accumulation in tobacco. Plant Cell Tiss Organ Cult 121, 389–396 (2015). https://doi.org/10.1007/s11240-015-0710-x

    Download citation

    • Received:

    • Accepted:

    • Published:

    • Issue Date:

    • DOI: https://doi.org/10.1007/s11240-015-0710-x

    Keywords

    Search

    Navigation