Absorption of zinc ions dissolved from zinc oxide nanoparticles in the tobacco callus improves plant productivity

  • Shizue Yoshihara
  • Kasumi Yamamoto
  • Yoshino Nakajima
  • Satomi Takeda
  • Kensuke Kurahashi
  • Hayato TokumotoEmail author
Original Article


Zinc oxide (ZnO) nanoparticles (NPs) are soluble in water and can release Zn2+, an essential mineral that promotes the growth of plant cells. When ZnO NPs were administered to tobacco (Nicotiana tabacum cv. Samusun-NN) callus under white light irradiation, a concentration-dependent increase in weight was observed. Specifically, an increase in chlorophyll levels was triggered by blue light and induced by ZnO NPs. mRNA-seq analysis showed that during the early stages of tobacco callus exposure to ZnO NPs and white light irradiation, there was considerable fluctuation in the expression of genes related to salt stress. After 24 h, the expression of cellular component and growth-related genes also fluctuated. Analysis by RT-qPCR revealed that, after 1 day of ZnO NPs exposure, the expression levels of photosynthesis-related genes were enhanced. The Zn content of control-treated callus was 0.19 mg g−1 dry weight, whereas that of callus cultured with the ZnO bulk particles (BPs), with a particle diameter of 2000 nm, was 2.59 mg g−1 dry weight, and for callus cultured with ZnO NPs, with a particle diameter 34 nm, the Zn content was 3.37 mg g−1 dry weight. These results indicate that ZnO particles supplied large amounts of Zn to the callus, suggesting that the smaller the particle size, the larger the surface area of particles dissolve zinc ions more efficiently and the more ions are supplied to tobacco callus cells, and resulting in an increase in plant productivity.

Key message

Under the light illumination, incubation of tobacco callus with zinc oxide nanoparticle dispersion resulted in supply of much zinc ions into cell, and induction of chlorophyll accumulation and cell proliferation.


Zn absorption Chlorophyll increase Photosynthesis 



We thank Ms. Junko Muramatsu for her advice on the experimental design. This research was supported by Grants-in-Aid for Scientific Research (No. 18K19882) from the Japanese Society for the Promotion of Science.

Author contributions

SY: Study conception and design, Acquisition of data, Analysis and interpretation of data, Drafting manuscript. KY: Acquisition of data, Analysis and interpretation of data. YN: Acquisition of data, Analysis and interpretation of data. ST: Analysis and interpretation of data. KK: Analysis and interpretation of data. HT: Study conception and design, Analysis and interpretation of data, Drafting manuscript.


This study was funded by Grants-in-Aid for Scientific Research from the Japanese Society for the Promotion of Science (no. 18K19882).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.


  1. Aravind P, Prasad MNV (2004) Zinc protects chloroplasts and associated photochemical functions in cadmium exposed Ceratophyllum demersum L., a freshwater macrophyte. Plant Sci 166:1321–1327CrossRefGoogle Scholar
  2. Aruoja V, Dubourguier HC, Kasemets K, Kahru A (2009) Toxicity of nanoparticles of CuO, ZnO and TiO2 to microalgae Pseudokirchneriella subcapitata. Sci Total Environ 407:1461–1468CrossRefGoogle Scholar
  3. Broadley MR, White PJ, Hammond JP, Zelko I, Lux A (2007) Zinc in plants: Tansley review. New Phytol 173:677–702CrossRefGoogle Scholar
  4. Chen P, Powell BA, Mortimer M, Ke PC (2012) Adaptive interactions between zinc oxide nanoparticles and Chlorella sp. Environ Sci Technol 46:12178–12185CrossRefGoogle Scholar
  5. Dennis G Jr, Sherman BT, Hosack DA, Yang J, Gao W, Lane HC, Lempicki RA (2003) DAVID: database for annnotation, visulalization, and integrated discovery. Genome Biol 4:P3CrossRefGoogle Scholar
  6. Ghosh M, Bandyopadhyay M, Mukherjee A (2010) Genotoxicity of titanium dioxide (TiO2) nanoparticles at two trophic levels: plant and human lymphocytes. Chemosphere 81:1253–1262CrossRefGoogle Scholar
  7. Hosack DA, Dennis G Jr, Sherman BT, Lane HC, Lempicki RA (2003) Identifying biological themes within lists of genes with EASE. Genome Biol 4:R70CrossRefGoogle Scholar
  8. Ismond KP, Dolferus R, Pauw MD, Dennis ES, Good AG (2003) Enhanced low oxygen survival in Arabidopsis through increased metabolic flux in the fermentative pathway. Environ Stress Adapt 132:1292–1302Google Scholar
  9. Javed R, Mohamed A, Yücesan B, Gürel E, Kausar R, Zia M (2017) CuO nanoparticles significantly influence in vitro culture, steviol glycosides, and antioxidant activities of Stevia rebaudiana Bertoni. Plant Cell Tiss Organ Cult 131:611–620CrossRefGoogle Scholar
  10. Jiang W, Mashayekhi H, Xing B (2009) Bacterial toxicity comparison between nano- and micro-scaled oxide particles. Environ Pollut 157:1619–1625CrossRefGoogle Scholar
  11. Jiao Y, Lau OS, Deng XW (2007) Light-regulated transcriptional networks in higher plants. Nat Rev Genet 8:217–230CrossRefGoogle Scholar
  12. Landa P, Prerostova S, Petrova S, Knirsch V, Vankova R, Vanek T (2015) The transcriptomic response of Arabidopsis thaliana to zinc oxide: a comparison of the impact of nanoparticle, bulk, and ionic zinc. Environ Sci Technol 49:14537–14545CrossRefGoogle Scholar
  13. Le VN, Rui Y, Gui X, Li X, Liu S, Han Y (2014) Uptake, transport, distribution and bio-effects of SiO2 nanoparticles in Bt-transgenic cotton. J Nanobiotechnol 12:50CrossRefGoogle Scholar
  14. Lee CW, Mahendra S, Zodrow K, Li D, Tsai YC, Braam J, Alvarez PJJ (2010) Development phytotoxicity of metal oxide nanoparticles to Arabidopsis thaliana. Environ Toxicol Chem 29:669–675CrossRefGoogle Scholar
  15. Lin D, Xing B (2007) Phytotoxicity of nanoparticles: inhibition of seed germination and root growth. Environ Pollut 150:243–250CrossRefGoogle Scholar
  16. Lin D, Xing B (2008) Root uptake and phytotoxicity of ZnO nanoparticles. Environ Sci Technol 42:5580–5585CrossRefGoogle Scholar
  17. Lunn D, Gaddipati SR, Tucker GA, Lycett GW (2013) Null mutants of individual RABA genes impact the proportion of different cell wall components in stem tissue of Arabidopsis thaliana. PLoS ONE 8:e75724CrossRefGoogle Scholar
  18. Ma L, Li J, Qu L, Hager J, Chen Z, Zhao H, Deng XW (2001) Light control of Arabidopsis development entails coordinated regulation of genome expression and cellular pathways. Plant Cell 13:2589–2607CrossRefGoogle Scholar
  19. Mahajan P, Dhoke SK, Khanna AS (2011) Effect of nano-ZnO particle suspension on growth of mung (Vigna radiata) and gram (Cicer arietinum) seedlings using plant agar method. J Nanotechnol. Google Scholar
  20. Maszkowska J, Debski J, Kulik A, Kistowski M, Bucholc M, Lichocka M, Klimecka M, Sztatelman O, SzymanskaKP DM, Dobrowolska G (2018) Phosphoproteomic analysis reveals that dehydrins ERD10 and ERD14 are phosphorylated by SNF1-related protein kinase 2.10 in response to osmotic stress. Plant, Cell Environ 2018:1–16Google Scholar
  21. Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant 15:473–497CrossRefGoogle Scholar
  22. Nakamura H, Muramatsu M, Hakata M, Ueno O, Nagamura Y, Hirochika H, Takano M, Ichikawa H (2009) Ectopic overexpression of the transcription factor OsGLK1 induces chloroplast development in non-green rice cells. Plant Cell Physiol 50:1933–1949CrossRefGoogle Scholar
  23. Nhat PVH, Ngo HH, Guo WS, Chang SW, Nguyen DD, Nguyen PD, Bui XT, Zhang XB, Guo JB (2018) Can algae-based technologies be an affordable green process for biofuel production and wastewater remediation? Bioresour Technol 256:491–501CrossRefGoogle Scholar
  24. Podlesáková K, Ugena L, Spíchal L, Dolezal Diego ND (2019) Phytohormones and polyamines regulate plant stress responses by altering GABA pathway. New Biotechnology 48:53–65CrossRefGoogle Scholar
  25. Porra RJ, Thompson WA, Kriedemann PE (1989) Determination of accurate extinction coefficients and simultaneous equations for assaying chlorophylls a and b extracted with four different solvents: verfication of the concentration of chlorophyll standards by atomic absorption sepctroscopy. Biochim Biophys Acta 975:384–394CrossRefGoogle Scholar
  26. Salama ES, Hwang JH, El-Dalatony MM, Kurade MB, Kabra AN, Abou-Shanab RAI, Kim KH, Yang IS, Govindwar SP, Kim S, Jeon BH (2018) Enhancement of microalgal growth and biocomponent-based transformations for improved biofuel recovery: a review. Bioresour Technol 258:365–375CrossRefGoogle Scholar
  27. Shi H, Liu W, Yao Y, Wei Y, Chan Z (2017) Alcohol dehydrogenase 1 (ADH1) confers both abiotic and biotic stress resistance in Arabidopsis. Plant Sci 262:24–31CrossRefGoogle Scholar
  28. Singh NB, Amist N, Yadav K, Singh D, Pandey JK, Singh SC (2013) Zinc oxide nanoparticles as fertilizer for the germination, growth and metabolism of vegetable crops. J Nanoeng Nanomanuf 3:353–364CrossRefGoogle Scholar
  29. Sivakumar G, Xu J, Thompson RW, Yang Y, Randol-Smith P, Weathers PJ (2012) Integrated green algal technology for bioremediation and biofuel. Bioresour Technol 107:1–9CrossRefGoogle Scholar
  30. Trapnell C, Pachter L, Salzberg SL (2009) TopHat: discovering splice junctions with RNA-seq. Bioinformatics 25:1105–1111CrossRefGoogle Scholar
  31. Trapnell C, Williams BA, Pertea G, Mortazavi A, Kwan G, van Baren MJ, Salzberg SL, Wold BJ, Pachter L (2010) Transcript assembly and quantification by RNA-Seq reveals unannotated transcripts and isoform switching during cell differentiation. Nat Biotechnol 28:511–515CrossRefGoogle Scholar
  32. Usami T, Mochizuki N, Kondo M, Nishimura M, Nagatani A (2004) Cryptochromes and phytochromes synergistically regulate Arabidopsis root greening under blue light. Plant Cell Physiol 45:1798–1808CrossRefGoogle Scholar
  33. Waters MT, Wang P, Korkaric M, Capper RG, Saunders NJ, Langdale JA (2009) GLK transcription factors coordinate expression of the photosynthetic apparatus in Arabidopsis. Plant Cell 21:1109–1128CrossRefGoogle Scholar
  34. Wilson RS, Swatek KN, Thelen JJ (2016) Regulation of the regulators: post-translational modifications, subcellular, and spatiotemporal distribution of plant 14-3-3 proteins. Front Plant Sci 7:611CrossRefGoogle Scholar
  35. Yang CY, Hsu FC, Li JP, Wang NN, Shih MC (2011) The AP2/ERF transcription factor AtERF73/HRE1 modulates ethylene responses during hypoxia in Arabidopsis. Plant Physiol 156:202–212CrossRefGoogle Scholar
  36. Yang Z, Chen J, Dou R, Gao X, Mao C, Wang L (2015) Assessment of the phytotoxicity of metal oxide nanoparticles on two crop plants, maize (Zea mays L.) and rice (Oryza sativa L.). Int J Environ Res Public Health 12:15100–15109CrossRefGoogle Scholar
  37. Zhai J, Tao X, Pu Y, Zeng XF, Chen JF (2010) Core/shell structured ZnO/SiO2 nanoparticles: preparation, characterization and photocatalytic property. Appl Surf Sci 257:393–397CrossRefGoogle Scholar

Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  • Shizue Yoshihara
    • 1
  • Kasumi Yamamoto
    • 2
  • Yoshino Nakajima
    • 2
  • Satomi Takeda
    • 1
  • Kensuke Kurahashi
    • 3
  • Hayato Tokumoto
    • 1
    Email author
  1. 1.Department of Biological ScienceOsaka Prefecture UniversitySakaiJapan
  2. 2.Department of Chemical EngineeringOsaka Prefecture UniversitySakaiJapan
  3. 3.Osaka Prefecture University College of TechnologyNeyagawaJapan

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