Inhibition Effect of Different Propolis Extracts against Fusarium solani in vitro

Abstract

Propolis is a natural brownish by-product created by honey bees. Due to the antimicrobial components it contains, propolis has been widely tested against important fungal pathogens that cause agricultural damage. The diluent used in the extraction of propolis causes differences in the total phenolic compounds and antimicrobial properties of propolis. Fusarium solani, division Ascomycota, is infectious to many cultivated crops such as peas, beans, potatoes, cucurbits, and maize, resulting in significant crop losses. In most plants, it generally leads to rot, wilt and necrotic spots and eventual plant death. In this study, the antifungal effect of propolis collected from Bingöl province of Turkey was evaluated against, with a total of 9 groups of applications. In order to determine the differences between preparations and the critical dose levels, 3 different preparations (Ethanol, (Dimethyl sulfoxide (DMSO), and pure propolis purified by supercritical fluid extraction method) were applied in 3 different concentrations (100µl, 200µl, and 600µl) in PDA medium. The statistical results of the study showed that pure extracts of propolis are more effective at increasing doses (around 30% for 600 µl) against the mycelial growth of pathogen. Furthermore, ethanol extracts of propolis showed a moderate antifungal effect (around 33% for 600 µl), while the DMSO extracts were less effective (around 58% for 600 µl). This study is the first report showing the fungicidal activity of pure propolis purified by the SFE method using CO2 on F. solani. The results of this study may shed light on the development of drug-based strategies against fungal-borne phytopathogens in the future studies.

Keywords

• F. solani
• DMSO
• ethanol
• pure propolis
• Turkey

Farklı Propolis Ekstraktlarının Fusarium solani'ye Karşı Antifungal Etkisi

Abstract

Propolis, bal arıları tarafından oluşturulan doğal kahverengimsi bir yan üründür. Propolis, içerdiği antimikrobiyal bileşenler nedeniyle tarımsal hasara neden olan önemli fungal patojenlere karşı yaygın olarak test edilmiştir. Propolisin ekstraksiyonunda kullanılan seyreltici, propolisin toplam fenolik bileşikleri ve antimikrobiyal özelliklerinde farklılıklara neden olur. Ascomycota şubesinde yer alan Fusarium solani, bezelye, fasulye, patates, kabakgiller ve mısır gibi birçok bitki türü için infeksiyözdür ve önemli ürün kayıplarına neden olur. Çoğu bitkide genellikle çürümeye, solgunluğa ve nekrotik lekelere ve sonunda bitki ölümüne yol açar. Bu çalışmada, Türkiye'nin Bingöl ilinden toplanan propolisin antifungal etkisi toplam 9 uygulama grubu ile değerlendirilmiştir. Preparatlar ve kritik doz seviyeleri arasındaki farkları belirlemek için 3 farklı preparasyon (Etanol, (Dimetil sülfoksit (DMSO) ve süperkritik sıvı ekstraksiyon yöntemi ile saflaştırılmış saf propolis) 3 farklı konsantrasyonda (100µl, 200µl ve 600µl) uygulanmıştır. ) PDA ortamında. Çalışmanın istatistiksel sonuçları, saf propolis ekstraktlarının artan dozlarda (600 µl için yaklaşık %30) patojenin misel büyümesine karşı daha etkili olduğunu gösterdi. Ayrıca, propolisin etanol özleri orta düzeyde bir antifungal etki gösterirken (600 µl için yaklaşık %33), DMSO özleri daha az etkiliydi (600 µl için yaklaşık %58). Bu çalışma, F. solani üzerinde CO2 kullanılarak SFE yöntemiyle saflaştırılmış saf propolisin fungisidal aktivitesini gösteren ilk rapordur. Bu çalışmanın sonuçları, gelecekteki çalışmalarda fungal kaynaklı fitopatojenlere karşı ilaç bazlı stratejilerin geliştirilmesine ışık tutabilir.

Keywords

• F. solani
• DMSO
• etanol
• saf propolis
• antifungal aktivite

References

1. Abd-El-Kareem, F., Saied, N.M. & El-Mohamedy, R.S.R., 2018. Seed treatment with chitosan and ethanol-extracted propolisfor suppression bean root rot disease under greenhouse conditions. J. Mater. Environ. Sci. 9(8), 2356–2361.
2. Ahmed, S.D., Mohanad, A.K. & Zaid, N.H., 2008. Study antifungal activity of ethanol extract propolis against Fusarium oxysporum fungi. J. Res. Diyala. Humanit. 31, 93–105.
3. AL-Ani, I., Zimmermann, S., Reichling, J. & Wink, M., 2018. Antimicrobial activities of european propolis collected from various geographic origins alone and in combination with antibiotics. Medicines. 5, 2. doi:10.3390/medicines5010002.
4. Alenezi, S., Manal, NA., Natto, J., De., Koning, HP., Igoli, J., Watson, DG., 2018. Chemical Profiling of Papua New Guinea Propolis and Assay its Antiprotozoal Activity. JAN. 35.
5. Al-Waili, N., Al-Ghamdi, A., Ansari, M., Al-Attal, Y. & Salom, K., 2012 Synergistic effects of honey and propolis toward drug multi-resistant Staphylococcus aureus, Escherichia coli and Candida albicans isolates in single and polymicrobial cultures. Int. J. Med. Sci. 9(9), 793-780.
6. Anjum, S.I., Ullah, A., Khan, K.A., Attaullah, M., Khan, H., Ali, H., Bashir, M.A., Tahir, M., Ansari, M.J., Ghramh, H.A., Adgaba, N. & Dash, C.K., 2019. Composition and functional properties of propolis (bee glue): A review. Saudi J. Biol. Sci. 26(7), 1695-1703.
7. Araujo, M.J.A.M., Bosco, S.M.G. & Sforcin, J.M., 2016. Pythium insidiosum: inhibitory effects of propolis and geopropolis on hyphal growth. Braz. J. Microbiol. 47, 863–869.
8. Benjilali, B., Tantaoui, E.A., Ayadi, A. & Ihlal, M., 1984. Method to study antimicrobial effects of essential oils: application to theantifungal activity of six Moroccan essences. J. Food Prot. 47, 748–752.

9. Buchta, V., Černý, J. & Opletalová, V,. 2011. In vitro antifungal activity of propolis samples of Czech and Slovak origin. Cent. Eur. J. Biol. 6(2), 160-166.
10. Curifuta, M., Vidal, J., Sánchez-Venegas, J., Contreras, A., Salazar, L.A. & Alvear, M., 2012. The in vitro antifungal evaluation of a commercial extract of Chilean propolis against six fungi of agricultural importance. Cien. Inv. Agr. 39(2), 347–359.
11. Çakar, G., Saraç Sivrikaya, I., Karakara, E., Güller, A. 2021. Evaluation of the In Vitro Fungicidal Activity of Summer Savory and Lavender Essential Oils Against Fusarium solani. ISPEC Journal of Agr. Sciences. 5(4): 795-805
12. Deans, S.G., Sobada, K.P., 1990. Antimicrobial Properties of Marjoram (Origanum marjorana L.) Volatile Oil. Flavour Fragr. J. 5, 187-190.
13. Dias, L.G., Pereira, A.P. & Estevinho, L.M., 2012. Comparative study of different Portuguese samples of propolis: Pollinic, sensorial, physicochemical, microbiological characterization and antibacterial activity. Food. Chem. Toxicol. 50(12), 4246-4253.
14. Drescher, N., Klein, A.M., Neumann, P., Yañez, O. & Leonhardt, S.D., 2017. Inside honeybee hives: impact of natural propolis on the ectoparasitic mite Varroa destructor and viruses. Insects. 8(1), 5 doi: 10.3390/insects8010015.
15. Er, Y., 2021. In vitro and in vivo antimicrobial activity of propolis extracts against various plant pathogens. J.Plant Dis. Protect. https://doi.org/10.1007/s41348-021-00437-y.
16. Erdoğan, O., Çelik, A., Yıldız, Ş., & Kökten, K. 2014. Pamukta fide kök çürüklüğü etmenlerine karşı bazı bitki ekstrakt ve uçucu yağlarının antifungal etkisi. Türk Tarım ve Doğa Bilimleri Dergisi 1(3): 398-404.
17. Erdoğan, O., Celik, A., & Zeybek, A. 2016. In Vitro Antifungal Activity of Mint, Thyme, Lavender Extracts and Essential Oils on Verticillium dahliae Kleb. Fresenius Environmental Bulletin, 25(11), 4856-4862.
18. Ertürk, Ö., Yavuz, C. & Sıralı, C., 2011. The Antimicrobial Activity of Propolis from Ordu Province of Turkey. HARUM. 28, 11-16. (in Turkish).
19. Ezazi, R., & Davari, M., 2018. Antifungal activity of ethanolic extract of propolis (EEP) against some postharvest fungi. Biological Control of Pests and Plant Diseases. 1(1), 103-107.
20. Falcão, SI., Vale, N., Cos, P., Games, P., Freire,C., Maes, L. & Vilas-Boas, M., 2014. In Vitro Evaluation of Portuguese Propolis and Floral Sources for Antiprotozoal, Antibacterial and Antifungal Activity. Phytother. Res. Res. 28, 437–443.
21. Farnesi, A.P., Aquino-Ferreira, R., De Jong, D. & Bastos, J.K., 2009. Effects of stingless bee and honey bee propolis on four species of bacteria. Genet. Mol. Res. 8, 635-640.
22. Ganesh P., Dwivedi S.K., 2018. Biological and Chemical Management of Fusarium solani Causing Wilt in Eggplants. IJSRR. 7(4), 1914-1925.
23. Gardana, C., Scaglianti, M., Pietta, P. & Simonetti, P., 2007. Analysis of the polyphenolic fraction of propolis from different sources by liquid chromatography–tandem mass spectrometry. J. Pharm. Biomed. Anal. 45, 390–399.
24. Ghasemi, F.S., Eshraghi, S.S., Andalibi F., Hooshyar, H., Kalantar- Neyestanaki, D., Samadi A. & Fatahi-Bafghi, M., 2017. Anti-Bacterial Effect of Propolis Extract in Oil Against Different Bacteria. Zahedan J. Res. Med. Sci. 19(3),7225.
25. Gregolin, F.S., Bonaldo, S.M., Sinhorin, A.P. & Banderó, J.L., 2019. The in vitro control of Fusarium proliferatum by propolis ethanolic extracts. Revista de Ciências Agrárias. 42(2), 456-463.
26. Guginski-Piva, C.A., Stantos, I.D. & Junior, A.W., 2015. Propolis for the control of Powdery mildew and the induction of phytoalexins in cucumber. IDESIA (Chile). 33(1), 39–47.
27. Gül, Y., 2019. Determination of chemical composition of propolis extracts obtained from different regions and antifungal efficiencies against soilborne disease agents of tomato. Master Thesis. University of Mustafa Kemal. Plant Protection Department. Hatay/Turkey. (in Turkish).
28. Gür, N., Bayrak, N. & Topdemir, A., 2020. Determination of antimicrobial activity and some biochemical properties of honey and propolis in Turkish markets. Prog. Nutr. 22, 3 DOI: 10.23751/pn.v22i3.9166.
29. Huang, S., Zhang, C., Wang, K., Li, G. & Hu, F., 2014. Recent advances in the chemical composition of Propolis. Molecules. 19, 19610-19632.
30. Kim, S.K., Woo, S.O., Han, S.M., Bang, K.W., Kim, S.G., Choi, H.M., Moon, H.J. & Lee, S.W., 2019. Antibacterial and antifungal effects of Korean propolis against ginseng disease. Int. J. Indust. Entomol. 39(2), 82-85.
31. Koç, A.N., Silici, S., Kasap, F., Hörmet-Öz, H.T., Mavus-Buldu, H. & Ercal, B.D., 2011. Antifungal Activity of the Honeybee Products Against Candida spp. and Trichosporon spp. Journal of Medicinal Food. 14(1-2), 128-134.
32. Koç, İ., Yardım, E. N., Çelik, A., Mendeş, M., Mirtagioğlu, H., & Namlı, A. (2018). Fındık Kabuklarından Elde Edilmiş Odun Sirkesi’nin In-Vitro Şartlarında Antifungal Etkisinin Belirlenmesi. Bitlis Eren Üniversitesi Fen Bilimleri Dergisi, 7(2), 296-300.
33. Kriaa, M., Hammami, I., Sahnoun, M, Azebou, MC., Triki, M.A. & Kammoun, R., 2015. Biocontrol of tomato plant diseases caused by Fusarium solaniusing a new isolated Aspergillus tubingensis CTM 507 glucoseoxidase. C. R. Biologies. 338: 666-677.
34. Kurt, Ş. & Şahinler, N., 2003. Antifungal Activity of Propolis Extract Against Certain Phytopathogenic Fungi. Uludag Bee Journal. 3(3), 35-37.
35. Kustiawan, P.M., Lirdprapamongkol, K., Palaga, T., Puthong, S., Phuwapraisirisan, P., Svasti J. & Chanchao, C., 2017. Molecular mechanism of cardol, isolated from Trigona incisa stingless bee propolis, induced apoptosis in the SW620 human colorectal cancer cell line. BMC Pharmacol. Toxicol. 18, 32. doi: 10.1186/s40360-017-0139-4.
36. La Torre, A., Guccione, M. & Imbroglini, G., 1990. Indaginepreliminare sull’azione di preparati a base di propolinei confronti di Botrytis cinerea Pers della fragola. Apicoltura. 6, 169–177.
37. Loebler, M., Sánchez, C., Muchagato Maurício, E., Diogo, E., Santos, M., Vasilenko, P., Cruz, A.S., Mendes, B., Gonçalves, M. & Duarte, M.P., 2020. Potential Application of Propolis Extracts to Control the Growth of Stemphylium vesicarium in “Rocha” Pear. Appl. Sci. 10, 1990. doi:10.3390/app10061990.
38. Marcucci, M.C., 1995. Propolis: chemical composition, biological properties and therapeutical activity, Apidologie. 26, 83-99.
39. Marini, D., Mensch, R., Freiberger, M.B., Dartora, J., Franzener, G., Gracia, R.C. & Stangarlin, J.R., 2012. Efeito antifúngico de extratos alcoólicos de própolis sobre patógenos da videira. Arquivos do Instituto Biológico. 79(2), 305-308. http://dx.doi.org/10.1590/S1808-16572012000200023.
40. Mavri, A., Abramovič, A., Polak, T., Bertoncelj, T., Jamnik, Smole Možina, S. & Jeršek, L., 2012. Chemical Properties and Antioxidant and Antimicrobial Activities of Slovenian Propolis. Chem. Biodivers. 9, 1545–1558.
41. Meneses, E.A., Durango, D.L. & García, C.M., 2009. Antifungal Activity Against Postharvest Fungi by Extracts from Colombian Propolis. Quim Nova. 32(8), 2011-2017. (in Colombian).
42. Netíkóva, L., Bogusch, P. & Heneberg, P., 2013. Czech Ethanol-Free Propolis Extract Displays Inhibitory Activity against a Broad Spectrum of Bacterial and Fungal Pathogens. J. Food Sci. 78(9). doi: 10.1111/1750-3841.12230.
43. Oliveira, A.C.P., Shinobu, C., Suemi Longhini, R., Franco, S.L. & Svidzinsk, T.I.E., 2006. Antifungal activity of propolis extract against yeasts isolated from onychomycosis lesions. Mem. Inst. Oswaldo Cruz. 101(5), 493-497.
44. Ota, C., Unterkircher, C., Fantinato, V. & Shimizu, M. T., 2002. Antifungal activity of propolis on different species of Candida. Mycoses. 44(9‐10), 375-378.
45. Ôzcan, M., 1999. Antifungal properties of propolis. Grasas Aceites. 50, 395–398.
46. Özdemir, A.E., Çandır, E.E., Kaplankıran, M., Soylu, E.M., Fiahinler, N. & Gül, A., 2010. The effects of ethanol dissolved propolis on the storage of grapefruit cv. Star Ruby Turk J Agric For. 34,155–162.
47. Özyiğit, Ç., 2020. Evaluation of effectiveness of propolis extracts, collected from different regions of Turkey, against mold agents. Master Thesis. University of Gaziosmanpaşa. Plant Protection Department. Tokat/Turkey. (in Turkish).
48. Pazin, W.M., Santos, S.N.D., Queiroz, S.C.N., Bagatolli, L.A., Soares, A.E.E., Melo, I.S. & Ito, A.S., 2019. Bioactivity and action mechanism of green propolis against Pythium aphanidermatum. An. Acad. Bras. Ciênc. 91(2), e20180598.
49. Pereira, C.S., Rempel, D., Sinhorin, A.P., Fernandes, H. & Fiorini, I.V.A., 2017. Aplicação de extrato etanólico de própolis em doenças da cultura da soja. Revista de Ciências Agrárias. 40(4), 854-862. http://dx.doi. org/10.19084/RCA17029.
50. Pérez-Hernández, A., Liliana, R., Elena, P., Brett, S., Liew, E.C.Y. & Julio, G.V., 2020. Pathogenic, Morphological, and Phylogenetic Characterization of Fusarium solani f. sp. cucurbitae Isolates From Cucurbits in Almería Province, Spain. Plant Dis. 104. PDIS09191954RE. 10.1094/PDIS-09-19-1954-RE.
51. Pérez-Hernández, A., Serrano-Alonso, Y., Aguilar-Pérez, M.I., Gómez-Uroz, R. & Gómez-Vázquez, J., 2014. Damping-Off and Root Rot of Pepper Caused by Fusarium oxysporum in Almería Province, Spain. Plant Dis. 2014 Aug, 98(8):1159. doi: 10.1094/PDIS-02-14-0212-PDN. PMID: 30708819.
52. Petruzzi, L., Corbo, MR., Campaniello, D., Speranza, B., Sinigaglia, M. & Bevilacqua, A., 2020. Antifungal and Antibacterial E ect of Propolis: A Comparative Hit for Food-Borne Pseudomonas, Enterobacteriaceae and Fungi. Foods. 9, 559; doi:10.3390/foods9050559.
53. Porta, G.D., Porcedda, S., Marongiu, B. & Reverchon, E., 1999. Isolation of Eucalyptus oil by Supercritical fluid extraction. Flavour Fragr. J.14, 214-218.
54. Porto, M.A.F., Ambrósio, M.M.Q., Nascimento, S.R.C., Cruz, B.L.S. & Torres, T.M., 2019. Interaction of Fusarium solani, Macrophomina phaseolina and Rhizoctonia solani as root rot pathogens of Cucumis melo. Summa Phytopathol. 45(4), 355-360.
55. Pourmortazavi, S.M., Baghaee, P. & Mirhosseini, M.A., 2004. Extraction Of Volatile Compounds From Juniperus communis L. Leaves With Supercritical Fluid Carbon Dioxide:Comparison With Hydrodistillation. Flavour Fragr. J.19, 417-420.
56. Quiroga, E.N., Sampietro, D.A., Soberón, J.R., Sgariglia, M.A. & Vattuon, M.A., 2006. Propolis from the northwest of Argentina as a source of antifungal principles. J. Appl. Microbiol. 101, 103–110.
57. Rufatto, L.C., Dos Santos, D.A., Marinho, F., Henriques, J.A.P., Ely, M.R. & Moura, S., 2017. Red propolis: chemical composition and pharmacological activity. Asian Pac. J. Trop. Biomed. 7(7), 591-598.
58. Shehu, A., Ismail, S., Rohin, M.A.K., Harun, A., Aziz, A.A. & Haque, M., 2016. Antifungal Properties of Malaysian tualang honey and stingless bee propolis against Candida albicans and Cryptococcus neoformans. J. Appl. Pharm. 6, 44–50.
59. Silici, S., Kutluca, S., 2005. Chemical composition and antibacterial activity of propolis collected by three different races of honeybees in the same region. J. Ethnopharmacol. 99(1), 69-73.
60. Summerell, B., Laurence, Matthew H., Liew, E.C.Y., Leslie, J.F. 2010. "Biogeography and phylogeography of Fusarium: a review". Fungal Diversity. 44 (1): 3–13.
61. Türk, M.U., 2017. Chemical structures and antifungal properties of propolis produced in Aegean region. Master Thesis. University of Uşak. Zootechnology Department. Uşak/Turkey. (in Turkish).
62. Ugur, A., Arslan, T., 2004. An In Vitro Study on Antimicrobial Activity of Propolis from Mugla Province of Turkey. J. Med. Food. 7(1), 90-94.
63. Velazquez, C., Navarro, M., Acosta, A., Angulo, A., Dominguez, Z., Roble, R., Robles-Zepeda, R., Lugo, E., Goycoolea, F.M., Velazquez, E.F., Astiazaran H. & Hernandez, J., 2007. Antibacterial and free-radical scavenging activities of Sonoran propolis. J. Appl. Microbiol. 103, 1747–1756.
64. Velikova, M., Bankova, V., Marcucci, M., Tsvetkova, I. & Kujungiev, A., 2000. Chemical Composition and Biological Activity of Propolis from Brazilian Meliponinae. Z. Naturforsch C. 55, 785-789. (in Brazilian).
65. Yamani,Y., Khajeh, M., Ghasemi, E., Mirza, M. & Javidnia, K., 2007. Comparison Of Essential Oil Compositions Of Salia Mirzayanii Obtained By Supercritical Carbondioxide Extraction And Hydrodistilaltion Methods. Food Chem. 108, 341-346.
66. Yanar, Y., Yanar, D. & Servet, A., 2005. Antifungal Activity of Turkish Propolis Against Phytophthora Species. Plant Pathol. 4(1), 118- 124.