Skip to main content
SpringerLink
Log in

Molecular cloning of class III chitinase gene from Avicennia marina and its expression analysis in response to cadmium and lead stress

  • Published:
Ecotoxicology Aims and scope Submit manuscript

Abstract

Mangrove species have high tolerance to heavy metal pollution. Chitinases have been widely reported as defense proteins in response to heavy metal stress in terrestrial plants. In this study, a full-length cDNA sequence encoding an acidic and basic class III chitinase (AmCHI III) was cloned by using RT-PCR and RACE methods in Avicennia marina. AmCHI III mRNA expression in leaf of A. marina were investigated under Cd, Pb stresses on using real-time quantitative PCR. The deduced AmCHI III protein consists of 302 amino acids, including a signal putative peptide region, and a catalytic domain. Homology modeling of the catalytic domain revealed a typical molecular structure of class III plant chitinases. Results further demonstrated that the regulation of AmCHI III mRNA expression in leaves was strongly dependent on Cd, Pb stresses. AmCHI III mRNA expressions were significantly increased in response to Cd, Pb, and peaked at 7 days Cd-exposure, 7 days Pb-exposure, respectively. AmCHI III mRNA expression exhibited more sensitive to Pb stress than Cd stress. This work was the first time cloing chitinase from A. marina, and it brought evidence on chitinase gene involving in heavy metals (Cd2+ and Pb2+) resistance or detoxification in plants. Further studies including the promoter and upstream regulation, gene over-expression and the response of mangrove chitinases to other stresses will shed more light on the role of chitinase in mangrove plants.

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

Similar content being viewed by others

References

  • Beintema JJ (1994) Structural features of plant chitinases and chitin-binding proteins. FEBS Lett 350:159–163

    Article  CAS  Google Scholar 

  • Békésiová B, Hraška Š, Libantová J, Moravčíková J, Matušíková I (2008) Heavy-metal stress induced accumulation of chitinase isoforms in plants. Mol Biol Rep 35:579–588

    Article  Google Scholar 

  • Boava LP, Cristofani-Yaly M, Stuart RM, Machado MA (2011) Expression of defense-related genes in response to mechanical wounding and Phytophthora parasitica infection in Poncirus trifoliata and Citrus sunki. Physiol Mol Plant Pathol 76:119–125

    Article  CAS  Google Scholar 

  • Caregnato FF, Koller CE, MacFarlane GR, Moreira JC (2008) The glutathione antioxidant system as a biomarker suite for the assessment of heavy metal exposure and effect in the grey mangrove, Avicennia marina (Forsk.) Vierh. Mar Pollut Bull 56:1119–1127

    Article  CAS  Google Scholar 

  • Cheng M-C, Liao P-M, Kuo W-W, Lin T-P (2013) The Arabidopsis ETHYLENE-RESPONSE-FACTOR1 regulates abiotic-stress-responsive gene expression by binding to different cis-acting elements in response to different stress signals. Plant Physiol 162:1566–1582

    Article  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  CAS  Google Scholar 

  • Collinge DB, Kragh KM, Mikkelsen JD, Nielsen KK, Rasmussen U, Vad K (1993) Plant chitinases. Plant J 3:31–40

    Article  CAS  Google Scholar 

  • DalCorso G (2012) Plants and heavy metals., Heavy metal toxicity in plantsSpringer, New York, pp 1–25

    Book  Google Scholar 

  • Dani V, Simon WJ, Duranti M, Croy RR (2005) Changes in the tobacco leaf apoplast proteome in response to salt stress. Proteomics 5:737–745

    Article  CAS  Google Scholar 

  • de A Gerhardt LB, Sachetto-Martins G, Contarini MG, Sandroni M, de P Ferreira R, de Lima VM, Cordeiro MC, de Oliveira DE, Margis-Pinheiro M (1997) Arabidopsis thaliana class IV chitinase is early induced during the interaction with Xanthomonas campestris. FEBS Lett 419:69–75

    Article  Google Scholar 

  • de las Mercedes Dana M, Pintor-Toro JA, Cubero B (2006) Transgenic tobacco plants overexpressing chitinases of fungal origin show enhanced resistance to biotic and abiotic stress agents. Plant Physiol 142:722–730

    Article  Google Scholar 

  • Gonzalez-Mendoza D, Moreno AQ, Zapata-Perez O (2007) Coordinated responses of phytochelatin synthase and metallothionein genes in black mangrove, Avicennia germinans, exposed to cadmium and copper. Aquat Toxicol 83:306–314

    Article  CAS  Google Scholar 

  • Grover A (2012) Plant chitinases: genetic diversity and physiological roles. Crit Rev Plant Sci 31:57–73

    Article  CAS  Google Scholar 

  • Henrissat B, Davies G (1997) Structural and sequence-based classification of glycoside hydrolases. Curr Opin Struct Biol 7:637–644

    Article  CAS  Google Scholar 

  • Hong JK, Hwang BK (2006) Promoter activation of pepper class II basic chitinase gene, CAChi2, and enhanced bacterial disease resistance and osmotic stress tolerance in the CAChi2-overexpressing Arabidopsis. Planta 223:433–448

    Article  CAS  Google Scholar 

  • Huang GY, Wang YS (2009) Expression analysis of type 2 metallothionein gene in mangrove species (Bruguiera gymnorrhiza) under heavy metal stress. Chemosphere 77:1026–1029

    Article  CAS  Google Scholar 

  • Huang G-Y, Wang Y-S, Ying G-G (2011) Cadmium-inducible BgMT2, a type 2 metallothionein gene from mangrove species (Bruguiera gymnorrhiza), its encoding protein shows metal-binding ability. J Exp Mar Biol Ecol 405:128–132

    Article  CAS  Google Scholar 

  • Kieffer P, Schröder P, Dommes J, Hoffmann L, Renaut J, Hausman J-F (2009) Proteomic and enzymatic response of poplar to cadmium stress. J Proteomics 72:379–396

    Article  CAS  Google Scholar 

  • Kikuchi T, Masuda K (2009) Class II chitinase accumulated in the bark tissue involves with the cold hardiness of shoot stems in highbush blueberry (Vaccinium corymbosum L.). Sci Hortic 120:230–236

    Article  CAS  Google Scholar 

  • Li H-Y, Li Y, Wei W, Zheng C-C, Shu H-R (2004) SA induction of a grapevine class III chitinase gene VCH3 promoter in transgenic tobacco vascular tissue. J Integr Plant Biol 46:148–153

    CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  • Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(T)(-Delta Delta C) method. Methods 25:402–408

    Article  CAS  Google Scholar 

  • MacFarlane G, Burchett M (1999) Zinc distribution and excretion in the leaves of the grey mangrove, Avicennia marina (Forsk.) Vierh. Environ Exp Bot 41:167–175

    Article  CAS  Google Scholar 

  • MacFarlane G, Burchett M (2002) Toxicity, growth and accumulation relationships of copper, lead and zinc in the grey mangrove Avicennia marina (Forsk.) Vierh. Mar Environ Res 54:65–84

    Article  CAS  Google Scholar 

  • MacFarlane G, Pulkownik A, Burchett M (2003) Accumulation and distribution of heavy metals in the grey mangrove, Avicennia marina (Forsk.) Vierh.: biological indication potential. Environ Pollut 123:139–151

    Article  CAS  Google Scholar 

  • Metwally A, Finkemeier I, Georgi M, Dietz KJ (2003) Salicylic acid alleviates the cadmium toxicity in barley seedlings. Plant Physiol 132:272–281

    Article  CAS  Google Scholar 

  • Peters EC, Gassman NJ, Firman JC, Richmond RH, Power EA (2009) Ecotoxicology of tropical marine ecosystems. Environ Toxicol Chem 16:12–40

    Article  Google Scholar 

  • Regalado AP, Pinheiro C, Vidal S, Chaves I, Ricardo CP, Rodrigues-Pousada C (2000) The Lupinus albus class-III chitinase gene, IF3, is constitutively expressed in vegetative organs and developing seeds. Planta 210:543–550

    Article  CAS  Google Scholar 

  • Rivera-Becerril F, Metwally A, Martin-Laurent F, Van Tuinen D, Dietz KJ, Gianinazzi S, Gianinazzi-Pearson V (2005a) Molecular responses to cadmium in roots of Pisum sativum L. Water Air Soil Pollut 168:171–186

    Article  CAS  Google Scholar 

  • Rivera-Becerril F, van Tuinen D, Martin-Laurent F, Metwally A, Dietz KJ, Gianinazzi S, Gianinazzi-Pearson V (2005b) Molecular changes in Pisum sativum L. roots during arbuscular mycorrhiza buffering of cadmium stress. Mycorrhiza 16:51–60

    Article  CAS  Google Scholar 

  • Straif K, Benbrahim-Tallaa L, Baan R, Grosse Y, Secretan B, El Ghissassi F, Bouvard V, Guha N, Freeman C, Galichet L (2009) A review of human carcinogens-part C: metals, arsenic, dusts, and fibres. Lancet Oncol 10:453–454

    Article  Google Scholar 

  • Sugimoto K, Matsui K, Ozawa R, Takabayashi J (2011) Characterization of the promoter sequence of chitinase gene from lima bean plant. J Plant Interact 6:163–164

    Article  CAS  Google Scholar 

  • Tapia G, Morales-Quintana L, Inostroza L, Acuña H (2011) Molecular characterisation of Ltchi7, a gene encoding a Class III endochitinase induced by drought stress in Lotus spp. Plant Biology 13:69–77

    Article  CAS  Google Scholar 

  • Terwisscha van Scheltinga AC, Kalk KH, Beintema JJ, Dijkstra BW (1994) Crystal structures of hevamine, a plant defence protein with chitinase and lysozyme activity, and its complex with an inhibitor. Structure 2:1181–1189

    Article  CAS  Google Scholar 

  • van Keulen H, Wei R, Cutright TJ (2008) Arsenate-induced expression of a class III chitinase in the dwarf sunflower Helianthus annuus. Environ Exp Bot 63:281–288

    Article  Google Scholar 

  • van Scheltinga AT, Hennig M, Dijkstra B (1996) The 1.8 angstrom resolution structure of hevamine, a plant chitinase/lysozyme, and analysis of the conserved sequence and structure motifs of glycosyl hydrolase family 18. J Mol Biol 262:243–257

    Article  Google Scholar 

  • Walliwalagedara C, Atkinson I, Van Keulen H, Cutright T, Wei R (2010) Differential expression of proteins induced by lead in the Dwarf Sunflower Helianthus annuus. Phytochemistry 71:1460–1465

    Article  CAS  Google Scholar 

  • Wu CT, Bradford KJ (2003) Class I chitinase and beta-1,3-glucanase are differentially regulated by wounding, methyl jasmonate, ethylene, and gibberellin in tomato seeds and leaves. Plant Physiol 133:263–273

    Article  CAS  Google Scholar 

  • Yadav S (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 

  • Yamamoto S, Nakano T, Suzuki K, Shinshi H (2004) Elicitor-induced activation of transcription via W box-related cis-acting elements from a basic chitinase gene by WRKY transcription factors in tobacco. Biochim Biophys Acta 1679:279–287

    Article  CAS  Google Scholar 

  • Yang S, Wu Q (2003) Effect of Cd on growth and physiological characteristics of Aegiceras corniculatum seedlings. Mar Environ Sci 22:38–42

    Google Scholar 

  • Zhang FQ, Wang YS, Lou ZP, Dong JD (2007) Effect of heavy metal stress on antioxidative enzymes and lipid peroxidation in leaves and roots of two mangrove plant seedlings (Kandelia candel and Bruguiera gymnorrhiza). Chemosphere 67:44–50

    Article  CAS  Google Scholar 

Download references

Acknowledgments

This research was supported by the key Projects in the National Science & Technology Pillar Program in the Eleventh Five-year Plan Period (No. 2012BAC07B0402), the Projects of Guangzhou Science and Technology (No. 201504010006), the National Natural Science Foundation of China (No. 41430966, No. 41076070 and No. 41176101) and the projects of knowledge innovation program of Chinese Academy of Sciences (No. KSCX2-SW-132).

Conflict of interest

The authors declare that they have no conflict of interest.

Author information

Authors and Affiliations

  1. State Key Laboratory of Tropical Oceanography, South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou, 510301, China

    Li-Ying Wang, You-Shao Wang & Jing-Ping Zhang

  2. Daya Bay Marine Biology Research Station, Chinese Academy of Sciences, Shenzhen, 518121, China

    You-Shao Wang

  3. Laboratory of Environmental Toxicology, Department of Ecology & Biodiversity, The University of Hong Kong, Pokfulam Road, Hong Kong, SAR, China

    Ji-Dong Gu

Authors
  1. Li-Ying Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. You-Shao Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jing-Ping Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Ji-Dong Gu
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to You-Shao Wang.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Wang, LY., Wang, YS., Zhang, JP. et al. Molecular cloning of class III chitinase gene from Avicennia marina and its expression analysis in response to cadmium and lead stress. Ecotoxicology 24, 1697–1704 (2015). https://doi.org/10.1007/s10646-015-1501-1

Download citation

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10646-015-1501-1

Keywords

Search

Navigation