Journal of Applied Botany and Food Quality 87, 249 - 255 (2014), DOI:10.5073/JABFQ.2014.087.035 1Department of Molecular Biology, Abbes LAGHROUR University, Khenchela, Algeria 2Laboratory of Natural Resources Valorization, Ferhat Abbas University, Setif, Algeria 3Blaise Pascal University, Clermont Ferrand, France 4LEXVA Analytique, Beaumont, France Characterization and chemosystematics of Algerian thuriferous juniper (Juniperus thurifera L.) Azzeddine Zeraib1, 2*, Messaoud Ramdani 2, Lamia Boudjedjou 2, Pierre Chalard 3, Gille Figuredo4 (Received December 15, 2013) * Corresponding author Summary Leaf essential oils (EO) of Juniperus thurifera L. collected at six lo- cates from Aures Mountains in Algeria, were analyzed by gas chro- matography (GC) and gas chromatography-mass spectrometry (GC/ MS). The main components identified were: sabinene (5.2-19.78 %), terpinene-4-ol (5.43-9.37 %), elemol (0.69-7.61 %), Δ-cadinene (3.26-6.11 %). Terpenoids data of our samples and those reported in other works realized by various authors were subjected to Principal Component Analysis (PCA), and Unweighted Pair Group Method with Arithmetic means (UPGMA) cluster was carried. This analysis revealed significant differences between Juniperus thurifera popula- tions, and confirmed the clear separation of Algerian populations to the European and Moroccan populations. Algerian thuriferous juni- per is more similar to J. thurifera from Moroccan populations, and different from that of essential oils obtained from European popula- tions. Introduction Thuriferous juniper (Juniperus thurifera L.), is an evergreen and a dioecious shrub or tree with scale leaves and bluish black berries at maturity, occurring from Algeria and Morocco over the Iberian Peninsula and the Pyrenees to the French and Italian Alps and to Corsica (GAMISANS et al., 1994; GAUQUELIN et al., 1988; GAUQUE- LIN et al., 2003). In Algeria, the thuriferous juniper is extremely rare and only localized in the Aures mountains with a number of scattered and often very large trees that are probably the remains of formerly more extensive stands (VELA and SCHÄFER, 2013). Juniperus thurifera is a morphologically variable species, perhaps as a result of long-term isolation of disjunct populations. MAIRE (1926) was the first to distinguish the North African and European popula- tions of J. thurifera. Later, based on morphometric characters such as the size of the cones and the number of seeds per cone. GAUQUE- LIN et al. (1988) recognized both entities as subspecies; J. thurifera subsp. africana (Maire) Gauquelin in North Africa, and J. thurifera subsp. thurifera in the European range of the species. Of the lat- ter, there are 3 varieties: var. thurifera on the Iberian Peninsula, var. gallica De Coincy in the Alps and var. corsicana Gauquelin in Corsica (GAUQUELIN et al., 2003). The distinction of North African and European populations of J. thurifera is validated by ROMO and BORATYNSKY (2007). Studies based on essential oils composition, random amplified polymorphic DNA (RAPD) and morphometric data (ADAMS, 1999; ADAMS et al., 2003; BARRERO et al., 2004; ACHAK et al., 2008; 2009; BAHRI et al., 2013; BORATYNSKY et al., 2013), supported the clear differentiation of J. thurifera subsp. africana from Morocco against the European populations, but without studying the Algerian ones, which are generally empirically assimilated to the Moroccan taxon. The study achieved by TERRAB et al. (2008), based on genetic poly- morphism (AFLP), has shown that The Algerian population was ge- netically more closely related to the European than to the Moroccan ones, probably due to dispersal events from Europe to Algeria, and the Moroccan populations should be recognized as a distinct subspe- cies (J. thurifera L. subsp. Africana (Maire) Romo and Boratynsky. A taxonomic synthesis, completed by a brief morphological study of fruits based on herbarium samples was performed by VELA and SCHÄFER (2013), concludes with the desirable distinction of a Mo- roccan taxon (subsp. africana) and an Algerian one (Juniperus thurifera var. aurasiaca Véla & P. Schäf). It is distinguishable as intermediate between subsp. thurifera and subsp. africana, more similar to the first one following molecular properties but sharing partial polymorphism with the second and more similar to the latter one following some morphological elements as fruit size average but with a variation amplitude partially sharing numerical values with the first. But above all, the polymorphism of Algerian populations is very poorly known and would be deeply studied in the future (VELA and SCHÄFER, 2013). The aim of the present study was to characterize the Algerian Thu- riferous juniper and sought its taxonomic status. We are based on the chemical composition of the essential oils isolated from the leaves of Juniperus thurifera L. collected at different locations in Aures mountains (Algeria), and compare these data with other studies on the chemical variability of essential oils from European and Moroc- can populations. Material and methods Plant material Leaves of juniperus thurifera L. were collected in March, 2010, from different localities around the Aures Mountains, in Algeria (Tkout1 at 1450 m of altitude, Tkout2 at 1700 m, Baâli at 1750 m, Tizi nerr- sas at 1500 m, Tibhirin at 1500 m, and Chelia at 1700 m of altitude). A voucher specimen is deposited in the herbarium of the Laboratory of Natural Resource Valorization, Faculty of Biology, Farhat Abbes University, Setif, Algeria. Isolation and analysis of the essential oils The plant material was submitted to hydro distillation for 3 h, ac- cording to TUMEN et al. (2010). The prepared volatile oils were de- hydrated over anhydrous sodium sulphate and stored in sealed glass vials at 4-5 °C prior to analysis. Yield based on dry weight of the sample was calculated. The essential oils were analyzed on a Hewlett-Packard gas chro- matograph Model 5890, coupled to a Hewlett-Packard model 5971, equipped with a DB5 MS column (30 m X 0.25 mm; 0.25 μm), pro- gramming from 50 °C (5 min) to 300 °C at 5 °C/min, with a 5 min hold. Helium was used as the carrier gas (1.0 mL/min); injection in split mode (1:30); injector and detector temperatures, 250 and 280 °C, respectively. The mass spectrometer worked in EI mode at 70 eV; electron multiplier, 2500 V; ion source temperature, 180 °C; MS data were acquired in the scan mode in the m/z range 33-450. The identification of the components was based on comparison of their mass spectra with those of NIST mass spectral library (NIST, 2002) and those described by ADAMS (2001) as well as on comparison of 250 A. Zeraib, M. Ramdani, L. Boudjedjou, P. Chalard, G. Figuredo their retention indices either with those of authentic compounds or with literature values (ADAMS, 2001). Statistical analysis To examine the phytochemical diversity based on the content (%) of chemical constituents in essential oil among the studied six popu- lations; these were first subjected to Principal Components Analy- sis (PCA) to examine the relationships among the compounds and identify the possible structure of the population. Cluster analysis (UPGMA) was carried out on the original variables and on the Man- hattan distance matrix to seek for hierarchical associations among the populations. Statistical analyses were carried out using STATIS- TICA 8 software. Results and discussion Variability of Algerian J. thurifera essential oils Volatile oil yield of the leaves of the investigated J. thurifera popu- lations is summarized in (Tab. 1). The yield varied from 0.40 to 0.53 % in different populations of J. thurifera. Maximum essential oil yield was noticed in Tkout1 population (0.53 %), followed by Tizi nerrsas (0.48 %), the minimum essential oils yield was noticed in Chelia population (0.40 %), which is very low compared to the yield obtained by ACHAK et al. (2008; 2009) and BAHRI et al. (2013) from the same species. Inter-population variation of essential oil yield is quite common phenomenon and encountered earlier in several other plant species. These variations might be due to climatic conditions of the grow- ing site, pedoclimatic variation or due to difference in the genetic makeup of the J. thurifera populations. The volatile oils of all six Juniperus thurifera populations were characterized and identified by gas chromatography and gas chroma- tography-mass spectrometry. The relative percentages of the constit- uents are listed in the Tab. 1. Seventy nine constituents, representing 90.2-98.7 % of the total oil composition, were identified. Seventeen constituents were monoterpene hydrocarbons accounting (18.6- 40.1 %) of the EO, 17 constituents were oxygenated monoterpenes (10.3-24 %), 22 constituents were sesquiterpene hydrocarbons (15.1-23.8 %), 18 constituents were oxygenated sesquiterpenes (18- 36.8 %), two constituents were diterpene hydrocarbons (0.2-1.7 %), and three constituents were oxygenated diterpenes (0.5-1.8 %). The main components identified were: Sabinen (5.2-20 %), ter- pinene-4-ol (5.4-9.4 %), Elemol (0.7-7.6 %), Δ-cadinene (3.3-6.1 %), linalyl acetate (0.9-6.2 %), γ- terpinene (2.6-3.9 %), α- pinene (2.6- 3.9 %) and myrcene (1.2-3.3 %). Leaf EO of Juniperus thurifera in deferent regions of Morocco are rich in sabinene (12.2 % to 45.8 %), α-pinene (4 % to 17.1 %) and terpinene-4-ol (2.6 % to 16.9 %) (ADAMS et al., 2003; ROMAN et al., 2008; 2009; BAHRI et al., 2013). The major constituent of the volatile oils obtained from branches of Moroccan Juniperus thurifera is β-pinene (36.26 %) (MANSOURI et al., 2010). Twenty-two compounds showed statistically significant varia- tions among the six locations, the components identified (sabinen, α-cadinol, valencen, elemol, linalyl acetate, and linalool) show a significant variability of terpenoid (Fig. 1). The principal component analysis (PCA) performed on the cor- relation matrix of the 79 variables showed that the first three axes explained 74.3 % of the observed variation. This analysis allowed recognizing two distinct EO types based on the content of sabinene, linalyl acetate, linalool, γ-terpinene, myrcene, bulnesol, valencene, γ-eudesmol, epi-α-cadinol, epi-α-muurolol, and 4-epi-abietal. The first group was represented by four populations (Tkout1, Baâ- li, Tizi nerrsas, Tibhirine), located on the positive part of axis one, characterized by high concentrations of sabinene, linalyl acetate, linalool, γ-terpinene, myrcene, and bulnesol. This group is opposed to the second group, formed by two populations (Chelia and Tk- out2), which is characterized by high concentrations of valencene, γ-eudesmol, epi-α-cadinol, epi-α-muurolol, and 4-epi-abietal (Fig. 2). Fig. 1: Chemical Variability of main compounds of Juniperus thurifera (Var 1 to Var 79 are mentioned in the Tab. 1). Fig. 2: Projection of Algerian Juniperus thurifera populations on the factor plane (1x 2). Characterization and chemosystematics of Algerian J. thurifera 251 Tab. 1: Composition of the leaf essential oils of J. thurifera from Aures mountains in Algeria. Compound Ki Tk1 Tk2 Tz Ba Tb Ch Var1 α-thujene 924 1.2 0.8 1.1 1.5 0.8 0.5 Var2 α-pinene 932 3.3 2.6 2.62 3.47 3.1 3.9 Var3 fenchene 946 0.1 tr 0.1 0.1 0.1 - Var4 camphene 947 0.1 0.1 0.1 0.1 0,1 tr Var5 sabinene 975 17.9 8.3 11.3 20.0 9.0 5.2 Var6 β-pinene 979 0.3 0.3 0.3 0.3 0.3 0.4 Var7 myrcene 991 2.8 1.7 2.0 3.3 1.7 1.2 Var8 Δ-3-carene 1011 0.2 1.5 0.9 0.5 1.0 0.8 Var9 α-terpinene 1018 1.6 1.3 1.7 1.8 1.9 1.2 Var10 p-cymene 1026 0.5 0.2 0.3 0.7 0.4 0.5 Var11 limonene 1031 1.4 3.2 2.5 2.0 1.2 0.7 Var12 β-phellandrene 1031 0.2 - 0.2 0.2 0.2 0.2 Var13 (Z)-β-ocimene 1040 0.10 - - - - - Var14 (E)-β-ocimene 1050 0.3 - 0.2 0.3 0.2 - Var15 γ-terpinene 1062 3.2 2.6 3.5 3.6 3.9 2.6 Var16 cis-sabinene hydrate 1068 0.8 0.8 1.6 0.6 2.3 0.6 Var17 terpinolene 1088 1.3 1.2 1.5 1.5 1.5 0.9 Var18 linalool 1096 4.6 0.3 0.4 2.5 1.6 0.4 Var19 cis-thujone 1100 0.1 0.4 - 0.3 0.6 - Var20 trans-thujone 1111 - 0.27 - - - 0.3 Var21 cis-p-menth-2-en-1-ol 1122 0.4 0.3 0.5 0.4 0.6 0.3 Var22 trans-menth-2-en-1-ol 1141 0.3 0.2 0.3 0.3 0.4 0.3 Var23 terpinene-4-ol 1177 7.2 5.4 7.6 7.5 9.4 7.1 Var24 α-terpineol 1195 1.6 0.6 1.1 1.1 1.1 0.9 Var25 verbanone 1205 0.2 0.2 0.3 0.3 0.4 0.3 Var26 trans piperitol 1209 0.1 - - - - - Var27 nerol 1 (80) 1224 0.3 - 0.2 0.2 0.2 - Var28 linalyl acetate 1249 6.2 1.2 3.4 3.1 3.7 0.9 Var29 pregeijerene 1280 0.12 - - - - - Var30 bornyl acetate 1284 0.1 - - 0.1 0.1 0.4 Var31 isobutyl benzene 1287 0.3 - 0.3 - 0.3 - Var32 decan-2,4-dien-1-ol 1312 0.3 0.2 - 0.5 0.1 - Var33 δ-elemene 1338 0.2 0.2 0.2 0.1 0.2 - Var34 α-terpinene acetate 1343 0.7 0.8 1.7 0.8 1.1 0.1 Var35 neryl acetate 1362 0.4 - 0.3 0.2 0.2 - Var36 geranyl acetate 1381 1.0 0.3 0.6 0.8 0.6 0.3 Var37 β-elemene 1391 0.3 0.4 0.3 0.4 0.4 0.2 Var38 β-caryophyllene 1419 0.9 1.1 0.9 1.0 1.2 0.5 Var39 γ-elemene 1429 0.4 0.4 0.2 0.1 0.4 - Var40 cadina-3,5-diene 1448 0.1 0.1 0.1 0.2 0.1 - Var41 α-humulene 1454 0.5 1.0 0.8 0.9 1.1 0.7 Var42 cis muurola-4(14),5-diene 1460 0.1 0.5 0.2 0.4 0.7 0.3 Var43 cadina-1(6),4-diène 1460 0.1 0.2 - 0.2 0.1 - Var44 γ-muurolene 1477 0.3 0.6 0.5 0.5 0.5 0.5 Var45 curcumene 1478 0.2 - 0.3 - 0.2 0.2 Var46 germacrene-D 1480 1.9 2.7 4.4 3.1 3.3 1.6 252 A. Zeraib, M. Ramdani, L. Boudjedjou, P. Chalard, G. Figuredo Var47 trans muurola-4(14),5-diene 1494 0.2 0.4 0.3 0.5 0.2 0.4 Var48 α-muurolene 1500 0.8 1.1 1.1 0.8 0.6 0.4 Var49 β-curcumene 1516 0.5 0.2 0.7 0.2 0.4 0.2 Var50 γ-cadinene 1514 0.7 1.8 1.0 1.5 1.2 1.4 Var51 Δ-cadinene 1523 3.3 6.1 4.1 5.6 3.9 4.1 Var52 α-cadinene 1539 0.2 0.4 0.2 0.3 0.3 0.3 Var53 elemool 1550 4.7 5.8 0.7 3.9 7.6 5.2 Var54 germacrene-B 1561 2.5 2.3 2.0 1.2 2.6 1.2 Var55 germacrene-D-4ol 1576 0.8 1.9 1.1 0.9 1.1 1.1 Var56 Caryophyllene Oxyde 1585 0.1 - 1.1 0.2 - - Var57 cedrol 1596 0.1 0.1 0.1 0.1 0.2 0.8 Var58 humulene epoxyde II 1607 0.2 0.2 0.1 - - 0.1 Var59 β-oplopenone 1608 0.5 1.2 0.7 0.6 0.5 0.9 Var60 epi-cedrol 1613 0.92 - - - - - Var61 valencen 1619 0.5 2.5 3.1 1.4 0.9 6.6 Var62 1,10-diepi-cubenol 1627 0.3 0.3 0.2 0.2 0.4 0.3 Var63 1-epi-cubenol 1629 1.1 1.3 1.0 1.8 0.8 1.6 Var64 γ-eudesmol 1632 1.3 2.1 2.4 1.1 2. 2.4 Var65 epi-α-cadinol 1640 0.9 2.9 1.1 0.8 1.2 3.0 Var66 epi-α-muurolol 1642 0.9 3.7 0.7 0.8 1.0 3.1 Var67 α-cadinol 1663 6.1 13.5 10.6 5.5 9.2 15.7 Var68 bulnesol 1666 3.2 1.8 0.9 1.9 2.0 0.6 Var69 gurjunene 1670 0.8 1.1 0.9 0.8 1.6 - Var70 γ-gurjunene 1678 0.3 0.6 0.6 0.8 0.4 0.7 Var71 epi-α-bisabolol 1686 0.3 0.4 0.4 0.1 0.4 0.6 Var72 2-pentadecanone 1698 0.3 0.3 - - 0.2 0.4 Var73 (Z,Z)-farnesol 1714 0.5 0.3 0.4 0.2 1.0 0.9 Var74 8-α-acetoxyl elemol 1789 0.1 0.2 - - 0.1 - Var75 manoyl oxyde 1989 0.4 0.2 - - 0.2 - Var76 13-epi-manoyl oxyde 1992 0.3 0.6 0.1 0.3 0.2 - Var77 Abietatriene 2090 0.3 0.5 0.4 0.2 0.3 1.7 Var78 phytol 2109 0.2 0.2 - - 0.1 - Var79 4-epi-abietal 2299 0.7 1.1 0.3 0.4 0.6 1.3 Monoterpene hydrocarbons 35.3 24.5 29.8 40.1 27.9 18.6 Oxygenated monoterpenes 24.0 10.3 16.7 18.3 20.6 12.3 Sesquiterpene hydrocarbons 15.1 23.8 21.9 20.2 20.4 19.5 Oxygenated sesquiterpenes 22.4 36.2 21.7 18.0 27.7 36.8 Diterpene hydrocarbons 0.5 0.7 0.4 0.2 0.4 1.7 Oxygenated diterpenes 1.4 1.8 0.5 0.6 1.0 1.3 Yield (%) 0.53 0.46 0.48 0.45 0.46 0.40 Total % 98.7 97.3 91.0 97.4 98.0 90.2 (Tk1: Tkout 1, Tk2: Tkout 2, Tz: Tizi nerrsas, Tb: Tibhirin, Ch: Chelia, Ba: Baâli). Mono and sesquiterpenoids variability reflects the heterogeneity of the genetic structure of population (DODD and POVEDA, 2003; LIMA et al., 2010; SHANJANI et al., 2010). HANNOVER (1992) provides evi- dence that terpene chemotypes are strongly controlled by genetic factors; he also reported instances of environmental variation in ter- pene expression under extreme habitat conditions. A prevalence of monoterpene hydrocarbons compared to other components was noted in Tkout1, Baâli, Tibhirin, and Tizi nerrsase populations, while its reverse trend could be seen in Tkout2 and Chelia populations, which were dominated by sesquiterpene hydro- carbons (Tab. 1). Monoterpenes and sesquiterpene hydrocarbons were strongly related to chemical balance in soils (organic matter, phosphor and base saturation). The chemovariation observed ap- pears to be environmentally determined (LESJAK et al., 2013). Characterization and chemosystematics of Algerian J. thurifera 253 Influence of environmental factors in the chemical composition of essential oils have also been reported in the genus Juniperus (DODD and POVEDA, 2003; LIMA et al., 2010; SHANJANI et al., 2010; LOŽIENE and LABOKAS, 2012; LESJAK et al., 2013), Cupressaceae family (OTTVIOLI, 2009), and are well known for other family (HAIDER et al., 2004; KAROUSOU et al., 2005; CUARDO et al., 2006; LEI et al., 2010; DJABOU et al., 2012). The dendrogram based on UPGMA clustering (Manhattan distance), shows the presence of two groups (Fig. 3) that confirms result obtained from PCA analyses. The first cluster is divided into two sub-groups based on the content of terpinene-4-ol, germacrene-D, α-terpinene acetate, cis-sabinene hydrate. The first sub-group formed by Tkout1 and Baâli, characterized by low concentration of these constituents unlike for Tizi nerrsas and Tibhirin populations. Fig. 5: Dendrogram of Juniperus thurifera populations, based on Manhat- tan Similarity distance (ADAMS, 1999: Spain S3 = 2 Km e Ruidera); (ADAMS et al., 2003: Spain SC = Consuegra, Spain S1 and S2 = Ruidera; Morocco M1 and M2 = Atlas Mts, Morocco TM = Tizi-n-Tichka, Morocco OM = Oukaimeden; Pyrenees P1 and P2 = Pyrenees, France; Corsica CR = Corse, Island); (ACHAK et al., 2009: Morocco FL = fresh leaves; Morocco DL = dried leaves, Ait Lkak Oukaimeden, Atlas Mts), (OT- TAVIOLI, 2009: Corsica Cor 1-16 = Corse) and (BAHRI et al., 2013: Morocco VHD and MW= Morocco) Fig. 3: Dendrogram of Algerian Juniperus thurifera populations, based on Manhattan Similarity distance. Fig. 4: Projection of Juniperus thurifera populations on the factor plane (1x 2). Characterization and chemosystematics Leaf essential oils are extremely useful for the analyses of popula- tional differentiation, hybridization and introgression and in assign- ing individual plants to a species (ADAMS, 2010). To compare the Algerian thuriferous juniper with other populations, terpenoids data of our samples and those reported in the work of ADAMS (1999), ADAMS et al. (2003), ACHAK et al. (2009), OTTAVIOLI (2009) and BAHRI et al. (2013) were subjected to PCA, and UPGMA cluster was carried. This analysis revealed significant differences be- tween Juniperus thurifera populations. The PCA performed on the correlation matrix of the 96 variables based on the oil composition resulted in 38 factors of which the first three accounted for 47.4 % of the variance among the 39 popula- tions, shows that J. thurifera populations are divided into three dis- tinct groups (Fig. 4). The Algerian populations separated by 27.30 % of the variance in the chemical composition of essential oils. How- ever, the Moroccan populations accounted for about 12.66 % of the variance (Fig. 4). The UPGMA based on the Unweighted pair-group average distance and the City-block (Manhattan) (Fig. 5), has divided 39 populations of J. thurifera into two clads, confirmed the separation of the North African populations from the European populations. Then, some qualitative differences in chemical composition can be deduced. The leaf essential oil composition of European populations was higher in limonene (30-75 %) and lower in sabinene (0-9,7 %) (ADAMS et al., 2003; ACHAK et al., 2008; OTTAVIOLI, 2009). How- ever, the composition of the oils from Algerian populations was sim- ilar to those reported for oils of J. thurifera from Moroccan popula- tions (ADAMS et al., 2003; ACHAK et al., 2008; BAHRI et al., 2013). The concentration of α-pinene and sabinene were greater in the 254 A. Zeraib, M. Ramdani, L. Boudjedjou, P. Chalard, G. Figuredo oil of the Moroccan populations, conversely, the concentrations of Δ-cadinene and α-cadinol were greater in the oil of Algerian popu- lation. Should be recognized that Algerian populations as sub-group dis- tinctive from the European and Moroccan populations or do they merely represent geographical interspecific variation? Algerian Juniperus thurifera is distinguishable as intermediate be- tween subsp. africana and subsp. thurifera. More similar to the first one following some morphological elements as fruit size average but with a variation amplitude partially sharing numerical values with the latter (VELA and SCHÄFER, 2013). The results of the present study confirmed this similarity between Moroccan populations subsp. africana and Algerian populations called Juniperus thuriefera var. aurasiaca Véla & P. Schäf. In view of the data presented in this study, it is apparent that Algerian populations is not related to subsp. thurifera (sensu stricto), but to subsp. africana. However, Terpenes are generally not as useful in making phylo- genetic decisions because several terpenes may be controlled by a single enzyme (ADAMS, 2010). FARJON (2005) considered species based on the chemistry of terpenes analysis as based on inconclusive evidence, but DNA sequence data certainly can (which is the main reason for their superiority). The study achieved by TERRAB et al. (2008), based on genetic poly- morphism, has shown that Algerian populations are distinct from the Moroccan ones and more related to European populations. Intra- and inter-populational morphological variability through- out this vast territory fragmented (Spain, Pyrenees, Alps, Corsica and Morocco) has recently been investigated by BORATYŃSKI et al. (2013) and gives results congruous with genetic pattern obtained on the same whole Europe/Morocco (TERRAB et al., 2008), the Algerian population excluded. Finally, it is true that adding up the number of terpene differences may or may not give a good estimation of divergence. But, the num- ber and scope of terpene differences between the Algerian and Eu- ropean populations indicate considerable differentiation. Additional research, using morphological variability, should help in elucidating these relationships. Conclusions In brief, essential oils analysis carried out on six populations of J. thurifera showed both inter-population variability in their terpe- noid content, with abundance of sabinene, terpinene-4-ol, elemol… The PCA and UPGMA analysis allowed recognizing two distinct EO types based on the content of monoterpene and sesquiterpene hydro- carbons, the chemovariation observed appears to be environmentally determined. VELA and SCHÄFER (2013), proposed to call the Algerian population by Juniperus thuriefera var. aurasiaca Véla & P. Schäf, in the varietal rank, which will allow us to consider now as belong- ing European subset (subsp. thurifera) or Moroccan subset (subsp. africana) or equal with both. In view of the results of this study, it seems that the Algerian populations are much more similar to the Moroccan populations, but it is still early to specify its taxonomic status. Therefore, we support the proposition of VELA and SCHÄFER (2013), and the conflict between chemical and genetical data should be resolved on the morphological level. Acknowledgments The authors are thankful to Mr. Fercha Azzeddine, teacher in the Department of Biology, Abbès Laghrour University, Khenchela, Al- geria, for the preliminary examination of this work. This study was supported, in part, by the chemistry of heterocyclic compounds and carbohydrates laboratory, higher National School of Chemistry, Clermont Ferrand, France. And the Ministry of Higher Education and Scientific Research of the Algerian People’s Demo- cratic Republic. 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SHANJANI, P.S., MIRZA, M., CALAGARI, M., ADAMS, R.P., 2010: Effects dry- ing and harvest season on the essential oil composition from foliage and berries of Juniperus excelsa. Industrial Crops and Products 32(2), 83- 87. TERRAB, A., SCHÖNSWETTER, P., TALAVERA, S., VELA, E., STUESSY, T.F., 2008: Range-wide phylogeography of Juniperus thurifera L., a presump- tive keystone species of western Mediterranean vegetation during cold stages of the Pleistocene. Molecular Phylogenetics and Evolution 48, 94-102. TUMEN, I., HAFIZOGLU, H., PRANOVICH, A., REUNANEN, M., 2010: Chemi- cal constituents of cones and leaves of cypress (Cupressus sempervirens L.) grown in Turkey. Fresenius Environmental Bulletin 19 (10), 2268- 2276. VELA, E., SCHÄFER, P.A., 2013: Typification de Juniperus thurifera var. afri- cana Maire, délimitation taxonomique et conséquences nomenclaturales sur le Genévrier thurifère d’Algérie. Ecologia mediterranea 39(1), 69- 80. Address of the corresponding author: A. Zeraib, Department of Molecular Biology, Abbes LAGHROUR Universi- ty, 40000 Khenchela, Algeria. Laboratory of Natural Resources Valorization, Ferhat Abbas University, 19000 Setif, Algeria. E-mail: azzeraib@yahoo.fr