Green Synthesis of Chitosan Capped-Copper Nano Biocomposites: Synthesis, Characterization, and Biological Activity against Plant Pathogens

Abstract

Metallic nanoparticles have been reported to have wide spread applications in the field of agricultural with a potential to enhance the activity natural substances to replace hazardous chemical fungicides and pesticides. Higher size to volume ratio, greater surface area, optimum efficacy, and immense precision are some of the benefits of employing nanotechnology in agricultural formulations. In this study, copper nano-biocomposites have been successfully synthesized using chitosan, a naturally occurring polymer and an organically available ascorbic acid. We report the appearance of the characteristic brick-brown color in aqueous solution that indicates the formation of copper nanoparticles. These particles were subjected to various physico-chemical characterizations using UV-Vis spectrophotometer, DLS, Zeta Potential, TEM, TGA, and FTIR to understand the morphology, chemical, and physical profiling. It was inferred that ascorbic acid acted as reducing agent while chitosan as capping agent to stabilize the colloidal copper nanoparticles of 20–120 nm size. In aqueous solution, the particles were observed to be monodispersed with minimum aggregation. The synthesized nanoparticles showed significant antifungal and antibacterial activity against common notorious agricultural plant pathogens, viz., Fusarium sp., Aspergillus sp., Alternaria alternata, Pythium sp., and Bacillus cereus, assayed using poisoned food method. The antimicrobial activity was showcased at a minimum concentration of 0.1–0.7% for different organisms. Furthermore, filed trial studies were performed on ginger (Zingiber officinale) plantations for a period of 8 weeks which substantiated with our lab results. These studies validated our claim of antifungal activity and simultaneously furnished us with toxicity results. The use of eco-friendly, nontoxic, biodegradable, and environmentally safe raw materials and the resultant nanoparticle solution ensure that the finished product does not induce any threat to the environment or human and shows maximum efficacy at minimum concentration.

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Abbreviations

°C:

Degree celsius

CCNPs:

Chitosan-copper nanoparticles

CS:

Chitosan

Cu-NPs:

Copper nanoparticles

Cu2O:

Cuprous oxide

CuO:

Copper oxide

DLS:

Dynamic light scattering

EDS:

Energy-dispersive X-ray spectroscopy

FTIR:

Fourier-transform infrared spectroscopy

hr./h:

Hour/hours

KBr:

Potassium bromide

lit:

Liter

M:

Molar

ml:

Milliliter

mV:

Milli-volt

nm:

Nanometer

SPR:

Surface plasmon resonance

TEM:

Transmission electron microscopy

TGA:

Thermogravimetric analysis

v/v:

Volume/volume

w/v :

Weight/volume

μl:

Micro-liter

References

  1. 1.

    Laura Orzali, Beatrice Corsi, Cinzia Forni, and Luca Riccioni., (2016). Chitosan in agriculture: a new challenge for managing plant disease (ed). Chapter 2 – Biological activities and application of Marine Polysaccharides, Intech.

  2. 2.

    Ramesh Chand Kasana, Nav Raten Panwar, Ramesh Kumar Kaul, and Praveen Kumar., 2016. Copper nanoparticles in agriculture_biological synthesis and antimicrobial activity (ed). Nanoscience in Food and Agriculture 3, Sustainable Agriculture Reviews 23.

  3. 3.

    Malerba, M., & Cerana, R. (2018). Recent advances of chitosan applications in plants. Polymers, 10, 118.

    Article  Google Scholar 

  4. 4.

    D. P. Chattopadhyay and Milind S. Inamdar., 2010. Aqueous behaviour of chitosan. Hindawi Publishing Corporation International Journal of Polymer Science Volume 2010, Article ID 939536, 7 pages.

  5. 5.

    Adamares M. Silva, Thayza C. M. Stamford, Patrícia M. Souza, Lucia R.R. Berger, Marcela V. Leite, Aline E. Nascimento and Galba M. Campos-Takaki., 2015. Antifungal Activity of Microbiological Chitosan and Coating Treatment on Cherry Tomato (Solanum lycopersicum var. cerasiforme) to Post-Harvest Protection. ISSN: 2319–7706 Volume 4 Number 9, pp. 228–240.

  6. 6.

    Bellich, B., D’Agostino, I., Semeraro, S., Gamini, A., & Cesàro, A. (2016). “The Good, the Bad and the Ugly” of Chitosans. Mar. Drugs, 14, 99.

    Article  Google Scholar 

  7. 7.

    Zain, N. M., Stapley, A. G. F., & Shama, G. (2014). Green synthesis of silver and copper nanoparticles using ascorbic acid and chitosan for antimicrobial applications. Carbohydrate Polymers, 112, 195–202.

    Article  Google Scholar 

  8. 8.

    Lamichhane, J. R., Osdaghi, E., Behlau, F., Köhl, J., Jones, J. B., & Aubertot, J.-N. (2018). Thirteen decades of antimicrobial copper compounds applied in agriculture. A review. Copper Nanoparticles Mediated by Chitosan: Synthesis and Characterization via Chemical Methods Agronomy for Sustainable Development, 38, 28.

    Google Scholar 

  9. 9.

    Usman, M. S., El Zowalaty, M. E., Shameli, K., Zainuddin, N., Salama, M., & Ibrahim, N. A. (2013). Synthesis, characterization, and antimicrobial properties of copper nanoparticles. International Journal of Nanomedicine, 2013(8), 4467–4479.

    Google Scholar 

  10. 10.

    Manikandan and Sathiyabama. (2015). Green Synthesis of Copper-Chitosan Nanoparticles and Study of its Antibacterial Activity. J Nanomed Nanotechnol, 6, 1.

    Google Scholar 

  11. 11.

    Balouiri, M., Sadiki, M., & Ibnsouda, S. K. (2016). Methods for in vitro evaluating antimicrobial activity: A review. Journal of Pharmaceutical Analysis, 6, 71–79.

    Article  Google Scholar 

  12. 12.

    Stevenson, K., McVey, A. F., Clark, I. B. N., Swain, P. S., & Pilizota, T. (2016). General calibration of microbial growth in microplate readers. Sci Rep, 6, 38828. https://doi.org/10.1038/srep38828.

    Article  Google Scholar 

  13. 13.

    Szymańska, E., & Winnicka, K. (2015). Stability of Chitosan—A Challenge for Pharmaceutical and Biomedical Applications. Mar. Drugs, 13, 1819–1846.

    Article  Google Scholar 

  14. 14.

    Gianfranco Romanazzi, Franka Mlikota Gabler, Dennis Margosan, Bruce E. Mackey, and Joseph L. Smilanick., 2009. Effect of chitosan dissolved in different acids on its ability to control postharvest gray mold of table grape. The American Phytopathological Society. Vol. 99, No. 9.

  15. 15.

    Tamilselvan Abiraman, and Sengottuvelan Balasubramanian., 2017. Synthesis and characterization of large scale, (< 2 nm) chitosan decorated copper nanoparticles and their application in anti-fouling coating. Ind. Eng. Chem. Res.

  16. 16.

    Qing-Ming, L., Yasunami, T., Kuruda, K., & Okido, M. (2012). Preparation of Cu nanoparticles with ascorbic acid by aqueous solution reduction method. Trans. Nonferrous Met. Soc. China, 22(2012), 2198–2203.

    Google Scholar 

  17. 17.

    Hina Khalid, S., & Shamaila, N. (2015). Zafar., 2015. Synthesis of copper nanoparticles by chemical reduction method. Sci.Int.(Lahore), 27(4), 3085–3088.

    Google Scholar 

  18. 18.

    Asim Umer, Shahid Naveed, Naveed Ramzan, Muhammad Shahid Rafique, Muhammad Imran., 2014. A green method for the synthesis of Copper Nanoparticles using L ascorbic acid. ISSN 1517–7076 artigo 11547, pp.197–203, 2014.

  19. 19.

    Khlebtsov, B. N., & Khlebtsov, N. G. (2011). On the Measurement of Gold Nanoparticle Sizes by the Dynamic Light Scattering Method. Colloid Journal, 73, 118–127.

    Article  Google Scholar 

  20. 20.

    Emilia Tomaszewska, Katarzyna Soliwoda, Kinga Kadziola, Beata Tkacz-Szczesna, Grzegorz Celichowski, Michal Cichomski, Witold Szmaja and Jaroslaw. Detection Limits of DLS and UV-Vis Spectroscopy in Characterization of Polydisperse Nanoparticles Colloids. Hindawi Publishing Corporation Journal of Nanomaterials Volume 2013, Article ID 313081, 10 pages. https://doi.org/10.1155/2013/313081.

  21. 21.

    Katarzyna Tokarek, Jose L Hueso, Piotr Kus’trowski, Grazyna Stochel, and Agnieszka Kyziol., 2013. Green Synthesis of Chitosan-Stabilized Copper Nanoparticles. Eur. J. Inorg. Chem..

  22. 22.

    Umer, A., Naveed, S., Ramzan, N., Rafique, M. S., & Imram, M. (2014). A green method for the synthesis of copper nanoparticles using L-ascorbic. Acidrevista Matéria, 19(3), 197–203.

    Google Scholar 

  23. 23.

    P. Narayanasamy., 2011. Detection of Fungal Pathogens in Plants (ed). Microbial Plant Pathogens-Detection and Disease Diagnosis: Fungal Pathogens, Vol. 1.

  24. 24.

    Cristian Covarrubias, Diego Trepiana And Camila Corral. (2018). Synthesis Of Hybrid Copper-Chitosan Nanoparticles With Antibacterial Activity. Dental Materials Journal, 37(3), 379–384.

    Article  Google Scholar 

  25. 25.

    Susanta Banik and Alejandro Pérez-de-Luque., 2017. In vitro effects of copper nanoparticles on plant pathogens, beneficial microbes and crop plants. Spanish Journal of Agricultural Research 15(2), e1005, 15 pages (2017) eISSN: 2171–9292.

  26. 26.

    Pham Van Viet, Hai Thi Nguyen, ThiMinh Cao and Le Van Hieu., 2016. Fusarium Antifungal Activities of Copper Nanoparticles Synthesized by a Chemical Reduction Method. Hindawi Publishing Corporation Journal of Nanomaterials Volume, Article ID 1957612, 7 pages.

  27. 27.

    Franklin Laemmlen, Alternaria Disease: University of California, Agriculture and Natural Resource. Publication 8040. ISBN 978–1–60107-218-4.

  28. 28.

    Meenu, G., & Manisha, K. (2017). 2017. Diseases infecting ginger (Zingiber officinale Roscoe). A review. Agricultural Reviews, 38(1), 15–28.

    Google Scholar 

  29. 29.

    Ling Yien Ing, Noraziah Mohamad Zin, Atif Sarwar, and Haliza Katas., 2012. Antifungal Activity of Chitosan Nanoparticles and Correlation with Their Physical Properties. Hindawi Publishing Corporation. International Journal of Biomaterials. Article ID 632698, 9 pages.

  30. 30.

    Rai, M., Ingle, A. P., Paralikar, P., Anasane, N., Gade, R., & Ingle, P. Effective management of soft rot of ginger caused by Pythium spp. and Fusarium spp.: emerging role of nanotechnology. Applied Microbiology and Biotechnology. https://doi.org/10.1007/s00253-018-9145-8.

Download references

Acknowledgments

The authors are thank the Agharkar Research Institute, Pune, India, CSIR-National Chemical Laboratory, Pune, India for providing facility to perform some of the characterization of nanoparticles samples. The authors are thankful to the management team of Netsurf communication Pvt. Ltd. Pune.

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Correspondence to Amol U. Hivrale.

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Mehta, M.R., Mahajan, H.P. & Hivrale, A.U. Green Synthesis of Chitosan Capped-Copper Nano Biocomposites: Synthesis, Characterization, and Biological Activity against Plant Pathogens. BioNanoSci. (2021). https://doi.org/10.1007/s12668-021-00823-8

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Keywords

  • Chitosan
  • Copper
  • Nanoparticles
  • Ch-cu nanoparticles
  • Biopesticides
  • Biocontrol