Antioxidative Responses of Eucalyptus camaldulensis to Different Concentrations of Copper

Document Type : Research Paper


1 Biotechnology Research Department, Research Institute of Forests and Rangelands, Tehran, Iran

2 Seed and Plant Certification and Registration Institute, Bolvar Nabovvat, Karaj, Iran


Anthropogenic activities have caused important increases in soil Cu levels not only in urban areas but also in croplands. This study was designed to find out the effect of different concentration of copper on physiological and biochemical changes in Eucalyptus camaldulensis seedlings. Seeds of Eucalyptus camaldulensis were grown in marble chips and irrigated with nutrient solution mixed with copper (control, 5, 10, 20 mM) for 10 months and after this period, leaf, stem and root tissues were harvested. Copper content was determined by ICP-OES and some characters such as proline, pigments, catalase (CAT), peroxidase (POX), superoxid desmotase (SOD) and weight of different tissues were measured. The concentrations of copper in root tissue were higher than leaf and stem tissues and stem concentration was lower than the concentration of leaf. The proline content was raised by increasing metal concentrations, but the content of pigments decreased. The activity of antioxidative enzymes, CAT, POX and SOD positively increased up to 10 mM Cu treatment and then slightly decreased in both leaf and root tissues. These results suggest that eucalypts have efficient mechanism to tolerate Cu excess, as evidenced by accumulating of osmoprotectants and antioxidative enzymes. Also eucalypts under stress can accumulate copper four times more than the control treatment without serious symptoms in growth, therefore it is a feasible plant for hyperaccumulation of copper and declining the environmental pollution.


Article Title [فارسی]

عکس‌العمل فیزیولوژیک و آنتی اکسیدانی Eucalyprtus camaldulensis در برابر غلظت‌های سمی مس

Abstract [فارسی]

فعالیت­های انسانی نه تنها در مناطق شهری بلکه در اراضی کشاورزی منجر به افزایش قابل توجه مس شده است. این مطالعه به منظور بررسی تغییرات فیزیولوژیکی و بیوشیمیایی نهال‏های Eucalyptus camaldulensis در پاسخ به تنش عنصر مس طراحی شد. بذرهای اکالیپتوس در گلدان‏های حاوی سیلت کشت و توسط محلول‏ غذایی به همراه غلظت‏های مس (0، 5، 10 و 20 میلی مولار) به مدت 10 ماه آبیاری و پس از این مدت برگ، ساقه و ریشه‏ها برداشت شدند. مقدار مس با دستگاه ICP-OES تعیین گردید و برخی از معیار­های فیزیولوژیک، مورفولوژیک و بیوشیمیایی مانند قندهای محلول، پرولین، رنگدانه‏ها، پتانسیل آب برگ، محتوای نسبی آب برگ، کاتالاز (CAT)، پراکسیداز (POX)، سوپراکسیددسموتاز (SOD)، تعداد روزنه‏های زیرین و زبرین برگ، وزن اندام‏ها و شاخص انتقال تعیین شدند. غلظت مس در بافت ریشه بیشتر از برگ و ساقه بود و در بافت ساقه نیز غلظت مس کمتر از برگ بود (ریشه >برگ>ساقه). با افزایش غلظت مس محتوای پرولین افزایش یافت ولی محتوای رنگدانه‏ها کاهش پیدا کرد. در بافت‏های برگ و ریشه فعالیت آنزیم‏های آنتی اکسیدانی CAT، POX و SOD تا غلظت mM 10 مس افزایش و پس از آن اندکی کاهش یافت. این نتایج بیانگر آن است که اکالیپتوس دارای سازوکارهای موثر در برابر سمیت مس از جمله تجمع حفاظت کننده­های اسمزی و آنزیم‏های اکسیدانی است. همچنین اکالیپتوس در شرایط تنش مس تا چهار برابر بیشتر از شاهد توان انباشت این فلز را داشت بدون آن که علایم جدی در رشد نشان دهد. بنابراین کاشت آن به عنوان یک گیاه انباشت کننده فلزات سنگین و موثر در کاهش آلاینده‏های محیط زیست توصیه می­شود.

Keywords [فارسی]

  • اکالیپتوس
  • آنزیم­های اکسیداتیو
  • پرولین
  • قندهای محلول
Ambrosini VG, Rosa DJ, Corredor Prado JP, Borghezan M, Bastos de Melo GW, Fonsêca de Sousa Soares CR and Brunetto G, 2015. Reduction of copper phytotoxicity by liming: a study of the root anatomy of young vines (Vitis labrusca L.). Plant Physiology and Biochemistry 96: 270–280.
Assareh MH and Shariat A, 2008. Seedling response of three Eucalyptus species to copper and zinc toxic concentrations. Caspian Journal of Environmental Sciences 6: 97–103.
Ausubel FM, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA and Struhl K (Eds), 1987. Current Protocols in Molecular Biology. John Wiley and Sons, Inc., New York.
Bates IS, Waldern RP and Teare ID, 1973. Rapid determination of free proline for water stress studies. Plant and Soil 39: 205-207.
Beauchamp C and Fridovich I, 1971. Superoxide dismutase: improved assays and an assay applicable to acrylamide gels. Analytical Biochemistry 44: 276–287.
Beers RR and Sizer IW, 1952. A spectrophotometric method for measuring the breakdown of hydrogen peroxide by catalase. Journal of Biological Chemistry 195: 133-140.
Bojarczuk K, 2004. Effect of toxic metals on the development of poplar (Populus tremula L. x P. alba L.) cultured in vitro. Polish Journal of Environmental Studies 13 (2): 115 – 120.
Boyer JS, 1968. Measurement of the water status of plants. Annual Review of Plant Physiology 9: 351-363.
Bradford MM. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry 72: 248–254.
Brunet J, Repellin A, Varrault G, Terrync N and Zuily-Fodil Y, 2008. Lead accumulation in the roots of grass pea (Lathyrus sativus L.): a novel plant for phytoremediation systems? Comptes Rendus Biologies 331: 859–864.
Cambrolle J, García JL, Figueroa ME and Cantos M, 2015. Evaluating wild grapevine tolerance to copper toxicity. Chemosphere 120: 171-178.
Cosio C and Dunand C, 2009. Specific functions of individual class III peroxidase genes. Journal of Experimental Botany 60: 391–408.
Demirevska-Kepova K, Simova-Stoilova L, Stoyanova Z, Hölzer R and Feller U, 2004. Biochemical changes in barley plants after excessive supply of copper and manganese. Environmental and Experimental Botany 52: 253–266.
Dey S, Mazumder PB and Paul SB, 2014. Effect of copper on growth and chlorophyll content in tea plants (Camellia sinensis (L.) O. Kuntze). IMPACT: International Journal of Research in Applied, Natural and Social Sciences 2: 223–230.
Ducic Tand Polle A, 2005. Transport and detoxification of manganese and copper in plants. Brazilian Journal of Plant Physiology 17: 103–112.
El-Khatib AA, Faheed FA and Azooz MM, 2015. Physiological response of Eucalyptus rostorata to heavy metal air pollution. El-Minia Science Bulletin 15: 429–451.
Elobeid M and Polle A, 2010. Response of grey poplar (Populus x canescens) to copper stress. Plant Stress 4 (special issue 1): 82–86.
Fariduddin Q, Yusuf M, Hayat S and Ahmad A, 2009. Effect of 28-homobrassinolide on antioxidant capacity and photosynthesis in Brassica juncea plants exposed to different levels of copper. Environmental and Experimental Botany 66: 418–424.
Fengtao LI, Jianmin QI, Gaoyang Z, Lihui L, Pingping F, Fen TA and Jiantang XU, 2013. Effect of cadmium stress on the growth antioxidative enzymes and lipid peroxidation in two kenaf (Hibiscus cannabinus L.) plant seedlings. Journal of Integrative Agriculture 12: 610–620.
Filippou P, Bouchagier P, Skotti E and Fotopoulos V, 2014. Proline and reactive oxygen/nitrogen species metabolism is involved in the tolerant response of the invasive plant species Ailanthus altissima to drought and salinity. Environmental and Experimental Botany 97: 1–10.
Flores-Cáceres ML, Hattab S, Hattab S, Boussetta H, Banni M and Hernández LE, 2015. Specific mechanisms of tolerance to copper and cadmium are compromised by a limited concentration of glutathione in alfalfa plants. Plant Science 233: 165–173.
Fong JDM, Masunaga T and Sato K, 2015. Control of micronutrients availability in soil and concentration in rice grain through field water management. Journal of Agricultural Science 7: 163–174.
Gharbi F, Rejeb S, Ghorbal MH and Morel JL, 2008. Plant response to copper toxicity as affected by plant species and soil type. Journal of Plant Nutrition 28: 379-382.
Ghosh M and Singh SP, 2005. A review on phytoremediation of heavy metals and utilization of it’s by products. Asian JournalonEnergyandEnvironment 6: 214–231.
Giannopolitis CN and Ries SK, 1977. Superoxide dismutases. Plant Physiology 59: 309–314.
Hames, BD and Richwood M, 1990. Gel Electrophoresis of Proteins, a Practical Approach. Oxford University Press, UK.
Irigoyen JJ, Einerich DW and Sanchez-Diaz M, 1992. Water stress induced changes in concentrations of proline and total soluble sugars in nodulated alfalfa (Medicago sativa) plants. Physiologia Plantarum 84 (1): 58-60.
Janas KM, Zielińska-Tomaszewska J, Rybaczek D, Maszewski J, Posmyk MM, Amarowicz R and Kosińska A, 2010. The impact of copper ions on growth, lipid peroxidation, and phenolic compound accumulation and localization in lentil (Lens culinaris Medic.) seedlings. Journal of Plant Physiology 167: 270–276.
Jenssen A, 1978. Chlorophyll and carotenoids. In: Hellebust JA and Craigie JS (Eds). Handbook of Phycological Methods. Physiological & Biochemical Methods. Pp. 59-65. Cambridge University Press, Cambridge, New York.
Kim YH, Lee HS and Kwak SS, 2010. Differential responses of sweet potato peroxidases to heavy metalsChemosphere 81: 7985.
Lequeux H, Hermans C, Lutts S and Verbruggen N, 2010. Response to copper excess in Arabidopsis thaliana: impact on the root system architecture, hormone distribution, lignin accumulation and mineral profile. Plant Physiology and Biochemistry 48: 673-682.
Liu J, Shi X, Qian M, Zheng L, Lian C, Xia Y and Shen Z, 2015. Copper-induced hydrogen peroxide upregulation of a metallothionein gene, OsMT2c, from Oryza sativa L. confers copper tolerance in Arabidopsis thaliana. Journal of Hazardous Materials 294: 99–108.
Lombardi L and Sebastian L, 2005. Copper toxicity in Prunus cerasifera: growth and antioxidant enzymes responses of in vitro grown plants. Plant Science 168: 797–802.
Lu Z, 1988. The sensitivity of adaxial and abaxial stomatal resistance in wheat leaf to soil water stress. Acta Phytophysiol Sinica 14: 223–227.
Lukatkin A, Egorova I, Michailova I, Malec P and Strzałka K, 2014. Effect of copper on pro- and antioxidative reactions in radish (Raphanus sativus L.) in vitro and in vivo. Journal of Trace Elements in Medicine and Biology 28(1): 80–86.
Malar S, Shivendra-Vikram S, JC-Favas P and Perumal V, 2014. Lead heavy metal toxicity induced changes on growth and antioxidative enzymes level in water hyacinths [Eichhornia crassipes (Mart.)]. Botanical Studies 55: 1–11.
Merlin TPA, Lima GPP, Leonel S and Vianello F, 2012. Peroxidase activity and total phenol content in citrus cuttings treated with different copper sources. South African Journal of Botany 83: 159–164.
Michel BE, 1972. Solute potentials of sucrose solutions. Plant Physiology 50: 196–198.
Moor RH, 1960. Laboratory Guide for Elementary Plant Physiology. Burgess Pub, Minneapolis.
Noctor G and Foyer CH, 1998. Ascorbate and glutathione: keeping active oxygen under control. Annual Review of Plant Physiology and Plant Molecular Biology 49: 249–279.
Passardi F, Cosio C, Penel C and Dunand C, 2005. Peroxidases have more functions than a Swiss army knife. Plant Cell Reports 24: 255–265.
Sharma SS and Dietz KJ, 2008. The relationship between metal toxicity and cellular redox imbalance. Trends in Plant Sciences 14: 43–50.
Sonmez S, Kaplan M, Sonmez NK, Kaya H and Uz I, 2006. High level of copper application to soil and leaves reduce the growth and yield of tomato plants. Scientia Agricola 63(3): 213–218.
Su Y, Han FX, Sridhar BBM and Monts DL, 2005. Phytotoxicity and phytoaccumulation of trivalent and hexavalent chromium in brake fern. Environmental Toxicologyand Chemistry 24: 2019–2026.
Verslues PE, Agrawal M, Katiyar-Agrawal S, Zhu JH and Zhu JK, 2006. Methods and concepts in quantifying resistance to drought, salt and freezing, abiotic stresses that affect plant water status. Plant Journal 45: 523-539.
Wang XQ, Wu WH and Assmann SM, 1998. Differential responses of abaxial and adaxial guard cells of broad bean to abscisic acid and calcium. Plant Physiology 118:1421–1429.
Westerma REL, 1990. Soil Testing and Plant Analysis. SSSA, Madison Wisconsin, USA. 571 pages.
White DA, Turner NC and Galbraith JH, 2000. Leaf water relations and stomatal behavior of four allopatric Eucalyptus species planted in Mediterranean southwestern Australia. Tree Physiology 20: 1157-1165.
Wintermans JFGM and Motes AD, 1965 Spectrophotometric characteristics of chlorophyll a and b and their pheophitin in ethanol. Biochimica et Biophysica Acta 109: 440-452.
Xiong ZT and Wang H, 2005. Copper Toxicity and Bioaccumulation in Chinese cabbage (Brassica pekinensis Rupr.). Environmental Toxicology 20 (2): 188-194.
Zhang C, Zhou W and Zhu D, 2007. Physiological responses induced by copper bioaccumulation in Eichhornia crassipes (Mart.). Hydrobiologica 579: 211–218.