The Application of Essential Oils and Silver Nanoparticles for Sterilization of Bermudagrass Explants in In Vitro Culture

Document Type : Research paper

Authors

Department of Horticultural Science, Faculty of Agriculture and Natural Resources, Arak University, Arak, Iran

Abstract

An important part of plant in vitro techniques is the sterilization of explants and the maintenance of aseptic conditions. Ideally, sterilizing materials should be effective on a vast range of microorganisms at low density. Nowadays, the use of compounds such as essential oils (EOs) and nanoparticles is applicable in microbiology studies. The main objective of this experiment was to study the substitution probability of silver nanoparticles (SNPs), thymol and carvacrol as novel sterilization agents in the tissue culture of Cynodon dactylon. Explants were sterilized with 70% ethanol for 2 min, and then 30% Clorox for 15 min. Sterilization complementary treatments (SNPs, thymol and carvacrol) were applied at different concentrations (100 and 200 mg l-1) with exposure times of 30, 60 and 120 min. According to the results, infection of bermudagrass explants was controlled successfully by SNPs, thymol and carvacrol. Examination of various concentrations in different exposure times showed that 200 mg L-1 SNPs in combination with 100 mg L-1 thymol in 60 min inhibited microbial growth. Thymol and carvacrol were more effective than SNPs in controlling bacteria and fungi contaminations. Finally, these novel agents could be used as an alternative to common chemical treatments for elimination and control of microbial population explants in in vitro conditions.

Keywords


1. Abdi, G., H. Salehi and M. Kush-Khui. 2008. Nano Silver: a Novel Nanomaterial for Removal of Bacterial Contaminants in Valerian (Valeriana officinalis L.) Tissue Culture. Acta Physiol. Plant. 30 (5):709-714.
2. Botelho, M.A., N.A.P. Nogueira, G.M. Bastos, S.G.C. Fonseca, T.L.G. Lemos, F.J.A. Matos, D. Montenegro, J. Heukelbach, V.S. Rao and G.A.C. Brito. 2007. Antimicrobial Activity of the Essential Oil From Lippia sidoides, Carvacrol and Thymol Against Oral Pathogens. Brazilian J. of Med. and Biol. Res. 40:349-356.
3. Bounatirou, S., S. Simitis, M.G. Miguel, L. Faleiro, M.N. Rejeb, M. Neffati, M.M. Costa, A.C. Figueiredo, J.G. Barroso and L.G. Pedro. 2007. Chemical Composition, Antioxidant And Antibacterial Activities of the Essential Oils Isolated from Tunisian Thymus capitatus Hoff. et link. Food Chem. 105:146-155.
4. Braga, P.C., M. Culici, M. Alferi and M. Dal Sasso. 2008. Thymol Inhibits Candida Albicans Biofilm Formation and Mature Biofilm. Int. J. of Antimic. Agents 31:472-477.
5. Carson, C.F., B.J. Mee and T.V. Riley. 2002. Mechanism of Action of Melaleuca alternifolia (Tea Tree) Oil on Staphylococcus Aurous Determined by Time-Kill, Lysis, Leakage and Salt Tolerance Assays and Electron Microscopy. Antimic. Agents Chemotherap. 46 (6):1914–1920.
6. Deein, W., C. Thepsithar, and A. Thongpukdee. 2013. In vitro Culture Medium Sterilization by Chemicals and Essential Oils without Autoclaving and Growth of Chrysanthemum Nodes. World Academy of Science, Engin. Technol. 78:1038-1041.
7. Fakhrfeshani, M., A. Bagheri and A. Sharifi. 2012. Disinfecting Effects of Nano Silver Fluids in Gerbera (Gerbera jamesonii) Capitulum Tissue Culture. J. Biol. Environ. Sci. 6(17): 121-127.
8. Feng, W. and X. Zheng. 2007. Essential oils to Control Alternaria alternate in vitro and in vivo. Food Control (18):1126–1130.
9. Gharati, S., R. Zarghami and M. Amiri. 2010. Evaluation of Nano silver Application for Eliminate Contamination and Viability Cultured 
Segment node of Persian Walnut in in vitro Condition. The 5st Conference of New Idea in Agriculture, Islamic Azad University, Khorasgan, Iran, 16-17 Feb, 2010.
10. Gholamhoseinpour Anvari, Sh., J. Carapetian and J. Dejampour. 2012. Effects of Nanosilver and Vancomycin in sterilization of Peach ×Almond hybrids in the in vitro cultures. Intl. J. Agr. Sci. 2(5): 457-465.
11. Juven, B.J., J. Kanner, F. Schved and H. Weisslowicz. 1994. Factors that Interact with the Antibacterial Action of Thyme Essential Oil and its Active Constituents. J. Appl. Bacteriol. 76:626-631.
12. Kim, J.S., E. Kuk, K.N. Yu, J. Kim, S.J. Park, H.J. Lee, S.H. Kim, Y.K. Park, Y.H. Park, C.Y. Hwany, Y.K. Kim, S.Y. Lee, D.H. Jeong and M.H. Cho. 2007. Antimicrobial Effects of Silver Nanoparticles. Nanomed.:Nanotechnol., Biol. Med. 3:95-101.
13. Kiskó, G. and S. Roller. 2005. Carvacrol and p-cymene Inactivate Escherichia coli O157:H7 in Apple Juice. BMC Microbiol. 5:36.
14. Knobloch, K., A. Pauli, B. Iberl, N. Weis and H. Weig. 1988. Antibacterial Activity and Antifungal Properties of Essential Oil components. J. Essen. Oils Res. 1:119–128.
15. Knowles, J.R., S. Roller, D.B. Murray and A.S. Naidu. 2005. Antimicrobial Action of Carvacrol at Different Stages of Dual-Species Biofilm Development by Staphylococcus aureus and Salmonella enteric Sero var. Typhimurium. Appl. Environ. Microbiol. 71:797-803.
16. Kumar, U. (ed.). 2001. Methods in Plant Tissue Culture. Second Resived and enlarged edition. Agrobios, Jodhpur, India.
17. Lambert, R.J.W., P.N. Skandamis, P. Coote and G.J.E. Nychas. 2001. A Study of the Minimum Inhibitory Concentration and Mode of Action of Oregano Essential Oil, Thymol and Carvacrol. J. Appl. Microbiol. 91: 453–462
18. Loziene, K., P.R. Venkutonis, A. Sipailiene and J. Labokas. 2007. Radical Scavenging and Antibacterial Properties of Extracts from Different Thymus pulegioidesL. Chemo types. Food Chem.. 103:546–559.
19. Lu, S., Z. Wang, X. Peng, Z. Guo, G. Zhang and L. Han. 2006. An Efficient Callus Suspension Culture for Triploid Bermudagrass (Cynodon transvaalensis ×C. dactylon) and Somaclonal Variations. Plant Cell Tissue Organ Cult. 87:77-84.
20. Macheboeuf, D., Y. Morgavi, D.P. Papon, J.L. Mousset and M. Arturo-Schaan. 2008. Dose–Response Effects of Essential Oils on in Vitro Fermentation Activity of the Rumen Microbial Population. Animal Feed Sci. Technol. 145: 335–350.
21. Maneerung, T., S. Tokura and Rujiravanit, R. 2008. Impregnation of Silver Nanoparticles into Bacterial Cellulose for Antimicrobial Wound Dressing. Carbohydrate Polymers. 72:43–51.
22. Martinez-Romer, D., F. Guillén, J. M. Valverde, G. Bailén, P. Zapata, M. Serrano, S. Castillo and D. Valero. 2007. Influence of Carvacrol on Survival of Botrytis Cinerea Inoculated in Table Grapes. Int. J. Food Microbiol. 115:144-148.
23. Morones, J.R., J.L. Elechiguerra, A. Camacho, K. Holt, J.B. Kouri, J.T. Ramirez and M.J. Yacaman. 2005. The Bactericidal Effect of Silver Nanoparticles. Nanotechnol. 16:2346-2353.
24. Morrones, J.R., J.L. Elechiguerra, A. Camacho, K. Holt, J. Kouri, J.T. Ramirez and M.J. Yacamoo. 2005. The Bactericidal Effect of Silver Nanoparticles. Nanotechnology 16: 2346-2353.
25. Navarro, E. A. Baun, R. Behra, N.B. Hartman, J. Filser, A.J. Miao, A. Quiagg, P.H. Santschi and L. Sigg. 2008. Environmental Behavior and Ecotoxicity of Engineered Nano Particles to Algae, Plants, and Fungi. Ecotoxicol. 17:372-386.
26. Olasupo, N.A., D.J. Fitzgerald, M.J. Gasson and A. Narbad. 2003. Activity of Natural Antimicrobial Compounds Against Escherichia coli and Salmonella Entericasero vartyphimurium. Lett. Appl. Microbiol. 36:448-451.
27. Rostami, A.A. and A. Shahsavar. 2009. Nano-Silver Particles Eliminate the in Vitro Contaminations of Olive ‘Mission’ Explants. Asian J. Plant Sci. 8:505-509.
28. Safavi, K., F. Mortazaeinezahad, M. Esfahanizadeh and M.J. Asgari. 2011. In Vitro Antibacterial Activity of Nanomaterial for using in Tobacco Plants Tissue Culture. World Academy of Sci., Eng. Technol. 55
29. Sharififar, F., M.H. Moshafi, S.H. Mansouri, M. Khodashenas and M. Khoshnoodi. 2007. In Vitro Evaluation of Antibacterial and Antioxidant Activities of The Essential oil and Methanol Extract of Endemic Zataria multiflora Boiss. Food Control 18:800-805.
30. Shokri, S., A. R. Babaei, M. Ahmadian, S. H. and M. Mehdi Arab. 2014. The Effects of Different Concentrations of Nano-Silver on Elimination of Bacterial Contaminations and Phenolic Exudation of Rose (Rosa hybrida L.) in Vitro Culture. Inter. J. Farming App. Sci. 1(3):50-54.
31. Sikkema, J., J.A.M. De Bont and B. Poolman. 1994. Interactions of Cyclic Hydrocarbons with Biological Membranes. J. Biologic. Chem. 269 (11): 8022-8028.
32. Skandamis, P.N. and G.J.E. Nychas. 2001. Effect of oregano essential oil on microbiological and physico-chemical attributes of minced meat stored in air and modified atmospheres. J. Appl. Microbiol. 91:1011-1022.
33. Solgi, M., M. Kafi, T.S. Taghavi and R. Naderi. 2009. Essential oils and Silver Nanoparticles (SNP) as Novel Agents to Extend Vase-Life of Gerbera (Gerbera jamesonii Cv. Dune) Flowers. Postharvest Biol. Technol. 53:155-158.
34. Sondi, I. and B. Salopek-Sondi. 2004. Silver Nanoparticles as Antimicrobial Agent: a Case Study as a Model for Gram-Negative Bacteria. J. Colloid Interface Sci. 275:177-182.
35. Taghizadeh, M. 2004. Direct and Indirect Regeneration of Adventitious Shoots in Ornamental Tulipa gesneriana L. 'Apeldoorn' by using in Vitro Culture Method. Tehran University, M.S.c thesis.
36. Taghizadeh, M. 2012. Assessment of Turfgrass Potential for Lead Phytoremediation, in Vitrocally Inducing and Molecular Tracing. Tehran University, Ph.D thesis.
37. Torres, K.C. (ed.) 1989. Tissue Culture Techniques for Horticultural Crop. Van No strand Reinhold. New York, 285 pp.
38. Yahyazadeh, M., R. Omidbaigi, R. Zare and H. Taheri. 2008. Effect of some Essential Oils on Mycelia Growth of Penicillium digitatum Sacc. World J. Microbiol. Biotechnol. 24(8): 1445-1450.
39. Zambonelli, A., A.Z. D'aulerio, A. Severi, S. Benvenuti, L. Maggi and A. Bianchi. 2004. Chemical Composition and Fungicidal Activity of Commercial Essential Oils of Thymus vulgaris L. J Essen. Oil Res. 16:69-74.