Investigation of Chemical Properties of Green Tea Ethanolic Extract and Its Inhibitory and Lethal Effects on Aspergillus niger, Botrytis cinerea and Rhizopus stolonifer (Causing Rot in Strawberry and Grapes)

Document Type : Research Article

Authors

• Mostafa Rahmati-Joneidabad ¹
• Behrooz Alizadeh Behbahani ²
• Mohammad Noshad ²

¹ Department of Horticultural Science, Faculty of Agriculture, Agricultural Sciences and Natural Resources University of Khuzestan, Mollasani, Iran

² Department of Food Science and Technology, Faculty of Animal Science and Food Technology, Agricultural Sciences and Natural Resources University of Khuzestan, Mollasani, Iran

https://doi.org/10.22067/ifstrj.2022.76184.1163

Abstract

Introduction
Strawberry and grapes are generally infected with pathogenic fungi (e.g., Aspergillus niger, Botrytis cinerea, Rhizopus stolonifera, etc.). Synthetic fungicides are commonly used as the first line of defense against post-harvest pathogens on packaging lines. However, disposal of toxic waste is a costly process and the hazardous waste causes serious environmental problems. In addition, fungal pathogens have shown a worrying trend of resistance to these fungicides, thus shortening the shelf life of products. Compounds that can be equally effective in controlling pathogens, but preventing or minimizing the waste problems will be inevitable. The large volume of internationally processed agricultural products, as well as the increasing demand for organically produced fruits, emphasizes the need to replace synthetic fungicides with safer and biodegradable alternatives. Natural plant-derived products effectively meet this criterion and have great potential to influence modern agricultural research. Catechins and other polyphenols in green tea show strong antioxidant activity. Also, the antimicrobial activity of green tea extract against Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, and Candida albicans has been reported. Therefore, the present study was performed to prepare the ethanolic extract of green tea and to determine the content of total phenol, total flavonoids, antioxidant activity, and its antifungal effect against Aspergillus niger, Botrytis cinerea, and Rhizopus stolonifer (causing rot in strawberry and grapes).
 
Materials and Methods
Fresh green tea leaves were dried at room temperature and then powdered. Then, ethanol (70%) was added to the powdered leaves (solvent to powder ratio of 10:1 v/w) and the mixture was refluxed for 120 min. The resulting mixture was filtered through a filter paper and then concentrated under vacuum and finally dried in an oven.
Total phenol content (by Folin-Ciocalteu reagent at 756 nm), total flavonoid content (spectrophotometrically at 510 nm), antioxidant activity (by DPPH and ABTS radical scavenging methods), and antifungal effect (by disk diffusion agar, well diffusion agar, minimum inhibitory concentration, and minimum fungicidal concentration) of the extract were evaluated.
 
Results and Discussion
The extract contained 175.60 mg GAE /g total phenol and 47.53 mg QE/g total flavonoids and its antioxidant activity using DPPH and ABTS free radical assays was 78.89% and 86.57%, respectively. The results of antifungal activity showed that the diameter of the growth inhibition zone increased significantly with increasing the concentration of the extract, and Botrytis cinerea and Rhizopus stolonifer were the most sensitive and resistant fungal strains to the extract, respectively. The minimum fungicidal concentrations for the strains of Botrytis cinerea and Rhizopus stolonifer were 64 and 512 mg/ml, respectively.
 
Conclusion
The results of the present study showed that the ethanolic extract of green tea could be considered as potential source of natural antioxidant and antifungal agents. The presence of phenolic and flavonoid compounds may be responsible for the antifungal and antioxidant effects of the extract. However, due to the fact that this study was performed with the crude extract of green tea, it is difficult to identify compounds responsible for antifungal and antioxidant activity. On this point, only the separation of the components of the extract allows the detection of antifungal and antioxidant compounds. This study provides a basis for further researches, in particular the use of these antioxidants and antifungal compounds. Green tea extract is especially suitable for products with high sensitivity to lipid oxidation and infection with molds.

Keywords

• Antifungal activity
• Antioxidant activity
• Bioactive extract
• Green tea
• Natural preservative

Main Subjects

• Food Biotechnology

©2023 The author(s). This is an open access article distributed under Creative Commons Attribution 4.0 International License (CC BY 4.0), which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source.

References

1. Ahmed, A.F., Attia, F.A., Liu, Z., Li, C., Wei, J., & Kang, W. (2019). Antioxidant activity and total phenolic content of essential oils and extracts of sweet basil (Ocimum basilicum) plants. Food Science and Human Wellness, 8(3), 299-305. https://doi.org/10.1016/j.fshw.2019.07.004
2. Alghooneh, A., Alizadeh Behbahani, B., Noorbakhsh, H., & Tabatabaei Yazdi, F. (2015). Application of intelligent modeling to predict the population dynamics of Pseudomonas aeruginosa in Frankfurter sausage containing Satureja bachtiarica extracts. Microbial Pathogenesis, 85, 58-65. https://doi.org/10.1016/j.micpath.2015.06.003
3. Alizade Behbahani, B., Tabatabaei Yazdi, F., Heidari Sureshjani, M., Mortazavi, A., & Tabatabaei Yazdi, F. (2014). Antimicrobial effect of the aqueous and ethanolic Satureja bachtiarica extracts “in vitro”. Iranian Journal of Infectious Diseases and Tropical Medicine, 19(64), 13-19.
4. Alizadeh Behbahani, B., Falah, F., Vasiee, A., & Tabatabaee Yazdi, F. (2021). Control of microbial growth and lipid oxidation in beef using a Lepidium perfoliatum seed mucilage edible coating incorporated with chicory essential oil. Food Science & Nutrition, 9(5), 2458-2467. https://doi.org/10.1002/fsn3.2186
5. Alizadeh Behbahani, B., Noshad, M., & Falah, F. (2019). Study of chemical structure, antimicrobial, cytotoxic and mechanism of action of Syzygium aromaticum essential oil on foodborne pathogens. Potravinarstvo Slovak Journal of Food Sciences, 13(1), 875-883.
6. Alizadeh Behbahani, B., Shahidi, F., Yazdi, F.T., & Mohebbi, M. (2013a). Antifungal effect of aqueous and ethanolic mangrove plant extract on pathogenic fungus" in vitro". International Journal of Agronomy and Plant Production, 4(7), 1652-1658.
7. Alizadeh Behbahani, B., Tabatabaei Yazdi, F., Mortazavi, S.A., Zendeboodi, F., Gholian, M.M., & Vasiee, A. (2013b). Effect of aqueous and ethanolic extract of Eucalyptus camaldulensis on food infection and intoxication microorganisms “in vitro”. Journal of Paramedical Sciences, 4(3), 89-99.
8. Alizadeh Behbahani, B., Tabatabaei Yazdi, F., Noorbakhsh, H., Riazi, F., Jajarmi, A., & Tabatabaei Yazdi, F. (2016b). Study of the antibacterial activity of methanolic and aqueous extracts of Myrtus communis on pathogenic strains causing infection. Zahedan Journal of Research in Medical Sciences, 18(2), e5989.
9. Alizadeh Behbahani, B., Tabatabaei Yazdi, F., Shahidi, F., & Riazi, F. (2016a). Antifungal effect of the aqueous and ethanolic Avicennia marina extracts on Alternaria citri and Penicillium digitatum. Zahedan Journal of Research in Medical Sciences, 18(2), e5992.
10. Almajano, M.P., Carbo, R., Jiménez, J.A.L., & Gordon, M.H. (2008). Antioxidant and antimicrobial activities of tea infusions. Food Chemistry, 108(1), 55-63. https://doi.org/10.1016/j.foodchem.2007.10.040
11. Al-Samarrai, G., Singh, H., & Syarhabil, M. (2012). Evaluating eco-friendly botanicals (natural plant extracts) as alternatives to synthetic fungicides. Annals of Agricultural and Environmental Medicine, 19(4), 673-676.
12. Baena, R., Araujo, E.S., Souza, J.P., Bischoff, A.M., Zarbin, P.H., Zawadneak, M.A., & Cuquel, F.L. (2022). Ripening stages and volatile compounds present in strawberry fruits are involved in the oviposition choice of Drosophila suzukii (Diptera: Drosophilidae). Crop Protection, 153, 105883. https://doi.org/10.1016/j.cropro.2021.105883
13. Barzegar, H., Alizadeh Behbahani, B., & Mehrnia, M.A. (2020). Quality retention and shelf life extension of fresh beef using Lepidium sativum seed mucilage-based edible coating containing Heracleum lasiopetalum essential oil: an experimental and modeling study. Food Science and Biotechnology, 29(5), 717-728. https://doi.org/10.1007/s10068-019-00715-4
14. Bharti, R., & Singh, B. (2020). Green tea (Camellia assamica) extract as an antioxidant additive to enhance the oxidation stability of biodiesel synthesized from waste cooking oil. Fuel, 262, 1, 16658. https://doi.org/10.1016/j.fuel.2019.116658
15. Camargo, L., Pedroso, L., Vendrame, S., Mainardes, R., & Khalil, N. (2016). Antioxidant and antifungal activities of Camellia sinensis (L.) Kuntze leaves obtained by different forms of production. Brazilian Journal of Biology, 76, 428-434.
16. Chou, -C., Lin, L.-L., & Chung, K.-T. (1999). Antimicrobial activity of tea as affected by the degree of fermentation and manufacturing season. International Journal of Food Microbiology, 48(2), 125-130. https://doi.org/10.1016/S0168-1605(99)00034-3
17. du Plooy, W., Regnier, T., & Combrinck, S. (2009). Essential oil amended coatings as alternatives to synthetic fungicides in citrus postharvest management. Postharvest Biology and Technology, 53(3), 117-122. https://doi.org/10.1016/j.postharvbio.2009.04.005
18. Dufresne, C.J., & Farnworth, E.R. (2001). A review of latest research findings on the health promotion properties of tea. The Journal of Nutritional Biochemistry, 12(7), 404-421. https://doi.org/10.1016/S0955-2863(01)00155-3
19. Ebrahimi Hemmati Kaykha, M., Jooyandeh, H., Alizadeh behbahani, B., & Noshad, M. (2020). Antimicrobial potential of Cordia myxa fruit on pathogenic bacteria: A study “in vitro” laboratory conditions. Food Science and Technology, 17(101), 71-80.
20. Frei, B., & Higdon, J.V. (2003). Antioxidant activity of tea polyphenols in vivo: evidence from animal studies. The Journal of nutrition, 133(10), 3275S-3284S. https://doi.org/10.1093/jn/133.10.3275S
21. Gramza, A., & Korczak, J. (2005). Tea constituents (Camellia sinensis) as antioxidants in lipid systems. Trends in Food Science & Technology, 16(8), 351-358. https://doi.org/10.1016/j.tifs.2005.02.004
22. Gupta, D., & Kumar, M. (2017). Evaluation of in vitro antimicrobial potential and GC–MS analysis of Camellia sinensis and Terminalia arjuna. Biotechnology Reports, 13, 19-25. https://doi.org/10.1016/j.btre.2016.11.002
23. Ho, C., Chen, C., Wanasundara, U., & Shahidi, F. (1997). Natural antioxidants from tea. Natural Antioxidants, 213-223.
24. Hojjati, M., & Alizadeh Behbahani, B. (2021). Evaluation of the effect of aqueous and methanolic extraction methods on the antioxidant and antimicrobial characteristics of Allium jesdianum extract: in vitro study. Iranian Food Science and Technology Research Journal, 17(1), 83-91. https://doi.org/10.22067/ifstrj.v17i1.85992
25. Katalinic, V., Milos, M., Kulisic, T., & Jukic, M. (2006). Screening of 70 medicinal plant extracts for antioxidant capacity and total Food Chemistry, 94(4), 550-557. https://doi.org/10.1016/j.foodchem.2004.12.004
26. Kiarsi, Z., Hojjati, M., Alizadeh Behbahani, B., & Noshad, M. (2020). In vitro antimicrobial effects of Myristica fragrans essential oil on foodborne pathogens and its influence on beef quality during refrigerated storage. Journal of Food Safety, 40(3), e12782. https://doi.org/10.1111/jfs.12782
27. Koech, K., Wachira, F.N., Ngure, R., Orina, I., Wanyoko, J., Bii, C., & Karori, S. (2013). Antifungal activity of crude tea extracts. African Journal of Agricultural Research, 8(19), 2086-2089.
28. Łuczaj, W., & Skrzydlewska, E. (2005). Antioxidative properties of black tea. Preventive Medicine, 40(6), 910-918. https://doi.org/10.1016/j.ypmed.2004.10.014
29. Marquenie, D., Michiels, C., Geeraerd, A., Schenk, A., Soontjens, C., Van Impe, J., & Nicolaı, B. (2002). Using survival analysis to investigate the effect of UV-C and heat treatment on storage rot of strawberry and sweet cherry. International Journal of Food Microbiology, 73(2-3), 187-196. https://doi.org/10.1016/S0168-1605(01)00648-1
30. Mohseni, P., Borghei, A.M., & Khanali, M. (2018). Coupled life cycle assessment and data envelopment analysis for mitigation of environmental impacts and enhancement of energy efficiency in grape production. Journal of Cleaner Production, 197, 937-947. https://doi.org/10.1016/j.jclepro.2018.06.243
31. Namita, P., Mukesh, R., & Vijay, K.J. (2012). Camellia sinensis (green tea): a review. Global Journal of Pharmacology, 6(2), 52-59.
32. Nazer, A., Kobilinsky, A., Tholozan, J.-L., & Dubois-Brissonnet, F. (2005). Combinations of food antimicrobials at low levels to inhibit the growth of Salmonella Typhimurium: a synergistic effect? Food Microbiology, 22(5), 391-398. https://doi.org/10.1016/j.fm.2004.10.003
33. Negi, P., Jayaprakasha, G., & Jena, B. (2003). Antioxidant and antimutagenic activities of pomegranate peel extracts. Food Chemistry, 80(3), 393-397. https://doi.org/10.1016/S0308-8146(02)00279-0
34. Noshad, M., Alizadeh Behbahani, B., Jooyandeh, H., Rahmati‐Joneidabad, M., Hemmati Kaykha, M.E., & Ghodsi Sheikhjan, M. (2021). Utilization of Plantago major seed mucilage containing Citrus limon essential oil as an edible coating to improve shelf‐life of buffalo meat under refrigeration conditions. Food Science & Nutrition, 9(3), 1625-1639. https://doi.org/10.1002/fsn3.2137
35. Persaud, R., Khan, A., Isaac, W.-A., Ganpat, W., & Saravanakumar, D. (2019). Plant extracts, bioagents and new generation fungicides in the control of rice sheath blight in Guyana. Crop Protection, 119, 30-37. https://doi.org/10.1016/j.cropro.2019.01.008
36. Perva-Uzunalić, A., Škerget, M., Knez, Ž., Weinreich, B., Otto, F., & Grüner, S. (2006). Extraction of active ingredients from green tea (Camellia sinensis): Extraction efficiency of major catechins and caffeine. Food Chemistry, 96(4), 597-605. https://doi.org/10.1016/j.foodchem.2005.03.015
37. Rahmati-Joneidabad, M., & Alizadeh Behbahani, B. (2021). Identification of chemical compounds, antioxidant potential, and antifungal activity of Thymus daenensis essential oil against spoilage fungi causing apple rot. Iranian Food Science and Technology Research, 17(5), 691-700. https://doi.org/10.22067/ifstrj.v18i1.87595
38. Rahmati-Joneidabad, M., Alizade Behbahani, B., & Noshad, M. (2021). Antifungal effect of Satureja khuzestanica essential oil on Aspergillus niger, Botrytis cinerea, and Rhizopus stolonifer causing strawberry’s rot and mold. Food Science and Technology, 18(115), 171-180. https://doi.org/10.52547/fsct.18.115.13
39. Sabaghi, M., Maghsoudlou, Y., Khomeiri, M., & Ziaiifar, A.M. (2015). Active edible coating from chitosan incorporating green tea extract as an antioxidant and antifungal on fresh walnut kernel. Postharvest Biology and Technology, 110, 224-228. https://doi.org/10.1016/j.postharvbio.2015.08.025
40. Saki, A., Mozafari, H., Asl, K. K., Sani, B., & Mirza, M. (2019). Plant yield, antioxidant capacity and essential oil quality of Satureja mutica supplied with cattle manure and wheat straw in different plant densities. Communications in Soil Science and Plant Analysis, 50(21), 2683-2693. https://doi.org/10.1080/00103624.2019.1670835
41. Senanayake, S.N. (2013). Green tea extract: Chemistry, antioxidant properties and food applications–A review. Journal of Functional Foods, 5(4), 1529-1541. https://doi.org/10.1016/j.jff.2013.08.011
42. Si, W., Gong, J., Tsao, R., Kalab, M., Yang, R., & Yin, Y. (2006). Bioassay-guided purification and identification of antimicrobial components in Chinese green tea extract. Journal of Chromatography A, 1125(2), 204-210. https://doi.org/10.1016/j.chroma.2006.05.061
43. Solairaj, D., Legrand, N.N.G., Yang, Q., & Zhang, H. (2020). Isolation of pathogenic fungi causing postharvest decay in table grapes and in vivo biocontrol activity of selected yeasts against them. Physiological and Molecular Plant Pathology, 110, 101478. https://doi.org/10.1016/j.pmpp.2020.101478
44. Taguri, T., Tanaka, T., & Kouno, I. (2004). Antimicrobial activity of 10 different plant polyphenols against bacteria causing food-borne disease. Biological and Pharmaceutical Bulletin, 27(12), 1965-1969.
45. Tang, Y., Ma, X., Li, M., & Wang, Y. (2020). The effect of temperature and light on strawberry production in a solar greenhouse. Solar Energy, 195, 318-328. https://doi.org/10.1016/j.solener.2019.11.070
46. Tehranifar, A., & Sarsaefi, M. (2000). Strawberry growing in Iran. IV International Strawberry Symposium 567.
47. Wang, H., Provan, G.J., & Helliwell, K. (2000). Tea flavonoids: their functions, utilisation and analysis. Trends in Food Science & Technology, 11(4-5), 152-160. https://doi.org/10.1016/S0924-2244(00)00061-3
48. Xi, D., Liu, C., & Su, Y.-C. (2012). Effects of green tea extract on reducing Vibrio parahaemolyticus and increasing shelf life of oyster meats. Food Control, 25(1), 368-373. https://doi.org/10.1016/j.foodcont.2011.11.002
49. Yang, Z., Tu, Y., Baldermann, S., Dong, F., Xu, Y., & Watanabe, N. (2009). Isolation and identification of compounds from the ethanolic extract of flowers of the tea (Camellia sinensis) plant and their contribution to the antioxidant capacity. LWT - Food Science and Technology, 42(8), 1439-1443. https://doi.org/10.1016/j.lwt.2009.03.017
50. Yeganegi, M., Tabatabaei Yazdi, F., Mortazavi, S.A., Asili, J., Alizadeh Behbahani, B., & Beigbabaei, A. (2018). Equisetum telmateia extracts: Chemical compositions, antioxidant activity and antimicrobial effect on the growth of some pathogenic strain causing poisoning and infection. Microbial Pathogenesis, 116, 62-67. https://doi.org/10.1016/j.micpath.2018.01.014