Journal of Experimental Biology and Agricultural Sciences, March - 2014; Volume – 2(1S)
Journal of Experimental Biology and Agricultural Sciences
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ISSN No. 2320 – 8694
CORRELATION BETWEEN THE CHEMOTAXONOMIC CLASSIFICATIONS OF THE ESSENTIAL OILS OF 48 Eucalyptus SPECIES HARVESTED FROM TUNISIA AND THEIR PHYLOGENETIC CLASSIFICATION
Elaissi Ameur1*, Medini Hanene 1, 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 – January 24, 2014; Revision – February 19, 2014, Accepted – March 16, 2014 Available Online - March 31, 2014
KEYWORDS ABSTRACT Eucalyptus Various chemical classes (monoterpenes hydrocarbons, oxygenated monoterpenes, sesquiterpenes Essential oils hydrocarbons, oxygenated sesquiterpenes, esters, ketones, non classified coumpounds and non identified compounds) and twenty five of the main components from the essential oils of 48 Tunisian Eucalyptus 1, 8-cineole species has been reported. The compounds includes 1,8-cineole, torquatone, p-cymene, spathulenol, trans-pinocarveol, α-pinene, borneol, cryptone, 4-methyl-2-pentyl acetate, globulol, isoamyl isovalerate, ACP; HCA α-terpineol, (E,E)-farnesol, viridiflorol, aromadendrene, terpinen-4-ol, β-eudesmol, α-eudesmol, limonene, D-piperitone, caryophyllene oxide, β-phellandrene, bicyclogermacrene, α-phellandrene and GC-MS benzaldehyde, as a principal component when analysed by GC-MS.. The comparison of this classification to the phylogenetic classification showed a divergence for the majority of the species, Chemotype, taxonomie however some concordance was found.
* Corresponding author E-mail: [email protected] (Ameur Elaissi)
Peer review under responsibility of Journal of Experimental Biology and Agricultural Sciences.
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1 Introduction i.d., film thickness 0.25 μm) as described by Elaissi et al. (2010). Relative concentrations were calculated using the The genus Eucalyptus comprises more than 600 species software HP Chemstation, which allows assimilating the (Chippendale, 1988). More than 300 species of this genus percentages of the peak areas to the percentages of the various contain volatile oil in their leaves. However, less than 20 constituents. Retention indices were obtained by running a species have ever been exploited commercially for the series of aliphatic hydrocarbons (C9 - C28) in increasing the production of essential oil rich in 1, 8-cineole which is number of carbon atoms in the Carbowax GC column. essentially used in the pharmaceutical and cosmetic industries (Pino et al., 2002). In Tunisian folk medicine, inhalation of 2.3.1 GC/MS analysis Eucalyptus sp. essential oil has traditionally been used to treat respiratory tract disorders such as pharyngitis, bronchitis, and Analyses of the composition of the essential oils were carried sinusitis (Boukef, 1986). Many studied were demonstrated out using a Hewlett-Packard (HP) 5890 series II gas their antibacterial, antifungal and antivirus activities against a chromatography apparatus equipped with a polar column wide range of microorganisms (Su et al., 2006; Cermelli et al., Carbowax (30m x 0.32 mm i.d., film thickness 0.25 μm) and 2008; Martin et al., 2010) furthermore, allelopathic effect of 5972 mass selective detectors. Helium was used as the carrier oil was also reported against many weeds (Batish et al. 2004; gas. The mass spectrometer operating conditions were: Verdeguer et al., 2009). In 1957, total 117 Eucalyptus species ionisation voltage, 70 eV, ion source 230°C. The GC/MS of Eucalyptus has been introduced in Tunisia. Local population parameters were identical to those for the GC analysis. were used tree for fire wood, production of mine wood, and in the fight against erosion (Khouja et al., 2001). The Eucalyptus 2.3.2 Compound Identification species were grouped by Pryor & Johnson (1971) into seven subgenera which followed by various sections, series, subseries The identification of the compounds was based on a and super species. Brooker (2000) presented a new comparison of retention indices (determined relatively to the classification that divided genus Eucalyptus into seven and six retention time of aliphatic hydrocarbons (C9 - C28), of the monotypic polytypic subgenera. Current study evaluates mass spectra with those of authentic compounds by means of whether the chemical composition of essential oils from 48 NBS75K.L. and Wiley 275 databases (Wiley & Sons, 1998) species of Eucalyptus from Tunisia align with the grouping of and with data in the literature. species proposed by Brooker (2000). 2.4 Statistical analysis 2 Materials and methods Average twenty-five compounds were detected from each oil 2.1 Plant Material sample at a concentration greater than 4.4%. The differences in their mean percentages were analysed using Duncan's Multiple Leaf samples were collected from total 48 species of Range. These compounds were also subjected to a Principal Eucalyptus trees acclimated in five arboreta throughout the Component Analysis (PCA) and a Hierarchical Cluster Centre and the North of Tunisia in June 2006 and in June 2007 Analysis (HCA) using SPSS 12.0 software (SPSS Inc. (Table 1). Botanical voucher specimens of all selected species Chicago, IL, USA) to evaluate whether the composition of the have been deposited in the Pharmacognosy Laborotary essential oils could be used to classify the species into groups Herbarium in the Faculty of Pharmacy, Monastir, Tunisia and whether these groups reflected those proposed by Brooker (Table1). (2000).
2.2 Sample preparation and extraction of essential oils 3 Results and Discussion
Dried leaves from each sample were removed and stored in 3.1 Chemical composition brown paper bags. Three samples of boorishly crushed leaves (3 x 100 g) from each tree were hydro-distilled for 4 h in a The chemical composition of essential oils obtained from 48 standard apparatus recommended in the European Eucalyptus species from Tunisia can be grouped together into Pharmacopoeia (2000). Essential oils were collected, dried eight classes (Table 2) on the basis of their chemical over anhydrous Na2SO4 and stored at 4°C until analysis. properties. In most species, the essential oils were dominated by oxygenated monoterpenes (6.9 to 86.7 %), especially 1,8- 2.3 Chemical analysis cineole (2.3 to 70.4 %), which was most abundant compound and reported from the oil sample of 32 species (Table 3), This Quantitative and qualitative data for all essential oils were concentration was followed by the trans-pinocarveol (0.0 – determined for triplicate by GC and GC/MS, respectively. GC 25.7 ± 0.4%) which was reported from the essential oils of was carried out using HP 6890 chromatography apparatus three species. equipped with FID and Carbowax column (30 m x 0.32 mm
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Correlation between the Chemotaxonomic Classifications of the essential oils of 48 Eucalyptus species harvested . . . 100
Table 1 List of studied Eucalyptus species along with their source, abbreviation and voucher specimen numbers.
N° Species Abbreviation Provenance Voucher number 1. E. ficifolia F.Muell. fic K5 0136 2. E. gomphocephala gom K6 0137 3a E. botryoides var. botryoides Sm. Morocco bot m Z1 0143 4. E. exserta F.Muell. exe S3 0121 5. E. gunnii F. Muell. gun S5 0123 6. E. brockwayi C.A.Gardner. broc H1 0107 7. E. fasciculosa F. Muell. fasc Z6 0148 8. E. macrorrhyncha F.Muell. macro S7 0125 9. E. diversifolia Bonpl. divfo K3 0134 10. E. kitsoniana Maiden kit JBA3 0153 11. E. pauciflora Sieber ex Sprengel. pau S10 0128 12. E. populifolia Hook. pop JBA5 0155 13. E. falcata Turcz. falc K4 0135 14. E. leucoxylon F.Muell. leuc JBA4 0154 15. E. ovata Labill. ova Z8 0150 16. E. occidentalis Endl occ H6 0112 17. E. camaldulensis Dehnh. cam K2 0133 3b E. botryoides var. botryoides Sm. Vilmorin bot v Z2 0144 18. E. largiflorens F. Muell. larg H4 0110 19. E. polyanthemos Schauer poly K10 0141 20. E. dundasii Maiden. dun JBA1 0151 21. E. tereticornis Sm. ter S12 0130 22. E. odorata Behr odo S9 0127 23. E. diversicolor F.Muell. divco Z5 0147 24. E. rudis Endl. rud K11 0142 25. E. gigantea Dehnh. gig S4 0122 26. E. cladocalyx F.Muell. cla Z3 0145 27. E. platypus Hook. plat K9 0140 28. E. strciklandii Maiden. str H10 0116 29. E. macarthurii Dean & Maiden. macar S6 0124 30. E. viminalis Labill. vim S13 0121 31. E. maculate Hook. macu K8 0139 32. E. grandis W. Hill gran Z7 0149 33. E. Sideroxylon A.Cunn. ex Schauer. sid S11 0129 34. E. woodwardii Maiden. wood H12 0118 35. E. bicostata Maiden. Blakely & Simmonds bic S1 0.119 36. E. torquata Luehm. torq H11 0117 37. E. salmonophloia F.Muell. salm H8 0114 38. E. maidenii Maiden. maid S8 0126 39. E. gillii Maiden gill H3 0109 40. E. citriodora Hook. citr Z4 0146
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41. E. lehmannii (Schauer) Benth. leh K7 0138 42. E. globulus Labill. glob JBA2 0152 43. E. astringens Maiden ast K1 0132 44. E. loxophleba Benth. lox H5 0111 45. E. cinerea F.Muell. ex Benth. cin S2 0120 46. E. oldfieldii Muell. old H7 0113 47. E. gracilis F.Muell. grac H2 0108 48. E. sargentii Maiden. sarg H9 0115 a) and b) E. Botryoides var. botryoides from two provenances. K): Korbous (North East of Tunisia). Z): Zerniza (Norh West of Tunisia). S): Souiniat (North of Tunisia). H): Hajeb Laayoun (Centre of Tunisia). JBA): Jbel Abderrahaman (North East of Tunisia).
Table 2 The chemical classes of the essential oils extracted from leafs of 48 Eucalyptus species.
Compounds
classes
erpene
Monot hydrocarbons Oxygentaed monoterpenes Sesquiterpene hydrocarbons Oxygentaed sesquiterpenes Esters ketones Others Not identifid N° Abbreviation m-hyd oxy-m ses-hyd oxy-ses est ket oth n-ide Species Percentage (%) 1. E. maidenii 20.0±4.6 66.4±2.9 3.1±1.0 8.9±2.3 0.2±0.1 0.1±0.0 0.0 1.3±0.4 2. E. cinerea 9.9±1.2 86.7±1.9 0.8±0.2 1.5±0.6 0.1±0.0 0.1±0.0 0.0 1.0±0.1 3. E. pauciflora 5.4±2.3 15.5±1.6 5.1±0.2 62.7±4.9 1.7±01.3 0.3±0.0 0.2±0.0 9.2±3.7 4. E. macrorrhyncha 8.3±5.4 33.9±8.5 2.2±0.8 47.8±6.3 0.4±0.5 0.7±0.5 tr 6.6±2.8 5. E. macarthurii 12.2±2.4 67.2±4.5 4.2±0.5 11.6±2.5 0.5±0.1 0.5±0.4 0.3±0.1 3.4±0.7 6. E. gunnii 9.5±5.4 6.9±0.4 10.9±4.1 51.7±0.4 1.8±0.3 1.1±0.7 0.0 18.3±1.7 7. E. sideroxylon 12.5±1.0 79.4±0.6 2.9±0.6 3.8±0.8 0.3±0.1 0.1±0.0 0.0 1.1±0.1 8. E. odorata 19.2±6.6 34.1±0.4 2.3±0.2 9.6±2.0 0.5±0.1 21.8±1.5 0.0 12.6±3.0 9. E. bicostata 6.5±2.2 79.6±5.7 3.4±1.1 8.1±1.9 0.3±0.2 0.2±0.0 0.0 2.0±0.9 10. E. exserta 7.1±2.7 20.7±2.2 5.3±2.7 52.7±6.9 1.3±0.5 0.2±0.2 0.2±0.1 12.6±4.1 11. E. tereticornis 31.3±8.0 24.1±5.0 1.9±1.0 27.8±9.1 0.2±0.2 9.6±12.6 0.0 5.1±0.3 12. E. viminalis 6.1±1.0 70.5±10.4 6.1±4.4 13.3±2.5 0.7±0.5 0.2±0.1 0.3±0.2 2.9±1.9 13. E. gigantea 8.1±2.1 59.0±6.4 6.3±1.2 19.1±1.9 0.3±0.2 0.9±0.1 0.1±0.1 6.2±0.8 14. E. maculata 7.5±0.7 40.6±1.8 18.8±0.6 24.0±0.8 0.4±0.5 1.0±0.1 0.0 7.7±1.6 15. E. diversifolia 5.8±2.9 57.9±14.9 5.0±2.0 20.8±5.3 0.2±0.3 0.6±0.4 0.1±0.1 9.6±7.9 16. E. polyanthemos 6.1±3.4 73.4±14.4 2.8±1.5 12.0±6.2 1.2±0.1 1.6±1.7 tr 3.0±1.4 17. E. falcata 7.2±1.8 70.7±3.5 3.5±0.1 13.7±3.6 0.3±0.3 0.4±0.1 t 4.3±1.3 18. E. platypus 19.3±3.4 40.6±7.5 4.1±0.9 30.6±4.9 0.8±0.2 0.2±0.1 0.0 4.6±1.6 19. E. lehmannii 25.6±7.9 69.4±7.2 1.1±0.2 3.1±.0.4 0.1±t0.0 0.1±t0.0 0.0 0.6±0.2 20. E. camaldulensis 12.8±4.6 23.2±1.0 2.0±0.7 38.5±3.7 0.5±0.3 13.6±1.0 0.2±0.1 9.0±1.3 21. E. gomphocephala 4.3±2.1 45.1±5.9 4.1±1.0 16.4±1.9 2.7±0.4 1.4±0.0 0.1±0.1 26.0±5.9 22. E. ficifolia 14.6±6.7 15.7±1.7 14.5±4.3 43.6±5.4 0.7±0.2 1.3±0.7 0.1±0.1 9.6±2.0 23. E. astringens 24.3±4.9 57.8±7.7 4.9±1.5 9.7±1.8 0.3±0.0 0.2±0.0 0.0 2.9±0.9 24. E. rudis 26.5±5.8 21.6±3.3 4.0±0.7 27.6±4.9 0.2±0.1 7.5±3.1 0.0 12.6±0.8 25. E. kitsoniana 18.8±12.8 51.3±9.2 3.9±1.0 18.3±12.0 0.2±0.2 1.2±1.3 0.1±0.2 6.2±3.3 26. E. populifolia 4.0±1.6 65.2±0.8 4.7±1.0 19.2±2.1 0.7±0.3 0.5±0.1 0.1±0.0 5.6±0.9
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Correlation between the Chemotaxonomic Classifications of the essential oils of 48 Eucalyptus species harvested . . . 102
27. E. globulus 15.9±4.3 66.1±3.0 6.0±2.8 10.1±3.6 0.3±0.1 0.3±0.1 tr 1.3±0.3 28. E. leucoxylon 13.6±1.4 70.4±9.0 3.7±1.9 9.1±4.3 0.6±0.3 0.3±0.1 tr 2.4±1.1 29. E. dundasii 29.7±3.9 53.1±2.5 3.2±1.7 8.9±3.5 1.9±0.4 0.2±0.2 0.1±0.0 2.9±1.4 30a) E. botryoides v 41.4±2.8 30.1±3.3 4.4±0.7 13.2±4.3 0.9±0.2 3.0±1.1 tr 7.5±3.9 31. E. fasciculosa 16.2±0.0 63.9±3.0 3.1±0.1 13.2±2.3 0.3±0.1 0.7±0.1 0.0 2.7±0.5 30b E. botryoides m 23.8±7.2 43.2±12.9 1.8±0.6 15.9±11.5 0.6±0.3 6.2±1.5 4.6±4.5 4.0±2.1 32. E. diversicolor 37.5±1.3 22.3±2.7 2.6±1.3 13.7±8.5 0.6±0.1 17.3±6.0 0.2±0.0 5.8±0.4 33. E. ovata 16.7±2.5 64.6±4.9 2.6±1.3 12.9±2.3 0.3±0.1 0.2±0.1 0.1±0.0 2.8±1.6 34. E. grandis 27.5±6.7 26.9±3.2 11.3±4.5 22.6±7.1 0.3±0.1 6.1±1.9 0.2±0.0 5.2±0.6 35. E. cladocalyx 34.6±1.7 56.1±2.1 1.2±0.1 4.1±0.6 1.6±0.1 0.4±0.0 tr 2.0±0.2 36. E. citriodora 32.1±4.8 64.8±5.3 0.5±0.1 1.3±0.5 0.6±0.3 0.2±0.1 0.0 0.6±0.1 37. E. woodwardii 23.8±2.0 33.6±2.9 5.5±0.4 15.7±0.8 0.5±0.2 18.1±1.8 0.0 2.8±0.1 38. E. stricklandii 13.0±4.7 34.3±11.0 3.5±0.9 10.0±0.1 1.3±0.4 30.9±1.7 0.0 7.0±4.2 39. E. occidentalis 9.9±10.3 32.3±7.0 17.3±4.1 32.3±9.8 0.7±0.1 0.4±0.3 0.0 7.1±3.0 40. E. brockwayii 2.8±0.2 35.0±2.6 8.3±0.2 26.6±3.7 19.6±5.3 0.6±0.7 0.0 7.2±0.3 41. E. almonopholoia 35.1±10.7 48.4±6.2 2.2±1.9 4.3±1.4 1.1±0.7 5.4±5.5 0.00 3.4±1.5 42. E. gillii 34.7±11.2 30.2±8.7 4.9±1.9 23.0±2.7 0.3±0.1 2.6±2.2 0.00 4.4±1.1 43. E. oldfieldii 10.3±2.8 70.3±0.9 5.9±0.9 10.4±1.3 0.8±0.2 0.3±0.0 0.0 2.1±0.1 44. E. largiflorens 8.60±2.1 72.6±0.2 3.8±0.1 11.9±1.2 0.4±0.0 0.2±0.1 tr 2.4±1.2 45. E. loxophleba 6.2±0.1 39.7±1.0 3.8±0.8 22.9±1.1 21.9±1.6 0.2±0.0 tr 5.3±0.6 46. E. sargentii 22.3±1.4 64.0±4.7 4.2±0.3 7.7±2.6 0.4±0.1 0.2±0.1 0.0 1.2±0.1 47. E. gracilis 16.6±4.2 64.3±2.3 2.5±0.2 9.2±3.7 0.5±0.5 2.4±0.4 0.0 4.6±2.0 48. E. torquata 13.4±2.8 20.2±5.0 2.1±0.2 19.1±2.8 0.1±0.1 43.2±0.9 0.0 1.9±0.1 a): E. Botryoides var. botryoides from Vilmorin. Italy (v). b): E. Botryoides var. botryoides from Morocco (m).
The concentration of oxygenated sesquiterpenes varied from The chemical variation between species of different series is 0.5 to 62.7 % with spathulenol (0.1 to 28.0 %) being the further summarized by the ACP in Figure 1 and Figure 2. This important component of five species (Table 2). Ketones were dendrogram indicates that E. botryoides from Morocco also reported from two species with mean percentage varied differed significantly from E. brockwayi and E. loxophleba from 0.1 to 43.2 % (Table 2). The monoterpenes hydrocarbons considered as being remote according to their morphological varied from 2.8 to 41.4 % with p-cymene being the most criteria. The leaf oil of E. botryoides from Morocco (series abundant compound in two species (Table 2 and Table 3). The Annulares) was characterized by a high mean percentage of esters varied from 0.1 to 19.6 % while the sesquiterpenes ketones and non classified compounds (others) (23.8±7.2, hydrocarbons varied from 0.1 to 18.8 %. 4.6±4.5%, respectively), however E. brockwayi (series Brockwayanae) and E. loxophleba (series Loxophlebae) oils 3.1.1 Principal components analysis (PCA) and hierarchical were characterized by their richness in esters (19.6±5.3, cluster analysis (HCA) of the chemical classes 21.9±1.6%, respectively) and in sesquiterpenes hydrocarbons (8.3±0.2, 3.8±0.8%, respectively). By principal components analysis (PCA) and hierarchical cluster analysis (HCA) several major chemical classes like The three species aligned together in their disposition an monoterpenes hydrocarbons, oxygenated monterpens, almost equal amount of oxygenated monoterpenes and sesquiterpenes hydrocarbons, oxygenated sesquiterpens, esters, sesquiterpenes, whereas Brooker (2000) classified them in ketones, and some other non identified components were three different series. E. botryoides from vilmorin classified reported from the various species of Eucalyptus (Table 1). The closer to E. botryoides from Morocco by Brooker (2000), PCA horizontal axis explained 31.50% of the total variance however their essential oils were characterized by completely while the vertical axis a further 19.09% (figure 1). Within a different chemotypes. In fact E. botryoides from vilmorin was dissimilarity >10, HCA (figure 2) indicated 8 groups of species very closer to E. gillii (subseries Decussatae) in the oils. Each one was considered as a chemotype. Group eight dendrogramm (Figure 1), their essential oils were characterized was divided into 5 subgroups within a dissimilarity > 3. by the highest mean percentage of the momonterpenes hydrocarbons (41.4±2.8, 34.7±11.2%, respectively).
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Table 3 Content (%) of 25 compounds selected for the principal component and the hierarchical cluster analyses in the essential oils extracted from the leafs of 48 Eucalyptus Species.
Species maidd) cin pau macro macar gun sid odo bic exe ter vim gig Compounds and abbreviations KIc) 1 2 3 4 5 6 7 8 9 10 11 12 13 α-Pinene (c1) 1053 7.3±0.7 4.5±0.7 0.2±0.0 5.7±4.3 6.3±1.5 3.5±2.2 6.9±1.1 1.0±0.7 3.7±1.2 0.3±0.1 1.2±0.1 1.7±0.8 1.0±0.0 4-Methyl-2- pentyl acetate (c2) 1114 -e) ------α-Phellandrene (c3) 1172 0.2±0.1 - trf) 0.3±0.3 - 0.5±0.4 0.2±0.1 0.1±0.1 - 0.2±0.1 0.2±0.0 - - Limonene (c4) 1209 3.1±0.2 3.7±0.5 0.2±0.1 0.6±0.2 2.7±0.7 0.3±0.0 4.1±0.1 0.4±0.2 0.9±0.5 0.4±0.2 10.2±12.5 3.1±2.1 4.1±2.0 β-Phellandrene (c5) 1215 - - - - 0.4±0.7 - - - - 0.3±0.4 - - - 1.8-Cineole (c6) 1218 57.8±1.9 70.4±2.5 2.4±0.9 13.0±4.1 54.8±4.2 2.6±0.8 69.2±0.6 4.5±1.6 68.0±5.3 2.9±3.2 3.8±1.1 62.5±6.2 40.8±4.8 p-Cymene (c7) 1282 7.4±2.9 1.2±0.1 4.7±2.2 1.4±0.6 1.8±1.0 4.0±1.4 0.8±0.1 16.7±5.2 1.4±0.5 5.7±2.3 17.5±2.2 0.9±0.6 2.4±0.2 Isoamyl isovalerate (c8) 1304 0.1±0.1 - - - 0.2±0.0 - 0.1±0.1 - tr - - 0.3±0.3 - Benzaldehyde (c9) 1541 ------0.2±0.3 - Terpinene-4-ol (c10) 1618 1.1±0.3 0.4±0.1 0.8±0.3 2.3±0.2 0.4±0.1 0.5±0.0 0.6±0.0 3.0±0.8 0.2±0.1 1.4±0.5 2.0±1.8 0.5±0.4 0.7±0.1 Aromadendrene (c11) 1625 1.6±0.8 0.1±0.0 1.6±0.3 0.4±0.2 2.8±0.3 1.0±0.35 0.5±0.2 0.4±0.0 2.0±0.8 1.4±0.7 0.2±0.1 3.7±3.2 0.3±0.0 trans-Pinocarveol (c12) 1675 2.0±0.8 1.0±0.2 0.1±0.1 0.1±0.0 1.2±0.20 0.2±0.1 1.2±0.1 0.8±0.2 4.6±0.6 0.1±0.0 0.6±0.7 2.4±3.8 0.3±0.0 Cryptone (c13) 1695 - - 0.1±0.1 0.5±0.6 0.3±0.5 0.1±0.0 tr 20.9±1.3 - 0.1±0.1 9.2±12.4 - 0.6±0.1 α-Terpineol (c14) 1713 2.2±0.2 10.3±1.1 1.3±0.3 3.6±1.4 1.9±0.8 - 5.4±0.9 0.8±0.2 0.6±0.3 1.1±0.4 0.6±0.4 1.8±1.1 11.5±2.1 Borneol (c15) 1720 0.1±0.1 0.2±0.1 tr 0.1±0.1 0.2±0.1 - 0.2±0.1 0.1±0.0 0.2±0.0 0.1±0.1 0.1±0.1 0.1±0.2 0.1±0.0 d-Piperitone (c16) 1751 0.1±0.2 tr 5.3±1.7 9.1±8.1 0.1±0.0 0.1±0.0 - 0.6±0.1 0.1±0.2 9.9±2.8 0.2±0.1 0.1±0.1 - Bicyclogermacrene (c17) 1758 tr tr 0.2±0.1 - - 1.6±0.6 0.6±0.2 tr - - - - - Caryophyllene oxide (c18) 2015 0.1±0.0 tr 0.2±0.1 1.3±1.5 0.1±0.1 0.8±0.2 0.1±0.1 1.7±0.2 tr 0.7±0.8 9.1±10.3 0.2±0.2 1.9±2.0 Globulol (c19) 2103 1.7±1.4 0.6±0.2 12.6±4.5 4.9±2.4 6.0±0.9 12.4±2.2 1.1±0.3 0.8±0.2 5.4±1.2 9.2±1.2 - 6.1±4.4 2.5±0.2 Viridiflorol (c20) 2113 0.7±0.3 0.2±0.1 14.1±2.1 2.7±2.0 0.8±0.1 11.5±2.4 0.4±0.1 1.0±0.3 0.8±0.2 7.7±3.3 0.5±0.1 1.1±0.8 0.7±0.1 Spathulenol (c21) 2151 0.1±0.1 0.2±0.1 18.0±0.9 5.3±2.0 2.0±2.9 16.3±6.2 0.5±0.3 3.2±0.9 0.1±0.0 12.3±2.9 12.7±2.5 0.5±0.3 4.6±1.5 α-Eudesmol (c22) 2252 1.1±0.6 - 0.7±1.3 10.8±4.3 0.1±0.0 - 0.4±0.1 - 0.1±0.0 4.5±2.6 - 0.2±0.1 1.6±0.3 β-Eudesmol (c23) 2262 3.0±1.9 0.2±0.2 2.3±2.1 11.9±4.7 0.2±0.1 1.3±0.2 0.6±0.1 0.1±0.2 tr 7.7±4.8 0.1±0.0 2.6±4.2 3.0±1.0 (E.E)-Farnesol (c24) 2376 - - 0.6±0.2 0.2±0.1 - 0.3±0.1 tr 0.2±0.1 - 0.4±0.5 0.6±0.7 - 0.2±0.1 Torquatone (c25) 2437 0.1±0.1 - tr - 0.1±0.1 0.3±0.1 - 0.5±0.2 0.2±0.1 - 0.2±0.3 0.1±0.1 0.1±0.0 c) KI: Kovats index determined on Carbowax GC column. d) The abbreviations of the Eucalyptus species are given inTable 4. e) –: Not detected. F): tr: Trace (<0.1%).
______Journal of Experimental Biology and Agricultural Sciences http://www.jebas.org
Correlation between the Chemotaxonomic Classifications of the essential oils of 48 Eucalyptus species harvested . . . 104
Tab le 3 cont.
Species macu divfo poly falc plat leh cam gom fic ast rud kit pop Compounds and abbreviations KI 14 15 16 17 18 19 20 21 22 23 24 25 26 α-Pinene (c1) 1053 1.3±0.0 3.5±3.4 1.5±1.0 5.9±1.6 9.4±0.7 17.6±7.5 0.3±0.1 0.7±0.2 9.0±4.5 21.3±4.4 0.6±0.2 9.5±8.0 1.9±1.0 4-Methyl-2- pentyl acetate (c2) 1114 ------α-Phellandrene (c3) 1172 - - 0.1±0.1 - 0.6±0.3 - tr - - 0.1±0.1 0.3±0.2 tr - Limonene (c4) 1209 3.4±0.3 0.8±0.1 1.8±0.4 0.6±0.2 0.5±0.1 4.4±0.3 0.4±0.2 0.3±0.1 0.9±0.5 1.3±0.2 0.8±0.4 1.0±0.8 1.2±0.7 β-Phellandrene (c5) 1215 ------7.7±3.2 - - 1.8-Cineole (c6) 1218 34.6±1.9 36.9±12.3 57.8±12.6 30.9±4.4 22.5±4.7 56.6±4.3 3.9±2.0 6.1±2.3 4.2±3.2 43.7±5.2 2.3±2.0 4.7±3.2 47.2±9.2 p-Cymene (c7) 1282 1.1±0.1 0.9±0.5 2.5±1.8 0.4±0.1 7.6±3.9 2.0±0.23 11.8±4.3 2.7±2.0 0.5±0.3 0.8±0.2 16.4±1.7 6.7±8.1 0.5±0.2 Isoamyl isovalerate (c8) 1304 - 0.1±0.2 0.8±0.1 - - - - 0.8±0.4 - - - - 0.3±0.3 Benzaldehyde (c9) 1541 ------Terpinene-4-ol (c10) 1618 0.9±0.0 0.6±0.2 0.8±0.5 0.1±0.0 1.3±0.6 0.3±0.1 1.9±0.4 3.6±1.4 0.9±0.2 0.3±0.1 2.2±0.1 0.5±0.4 0.5±0.1 Aromadendrene (c11) 1625 0.1±0.0 2.7±2.2 0.4±0.0 2.0±0.1 1.0±0.6 0.2±0.0 0.6±0.6 1.2±0.1 3.3±0.6 3.2±1.0 0.7±0.3 1.5±0.8 2.3±0.9 trans-Pinocarveol (c12) 1675 0.4±0.0 7.0±4.6 1.3±0.6 25.7±0.4 8.3±2.4 1.0±0.2 0.3±0.1 12.6±5.1 0.6±0.1 7.5±2.0 0.1±0.1 21.6±9.8 6.6±5.4 Cryptone (c13) 1695 0.1±0.0 tr 1.4±1.7 0.1±0.1 0.1±0.1 - 12.9±0.9 0.1±0.1 0.1±0.1 0.1±0.1 7.0±3.3 0.2±0.1 - α-Terpineol (c14) 1713 1.1±0.5 2.7±1.9 6.4±0.9 1.7±0.2 1.3±0.4 8.7±2.5 0.4±0.1 7.3±2.8 3.4±0.8 1.2±0.3 0.7±0.2 4.5±1.2 2.6±0.3 Borneol (c15) 1720 - 0.7±0.5 0.2±0.1 1.2±0.1 0.3±0.1 0.5±0.0 0.1±0.0 2.1±0.5 0.5±0.6 0.4±0.1 tr 4.4±0.8 0.8±0.7 d-Piperitone (c16) 1751 - 0.4±0.2 0.1±0.1 - - - 0.3±0.1 0.1±0.0 - - 0.2±0.1 - - Bicyclogermacrene (c17) 1758 - - 0.6±0.9 - 0.5±0.3 - tr - 2.2±1.6 - 0.2±0.1 - - Caryophyllene oxide (c18) 2015 1.6±0.1 0.1±0.1 2.2±1.4 0.1±0.1 1.1±0.8 0.1±0.1 3.9±4.1 0.3±0.1 0.5±0.1 0.²±0.0 0.2±0.1 0.1±0.1 0.1±0.1 Globulol (c19) 2103 1.8±0.1 6.4±5.6 2.2±0.1 6.7±1.9 6.3±2.1 0.6±0.2 1.1±0.8 7.8±1.0 6.4±2.3 5.7±1.0 1.0±0.5 9.1±7.9 12.8±1.8 Viridiflorol (c20) 2113 0.1±0.0 6.9±7.9 1.0±0.4 1.9±0.8 3.8±1.3 0.2±0.1 1.0±0.3 0.7±0.1 2.1±0.4 1.0±0.2 0.3±0.3 1.2±0.5 1.8±0.1 Spathulenol (c21) 2151 1.4±0.0 0.4±0.3 4.5±3.5 1.3±0.8 11.2±6.1 1.0±0.1 28.0±7.6 0.8±0.4 7.0±2.0 0.8±0.1 19.8±4.7 1.6±0.8 0.3±0.0 α-Eudesmol (c22) 2252 6.3±0.4 1.4±2.1 - 0.3±0.1 - 0.3±0.1 - 0.3±0.1 2.3±1.6 0.1±0.0 - 0.2±0.1 0.2±0.2 β-Eudesmol (c23) 2262 - - 0.2±0.1 0.7±0.4 1.3±0.3 0.3±0.1 - 0.9±0.2 2.1±1.4 0.1±0.0 0.2±0.0 0.1±0.1 0.2±0.2 (E.E)-Farnesol (c24) 2376 1.3±1.2 0.2±0.2 0.1±0.1 0.2±0.1 0.1±0.2 0.1±0.1 0.4±0.4 - 15.0±8.7 - 0.1±0.1 0.9±0.8 0.1±0.1 Torquatone (c25) 2437 0.2±0.1 0.5±0.4 0.1±0.1 0.2±0.1 0.1±0.1 tr 0.4±0.2 0.1±0.1 - 0.1±0.0 0.1±0.1 0.7±1.1 0.4±0.1 c) KI: Kovats index determined on Carbowax GC column. d) The abbreviations of the Eucalyptus species are given inTable 4. e) –: Not detected. F): tr: Trace (<0.1%).
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105 Elaissi et al
Table 3 cont.
Species glob leuc dun bot v fasc bot m divco ova gran cla citr wood str Compounds and abbreviations KI 27 28 29 30a) 31 30b) 32 33 34 35 36 37 38 α-Pinene (c1) 1053 12.1±3.0 7.8±2.3 23.6±4.3 27.5±11.5 1.6±0.1 9.0±3.6 0.9±0.2 13.5±2.8 3.3±1.8 0.8±0.2 23.6±3.9 21.2±2.2 11.3±4.5 4-Methyl-2- pentyl acetate (c2) 1114 ------0.2±0.0 - 0.4±0.3 1.3±0.4 α-Phellandrene (c3) 1172 - - - - 0.4±0.0 0.3±0.2 0.7±0.0 0.2±0.1 6.0±4.0 - 0.1±0.1 0.2±0.0 0.1±0.1 Limonene (c4) 1209 2.4±1.2 2.0±0.5 2.1±0.8 2.2±0.4 1.7±0.1 1.2±0.4 4.2±3.5 1.0±0.2 1.0±0.4 3.8±0.4 2.5±1.1 1.2±0.2 0.7±0.5 β-Phellandrene (c5) 1215 - - 0.9±1.6 0.0±0.0 - - 4.3±2.4 - 1.7±3.0 0.0 0.6±1.0 - - 1.8-Cineole (c6) 1218 53.8±3.6 59.2±10.1 44.5±2.5 14.5±3.6 55.3±4.4 22.6±7.3 2.5±0.3 41.6±4.2 15.9±4.9 3.0±0.4 54.1±3.1 25.3±0.1 20.4±0.0 p-Cymene (c7) 1282 0.7±0.2 2.9±3.4 1.0±0.4 9.5±8.5 11.2±0.1 11.5±2.6 24.8±2.5 1.4±0.6 11.7±2.8 24.1±1.7 2.8±0.1 0.9±0.2 0.7±0.1 Isoamyl isovalerate (c8) 1304 tr 0.3±0.2 1.2±0.5 0.1±0.0 0.1±0.1 - - tr - 0.2±0.0 0.4±0.4 0.1±0.1 - Benzaldehyde (c9) 1541 - - - - - 4.4±4.6 ------Terpinene-4-ol (c10) 1618 0.2±0.1 0.2±0.2 0.3±0.1 0.6±0.4 1.0±0.2 1.1±0.2 2.2±0.4 0.3±0.1 2.4±0.4 0.9±0.1 0.1±0.1 0.3±0.1 0.2±0.1 Aromadendrene (c11) 1625 3.4±1.9 2.1±1.4 1.4±0.6 0.3±0.2 0.3±0.1 0.2±0.1 0.1±0.0 0.7±0.9 0.6±0.9 0.1±0.0 0.0±0.1 3.1±0.4 1.9±0.5 trans-Pinocarveol (c12) 1675 3.7±1.8 4.3±1.0 3.5±2.2 0.0±0.0 0.0±01.0 5.4±5.8 0.1±0.1 13.8±1.9 0.3±0.2 0.2±0.1 2.3±0.9 4.9±2.8 7.5±7.6 Cryptone (c13) 1695 0.1±0.1 0.0 tr 0.2±0.2 0.6±0.1 6.0±1.5 17.0±6.0 0.1±0.1 5.5±2.4 0.1±0.1 0.1±0.1 0.1±0.0 0.1±0.1 α-Terpineol (c14) 1713 3.3±2.2 1.6±0.7 1.5±0.3 4.3±0.2 2.4±0.4 1.1±0.4 0.8±0.2 1.7±0.2 0.5±0.2 18.0±4.5 3.0±0.1 0.7±0.1 0.7±0.4 Borneol (c15) 1720 0.2±0.1 0.3±0.0 0.2±0.1 3.3±0.7 0.2±0.1 0.2±0.2 0.1±0.1 0.9±0.1 - 24.8±4.2 1.5±0.4 0.2±0.1 0.4±0.3 d-Piperitone (c16) 1751 - - - tr - 0.3±0.1 - 0.0 0.1±0.0 - - - - Bicyclogermacrene (c17) 1758 - - 0.6±0.7 - 0.7±0.1 - 0.1±0.1 tr 6.8±3.1 0.2±0.0 0.1±0.1 - 0.1±0.1 Caryophyllene oxide (c18) 2015 0.1±0.1 tr - 0.6±0.5 1.4±0.1 6.0±4.0 4.3±6.5 0.2±0.2 0.4±0.2 0.2±0.0 - 0.1±0.1 0.4±0.4 Globulol (c19) 2103 4.4±1.5 5.8±2.9 4.3±0.9 1.0±0.4 1.5±0.1 1.0±0.4 0.2±0.1 2.2±2.1 1.3±0.2 0.5±0.1 0.1±0.1 4.4±0.6 2.2±0.1 Viridiflorol (c20) 2113 0.8±0.2 0.8±0.4 1.6±0.9 0.7±0.1 0.8±0.0 0.7±0.6 0.2±0.2 1.5±1.3 0.8±0.2 0.2±0.0 tr 0.7±0.1 0.4±0.1 Spathulenol (c21) 2151 0.2±0.1 0.1±0.1 1.3±1.1 2.1±0.3 6.3±1.3 3.5±2.6 5.1±0.1 1.6±0.5 15.3±5.8 1.1±0.1 0.1±0.1 0.4±0.4 5.4±0.4 α-Eudesmol (c22) 2252 0.4±0.4 0.2±0.1 0.0 0.4±0.2 0.0±0.0 0.8±0.7 0.2±0.4 0.8±0.1 - 0.1±0.1 0.1±0.1 2.3±0.4 - β-Eudesmol (c23) 2262 0.1±0.2 0.1±0.2 0.1±0.0 0.6±0.3 0.2±0.1 0.5±0.8 tr 3.9±0.3 0.2±0.0 0.2±0.1 0.1±0.1 4.1±1.6 - (E.E)-Farnesol (c24) 2376 - - tr 1.9±0.3 0.1±0.0 0.3±0.3 0.4±0.7 0.03 0.1±0.1 - tr - 0.3±0.3 Torquatone (c25) 2437 0.1±0.0 0.2±0.1 0.2±0.3 0.2±0.2 - tr 0.1±0.2 0.03 0.1±0.1 - - 15.3±1.6 29.9±1.3 c) KI: Kovats index determined on Carbowax GC column. d) The abbreviations of the Eucalyptus species are given inTable 4. e) –: Not detected. F): tr: Trace (<0.1%).
______Journal of Experimental Biology and Agricultural Sciences http://www.jebas.org
Correlation between the Chemotaxonomic Classifications of the essential oils of 48 Eucalyptus species harvested . . . 106
Table 3 cont.
Species occ broc salm gill old larg lox sarg grac torq Compounds and abbreviations KI 39 40 41 41 43 44 45 46 47 48 α-Pinene (c1) 1053 7.0±9.3 1.6±0.1 2.4±3.0 15.1±3.2 8.2±2.3 6.2±1.8 3.9±0.6 18.5±1.5 7.1±2.6 10.5±3.9 4-Methyl-2- pentyl acetate (c2) 1114 - - - tr - - 20.7±1.6 - - - α-Phellandrene (c3) 1172 0.1±0.1 - 0.2±0.1 2.3±2.3 - 0.1±0.1 1.1±0.5 0.1±0.0 0.3±0.2 tr Limonene (c4) 1209 0.7±0.6 0.6±0.2 0.8±0.6 1.1±0.4 1.0±0.5 1.3±0.1 0.6±0.1 1.7±0.1 2.4±0.2 0.5±0.2 β-Phellandrene (c5) 1215 - 0.2±0.0 - 3.1±2.8 ------1.8-Cineole (c6) 1218 18.8±13.0 5.8±0.4 37.8±9.4 18.6±2.3 59.3±1.9 63.7±1.8 22.8±5.7 53.4±4.5 52.3±4.3 12.0±1.3 p-Cymene (c7) 1282 1.6±0.1 0.3±0.0 29.4±13.4 10.9±2.4 0.5±0.0 0.7±0.0 0.3±0.0 1.3±0.2 3.8±0.6 1.9±1.4 Isoamyl isovalerate (c8) 1304 - 18.7±5.2 1.0±0.6 - 0.5±0.1 0.1±0.0 0.5±0.2 0.3±0.1 0.3±0.4 tr Benzaldehyde (c9) 1541 ------Terpinene-4-ol (c10) 1618 1.8±0.3 0.3±0.0 1.0±1.3 1.3±0.2 0.2±0.0 0.2±0.1 12.2±4.5 0.1±0.0 1.0±0.1 tr Aromadendrene (c11) 1625 13.2±2.7 5.0±0.1 1.0±1.2 1.5±0.5 4.2±1.2 2.1±0.4 0.3±0.1 1.5±0.0 1.3±0.1 1.1±0.2 trans-Pinocarveol (c12) 1675 5.7±2.5 18.7±3.3 1.6±2.1 3.9±3.8 6.3±1.6 4.7±1.3 2.7±0.1 5.2±0.0 2.5±0.2 5.1±2.7 Cryptone (c13) 1695 - - 5.1±5.5 2.3±2.0 0.1±0.1 0.1±0.0 - 0.2±0.1 2.3±0.4 0.1±0.1 α-Terpineol (c14) 1713 0.9±0.4 1.1±0.2 0.6±0.3 0.5±0.3 0.5±0.2 0.4±0.1 0.2±0.0 1.9±0.1 1.8±0.8 0.2±0.1 Borneol (c15) 1720 0.2±0.0 0.3±0.0 0.2±0.2 0.2±0.2 0.2±0.1 0.2±0.0 0.1±0.1 0.2±0.1 0.1±0.0 0.2±0.1 d-Piperitone (c16) 1751 0.1±0.1 0.0 1.3±1.5 0.1±0.1 0.2±0.3 - - - 0.1±0.0 0.1±0.1 Bicyclogermacrene (c17) 1758 0.0 0.5±0.0 0.1±0.2 1.8±1.5 tr - - 0.5±0.4 - tr Caryophyllene oxide (c18) 2015 0.2±0.1 0.2±0.0 0.2±0.2 1.7±2.5 0.1±0.2 - - 0.1±0.1 0.2±0.1 0.1±0.0 Globulol (c19) 2103 19.9±5.5 15.1±1.1 0.5±0.1 2.9±0.9 6.8±0.9 7.9±0.9 14.1±2.5 2.0±0.1 4.1±0.8 1.9±0.1 Viridiflorol (c20) 2113 2.7±0.5 4.8±1.1 0.2±0.1 1.0±0.5 1.1±0.1 1.4±0.1 2.3±0.2 0.6±0.1 0.9±0.2 0.3±0.0 Spathulenol (c21) 2151 1.6±0.8 1.2±1.6 1.2±0.8 11.8±0.7 0.2±0.1 0.1±0.0 0.7±0.7 0.5±0.4 1.6±0.8 0.2±0.1 α-Eudesmol (c22) 2252 0.2±0.1 0.1±0.1 0.6±0.5 0.8±0.8 0.1±0.0 0.1±0.0 0.4±0.4 0.7±0.4 0.4±0.4 2.9±1.9 β-Eudesmol (c23) 2262 0.3±0.1 0.9±0.6 1.0±0.4 2.1±0.3 tr 0.2±0.0 0.9±1.3 2.7±1.2 0.7±0.8 10.1±0.4 (E.E)-Farnesol (c24) 2376 0.1±0.0 0.2±0.3 - 0.2±0.2 ------Torquatone (c25) 2437 0.3±0.1 0.6±0.7 0.1±0.1 0.1±0.1 0.1±0.1 0.1±0.0 0.2±0.0 0.0 0.1±0.1 42.0±0.8 c) KI: Kovats index determined on Carbowax GC column. d) The abbreviations of the Eucalyptus species are given inTable 4. e) –: Not detected. F): tr: Trace (<0.1%).
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107 Elaissi et al E. citriodora and E. maculta oils from the same series and the monterpens hydrocarbons (15.9±4.3 and 10.3±2.8%, (Maculatae) were characterized by essential oils with different respectively). Chemical composition of E. falcata was chemotypes. followed by E. bicostata (series Falcatae) in the HCA analysis, with a quantity of oxygenated monterpens (79.6±5.7, The leaf oil of E. citriodora differed significantly from that of 70.7±3.5%, respectively) much more important than that of E. E. maculata mainly because of a high percentage of globulus oil. E. maidenii approached E. sargentii (series monoterpenes hydrocarbons (32.1±4.8%) and oxygenated Micromembranae) by its equal content of monoterpene monoterpenes (64.8±5.3%) and a low percentage of hydrocarbons (20.0±4.6, 22.3±1.4%, respectively). This sesquiterpenes hydrocarbons (0.5±0.1%) and oxygentaed content is relatively higher than that of E. bicostata and E. sesquiterpenes (1.3±0.5%). However, it was observed that the globulus oils (6.5±2.2, 15.9±4.3%, respectively), while these 3 chemical characteristics of E. astringens and E. sargentii oils species share together a high mean percentage of oxygenated aligned with their morphological criteria classified by Brooker monterpenes. (2000). The species E. astringens and E. sargentii from the same series (Micromembranae) share together a relatively In present study chemical classification of the majority of equal high mean percentage in oxygenated monterpens species essential oils approached out lined by Brooker (2000). (57.8±7.7, 64.0±4.7%, respectively) and monterpenes In present classification oxygenated sesquiterpens form E. hydrocarbons (24.3±4.9, 22.3±1.4%, respectively). However E. pauciflora and E. exserta , ketones and monoterpenes occidentalis from the same series as E. astringens and E. hydrocarbons form E. odorata, E. stricklandii, E. torquata, E. sargentii (Brooker, 2000) was remote from the latter in their tereticornis and E. diversicolor were grouped in one one chemical classification, while it was closer to E. mauclata cluster. The presence of monoterpenes hydrocarbons and (series Maculata) and E. ficifolia (series Disjunctae), the oxygenated monoterpenes form E. cladocalyx and E. essential oils of which were characterized by the highest citriodora and oxygenated sesquiterpenes, oxygenated amount of sesquiterpenes hydrocarbons (24.0±0.8, 14.5±4.3%, monoterpenes and monoterpenes hydrocarbons were reported respectively) and by a relatively high quantity of oxygenated from E. camaldulensis and E. rudis, while the presence of sesquiterpenes (24.0±0.8, 43.6±5.4%, respectively). The oxygenated monoterpens was reported from E. cinerea and E. Globulares series was represented by E. globulus, E. bicostata sideroxylon. These chemicals separated others species and E. maidenii, while their essential oils displayed a belonging to the same series into different chemotypes, suchas significant difference of the mean percentages of their those of Globulares series (E. globulus, E. maidenii, E. oxygenated monterpenes and their monterpenes hydrocarbons. bicostata), Maculatae series (E. maculata and E. citriodora) E. globulus was closer to E. oldfieldii (subseries Xylocarpae), and Annulares series (E. botryoides origin vilmorin and E. the essential oils of which contained almost the same amount botryoides from Morocco). of oxygenated monterpens (66.1±3.0, 70.3±0.9%, respectively)
Figure 1 Principal component analysis of the eight chemical classes of essential oils in the leaves of 48 species of Eucalyptus. For the abbreviation of the Eucalyptus species (▲) and the classes (■), see Tables 1 and 2, respectively.
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Correlation between the Chemotaxonomic Classifications of the essential oils of 48 Eucalyptus species harvested . . . 108
Figure 2 Dendrogram obtained by hierarchical cluster analysis based on the Euclidean distances between groups of the leaf essential oils classes of 48 Tunisian Eucalyptus species. For the abbreviation of the Eucalyptus species and the provenance (Pro), see Table 1.
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109 Elaissi et al
Figure 3 Principal component analysis of 25 compounds in the essential oils of leaves in 48 species of Eucalyptus. For the abbreviation of the Eucalyptus species (▲) and the components (●), see Tables 1 and 3, respectively.
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Correlation between the Chemotaxonomic Classifications of the essential oils of 48 Eucalyptus species harvested . . . 110
Figure 4 Dendrogram obtained by hierarchical cluster analysis based on the Euclidian distances between groups of the leaf essential oils major components of 48 Tunisian Eucalyptus species. For the abbreviation of the Eucalyptus species and the provenance (Pro), see Table 1
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111 Elaissi et al 3.2 Principal components analysis (PCA) and hierarchical components. The comparison of the chemical classification of cluster analysis (HCA) of the individual components E. bicostata, E. globulus and E. maidenii to their hierarchical classification, showed that the three species, which belonged to The PCA horizontal axis explained 14.31% of the total the series Globulares (Blakeley, 1934b) were distant in both variance while the vertical axis a further 12.13% (figure. 3). ACP and HCA analysis. In fact E. bicostata was very close to With a dissimilarity > 7, HCA indicated 17 groups of species; E. largiflorens which belonged to the serie buxeales (Penfold the seventeenth group was divided into 8 subgroups with a and Willis, 1961), E. globulus was close to E. gracilis and E. dissimilarity < 6 (figure 4). viminalis belonging to two different series (Heterostemones Benth., Viminales Blakely resp.) (Benthman, 1867; Blakeley, The PCA and HCA have correctly grouped various chemical 1934c). Therefore they were described as being the same groups that have a commonality of essential oils (same chemotype. For many cases, the statistical analysis (PCA and chemotype) but they differ morphologically. Most recent HCA), permitted the grouping of species from different series botanical classification based on comparative morphology in one group representing one chemotype, such as those of the (Brooker, 2000) was supported by the results of the series Foveolatae by Maiden (1929); E. ovata, E. macarthurii mathematical analyses; the score of the principal components by Blakeley (1934c), E. bicostata, E. globulus and E. maidenii of all the oils was examined. A link between the two by Blakeley (1934b), E. gracilis, E. viminalis by Benthman classifications will be indicated to support the general (1867) and Blakeley (1934c), being all of them characterized approach. by similar major components (c6, c1, c12 and c11). The findings of present study are very similar to the findings of Li The comparison of the chemical classification to the et al. (1996) who studied the classification of seventeen species phylogenetic classification showed a significant divergence for Eucalyptus essential. The species which were very close in the many species which were very close in the hierarchic classification of Brooker (2000) were very distant within their classification (Brooker, 2000). E. botryoides from Morocco chemical classification in present study. In present and E. botryoides from Vilmorin belongs to the same series investigation E. ovata (series Triangulares) populations from but not to the same provenance, which showed a significant eastern Tasmania were significantly differed from the two west difference in their essential oils chemical composition. coast populations (Henty River and Montagu Road). Therefore they were classified into two different chemotypes. A classification based on their oils only might conclude that E. ovata populations from eastern Tasmania were these two provenances should be classified in two series. characterized by high levels of nerolidol, and linalool or p- Whereas the comparison of different species viz E. cymene and spathulenol. In contrast, other species populations occidentalis, E. astringens and E. sargentii which belongs to have high levels of 1,8-cineole and/or - pinene Li et al. the same series Micrombranae, showed a significant (1996). The same remark was observed with E. cinerea, E. discrimination in oil chemical composition between E. sideroxylon, E. lehmannii belonging to different series astringens and E. sargentii oils from that of E. occidentalis (Argyrophyllae Blakely, Solidae Brooker, Lehmannianae which was considered as a different chemotype. Findings of (Carr, D.G. & Carr S.M.G, 1980) but to the same chemotype the present study are in agreement with the findings of Dunlop within the chemical classification; however, it was noticed that et al. (2003). In present study, essential oils of E. pauciflora the separation of other species which were classified in the and E. gunni were characterized by same chemotype, while same series by Brooker (2000) such as E. citriodora and E. they were classified in two different series (Pauciflora, maculata of the series Maculatae (Blakely) Chippendale Orbiculares, respectively) by Brooker (2000). These results (Blakeley, 1988), which differed significantly in their chemical were confirmed by Li et al. (1995), who grouped together three composition oils. In fact E. citriodora oil was highlighted by 4 species from different series: E. pauciflora, E. reganas major components (c1, c6, c12 and c11), however, E. maculata (Reganates) and E. sieberi (Psathyroxylon) in the same oil was represented essentially by only one component, 1,8- chemotype, represented essentially by 3 major components: - cineole (c6) and was remote from the last species in a separate eudesmol (8.5±1.7-12.9±8.1%), -eudesmol (9.6±3.5- group within HCA and PCA annalysis. The comparison of the 18.7±3.3%) and β -eudesmol (9.2±2.6-15.0±4.1%). hierarchical classification of the genus Eucalyptus of Penfold and Willis (1961) with Brooker (2000) and present study The essential oils of E. pauciflora with other species belonging chemical classification, showed that some species such as E. to different series such as E. reganas (Reganates) and E. stricklandii and E. woodwardii which were grouped together in sieberi (Psathyroxylon). In contrast Li & Madden (1995) the same series Obliquae (Penfold and Willis, 1961) but demonstrated that E. nitens and E. denticulata of the same separated in to two series in that of Brooker (2000) series (Remanates) but these were characterised by different (Stricklandinae Brooker, Rufispermae Maiden, resp.) and major essential oils components (1,8-cineole and -pinene for grouped together in the same chemotype within our results, E. nitens and p-cymene, γ-terpinene for E. denticulata). however oils of E. tereticornis, E. exserta, E. rudis and E. However it was noticed a concordance of the phylogentic and camaldulensis of the series Exsertae (Penfold & Willis, 1961) the chemical classification of E. fasciculosa and E. were found discriminated in four different chemotypes and in polyanthemos which belonged to the series Heterophloiae four different series within Brooker (2000). Blakley (Blakeley, 1934a) and which had the same major
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Correlation between the Chemotaxonomic Classifications of the essential oils of 48 Eucalyptus species harvested . . . 112 Conclusions oils of twelve Eucalyptus Species harvested from Hajeb Layoun arboreta (Tunisia). Chemestry and Biodiversity 7: The Essential oils chemical classification approached many 705716. Eucalyptus species out lined by the phylogenetic classification, however we noticed a concordance of the two classifications European Pharmacopoeia, Third Edition, Supplement 2000. within some species. Edité par Council of Europe, Strasbourg, 1999.
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