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Journal of Experimental Biology and Agricultural Sciences, June - 2015; Volume – 3(3)

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Journal of Experimental Biology and Agricultural Sciences, June - 2015; Volume – 3(3) Journal of Experimental Biology and Agricultural Sciences, June - 2015; Volume – 3(3) Journal of Experimental Biology and Agricultural Sciences http://www.jebas.org ISSN No. 2320 – 8694 IMPACT OF GEOGRAPHIC’S VARIATION ON THE ESSENTIAL OIL YIELD AND CHEMICAL COMPOSITION OF THREE Eucalyptus SPECIES ACCLIMATED IN TUNISIA Elaissi Ameur1*, Medini Hanene1, Rouis Zied2, Khouja Mohamed Larbi3, Chemli Rachid1 and Harzallah-Skhiri Fethia1 1Laboratory of The Chemical, Galenic and pharmacological Drug Development, Faculty of Pharmacy, University of Monastir, Avenue Avicenne, 5019 Monastir, Tunisia 2Laboratory of Genetic, Biodiversity and Bio-resources Valorisation, Higher Institute of Biotechnology of Monastir, University of Monastir, Avenue Tahar Haddad, 5000 Monastir, Tunisia 3National Institute for Research on Rural Engineering, Water and Forestry, Institution of Agricultural Research and Higher Education, BP. N.2, 2080 Ariana, Tunisia Received – March 28, 2015; Revision – April 10, 2015; Accepted – June 15, 2015 Available Online – July 07, 2015 DOI: http://dx.doi.org/10.18006/2015.3(3).324.336 KEYWORDS ABSTRACT Eucalyptus Present study has been carried out to estimate the impact of geographical distribution on the yield and chemical constitute of three Eucalyptus verities viz E. cinerea F. Muell. ex Benth., E. astringens Essential oils Maiden and E. sideroxylon A.Cunn. ex Schauer-. These species were collected from six arboreta of 1,8-cineole Tunisia in January 2008. The essential oil was extracted by hydrodistillation method and estimated the essential oil yield which varies from 1.5±0.1% to 4.0±0.2%. Results of the study revealed that yield of ACP essential oil are not only depends on the Eucalyptus species but also depends on the origin of harvest. E. sideroxylon A. Cunn. exWoolls, cultivated in jbel abderrahman arboreta and E. cinerea F. Muell. ex HCA Benth. from choucha (sejnanae) arboreta provided the lowest and the highest percentage of essential oil amongst all the studied provenances, respectively. GC (RI) and GC/MS analysis showed the presence of GC-MS 163 components, representing 98.8 to 99.5% of the total oil. The contents of the different samples varied according to the species and the origin of harvest. The main components of the Eucalyptus essential oil were 1,8-cineole (39.1±0.0 – 79.4±0.0%), followed by α-pinene (2.1±0.0- 30.0±0.0), trans-pinocaveol All the article published by Journal of Experimental * Corresponding author Biology and Agricultural Sciences is licensed under a E-mail: [email protected] (Elaissi Ameur) Creative Commons Attribution-NonCommercial 4.0 International License Based on a work at www.jebas.org. Peer review under responsibility of Journal of Experimental Biology and Agricultural Sciences. Production and Hosting by Horizon Publisher (www.my- vision.webs.com/horizon.html). All _________________________________________________________rights reserved. Journal of Experimental Biology and Agricultural Sciences http://www.jebas.org 325 Ameur et al Chemotype (0.7±0.5-10.0±2.5%), α-terpineol (0.2±0.0 – 8.8±1.1%), globulol (0.3±0.0 – 5.7±1.3), aromadendrene (0.2±0.0 – 4.9±2.1%), limonene (0.6±0.0 – 4.3±0.8%), pinocarvone (0.2±0.0- 3.7±0.6%), Variation bicyclogermacrene (0.0 – 3.1±0.0%), α-terpinyl acetate (0.0-2.9±0.8%), p-cymene (0.7±0.0- 2.7±2.1%), β-carophyllene (0.3±0.0-1.9±0.0%), viridiflorol (0.2±0.0- 1.8±0.%), epiglobulol (traces – 1.3±0.6%) and spathulenol (0.2±0.0- 1.1±0.0%). The principal component and the hierarchical cluster analyses separated the Eucalyptus populations leaf essential oils into four groups, each constituting a chemotype. E. sideroxylon from Korbous and Sejnane arboreta afforded the highest mean percentage in 1,8-cineole. 1 Introduction best content in essential oil yield and 1,8-cineole, were select for this research work to identify the effect of the origin of Genus Eucalyptus comprises about 900 species and subspecies harvest on their essential oils yield and composition. (Pereira et al., 2014). More than 300 species of this genus contain volatile oil in their leaves. In 1957, total 117 2 Materials and Methods Eucalyptus have been introduced in Tunisia. They were used essentially as fire wood, for the production of mine wood and 2.1 Plant Materials against the erosion (Khouja et al., 2001). However, less than 20 species have ever been exploited commercially for the Clean and mature leaves of three Eucalyptus species L Hér., production of essential oil rich in 1,8-cineole (>70%) which is viz, E. astringens Maiden, E. cinerea F. Muell. ex Benth. and essentially used in the pharmaceutical and cosmetic industries E. sideroxylon A.Cunn. ex Schauer, were collected from six (Pino et al., 2002). In Tunisian folk medicine, inhalation of arboreta belonging to lower humid, higher humid and sub- Eucalyptus sp. essential oil has been traditionally used to treat humid bioclimatic stage of Tunisia in January 2008, Table 1. respiratory tract disorders such as pharyngitis, bronchitis and The leaves were dried in airy premises, shielded from the light, sinusitis (Boukef, 1986). Many studied have been then packed in paper bags, and kept in the shade. Botanical demonstrated their antibacterial, antifungal and antiviral voucher specimens of the collected species have been activities of Eucalyptus sp. essential oil against a wide range of deposited in the Herbarium of the Pharmacognosy Laborotary, microorganisms (Su et al., 2006;Cermelli et al., 2008; Gilles et Faculty of Pharmacy, Monastir, Tunisia (references 0156, al., 2010; Jha et al., 2014). Furthermore, allelopathic effect of 0157, 0158, 0159, 0160, 0.161, 0.162, 0.163, 0.164, 0.165, this essential oil against many weeds was also reported by 0166 and 0167). many researchers (Batish et al., 2004; Verdeguer et al., 2009; Rassaeifar et al., 2013). Eucalyptus essential oil was also 2.2 Sample preparation and extraction of Essential Oils reported as an effective anti-inflammatory, analgesic, antioxidant, antimutagenic, insecticide, nematocide and The essential oils were extracted by hydrodistillation method, acaricide oil (Batish et al., 2008; Bugarin et al., 2014; Rossi & for this 100 g of boorishly crushed Eucalyptus leaves for 4 h, Palacios, 2015).The Eucalyptus essential oils can be obtained using a standard apparatus recommended in the European by different methods, such as hydrodistillation, supercritical Pharmacopoeia. Hydrodistillations were performed in triplicate CO2 extraction, microwave-assisted extraction, and vacuum for each Eucalyptus species. The oil collected from each plant extraction by solvents. These methods affect the final yield of was dried (Na2SO4) and stored at 4°C until analysis. The yield the oil and it varied from 0.1± 0.1 to 5.7±0.5 7% (Elaissi et al., of essential oil was expressed in % (v/w) of the dry material 2010a; Elaissi et al., 2010b). High levels of 1,8-cineole (Elaissi et al., 2010a; Elaissi et al., 2010b). (87.8%) were found for the majority of the Eucalyptus species (Zrira et al., 2004; Elaissi et al., 2010c). Generally, the yields 2.3 Chemical analysis and composition of the oils varies and it depending on species, used part, plant origin zone, collection period, growth stage of 2.3.1 GC Analysis the plant as well extraction methods and storage conditions (Arango-Bedoya et al., 2012). Quantative and qualitative data of the essential oils were determined in triplicate by GC and GC/MS, respectively. GC In previous studies published on Eucalyptus species growing in Analysis was carried out with a Hewlett-Packard 6890 Tunisia, characterization of the leaves essential oils of 48 apparatus equipped with FID and a polar Carbowax column Eucalyptus species and their antibacterial activities were (30 m_0.32 mm i.d., film thickness 0.25mm). The oven carried out by the same authors. On the basis of this work three temperature was programmed isothermal at 35°C for 1 min, Eucalyptus species, E. cinerea F. Muell. ex Benth., E. rising from 35 to 250°C at 5°C/min, and then held isothermal astringens Maiden and E. sideroxylon A.Cunn. ex Schauer, at 250°C for 3 min; injector temp., 250°C; detector temp., which demonstrated the best antibacterial activity and also the 280°C; N2 used as carrier gas (1.2 ml/min). _________________________________________________________ Journal of Experimental Biology and Agricultural Sciences http://www.jebas.org Impact of Geographic’s variation on the essential oil yield and chemical composition of three Eucalyptus species acclimated in Tunisia. 326 Table 1 Provenances and Classification by Means of Duncan_s Multiple Range Test of the Average Essential Oil Yields of 3 Eucalyptus Species Eucalyptus Provenanace Latitude Longitude Altitude Annual Bioclimatique Soil type Abbreviation Yield [%] species (Arboreta) (m) rainfall stage (mm) E. cinerea Souiniat 35° 54' 8°48' 492 1140 humid sandstone cin Soui 3.7±0.7 inferior with hydromorphe (de)a) temperate winter E. cinerea Zerniza 30°94 9°7' 60 920 humid sandstone cin Zer 3.8±0.4 inferior with hydromorphe (de) warm winter E. cinerea Sejnane 37°3' 9° 14' 159 871 humid sandstone cin Sej 4.0±0.2 (e) inferior with temperate winter E. astringens Mrifek 37°07' 9°15' 60 950 humid Marl ast Mri 3.1±1.0(c) inferior with mild winter E. astringens korbous 36°50' 10°35' 180 540 sub-humid Sandy ast kor 3.2±0.1(c) with warm winter E. astringens Pryor 37°3' 9° 14' 159 871 humid sandstone ast Sej 3.3±0.3 Choucha inferior with (cd) (Sejnane) temperate winter E. Mrifek 37°07' 9°15' 60 950 humid Marl sid Mri 2.6±0.3 (b) sideroxylon inferior with mild winter E. Jbel 36°40' 10°40' 255 600 Sub-humid sandy clay sid JBA 1.5±0.1 (a) sideroxylon Abderrahmane with warm winter E.
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