Rezene ve Defne Uçucu Yağ Karışımlarının Kimyasal Bileşenlerinin ve Pestalotiopsis funerea’ya Karşı Antifungal Etkinliklerinin Belirlenmesi
Yıl 2022,
Cilt: 25 Sayı: 1, 113 - 126, 28.02.2022
Merve Kara
,
Musa Türkmen
,
Soner Soylu
Öz
Fungal hastalık etmeni Pestalotiopsis funerea, kozalaklı çam ağaçlarında sürgün uç yanıklığı olarak bilinen hastalığa neden olur. Bu çalışmada, farklı oranlarda harmanlanmış rezene (Foeniculum vulgare Mill.) ve defne (Laurus nobilis L.) uçucu yağ karışımlarının kimyasal bileşimleri ve P. funerea’ya karşı antifungal etkileri in vitro koşullarda araştırılmıştır. Uçucu yağların kimyasal bileşimleri, GC-MS analizi kullanarak belirlenmiştir. Uçucu yağların buhar fazındaki farklı konsantrasyonların misel gelişimi ve konidi çimlenmesinin engellemesi üzerine antifungal etkileri in vitro koşullarda belirlenmiştir. Uçucu yağ karışımlarının en etkili olduğu konsantrasyonlarda fungus hifi ve konidi morfolojisinde neden olduğu etkileri ışık mikroskobu altında incelenmiştir. GC-MS ile yapılan kimyasal analizler sonucunda eucalyptol (%52.88) ve trans-anethole (%81.55)’ün defne ve rezene uçucu yağlarının ana bileşenleri olduğu belirlenmiştir. Tek başlarına kullanıldığında rezene ve defne uçucu yağları P. funerea’nın misel gelişimini 20.0 ve 25.0 µl petri-1 konsantrasyonlarında tamamen engellemiştir. Rezene/defne uçucu yağlarının harmanlanması ile misel gelişimi ve konidi çimlenmesinin engellenmesinde sinerjik antifungal etki görülmüştür. Rezene:defne uçucu yağların 3:1 oranında harmanlanması antifungal etkinliği önemli düzeyde artırmış olup, nispeten düşük konsantrasyonda (10 µl petri-1) daha yüksek düzeyde antifungal etkinlik göstermiştir. Işık mikroskobu ile yapılan gözlemlerde uçucu yağ harmanlarının en etkili konsantrasyonuna maruz kalan hifler ve konidilerde, sitoplazmik pıhtılaşma, vakuolleşme ve protoplazmik içeriğin hücre dışına akıntısı şeklinde gerçekleşen önemli yapısal deformasyonları gözlenmiştir. Sonuçlar, sinerjik antifungal etki gösteren rezene ve defne uçucu yağ karışımının bitki fungal hastalık etmenlerle mücadelede etkili bir biyofungisit olarak kullanılabileceğini göstermiştir.
Kaynakça
- Acimovic M, Tesevic V, Todosijevic M, Djisalov J, Oljaca S 2015. Compositional characteristics of the essential oil of Pimpinella anisum and Foeniculum vulgare grown in Serbia. Botanica Serbica, 39: 9-14.
- Adams RP 2007. Identification of Essential Oils by Gas Chromatography / Mass Spectrometry. Allured Publ. Corp., Carol Stream.
- Ahmed AF, Shi MJ, Liu CY, Kang WY 2019. Comparative Analysis of Antioxidant Activities of Essential Oils and Extracts of Fennel (Foeniculum vulgare Mill.) Seeds From Egypt and China. Food Science and Human Wellness, 8: 67-72.
- Bajo J, Santamaria O, Diez JJ 2008. Cultural Characteristics and Pathogenicity of Pestalotiopsis funerea on Cupressus arizonica. Forest Pathology, 38: 263–274.
- Bakkali F, Averbeck S, Averbeck D, Waomar M 2008. Biological Effects of Essential Oils-A Review. Food and Chemical Toxicology, 46: 446-475.
- Chahal KK, Kaur M, Bhardwaj U, Singla N, Kaur A 2017. A Review on Chemistry and Biological Activities of Laurus nobilis L. Essential Oil. Journal of Pharmacognosy and Phytochemistry, 6: 1153-1161.
- Cheng SS, Lin CY, Gu HJ, Chang ST 2011. Antifungal Activities and Chemical Composition of Wood and Leaf Essential Oils from Cunninghamia konishii. Journal of Wood Chemistry and Technology, 31:204-217.
- Demir V, Gunhan T, Yagcioglu AK, Degirmencioglu A 2004. Mathematical Modelling and the Determination of Some Quality Parameters of Air-Dried Bay Leaves. Biosystems Engineering, 88: 325-335.
- Elkıran O, Akbaba E, Bagcı E 2018. Constituents of Essential Oils from Leaves and Seeds of Laurus nobilis L.: A Chemotaxonomic Approach. Bangladesh Journal of Botany, 47(4): 893-901.
- Espinosagarcia FJ, Saldivargarcia P, Langenheim, JH 1993. Dose-Dependent Effects In vitro Of Essential Oils on the Growth of 2 Endophytic Fungi in Coastal Redwood Leaves. Biochemical Systematics and Ecology, 21:185-194.
- Fujita K, Fujita T, Kubo I 2007. Anethole, a Potential Antimicrobial Synergist, Converts a Fungistatic Dodecanol to a Fungicidal Agent. Phytotherapy Research, 21:47-51
- Gordon TR, Swett CL, Wingfield MJ 2015. Management of Fusarium Diseases Affecting Conifers. Crop Protection, 73: 28-39.
- Ho CL, Hua KF, Hsu KP, Wang EIC, Su YC 2012. Composition and Antipathogenic Activities of the Twig Essential Oil of Chamaecyparis formosensis from Taiwan. Natural Product Communications, 7:933-936.
- Hongratanaworakit T, Srimuang P, Wichanpreechar A, Akarapattaranithi T, Taorang B, Rawdlaurh P 2017. Chemical Composition and Effects of Blended Essential Oil on Humans. Journal of Applied Pharmacy Science, 7 (01): 165-170.
- Huang Y, Zhao J, Zhou L, Wang J, Gong Y, Chen X, Sun Z-X, Wang Q, Jiang W 2010. Antifungal Activity of the Essential Oil of Illicium verum Fruit and Its Main Component trans-Anethole. Molecules, 15(11):7558-7569.
- Kachur K, Suntres Z 2020. The Antibacterial Properties of Phenolic Isomers, Carvacrol and Thymol. Critical Reviews in Food Science and Nutrition, 60:3042-3053.
- Kara M, Soylu S, Türkmen M, Kaya DA 2020. Determination and Antifungal Activities of Laurel and Fennel Essential Oils Against Fungal Disease Agents of Cypress Seedlings. Journal of Tekirdag Agricultural Faculty, 17: 264-275.
- Lucas GC, Alves E, Pereira RB, Perina FJ, de Souza RM 2012. Antibacterial Activity of Essential Oils on Xanthomonas vesicatoria and Control of Bacterial Spot in Tomato. Pesquisa Agropecuária Brasileira, 47(39): 351-359.
- Morcia C, Terzi V 2012. In vitro Antifungal Activity of Terpinen-4-ol, Eugenol, Carvone, 1,8-Cineole (Eucalyptol) and Thymol Against Mycotoxigenic Plant Pathogens. Food Additives and Contaminants - Part A Chemistry, Analysis, Control, Exposure and Risk Assessment, 29(3):415-22
- Mordue JEM 1976. Pestalotiopsis funerea. CMI Descriptions of Pathogenic Fungi and Bacteria, 514: 1-2.
- Nazzaro F, Fratıanni F, Martino LD, Coppola R, Feo VD 2013. Effect of Essential Oils on Pathogenic Bacteria. Pharmaceuticals, 6: 1451-1474.
- Nazzaro F, Fratianni F, Coppola R, Feo VD 2017. Essential Oils and Antifungal Activity. Pharmaceuticals, 10: 86.
- Ozcan MM, Chalchat JC, Arslan D, Ates A, Unver A 2006. Comparative Essential Oil Composition and Antifungal Effect of Bitter Fennel (Foeniculum vulgare ssp. piperitum) Fruit Oils Obtained During Different Vegetation. Journal of Medicinal Food, 9: 552-561.
- Ozcan M, Chalchat JC 2005. Effect of Different Locations on the Chemical Composition of Essential Oils of Laurel (Laurus nobilis L.) Leaves Growing Wild in Turkey. Journal of Medical Food, 8 (3): 408–411.
- Pinto E, Vale-Silva L, Cavaleiro C, Salgueiro L 2009. Antifungal Activity of the Clove Essential Oil From Syzygium aromaticum on Candida, Aspergillus and Dermatophyte Species. Journal of Medical Microbiology, 58: 1454–1462.
- Salem MZM, Elansary HO, Ali HM, El-Settawy AA, Elshikh MS, Abdel-Salam EM, Sklacka-Wozniak K 2018.
Bioactivity of Essential Oils Extracted From Cupressus macrocarpa Branchlets and Corymbia citriodora Leaves Grown in Egypt. BMC Complementary and Alternative Medicine, 18:23
- Sinclair WA, Lyon HH, Johnson WT 1993. Diseases of Trees and Shrubs. New York: Cornell University Press.
- Soylu EM, Soylu S, Kurt Ş 2006. Antimicrobial Activities of the Essential Oils of Various Plants Against Tomato Late Blight Disease Agent Phytophthora infestans. Mycopathologia, 161: 119-128.
- Soylu S, Yigitbas H, Soylu EM, Kurt S 2007. Antifungal Effects of Essential Oils from Oregano and Fennel on Sclerotinia sclerotiorum. Journal of Applied Microbiology, 103(4): 1021-1030.
- Soylu EM, Kurt S, Soylu S 2010. In vitro and in vivo Antifungal Activity of Essential Oils of Various Plants Against Tomato Grey Mould Disease Agent Botrytis cinerea. International Journal of Food Microbiology, 143(3): 183-189.
- Swamy MK, Akhtar MS, Sinniah UR 2016. Antimicrobial Properties of Plant Essential Oils Against Human Pathogens and Their Mode of Action: An Updated Review. Evidence-Based Complementary and Alternative Medicine, 2016: 3012462.
- Staub T 1991. Fungicide Resistance: Practical Experience with Antiresistance Strategies and the Role of Integrated Use. Annual Review of Phytopathology, 29:421-442.
- Taban A, Saharkhiza MJ, Niakousarıc M 2018. Sweet Bay (Laurus nobilis L.) Essential Oil and Its Chemical Composition, Antioxidant Activity and Leaf Micromorphology Under Different Extraction Methods. Sustainable Chemistry and Pharmacy, 9:12–18.
- Tadtong S, Suppawat S, Tintawee A, Saramas P 2012. Antimicrobial Activity of Blended Essential Oil Preparation. Natural Product Communications, 7(10):1401-1404.
- Türkmen M 2015. Farklı Yöntemlerle Elde Edilen Uçucu Yağ Karışımlarının Antifungal Etkinliklerinin Belirlenmesi. Mustafa Kemal Üniversitesi Fen Bilimleri Enstitüsü Tarla Bitkileri Ana Bilim Dalı. Yüksek Lisans Tezi, 55 sayfa.
- Türkmen M, Mert A 2020. Farklı Azot Dozlarının Kişniş (Coriandrum sativum L.) Yaş Herba Uçucu Yağ Bileşenleri Üzerine Etkisi. Mustafa Kemal Üniversitesi Tarım Bilimleri Dergisi, 25(3):309-315.
- Türkmen M, Kaya DA 2021. Antifungal effect of some essential oil combinations. International Journal of Chemistry and Technology, 5(1): 6-10.
- Türkmen M, Kara M, Maral, H, Soylu S 2021. Determination of chemical component of essential oil of Origanum dubium plants grown at different altitudes and antifungal activity against Sclerotinia sclerotiorum. Journal of Food Processing and Preservation, 45: e15787.
- Uysal A, Kurt Ş, Kara M, Soylu S, Soylu EM 2021. Bazı Fidanlıklarda Görülen Toprak Kökenli Fungusların Belirlenmesi. Yüzüncü Yıl Üniversitesi Tarım Bilimleri Dergisi, 31: 142-149.
- Wodnicka A, Huzar E, Dzieciol M, Krawczyk M 2019. Comparison of the Composition and Fungicidal Activity of Essential Oils from Fennel Fruits Cultivated in Poland and Egypt. Polish Journal of Chemical Technology, 21: 38-42.
- Woolf A 1999. Essential Oil Poisoning. Journal of Toxicology: Clinical Toxicology, 37: 721-727.
- Yılmaz B, Deniz İ 2017. The Effects of Cultivation Area and Altitude Variation on the Composition of Essential Oil of Laurus nobilis L. Grown in Eastern, Western and Central Karadeniz Region. International Journal Secondary Metabolite, 4: 187-194.
- Yong AL, Ooh KF, Ong HC, Chai TT, Wong FC 2015. Investigation of Antibacterial Mechanism and Identification of Bacterial Protein Targets Mediated by Antibacterial Medicinal Plant Extracts. Food Chemistry, 186: 32-36.
Determination of Chemical Composition and Antifungal Activities of Fennel and Laurel Essential Oil Mixtures Against Pestalotiopsis funereal
Yıl 2022,
Cilt: 25 Sayı: 1, 113 - 126, 28.02.2022
Merve Kara
,
Musa Türkmen
,
Soner Soylu
Öz
Fungal disease agent Pestalotiopsis funerea causes conifer seedlings shoot or tip blight disease. In this study, chemical compositions and antifungal activities of different blended essential oils obtained from fennel (Foeniculum vulgare Mill.) and laurel (Laurus nobilis L.) were investigated against P. funerea in vitro conditions. Chemical compositions of essential oils were determined using GC-MS analysis. Antifungal volatile phase effects of different concentrations of essential oils mixtures were determined on inhibition of mycelial growth and conidial germination in vitro conditions. The effect of most effective concentrations of essential oil mixtures on the morphology of fungal hypha and conidia were also investigated using light microscope. GC-MS analysis of essential oils revealed that eucalyptol (52.88%) and trans-anethole (81.55%) were the major components of laurel and fennel essential oils. Individual use of fennel and laurel essential oils were found to completely inhibit mycelial growth at 20.0 and 25.0 µl petri-1 concentrations, respectively. Dual blending fennel/laurel essential oil mixtures showed synergistic antifungal activity on inhibition of mycelial growth and conidial germination. By blending fennel and laurel essential oils at proportion of 3:1, antifungal activities were significantly increased that complete mycelial growth and conidial inhibition were recorded at relatively lower concentration (10 µl petri-1). Microscopic observations on hyphae, exposed to volatile phase of the most efficient concentrations of essential oil mixtures, revealed considerable structural deformations such as cytoplasmic coagulation, vacuolization and protoplast leakage from fungal hyphae and conidia. The results showed that blending of fennel and laurel essential oils, which have synergistic antifungal effects, might be used as an effective bio fungicide in management of fungal plant disease agents.
Kaynakça
- Acimovic M, Tesevic V, Todosijevic M, Djisalov J, Oljaca S 2015. Compositional characteristics of the essential oil of Pimpinella anisum and Foeniculum vulgare grown in Serbia. Botanica Serbica, 39: 9-14.
- Adams RP 2007. Identification of Essential Oils by Gas Chromatography / Mass Spectrometry. Allured Publ. Corp., Carol Stream.
- Ahmed AF, Shi MJ, Liu CY, Kang WY 2019. Comparative Analysis of Antioxidant Activities of Essential Oils and Extracts of Fennel (Foeniculum vulgare Mill.) Seeds From Egypt and China. Food Science and Human Wellness, 8: 67-72.
- Bajo J, Santamaria O, Diez JJ 2008. Cultural Characteristics and Pathogenicity of Pestalotiopsis funerea on Cupressus arizonica. Forest Pathology, 38: 263–274.
- Bakkali F, Averbeck S, Averbeck D, Waomar M 2008. Biological Effects of Essential Oils-A Review. Food and Chemical Toxicology, 46: 446-475.
- Chahal KK, Kaur M, Bhardwaj U, Singla N, Kaur A 2017. A Review on Chemistry and Biological Activities of Laurus nobilis L. Essential Oil. Journal of Pharmacognosy and Phytochemistry, 6: 1153-1161.
- Cheng SS, Lin CY, Gu HJ, Chang ST 2011. Antifungal Activities and Chemical Composition of Wood and Leaf Essential Oils from Cunninghamia konishii. Journal of Wood Chemistry and Technology, 31:204-217.
- Demir V, Gunhan T, Yagcioglu AK, Degirmencioglu A 2004. Mathematical Modelling and the Determination of Some Quality Parameters of Air-Dried Bay Leaves. Biosystems Engineering, 88: 325-335.
- Elkıran O, Akbaba E, Bagcı E 2018. Constituents of Essential Oils from Leaves and Seeds of Laurus nobilis L.: A Chemotaxonomic Approach. Bangladesh Journal of Botany, 47(4): 893-901.
- Espinosagarcia FJ, Saldivargarcia P, Langenheim, JH 1993. Dose-Dependent Effects In vitro Of Essential Oils on the Growth of 2 Endophytic Fungi in Coastal Redwood Leaves. Biochemical Systematics and Ecology, 21:185-194.
- Fujita K, Fujita T, Kubo I 2007. Anethole, a Potential Antimicrobial Synergist, Converts a Fungistatic Dodecanol to a Fungicidal Agent. Phytotherapy Research, 21:47-51
- Gordon TR, Swett CL, Wingfield MJ 2015. Management of Fusarium Diseases Affecting Conifers. Crop Protection, 73: 28-39.
- Ho CL, Hua KF, Hsu KP, Wang EIC, Su YC 2012. Composition and Antipathogenic Activities of the Twig Essential Oil of Chamaecyparis formosensis from Taiwan. Natural Product Communications, 7:933-936.
- Hongratanaworakit T, Srimuang P, Wichanpreechar A, Akarapattaranithi T, Taorang B, Rawdlaurh P 2017. Chemical Composition and Effects of Blended Essential Oil on Humans. Journal of Applied Pharmacy Science, 7 (01): 165-170.
- Huang Y, Zhao J, Zhou L, Wang J, Gong Y, Chen X, Sun Z-X, Wang Q, Jiang W 2010. Antifungal Activity of the Essential Oil of Illicium verum Fruit and Its Main Component trans-Anethole. Molecules, 15(11):7558-7569.
- Kachur K, Suntres Z 2020. The Antibacterial Properties of Phenolic Isomers, Carvacrol and Thymol. Critical Reviews in Food Science and Nutrition, 60:3042-3053.
- Kara M, Soylu S, Türkmen M, Kaya DA 2020. Determination and Antifungal Activities of Laurel and Fennel Essential Oils Against Fungal Disease Agents of Cypress Seedlings. Journal of Tekirdag Agricultural Faculty, 17: 264-275.
- Lucas GC, Alves E, Pereira RB, Perina FJ, de Souza RM 2012. Antibacterial Activity of Essential Oils on Xanthomonas vesicatoria and Control of Bacterial Spot in Tomato. Pesquisa Agropecuária Brasileira, 47(39): 351-359.
- Morcia C, Terzi V 2012. In vitro Antifungal Activity of Terpinen-4-ol, Eugenol, Carvone, 1,8-Cineole (Eucalyptol) and Thymol Against Mycotoxigenic Plant Pathogens. Food Additives and Contaminants - Part A Chemistry, Analysis, Control, Exposure and Risk Assessment, 29(3):415-22
- Mordue JEM 1976. Pestalotiopsis funerea. CMI Descriptions of Pathogenic Fungi and Bacteria, 514: 1-2.
- Nazzaro F, Fratıanni F, Martino LD, Coppola R, Feo VD 2013. Effect of Essential Oils on Pathogenic Bacteria. Pharmaceuticals, 6: 1451-1474.
- Nazzaro F, Fratianni F, Coppola R, Feo VD 2017. Essential Oils and Antifungal Activity. Pharmaceuticals, 10: 86.
- Ozcan MM, Chalchat JC, Arslan D, Ates A, Unver A 2006. Comparative Essential Oil Composition and Antifungal Effect of Bitter Fennel (Foeniculum vulgare ssp. piperitum) Fruit Oils Obtained During Different Vegetation. Journal of Medicinal Food, 9: 552-561.
- Ozcan M, Chalchat JC 2005. Effect of Different Locations on the Chemical Composition of Essential Oils of Laurel (Laurus nobilis L.) Leaves Growing Wild in Turkey. Journal of Medical Food, 8 (3): 408–411.
- Pinto E, Vale-Silva L, Cavaleiro C, Salgueiro L 2009. Antifungal Activity of the Clove Essential Oil From Syzygium aromaticum on Candida, Aspergillus and Dermatophyte Species. Journal of Medical Microbiology, 58: 1454–1462.
- Salem MZM, Elansary HO, Ali HM, El-Settawy AA, Elshikh MS, Abdel-Salam EM, Sklacka-Wozniak K 2018.
Bioactivity of Essential Oils Extracted From Cupressus macrocarpa Branchlets and Corymbia citriodora Leaves Grown in Egypt. BMC Complementary and Alternative Medicine, 18:23
- Sinclair WA, Lyon HH, Johnson WT 1993. Diseases of Trees and Shrubs. New York: Cornell University Press.
- Soylu EM, Soylu S, Kurt Ş 2006. Antimicrobial Activities of the Essential Oils of Various Plants Against Tomato Late Blight Disease Agent Phytophthora infestans. Mycopathologia, 161: 119-128.
- Soylu S, Yigitbas H, Soylu EM, Kurt S 2007. Antifungal Effects of Essential Oils from Oregano and Fennel on Sclerotinia sclerotiorum. Journal of Applied Microbiology, 103(4): 1021-1030.
- Soylu EM, Kurt S, Soylu S 2010. In vitro and in vivo Antifungal Activity of Essential Oils of Various Plants Against Tomato Grey Mould Disease Agent Botrytis cinerea. International Journal of Food Microbiology, 143(3): 183-189.
- Swamy MK, Akhtar MS, Sinniah UR 2016. Antimicrobial Properties of Plant Essential Oils Against Human Pathogens and Their Mode of Action: An Updated Review. Evidence-Based Complementary and Alternative Medicine, 2016: 3012462.
- Staub T 1991. Fungicide Resistance: Practical Experience with Antiresistance Strategies and the Role of Integrated Use. Annual Review of Phytopathology, 29:421-442.
- Taban A, Saharkhiza MJ, Niakousarıc M 2018. Sweet Bay (Laurus nobilis L.) Essential Oil and Its Chemical Composition, Antioxidant Activity and Leaf Micromorphology Under Different Extraction Methods. Sustainable Chemistry and Pharmacy, 9:12–18.
- Tadtong S, Suppawat S, Tintawee A, Saramas P 2012. Antimicrobial Activity of Blended Essential Oil Preparation. Natural Product Communications, 7(10):1401-1404.
- Türkmen M 2015. Farklı Yöntemlerle Elde Edilen Uçucu Yağ Karışımlarının Antifungal Etkinliklerinin Belirlenmesi. Mustafa Kemal Üniversitesi Fen Bilimleri Enstitüsü Tarla Bitkileri Ana Bilim Dalı. Yüksek Lisans Tezi, 55 sayfa.
- Türkmen M, Mert A 2020. Farklı Azot Dozlarının Kişniş (Coriandrum sativum L.) Yaş Herba Uçucu Yağ Bileşenleri Üzerine Etkisi. Mustafa Kemal Üniversitesi Tarım Bilimleri Dergisi, 25(3):309-315.
- Türkmen M, Kaya DA 2021. Antifungal effect of some essential oil combinations. International Journal of Chemistry and Technology, 5(1): 6-10.
- Türkmen M, Kara M, Maral, H, Soylu S 2021. Determination of chemical component of essential oil of Origanum dubium plants grown at different altitudes and antifungal activity against Sclerotinia sclerotiorum. Journal of Food Processing and Preservation, 45: e15787.
- Uysal A, Kurt Ş, Kara M, Soylu S, Soylu EM 2021. Bazı Fidanlıklarda Görülen Toprak Kökenli Fungusların Belirlenmesi. Yüzüncü Yıl Üniversitesi Tarım Bilimleri Dergisi, 31: 142-149.
- Wodnicka A, Huzar E, Dzieciol M, Krawczyk M 2019. Comparison of the Composition and Fungicidal Activity of Essential Oils from Fennel Fruits Cultivated in Poland and Egypt. Polish Journal of Chemical Technology, 21: 38-42.
- Woolf A 1999. Essential Oil Poisoning. Journal of Toxicology: Clinical Toxicology, 37: 721-727.
- Yılmaz B, Deniz İ 2017. The Effects of Cultivation Area and Altitude Variation on the Composition of Essential Oil of Laurus nobilis L. Grown in Eastern, Western and Central Karadeniz Region. International Journal Secondary Metabolite, 4: 187-194.
- Yong AL, Ooh KF, Ong HC, Chai TT, Wong FC 2015. Investigation of Antibacterial Mechanism and Identification of Bacterial Protein Targets Mediated by Antibacterial Medicinal Plant Extracts. Food Chemistry, 186: 32-36.