Document Type : Original Paper


1 1 Department of Plant, Cell and Molecular Biology, Faculty of Natural Sciences, University of Tabriz, Tabriz, Islamic Republic of Iran

2 2 Department of Inorganic Chemistry, Faculty of Chemistry, University of Tabriz, Tabriz, Islamic Republic of Iran


Copper (Cu) is an essential micronutrient for higher plants and is required for cell redox homeostasis, free radical scavenging, function of electron transport chains and cell wall lignification. Copper deficiency is a widespread nutritional disorder in plants and its adequate supply is necessary for an optimum crop production. In order to evaluate the efficacy of nano tetraammine copper (II) sulfate complex ([Cu(NH3)4]SO4) (NCu) in the meeting of plants Cu requirement, Cu-sufficient (+Cu) and Cu-deficient (–Cu) tobacco (Ncotiana rustica L.) plants were treated with 0.5 µM NCu complex through leaves. The shoot and root biomass and photosynthesis of –Cu plants were restored by foliar application of NCu complex, while in the +Cu plants the shoot biomass was repressed likely due to a supra optimal Cu level. Foliar application of NCu complex restored almost completely the activity of Cu-containing enzymes, superoxide dismutase, polyphenol oxidase and diamine oxidase. Iron (Fe) homeostasis was also significantly influenced by both Cu starvation and foliar application as could be confirmed by Fe concentration data and activity of Fe-enzymes, peroxidase and polyamine oxidase. The activity of phenylalanine ammonia lyase and the levels of phenolics and lignin were markedly decreased in the –Cu plants. These parameters, however, were completely restored or even exceeded that of the +Cu plants upon foliar application. Our results suggest that, foliar application of NCu is a feasible method for a rapid and efficient compensation of Cu deficiency symptoms due to a high penetration ability and a sufficient retranslocation of applied Cu in the phloem.


  1. Broadley M, Brown P, Cakmak I, Rengel Z, Zhao F. Function of nutrients: Micronutrients. In: Marschner P (ed.) Marschner’s mineral nutrition of higher plants, Academic Press, London. 2012:71-84.
  2. Mayer 2006 Mayer AM. Polyphenol oxidases in plants and fungi: going places? A review. Phytochemistry. 2006;67:2318–2331.
  3. Kopsell DE, Kopsell DA. Copper. In: Barker AV, Pilbeam DJ (eds.) Handbook of plant nutrition. CRC Press, Taylor & Francis Group, USA. 2007;293-328.
  4. Burkhead JL, Gogolin Reynolds KA, Abdel‐Ghany SE, Cohu CM, Pilon M. Copper homeostasis. New Phytol. 2009;182:799-816.
  5. Karamanos RE, Pomarenski Q, Gog TB, Flore NA. The effect of foliar copper application on grain yield and quality of wheat. Can J Plant Sci. 2004;840:47-56.
  6. Fouad A, Saad D, Kacem M, Abdelwahed M, Khalid D, Abderrahim R, Abdelhadi AH. Efficacy of copper foliar spray in preventing copper deficiency of rainfed wheat (Triticum aestivum L.) grown in a calcareous soil. J Plant Nutr. 2020;43:1617-1626.
  7. Drissi S, Houssa AA, Amlal F, Dhassi K, Lamghari M, Maataoui A. Barley responses to copper foliar spray concentrations when grown in a calcareous soil. J Plant Nutr. 2018;41:2266-2272.
  8. Ma J, Zhang M, Liu Z, Chen H, Li YC, Sun Y, Ma Q, Zhao C. Effects of foliar application of the mixture of copper and chelated iron on the yield, quality, photosynthesis, and microelement concentration of table grape (Vitis vinifera L.). Sci Hort. 2019;254:106-115.
  9. Eichert T, Fernandez V. Uptake and release of elements by leaves and other aerial plant. Evidence from measurement of diffusion potentials. Plant Physiol. 2012;92:103-109.
  10. Monreal CM, De Rosa M, Mallubhotla SC, Bindraban PS, Dimkpa C. The application of nanotechnology for micronutrients in soil-plant systems. VFRC (Virtual Fertilizer Research Center) Report, Washington, DC, USA. 2015;53.
  11. Mittal D, Kaur G, Singh P, Yadav K, Ali SA. Nanoparticle-based sustainable agriculture and food science: Recent advances and future outlook. Front Nanotechnol. 2020;2:10.
  12. López-Vargas ER, Ortega-Ortíz H, Cadenas-Pliego G, de Alba Romenus K, Cabrera de la Fuente M, Benavides-Mendoza A, Juárez-Maldonado A. Foliar application of copper nanoparticles increases the fruit quality and the content of bioactive compounds in tomatoes. Appl Sci. 2018;8:1020.
  13. Nair PM, Chung IM. Impact of copper oxide nanoparticles exposure on Arabidopsis thaliana growth, root system development, root lignificaion, and molecular level changes. Environ Sci Poll Res. 2014;21:12709-12722.
  14. Da Costa MV, Sharma PK. Effect of copper oxide nanoparticles on growth, morphology, photosynthesis, and antioxidant response in Oryza sativa. Photosynthetica 2016;54:110-119.
  15. Lee WM, An YJ, Yoon H, Kweon HS. Toxicity and bioavailability of copper nanoparticles to the terrestrial plants mung bean (Phaseolus radiatus) and wheat (Triticum aestivum): plant agar test for water‐insoluble nanoparticles. Environ Toxicol Chem. 2008;27:1915-1921.
  16. FAO (Food and Agricultural Organization of the United Nations). 2020; Available from:
  17. Bastani S, Hajiboland R, Khatamian M, Saket-Oskoui M. Nano iron (Fe) complex is an effective source of Fe for tobacco plants grown under low Fe supply. J Soil Sci Plant Nutr. 2018;18:524-541.
  18. Bahrami-Rad S, Hajiboland R. Effect of potassium application in drought-stressed tobacco (Nicotiana rustica L.) plants: Comparison of root with foliar application. Ann Agric Sci. 2017;62:121-130.
  19. Nakamoto K. Infrared and raman spectra of inorganic and coordination compounds: Part B: Applications in coordination, organometallic, and bioinorganic chemistry, 6th ed. John Wiley & Sons, Inc, USA. 2009.
  20. Pavia DL, Gary M, Lampman GM, Kriz GS, Vyvyan JA. Introduction to spectroscopy, 5th ed. Cengage Learning, USA. 2009.
  21. Giannopolitis CN, Ries SK. Superoxide dismutases: I. Occurrence in higher plants. Plant Physiol. 1977;59:309-314.
  22. Kampatsikas I, Bijelic A, Rompel A. Biochemical and structural characterization of tomato polyphenol oxidases provide novel insights into their substrate specificity. Sci Rep. 2019;9:1-3.
  23. Federico R, Angelini R, Cesta A, Pini C. Determination of diamine oxidase in lentil seedlings by enzymic activity and immunoreactivity. Plant Physiol. 1985;79:62-64.
  24. Asthir B, Duffus CM, Smith RC, Spoor W. Diamine oxidase is involved in H2O2 production in the chalazal cells during barley grain filling. J Exp Bot. 2002;53:677-682.
  25. Hajiboland R, Bastani S, Bahrami-Rad S. Photosynthesis, nitrogen metabolism and antioxidant defense system in B-deficient tea (Camellia sinensis (L.) O. Kuntze) plants. J Sci I R Iran. 2011;22:311-320.
  26. Hajiboland R, Farhanghi F. Remobilization of boron, photosynthesis, phenolic metabolism and anti-oxidant defense capacity in boron-deficient turnip (Brassica rapa L.) plants. Soil Sci Plant Nutr. 2010;56:427-437.
  27. Swain T, Hillis WE. The phenolic constituents of Prunus domestica. I. The quantitative analysis of phenolic constituents. J Sci Food Agric. 1959;10:63-68.
  28. Brinkmann K, Blaschke L, Polle A. Comparison of different methods for lignin determination as a basis for calibration of near-infrared reflectance spectroscopy and implications of lignoproteins. J Chem Ecol. 2002;28:2483-2501.
  29. Morrison IM. A semi‐micro method for the determination of lignin and its use in predicting the digestibility of forage crops. J Sci Food Agric. 1972;23:455-463.
  30. Fernández V, Eichert T. Uptake of hydrophilic solutes through plant leaves: current state of knowledge and perspectives of foliar fertilization. Crit Rev Plant Sci. 2009;28:36-68.
  31. Hill J, Robson AD, Loneragan JF. The effects of copper supply and shading on retranslocation of copper from mature wheat leaves. Ann Bot. 1979;43:449-457.
  32. Hajiboland R, Niknam V, Ebrahimzadeh H. Mozafari A. Uptake, transport and chelation of Cu and Zn at toxic levels in tolerant and sensitive species from North West of Iran. J Sci IR Iran. 2006;17:203-214.
  33. Li SZ, Zhu XK, Wu LH, Luo YM. Zinc, iron, and copper isotopic fractionation in Elsholtzia splendens Nakai: a study of elemental uptake and (re) translocation mechanisms. J Asian Earth Sci. 2020;192:104227.
  34. Garnett TP, Graham RD. Distribution and remobilization of iron and copper in wheat. Ann Bot. 2005;95:817-826.
  35. Bernal M, Casero D, Singh V, Wilson GT, Grande A, Yang H, Dodani SC, Pellegrini M, Huijser P, Connolly EL, Merchant SS. Transcriptome sequencing identifies SPL7-regulated copper acquisition genes FRO4/FRO5 and the copper dependence of iron homeostasis in Arabidopsis. The Plant Cell 2012;24:738-761.
  36. Bernal MI, Krämer U. Involvement of Arabidopsis multi-copper oxidase-encoding LACCASE12 in root-to-shoot iron partitioning: a novel example of copper-iron crosstalk. Front Plant Sci. 2021;11:1998.
  37. Hoopes JT, Dean JFD. Ferroxidase activity in a laccase-like multicopper oxidase from Liriodendron tulipifera. Plant Physiol Biochem. 2004;42:27-33.
  38. Abdel-Ghany SE, Pilon M. MicroRNA-mediated systemic down-regulation of copper protein expression in response to low copper availability in Arabidopsis. J Biol Chem. 2008;283:15932-15945.
  39. Delhaize E, Loneragan JF, Webb J. Development of three copper metalloenzymes in clover leaves. Plant Physiol. 1985;78:4-7.


  1. Schuetz M, Smith R, Ellis B. Xylem tissue specification, patterning, and differentiation mechanisms. J Exp Bot. 2013;64:11-31.