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Characterization and Expression of a Phytochelatin Synthase Gene in Birch-leaf Pear (Pyrus betulaefolia Bunge)

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Abstract

Phytochelatins play a major role in metal detoxification in plants. However, the molecular mechanisms of heavy metal detoxification remain unknown, and birch-leaf pear genes related to metal detoxification have not yet been identified. Here, we report the isolation of cDNA and genomic DNA sequences encoding a phytochelatin synthase (PCS) from birch-leaf pear (Pyrus betulaefolia Bunge). The PbPCS1 cDNA sequence contained 1,965 nucleotides of a 1,494 bp open reading frame (ORF), which encodes a 497-amino-acid protein with a molecular weight of 55.00 kDa. Its corresponding genomic DNA sequence consists of 3,820 nucleotides and eight exons separated by seven introns. The deduced amino acid sequence of PbPCS1 contains five Cys residues (56, 90, 91, 109 and 113 amino acids) that are highly conserved in the plant PCS1 family, and 12 cysteine residues putatively involved in heavy metal binding sites, which included three adjacent Cys–Cys components (331–332, 351–352 and 369–370 amino acids) in the C-terminal variable domain. Homology analysis of the deduced PbPCS1 amino acid sequence revealed that it shares a high sequence similarity amongst N-terminal amino acids and low similarity with C-terminal amino acids with plant PCS1 proteins deposited with NCBI. PbPCS1 was most related to PCS1 from legume plants Lotus japonicus (GenBank accession no. AAT80342) and soybean (Glycine max L.; AAL78384) as they were clustered into the same clade in a neighbor-joining phylogenetic tree. Quantitative real-time PCR (qPCR) expression analysis revealed that PbPCS1 had a very low basal expression level in untreated whole seedlings, and levels were higher in roots than in leaves and stems. After 24 h of exposure to 20 μM CdSO4, CuSO4 or ZnSO4, PbPCS1 expression increased significantly in different organs. In addition, L-buthionine-sulfo-ximine (BSO) can inhibit PbPCS1 expression in roots, stems and leaves, while L-glutathionereduced (GSH) stimulates PbPCS1 expression in different organs of birch-leaf pear.

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References

  • Alberich A, Diaz-Cruz JM, Arino C, Esteban M (2008) Combined use of the potential shift correction and the simultaneous treatment of spectroscopic and electrochemical data by multivariate curve resolution: analysis of a Pb(II)-phytochelatin system. Analyst 133:470–477

    Article  PubMed  CAS  Google Scholar 

  • Blum R, Meyer KC, Wünschmann J, Lendzian KJ, Grill E (2010) Cytosolic action of phytochelatin synthase. Plant Physiol 153:159–169

    Article  PubMed  CAS  Google Scholar 

  • Chaurasia N, Mishra Y, Rai LC (2008) Cloning expression and analysis of phytochelatin synthase (pcs) gene from Anabaena sp. PCC 7120 offering multiple stress tolerance in Escherichia coli. Biochem Biophys Res Commun 376:225–230

    Article  PubMed  CAS  Google Scholar 

  • Clemens S (2006) Evolution and function of phytochelatin synthases. J Plant Physiol 163:319–332

    Article  PubMed  CAS  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  CAS  Google Scholar 

  • Cobbett CS (2000) Phytochelatins and their roles in heavy metal detoxification. Plant Physiol 123:825–832

    Article  PubMed  CAS  Google Scholar 

  • Cobbett C, Goldsbrough P (2002) Phytochelatins and metallothioneins: roles in heavy metal detoxification and homeostasis. Annu Rev Plant Biol 53:159–182

    Article  PubMed  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  PubMed  CAS  Google Scholar 

  • Grill E, Loffler S, Winnacker EL, Zenk MH (1989) Phytochelatins, the heavy-metal-binding peptides of plants, are synthesized from glutathione by a specific gamma-glutamylcysteine dipeptidyl transpeptidase (phytochelatin synthase). Proc Natl Acad Sci USA 86:6838–6842

    Article  PubMed  CAS  Google Scholar 

  • Guan LL, Wang YB, Shen H, Hou K, Xu YW, Wu W (2012) Molecular cloning and expression analysis of genes encoding two microsomal oleate desaturases (FAD2) from safflower (Carthamus tinctorius L.). Plant Mol Biol Rep 30:139–148

    Article  CAS  Google Scholar 

  • Ha S, Smith AP, Howden R, Dietrich WM, Bugg S, O’Connell MJ, Goldsbrough PB, Cobbett CS (1999) Phytochelatin synthase genes from Arabidopsis and the yeast Schizosaccharomyces pombe. Plant Cell 11:1153–1163

    PubMed  CAS  Google Scholar 

  • He Z, Li J, Zhang H, Ma M (2005) Different effects of calcium and lanthanum on the expression of phytochelatin synthase gene and cadmium absorption in Lactuca sativa. Plant Sci 168:309–318

    Article  CAS  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  PubMed  CAS  Google Scholar 

  • Kaneyoshi J, Wabiko H, Kobayashi S, Tsuchiya T (2001) Agrobacterium tumefaciens AKE10-mediated transformation of an Asian pea pear, Pyrus betulaefolia Bunge: host specificity of bacterial strains. Plant Cell Rep 20:622–628

    Article  CAS  Google Scholar 

  • Liu XM, Anderson JM, Pijut PM (2010) Cloning and characterization of Prunus serotina AGAMOUS, a putative flower homeotic gene. Plant Mol Biol Rep 28:193–203

    Article  CAS  Google Scholar 

  • Maier T, Yu C, Küllertz G, Clemens S (2003) Localization and functional characterization of metal-binding sites in phytochelatin synthases. Planta 218:300–308

    Article  PubMed  CAS  Google Scholar 

  • Matsumoto K, Chun J, Tamura F, Kamamoto Y, Tanabe K (2006) Salt tolerance in Pyrus species is linked to levels of Na and Cl translocation from roots to leaves. J Japan Soc Hort Sci 75:385–391

    Article  CAS  Google Scholar 

  • Matsumoto K, Tamura F, Chun J, Ikeda T, Imanishi K, Tanabe K (2007) Enhancement in salt tolerance of Japanese pear by using Pyrus betulaefolia rootstock. Hortic Res (Japan) 6:47–52

    Article  Google Scholar 

  • Okubo M, Sakuratani T (2000) Effects of sodium chloride on survival and stem elongation of two Asian pear rootstock seedlings. Sci Hortic 85:85–90

    Article  CAS  Google Scholar 

  • Okubo M, Furukawa Y, Sakuratani T (2000) Growth, fowering and leaf properties of pear cultivars grafted on two Asian pear rootstock seedlings under NaCl irrigation. Sci Hortic 85:91–101

    Article  CAS  Google Scholar 

  • Ramos J, Clemente MR, Naya L, Loscos J, Perez-Rontome C, Sato S, Tabata S, Becana M (2007) Phytochelatin synthases of the model legume Lotus japonicus. a small multigene family with differential response to cadmium and alternatively spliced variants. Plant Physiol 143:1110–1118

    Article  PubMed  CAS  Google Scholar 

  • Ramos J, Naya L, Gay M, Abian J, Becana M (2008) Functional characterization of an unusual phytochelatin synthase, LjPCS3, of Lotus japonicus. Plant Physiol 148:536–545

    Article  PubMed  CAS  Google Scholar 

  • Rea PA (2006) Phytochelatin synthase, papain’s cousin, in stereo. Proc Natl Acad Sci USA 103:507–508

    Article  PubMed  CAS  Google Scholar 

  • Ruotolo R, Peracchi A, Bolchi A, Infusini G, Amoresano A, Ottonello S (2004) Domain organization of phytochelatin synthase. J Biol Chem 279:14686–14693

    Article  PubMed  CAS  Google Scholar 

  • Tamura F, Tanabe K, Katayama M, Itai A (1996) Effect of flooding on ethanol and ethylene production by pear rootstocks. J Japan Soc Hortic Sci 65:261–266

    Article  CAS  Google Scholar 

  • Tamura K, Dudley J, Nei M, Kumar S (2007) MEGA4: Molecular Evolutionary Genetics Analysis (MEGA) software version 4.0. Mol Biol Evol 24:1596–1599

    Article  PubMed  CAS  Google Scholar 

  • Tennstedt P, Peisker D, Bottcher C, Trampczynska A, Clemens S (2009) Phytochelatin synthesis is essential for the detoxification of excess Zn and contributes significantly to the accumulation of Zn. Plant Physiol 149:938–948

    Article  PubMed  CAS  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 USA 96:7110–7115

    Article  PubMed  CAS  Google Scholar 

  • Vestergaard M, Matsumoto S, Nishikori S, Shiraki K, Hirata K, Takagi M (2008) Chelation of cadmium ions by phytochelatin synthase: role of the cystein-rich C-terminal. Anal Sci 24:277–281

    Article  PubMed  CAS  Google Scholar 

  • Zhang H, Xu W, Guo J, He Z, Ma M (2005) Coordinated responses of phytochelatins and metallothioneins to heavy metals in garlic seedlings. Plant Sci 169:1059–1065

    Article  CAS  Google Scholar 

  • Zhao D, Zhou C, Sheng Y, Liang G, Tao J (2011) Molecular cloning and expression of phytoene synthase, lycopene beta-cyclase, and beta-carotene hydroxylase genes in persimmon (Diospyros kaki L.) fruits. Plant Mol Biol Rep 29:345–351

    Article  CAS  Google Scholar 

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Acknowledgments

This work was supported by the Jiangsu Agriculture Science and Technology Innovation Fund (China) through project CX(11)4050, and the National Natural Sciences Foundation of China (No. 31101529).

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Authors and Affiliations

  1. Institute of Horticulture, Jiangsu Academy of Agricultural Sciences, Nanjing, 210014, People’s Republic of China

    You Hong Chang, Hui Li, Jing Lin & Bao Long Sheng

  2. Efficient Horticulture Crop Genetic Improvement Laboratory, National Agricultural Science and Technology Jiangsu Innovative Center, Nanjing, 210014, People’s Republic of China

    You Hong Chang & Hui Li

  3. State Key Laboratory of Soil and Sustainable Agriculture, Institute of Soil Science, Chinese Academy of Sciences, Nanjing, 210008, People’s Republic of China

    Yu Cong

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  1. You Hong Chang
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  2. Hui Li
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Correspondence to Hui Li.

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Chang, Y.H., Li, H., Cong, Y. et al. Characterization and Expression of a Phytochelatin Synthase Gene in Birch-leaf Pear (Pyrus betulaefolia Bunge). Plant Mol Biol Rep 30, 1329–1337 (2012). https://doi.org/10.1007/s11105-012-0447-1

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