Chemical Composition and Antifungal and Antibacterial Activity of Mentha spicata L. Volatile Oil

Cien. Inv. Agr. 45(1):64-69. 2018 www.rcia.uc.cl phytochemistry DOI 10.7764/rcia.v45i1.1897 research paper Chemical Composition and Antifungal and Antibacterial Activity of Mentha spicata L. Volatile Oil Yusuf Bayan1, and Mustafa Küsek2 Ahi Evran University, Faculty of Agriculture, Department of Plant Protection, 40200 Kırşehir-Turkey 1 2 Kahramanmaras Sutcu Imam University, Faculty of Agriculture, Department of Plant Protection, 46100 Kahramanmaras-Turkey Abstract Y. Bayan, and M. Küsek. 2018. Chemical Composition and Antifungal and Antibacterial Activity of Mentha spicata L. Volatile Oil. Cien. Inv. Agr. 45(1): 64-69. In this study, we researched the chemical composition and the antifungal and antibacterial activity of volatile oil from Mentha spicata. The Gas chromatography/mass spectrometry (GC/MS) analysis of M. spicata showed that the main component was carvone (56.94%), followed by limonene (11.63%), sabinene hydrate (7.04%) and caryophyllene (4.06%). The antifungal activity of the volatile oil from M. spicata L. was determined with respect to plant pathogenic fungi, such as Fusarium oxysporum f. sp. radicis-lycopersici (Sacc.) W.C. Synder & H.N. Hans (FORL), Rhizoctonia solani J.G. Kühn. (R. solani), Alternaria solani (A. solani), and Verticillium dahliae Kleb (V. dahliae). The volatile oil was shown to have strong antifungal activity against plant pathogenic fungi. The result of the study was that at a dose of 12 μL petri-1, the volatile oil inhibited 100% of mycelium growth in V. dahliae, A. solani FORL and R. solani. Volatile oil exhibited remarkable activity against the selected bacterial strains of Xanthomonas spp. (ZI365, ZI366, ZI368, ZI370, ZI373, ZI375, ZI376, ZI378). Keywords: Antibacterial, Antifungal, Mentha spicata L., Volatile oils. Introduction The Lamiaceae family contains more than 4000 species in 200 genera. In the Lamiaceae family, mint is one of the most cultivated and aromatic plants. Mint is grown in temperate regions in many parts of the world (South America, Antarctica, Europe and Asia) (Chambers, 1992; Kanatt et al., 2007). Received Nov 14, 2017. Accepted Mar 05, 2018. Corresponding author: Mentha leaves have been traditionally used fresh and dried with various spices. Mentha species are comprised of biologically active components, which are used in traditional medicines. Additionally, mint species can be used in traditional medicine for common ailments, such as colds, coughs, sinusitis, fever, bronchitis, and nausea (Starburck, 2001; Dhifi et al., 2011). In addition, the mint plant has been reported to have insecticidal, antimicrobial, antispasmodic, antioxidant and antifungal activities (Papachristos and Stamopoulos, 2001; Shetty 2001; Nosrati, et al., 2011; Karagozlu et al., 2011). VOLUME 45 Nº1 JANUARY – APRIL 2018 The main components of the volatile oil of M. spicata L. are carvone, limonene, cis-carveol, 1,8 cineol and cis-sabinene hydrate, of which carvone is the most important constituent (Baser, 1993; Wyk and Wink, 2008). The antifungal and antibacterial activities of the volatile oil components have been defined in the literature. Volatile oil molecules are less-produced by the plant, but they act as defense mechanisms against predator attacks, such as pathogens and insects (Bayan and Aksit, 2016; Silva and Câmara, 2013). The objectives of this study were to determine the chemical components of volatile oils from M. spicata L. and to investigate their antibacterial and antifungal activities. Materials and methods Preparation of plant materials and volatile oil The M. spicata plant materials were collected from Kirsehir, Turkey in July 2016. The volatile oil was extracted from wet aerial parts by hydrodistillation in a Clevenger’s apparatus for 2 h. The volatile oil was obtained from plant materials and was then kept in the dark at 4 °C until it was used. Gas chromatography/mass spectrometry (GC/ MS) analysis GC/MS analyses were done in an Agilent Technologies 7890A GC System, 5975C using a TripleAxis Detector mass spectrometer with a built-in Autosampler, with the use of the HP-5MS capillary column (30 m × 0.25 mm × 0.25 mm). For GC/MS detection, an electron ionization system with an ionization energy of 70 eV was used. Helium was the transporter gas at a flow rate of 1 mL min-1. The column temperature program was the same as defined upstairs. As in the gas chromatography, 1.0 L split/splitless (10:1) of the sample, diluted with hexane, was transferred to the clone. Identification of oil components was successful by comparison of their mass spectral fragmentation model by the available mass library (WILLEY and NIST). 65 In vitro antifungal effect of the volatile oils The antifungal activities of volatile oil were determined using the agar well diffusion method (Tepe et al., 2005). The Potato Dextrose Agar (PDA) was autoclaved and cooled to 40 °C. Then, it was transferred to 60-mm petri dishes (10 ml petri-1). Next, 5-mm-diameter wells were opened on the PDA within the petri dishes. The plant volatile oils were applied at doses of 0.5, 1, 2, 4, 8 and 12 μl petri-1 into the wells. Mycelium disks of 5 mm were then placed at equal distances from these wells. The fungi transferred to the petri dishes were incubated at 22±2 °C. The inhibition in development was compared tothat in the control group, and percentile mycelial growth was calculated I=100 × (DC -DT)/DC I: Inhibition percentage compared to the control (mycelium development) DC: Mycelium development in the control DT: Mycelium development in volatile oil applications. Data were analyzed using the One-Way procedure of ANOVA (Windows version of SPSS, release 15.00). Differences among concentrations were compared using Duncan’s Multiple Range Test with a level of p<0.05. İn vitro antibacterial activity In the plant bacterial sample, eight different Xanthomonas spp. (ZI365, ZI366, ZI368, ZI370, ZI373, ZI375, ZI376, ZI378) strains were isolated from pepper fields in Kahramanmaras, Turkey. The bacteria cultures were grown in nutrient glucose agar solid medium, at 25 °C. After 24 h of growth, each bacteria strain, at a concentration of 0.1 OD density, was set with a spectrophotometer at 600 nm. Then, they were inoculated on the surface of nutrient glucose agar petri dishes with the aid of a drumstick. Later, filter paper discs (10 mm 66 CIENCIA E INVESTIGACIÓN AGRARIA in diameter) saturated with 10 µl of volatile oil were placed on the surface of each inoculated petri dish. The petri dishes were incubated at 25 °C for 48 h. Bacteria were evaluated by measuring the undeveloped zone. The experiment was performed with four duplicates and was repeated twice. Results and Discussion Chemical Composition results The compounds identified from M. spicata are shown in table 1. Thirty components, representing 100% of the total, were identified in M. spicata volatile oil. As the results of GC-MS analysis showed, the main component of M. spicata L. volatile oil was Car (...truncated)