Effect of biologically synthesized silver nanoparticles on Melissa officinalis L.: Evaluation of growth parameters, secondary metabolites and antioxidant enzymes

Document Type : Original Article


Department of Biology, Faculty of Basic Sciences, University of Zabol, Zabol, Iran



Using bionanoparticles plays an important role in increasing agricultural productivity. In recent years, the use of nanoparticles in plants has been considered as pesticides, protective agents and nutrients. The present study investigates the effects of different concentrations of bio-synthesized silver nanoparticles (AgNPs) on growth indices, secondary metabolites, proline, carbohydrates, and antioxidant enzymes activity of Melissa Officnalis L. The Plantlets were treated with different concentrations of AgNPs (0, 20, 60 and 100 ppm) at eight leaf stages. Seedlings performances in terms of growth, antioxidant defence and secondary metabolites content were studied under three different concentrations (20, 60, and 100 ppm) at different days of AgNPs showed growth promotory effect on M. officinalis. The maximum growth rate and photosynthesis pigments content were observed at 60 ppm AgNPs concentration on day 15. Proline and carbohydrate contents increased significantly compared to the control by all concentrations of AgNPs which exhibited time-dependent response. The AgNPs also enhanced secondary metabolites content in M. officinalis seedlings. The highest amount of rosmarinic acid (about 50 mg/g DW) was obtained for those plants treated with 60 and 100 ppm of AgNPs on day 15 which was about 3 fold higher than control. Up-regulation of antioxidant enzymes was observed with AgNPs which led to decrease in MDA content. Our findings confirmed for the first time that biologically synthesized AgNPs at specific levels has significant growth promotory effect as well as increased production of valuable secondary metabolites.


Abeles, F. B. and C. L. Biles, 1991. Characterization of peroxidases in lignifying peach fruit endocarp. Plant physiology. 95, (1) 269-273.
Anusuya, S. and K. N. Banu, 2016. Silver-chitosan nanoparticles induced biochemical variations of chickpea (Cicer arietinum L.). Biocatalysis and Agricultural Biotechnology. 8, 39-44.
Baskar, V., J. Venkatesh and S. W. Park, 2015. Impact of biologically synthesized silver nanoparticles on the growth and physiological responses in Brassica rapa ssp. pekinensis. Environmental Science and Pollution Research. 22, (22) 17672-17682.
Bates, L. S., R. P. Waldren and I. Teare, 1973. Rapid determination of free proline for water-stress studies. Plant and soil. 39, (1) 205-207.
 Bradford, M. M., 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical biochemistry. 72, (1-2) 248-254.
Chang, C.-C., M.-H. Yang, H.-M. Wen and J.-C. Chern, 2002. Estimation of total flavonoid content in propolis by two complementary colorimetric methods. Journal of food and drug analysis. 10, (3).
Cock, J., S. Yoshida and D. A. Forno. 1976. Laboratory manual for physiological studies of rice. Int. Rice Res. Inst.
Ghorbanpour, M., 2015. Major essential oil constituents, total phenolics and flavonoids content and antioxidant activity of Salvia officinalis plant in response to nano-titanium dioxide. Indian Journal of Plant Physiology. 20, (3) 249-256.
Ghorbanpour, M. and J. Hadian, 2015. Multi-walled carbon nanotubes stimulate callus induction, secondary metabolites biosynthesis and antioxidant capacity in medicinal plant Satureja khuzestanica grown in vitro. Carbon. 94, 749-759.
Ghorbanpour, M. and M. Hatami, 2015. Changes in growth, antioxidant defense system and major essential oils constituents of Pelargonium graveolens plant exposed to nano-scale silver and thidiazuron. Indian Journal of Plant Physiology. 20, (2) 116-123.
Giraldo, J. P., M. P. Landry, S. M. Faltermeier, T. P. Mcnicholas, N. M. Iverson, A. A. Boghossian, N. F. Reuel, A. J. Hilmer, F. Sen and J. A. Brew, 2014. Plant nanobionics approach to augment photosynthesis and biochemical sensing. Nature materials. 13, (4) 400-408.
Gruyer, N., M. Dorais, C. Bastien, N. Dassylva and G. 2013. Triffault-Bouchet. Interaction between silver nanoparticles and plant growth. Proc. International Symposium on New Technologies for Environment Control, Energy-Saving and Crop Production in Greenhouse and Plant 1037, 2013:795-800.
Gurunathan, S., K.-J. Lee, K. Kalishwaralal, S. Sheikpranbabu, R. Vaidyanathan and S. H. Eom, 2009. Antiangiogenic properties of silver nanoparticles. Biomaterials. 30, (31) 6341-6350.
Hatami, M. and M. Ghorbanpour, 2014. Defense enzyme activities and biochemical variations of Pelargonium zonale in response to nanosilver application and dark storage. Turkish Journal of Biology. 38, (1) 130-139.
Hatier, J. and K. S. Gould, 2008. Foliar anthocyanins as modulators of stress signals. Journal of Theoretical Biology. 253, (3) 625-627.
Homaee, M. B. and A. A. Ehsanpour, 2015. Physiological and biochemical responses of potato (Solanum tuberosum) to silver nanoparticles and silver nitrate treatments under in vitro conditions. Indian Journal of Plant Physiology. 20, (4) 353-359.
Jiang, H. S., M. Li, F. Y. Chang, W. Li and L. Y. Yin, 2012. Physiological analysis of silver nanoparticles and AgNO3 toxicity to Spirodela polyrhiza. Environmental Toxicology and Chemistry. 31, (8) 1880-1886.
Kamdem, J. P., A. Adeniran, A. A. Boligon, C. V. Klimaczewski, O. O. Elekofehinti, W. Hassan, M. Ibrahim, E. P. Waczuk, D. F. Meinerz and M. L. Athayde, 2013. Antioxidant activity, genotoxicity and cytotoxicity evaluation of lemon balm (Melissa officinalis L.) ethanolic extract: Its potential role in neuroprotection. Industrial Crops and Products. 51, 26-34.
Karimi, J. and S. Mohsenzadeh, 2017. Physiological effects of silver nanoparticles and silver nitrate toxicity in Triticum aestivum. Iranian Journal of Science and Technology, Transactions A: Science. 41, (1) 111-120.
Kowalska, I., L. Pecio, L. Ciesla, W. Oleszek and A. Stochmal, 2014. Isolation, chemical characterization, and free radical scavenging activity of phenolics from Triticum aestivum L. aerial parts. Journal of agricultural and food chemistry. 62, (46) 11200-11208.
Krishnaraj, C., E. Jagan, R. Ramachandran, S. Abirami, N. Mohan and P. Kalaichelvan, 2012. Effect of biologically synthesized silver nanoparticles on Bacopa monnieri (Linn.) Wettst. plant growth metabolism. Process Biochemistry. 47, (4) 651-658.
Lei, Z., S. Mingyu, W. Xiao, L. Chao, Q. Chunxiang, C. Liang, H. Hao, L. Xiaoqing and H. Fashui, 2008. Antioxidant stress is promoted by nano-anatase in spinach chloroplasts under UV-B radiation. Biological Trace Element Research. 121, (1) 69-79.
Ma, X., J. Geiser-Lee, Y. Deng and A. Kolmakov, 2010. Interactions between engineered nanoparticles (ENPs) and plants: phytotoxicity, uptake and accumulation. Science of the total environment. 408, (16) 3053-3061.
Michalak, A., 2006. Phenolic compounds and their antioxidant activity in plants growing under heavy metal stress. Polish Journal of Environmental Studies. 15, (4).
Miller, G., N. Suzuki, S. Ciftci‐Yilmaz and R. Mittler, 2010. Reactive oxygen species homeostasis and signalling during drought and salinity stresses. Plant, cell & environment. 33, (4) 453-467.
Movafeghi, A., A. Khataee, M. Abedi, R. Tarrahi, M. Dadpour and F. Vafaei, 2018. Effects of TiO2 nanoparticles on the aquatic plant Spirodela polyrrhiza: evaluation of growth parameters, pigment contents and antioxidant enzyme activities. Journal of Environmental Sciences. 64, 130-138.
Mulvaney, P., 1996. Surface plasmon spectroscopy of nanosized metal particles. Langmuir. 12, (3) 788-800.
Nair, P. M. G. and I. M. Chung, 2014. Physiological and molecular level effects of silver nanoparticles exposure in rice (Oryza sativa L.) seedlings. Chemosphere. 112, 105-113.
Nicolai, M., P. Pereira, R. F. Vitor, C. P. Reis, A. Roberto and P. Rijo, 2016. Antioxidant activity and rosmarinic acid content of ultrasound-assisted ethanolic extracts of medicinal plants. Measurement. 89, 328-332.
Nowack, B., H. F. Krug and M. Height. 2011. 120 years of nanosilver history: implications for policy makers. ACS Publications.
Pandey, A. C., S. S. Sanjay and R. S. Yadav, 2010. Application of ZnO nanoparticles in influencing the growth rate of Cicer arietinum. Journal of Experimental nanoscience. 5, (6) 488-497.
Pereira, R. P., A. A. Boligon, A. S. Appel, R. Fachinetto, C. S. Ceron, J. E. Tanus-Santos, M. L. Athayde and J. B. T. Rocha, 2014. Chemical composition, antioxidant and anticholinesterase activity of Melissa officinalis. Industrial Crops and Products. 53, 34-45.
Rani, P. U., J. Yasur, K. S. Loke and D. Dutta, 2016. Effect of synthetic and biosynthesized silver nanoparticles on growth, physiology and oxidative stress of water hyacinth: Eichhornia crassipes (Mart) Solms. Acta physiologiae plantarum. 38, (2) 58.
Salama, H. M., 2012. Effects of silver nanoparticles in some crop plants, common bean (Phaseolus vulgaris L.) and corn (Zea mays L.). Int Res J Biotechnol. 3, (10) 190-197.
Salari, S., S. Esmaeilzadeh Bahabadi, A. Samzadeh-Kermani and F. Yosefzaei, 2019. In-vitro Evaluation of Antioxidant and Antibacterial Potential of Green Synthesized Silver Nanoparticles Using Prosopis farcta Fruit Extract. Iranian journal of pharmaceutical research: IJPR. 18, (1) 430.
Sankar, B., C. A. Jaleel, P. Manivannan, A. Kishorekumar, R. Somasundaram and R. Panneerselvam, 2007. Effect of paclobutrazol on water stress amelioration through antioxidants and free radical scavenging enzymes in Arachis hypogaea L. Colloids and Surfaces B: Biointerfaces. 60, (2) 229-235.
Sathishkumar, G., P. K. Jha, V. Vignesh, C. Rajkuberan, M. Jeyaraj, M. Selvakumar, R. Jha and S. Sivaramakrishnan, 2016. Cannonball fruit (Couroupita guianensis, Aubl.) extract mediated synthesis of gold nanoparticles and evaluation of its antioxidant activity. Journal of Molecular Liquids. 215, 229-236.
Sharma, P., D. Bhatt, M. G. H. Zaidi, P. P. Saradhi, P. K. Khanna and S. Arora, 2012. Silver nanoparticle-mediated enhancement in growth and antioxidant status of Brassica juncea. Applied biochemistry and biotechnology. 167, (8) 2225-2233.
Siddiqui, M. H. and M. H. Al-Whaibi, 2014. Role of nano-SiO2 in germination of tomato (Lycopersicum esculentum seeds Mill.). Saudi journal of biological sciences. 21, (1) 13-17.
Singleton, V. L., R. Orthofer and R. M. Lamuela-Raventós, 1999. [14] Analysis of total phenols and other oxidation substrates and antioxidants by means of folin-ciocalteu reagent. Methods in enzymology. 299, 152-178.
Vannini, C., G. Domingo, E. Onelli, F. De Mattia, I. Bruni, M. Marsoni and M. Bracale, 2014. Phytotoxic and genotoxic effects of silver nanoparticles exposure on germinating wheat seedlings. Journal of plant physiology. 171, (13) 1142-1148.
Vurayai, R., V. Emongor and B. Moseki, 2011. Physiological responses of bambara groundnut (Vigna subterranea L. Verdc) to short periods of water stress during different developmental stages. Asian Journal of Agricultural Sciences. 3, (1) 37-43.
Wagner, G. J., 1979. Content and vacuole/extravacuole distribution of neutral sugars, free amino acids, and anthocyanin in protoplasts. Plant physiology. 64, (1) 88-93.
Yasur, J. and P. U. Rani, 2013. Environmental effects of nanosilver: impact on castor seed germination, seedling growth, and plant physiology. Environmental Science and Pollution Research. 20, (12) 8636-8648.