Leaf essential oils and their application in systematics of Juniperus excelsa complex in Iran

Biochemical Systematics and Ecology 84 (2019) 29–34

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Leaf essential oils and their application in systematics of Juniperus excelsa complex in Iran

T

Fatemeh Hojjatia,∗, Hassan Sereshtib, Marzieh Hojjatic a

Department of Plant Biology, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, 14115-154, Iran School of Chemistry, University College of Science, University of Tehran, Tehran, 14155-6455, Iran c Department of Chemistry, College of Science, Bu-Ali Sina University, Hamadan, 65174-4135, Iran b

ARTICLE INFO

ABSTRACT

Keywords: Juniperus excelsa Juniperus polycarpos Juniperus seravschanica Cupressaceae Chemotaxonomy GC-MS Essential oils Iran

The composition of the leaf essential oils of ten populations of Juniperus excelsa complex in Iran are reported and compared. Hydrodistillation yielded in 0.08%–3.78% dry weight (v/w) clear oils among populations studied. The oils are mostly dominated by α- Pinene, Limonene and β-Myrcene. The simplest oil was found in Qushchi (J. polycarpos var. polycarpos) with 25 components and the most complex oil was found in Lushan1 (J. polycarpos var. polycarpos) with 92 components. Quantitative data is presented for 164 components were found. The southern populations, Fasa and Khabr, show differences in composition of essential oils compared with other populations so that Borneol and isopinocamphone are diagnostic for them. These populations have been identified as Juniperus seravschanica in southeast of Iran and hybrid samples between J. polycarpos and J. seravschanica in southwest of Iran whereas other populations, as is confirmed in this study, are J. polycarpos. It is confirmed by essential oils data that there is no J. excelsa in Iran.

1. Introduction Juniperus (Cupressaceae) is composed of approximately 75 species in 3 monophyletic sections (Mao et al., 2010; Adams and Schwarzbach, 2013): Caryocedrus with one species, Juniperus with 14 species, and Sabina with 60 species. J. excelsa M. Bieb. (sect. Sabina) and its relatives (referred as the J. excelsa complex) make a taxonomically difficult group, so that these taxa are poorly distinguished morphologically. According to the latest study, this complex consists of three morphologically very similar species as: J. excelsa, J. polycarpos K. Koch. (var. polycarpos Linnaea and var. turcomanica R.P. Adams) and J. seravschanica Kom. (Hojjati et al., 2018). In Iran J. polycarpos is distributed in north, J. seravschanica is distributed in southeast and hybrid samples are distributed in southwest of the country and there is no J. excelsa (Hojjati et al., 2018). However there are different treatments of this group (Marschal von Bieberstein, 1800, 1808; Koch, 1849; Boissier, 1884; Fedtschenko et al., 1932; Komarov, 1932; Riedel, 1968; Farjon, 1992; Assadi, 1998; Adams, 1999, 2014). Hojjati et al. (2009) recognized 3 major population clusters as J. polycarpos (var. polycarpos and var. turcomanica) and J. seravschanica. of this complex in Iran using isozymes. Adams and Shanjani (2011) demonstrated that juniper from the Elburz Mtns. was typical J. polycarpos, not J. excelsa. Subsequently, Adams & Hojjati (2012) and Adams et al. (2014) based on four DNA ∗

regions (nrDNA, petN-psbM, trnD-trnT and trnS-trnG, 3,705 nucleotide site) concluded that samples from northwest of Iran are J. polycarpos var. polycarpos and samples from northeast and southwest of Iran are clearly J. polycarpos var. turcomanica. Samples from nearby south central Iran (Khabr protected area) are part of a clade including J. polycarpos var. seravschanica (Komarov) Kitamura. Here we performed an essential oil study on this complex in Iran to test the application of essential oils in systematics of this group. 2. Materials and methods 2.1. Plant materials Samples of J. excelsa complex were collected in November and December, 2006, from ten populations in Iran. Name of the population, herbarium number, location and locality altitudes for each specimen used in this study are given in Table 1. Voucher specimens were deposited at Tehran University Central Herbarium (TUH). Aerial parts of all samples were dried and stored at room temperature and subsequently cut into small pieces for distillation of essential oils.

Corresponding author. E-mail address: [email protected] (F. Hojjati).

https://doi.org/10.1016/j.bse.2019.03.004 Received 18 December 2018; Received in revised form 12 February 2019; Accepted 23 March 2019 0305-1978/ © 2019 Elsevier Ltd. All rights reserved.

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Table 1 Name of the population, herbarium number, location and locality altitude for accessions. Population

Species

Lushan1 Lushan2 Hashtjin Qushchi Shahmirzad Bajgiran Golestan Balade Fasa

J. J. J. J. J. J. J. J. J. J. J.

Khabr

polycarpos var. polycarpos var. polycarpos var. polycarpos var. polycarpos var. polycarpos var. polycarpos var. polycarpos var. polycarpos X seravschanica seravschanica

polycarps polycarps polycarps polycarps turcomanica turcomanica < turcomanica turcomanica

herbarium number

Location

Altitude(m)

33606 33607 33608 33609 33610 33611 33612 33613 33614

N of Iran. Prov. Gilan, Lushan to Jirandeh, 15 km to Jirandeh N of Iran. Prov. Gilan, 5 km after Jirandeh toward Yeilaqh and Amarloo N of Iran. Prov. Ardebil, 16 km toward Hashtjin after Khalkhal N of Iran. Prov. Azerbaijan, 28 km to Salmas after Orumieh N of Iran. Prov. Khorasan, 15 km after Shahmirzad toward Fooladmahalle N of Iran. Prov. Khorasan, 35 km to Bajgiran N of Iran. Prov. Golestan, Golestan National Park, Sharleqh, 9 km toward Azadshahr N of Iran. Prov. Mazandaran, 10 km after Baladeh toward Kojur S of Iran. Prov. Fars, 30 km after Fasa toward Neiriz

1100–1120 1670–1690 1590–1610 1730–1808 2422 1868 1055 2924 1607

33616

S of Iran. Prov. Kerman, Kuh-e Khabr

2418

2.2. Extraction and isolation of essential oils

compounds were identified accounting for 98.8–100% of the total oils. Essential oils from Lushan1, Lushan2, Hashtjin, Qushchi, Fasa, Khabr, Bajgiran, Golestan, Shahmirzad, Balade, were composed of 92 (99.2% of the total oil), 71 (99.4% of the total oil), 74 (98.8% of the total oil), 25 (100% of the total oil), 78 (99.9% of the total oil), 60 (100% of the total oil), 61 (100% of the total oil), 58 (100% of the total oil), 26 (99.9% of the total oil), 48 (100% of the total oil), components respectively. The main oil constituents of Lushan1 were α-pinene (66.4%), α-cedrol (3.3%), limonene (3.0%) and β-myrcene (3.0%); Lushan2 were α-pinene (61.0%), β-myrcene (4.0%), germacreneB (3.9%) and limonene (3.5%); Hashtjin were α-pinene (55.7%), α-cedrol (13.2%), δ-3-carene (6.3%), β-cedrene (3.1%) and β-myrcene (2.4%); Qushchi were α-pinene (90.9%) and β-myrcene (2.5%); Shahmirzad were α-pinene (82.0%) and trans-sobrerol (5.1%); Bajgiran were αpinene (46.8%), δ-3-carene (13.7%), δ-cadinene (4.1%), germacreneB (3.2%), α-amorphene (2.5%), germacreneD (2.4%), limonene (2.4%), hexyl isovalerate (2.4%) and β-myrcene (2.4%); Golestan were αpinene (80.1%) and β-myrcene (4.3%); Balade were α-pinene (79.7%), β-myrcene (4.2%) and limonene (3.9%); Fasa were α-pinene (28.6%), α-cedrol (29.5%), β-cedrene (4.3%), γ-cadinene (3.4%) and δ-cadinene (2.8%) and Khabr were α-pinene (12.5%), α-cedrol (9.6%), limonene (9.1%), trans caryophyllene (6.9%), δ-3-carene (6.2%), α-humulene (5.5%), β-myrcene (5.5%), β-pinene (5.4%), β-cedrene (4.7%) and germacreneB (2.6%). The quantitative cluster analysis performed (Fig. 1) and revealed two clusters: a cluster including the southern populations (named as cluster A including Fasa and Khabr populations) at the phenon line 56.03 based on 144 compounds and cluster B (Samples from N Iran).

Leaf samples from Lushan1 (189 g), Lushan 2 (124 g), Hashtjin (60.35 g), Qushchi (269.77 g), Shahmirzad (125.85 g), Bajgiran (118.90 g), Golestan (151.43 g), Balade (161.80 g), Fasa (278.24 g) and Khabr (157.81 g) were subjected to hydrodistillation for 5 h in a circulatory Clevenger type apparatus. The essential oils were collected and stored at −20 °C. 2.3. General experimental procedures GC-MS analyses were performed on an Agilent Technologies (USA) 6890 GC system coupled with a post column splitting 5973 network mass selective detector with a quadrupole analyzer and resolution of 0.1 amu and equipped with a HP5-MS capillary fused silica column (60 m × 0.25 mm i.d.), 0.25 μm film thickness, methyl 5% phenyl polysiloxane (Agilent Technologies, USA). The temperature program initiated at 40 °C, held for 1 min then raised at 3 °C min−1 to 250 °C, held for 20 min. Carrier gas, helium (99.999%); with a flow rate of 1 mL min−1; injector temperature, 250 °C; split ratio, 1:50. Mass spectra were taken at 70 eV. Mass range was from 20 to 500 amu. An enhanced ChemStation G1701 DA version D.00.01.27 was used for the data collection and processing. The injections of sample into GC and GC-MS were carried out using a 10 μL micro-syringe model ITO MS-E10 (Japan) with the needle tip of angled cut. 2.4. Identification of essential oil component Compounds in essential oils were identified by comparison to Wiley7n.l MS mass spectra data libraries. The Kovats’ retention indices of all the constituents were obtained by interpolating between bracketing n-alkanes (Oprean et al., 1998, 2001; Burt, 2004).

4. Discussion β-Myrcene, Limonene and α-Pinene with high concentrations were found to be in the main constituents of all population oils in addition to β- Cedrene and α-Cedrol for Khabr and Fasa populations. Most of these main components were reported previously for J. polycarpos (Adams, 2001), but the components considered as characteristic for J. excelsa (Adams, 2001), like decadienal isomer (KI 1312), trans-cadina-1 (6), 4diene, cubebol, 1-epi-cubenol, and KI 1666, were not observed in any population oil studied here. Therefore, the existence of J. excelsa in Iran, finds no support from the data presented here. These data also confirm the presence of J. polycarpos throughout Iran except for South of the country. One of the components known as characteristic for J. polycarpos according to Adams (2001) is δ-elemene that were not found in the southern populations, including Fasa and Khabr (cluster A). Moreover borneol and isopinocamphone, were diagnostic for cluster A populations. Some compounds were at highest levels in both Fasa and Khabr populations including linalool, β-cedrene, β-eudesmol, transcaryophyllene, widdrene and caryophyllene. Compound α-pinene was at the lowest levels in both Khabr and Fasa populations. Some compounds were observed only in Khabr population including manonyl oxide, cis-sabinol, α-eudesmol, fenchyl acetate, 2-carene, α-cubebene,

2.5. Statistical analyses Essential oil compositions were scored quantitatively so that the percentage of each composition was included in the matrix as a Euclidean coefficient. Clustering was performed using the Unweighted Paired Groups Arithmetic Average (UPGMA) with the NTSYS- pc version 2.02 k (Applied Biostatistics Inc., 1986–1998). 3. Results Hydrodistillation of samples from ten populations of J. excelsa complex yielded in 0.08%–3.78% dry weight (v/w) clear oils. The highest oil content was found in the sample from Bajgiran (3.8%), followed by Hashtjin (3.3%), Golestan (2.6%), Lushan1 (2.1%), Shahmirzad (1.2), Qushchi (0.7%), Khabr (0.6%), Fasa (0.1%), Baladeh (0.1%) and Lushan2 (0.1%). The chemical constituents of essential oils identified by GC-MS in combination with their retention indices (RI) and percentages are listed in Table 2. One hundred sixty four 30

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Table 2 Total oil component percentages of the leaf essential oils of ten populations of Juniperus excelsa complex in Iran: Lushan1 (L1), Lushan2 (L2), Hashtjin (H), Qushchi (Q) as J. polycarpos var. polycarpos, Shahmirzad (S), Bajgiran (BJ), Golestan (G), Baladeh (BL) as J. polycarpos var. turcomanica, Fasa (F) as J. polycarpos X J. seravschanica, and Khabr (K) as J. seravschanica. Component 3-Methyl butanal 1,2-Dimethyl-4-ethylene cyclopentene Hexanal 2-Methyl-3-buten2-ol 1-Pentene-3-ol 3-Methyl-1-butanol 1-Methoxy hexane 1-Hexanol Tricyclene α-Pinene Comphene Verbene m-Cymene Sabinene β-Pinene 2,3-Dehydro-1,8cineol Mentha-1,4,8-triene β-Myrcene Mentha-1,5,8-triene Phellandrene 4-(1-methylethenyl) δ-3-Carene Cymol α-Terpinene o-Cymene Cyclohexene,1methyl-4-(1methylethyl) β-Phellandrene p-Cymene Limonene cis-Ocymene Butanoic acid,3-methyl-, buthyl ester n-Buthyl isovalerate β-OcymeneY 1,3,6-Octatriene,3,7-dimethyl γ-Terpinene 1-Octanol Cyclooctane l-Fenchone Fencholenic aldehyde α-Thujone o-Allyltoluene α-Terpinolene Linalool Isoamyl valerianate d-Fenchyl alcohol 2,3,3-Trimethyl-3-cyclopentene acetaldehyde α-Campholene aldehyde Comphore trans-Pinocarveol cis-Sabinol Pinocarvone cis-Verbenol Limonen-10-ol 3-Pinanone Isopinocamphone Sopulegol p-Mentha-1,5-diene-8-ol α-Cedrene Borneol Unknown p-Acetoplenone 4-Terpineol p-Cymene-8-ol Myrtenal β-Fenchyl alcohol

RI

L1

L2

H

Q

S

BJ

G

BL

F

K

t t t _

_ _ t _

_ _ t _

_ _ 0.1 _

_ _ _ 1.0

_ _ t _

_ _ t _

_ _ t _

_ _ t _

_ _ _ _

741 782 846 868 883 884 895 898 902 912

_ _ 0.2 t 0.2 66.7 0.8 0.3 _ _ 1.6 t

_ _ 0.1 _ 0.4 61.0 0.9 0.2 _ 0.1 2.0 _

_ _ t t 0.2 55.7 0.8 t t 0.2 0.9 _

_ _ 0.6 0.1 0.7 90.9 1.0 0.2 _ 0.3 1.5 _

0.5 0.3 0.2 1.3 0.4 82.9 0.5 0.4 _ _ 1.0 _

_ _ 0.8 0.1 0.1 46.8 0.8 0.2 0.2 _ 1.2 _

_ _ 0.1 0.7 0.2 80.1 0.9 0.2 _ _ 1.9 _

_ _ 0.7 _ 0.1 79.7 1.6 0.4 _ _ 2.0 _

_ _ t _ 0.2 28.6 0.6 0.1 0.1 _ 1.0 _

_ _ _ _ 0.4 12.5 2.3 0.3 0.1 _ 5.4 _

914 918 923 925 929 934 935 937 940 945

0.1 3.0 0.1 _ t 0.1 _ 0.1 _ t

_ 4.0 t 0.1 _ 0.1 _ 0.1 0.5 _

_ 2.4 _ t _ 6.3 _ 0.1 0.2 _

_ 2.5 _ _ _ 0.1 _ t 0.2 _

_ 1.2 _ _ _ 0.6 0.4 _ _ _

t 2.4 _ _ _ 13.7 _ _ _ _

t 4.3 _ _ _ 0.1 _ 0.1 1.3 _

0.1 4.2 0.1 _ _ 0.1 _ 0.2 0.5 _

_ 0.6 _ _ _ 2.0 _ t 0.2 _

0.1 5.5 _ 0.2 _ 6.2 _ 0.1 1.6 _

946 949 950 958 961 962 964 967 974 983 985 989 992

0.1 0.8 3.0 0.1 0.1 _ _ t 0.8 _ t 0.7 0.1

0.2 _ 3.5 0.1 _ t t _ 1.0 _ _ 0.2 0.1

_ _ 1.5 t _ t t _ 0.7 t _ 0.1 t

0.1 _ 1.1 _ _ _ t _ 0.1 _ _ 0.1 _

_ _ 1.1 _ _ _ _ _ 0.2 _ _ _ _

0.3 0.5 2.4 t _ 0.1 t _ 0.6 _ _ _ 0.1

0.1 _ 2.1 t _ 0.1 _ _ 0.7 _ _ _ t

0.1 _ 3.9 t _ 0.6 _ _ 1.0 _ _ _ t

0.3 _ 0.6 _ _ _ _ _ 0.2 0.1 _ _ 0.1

0.8 _ 9.1 _ _ _ _ _ 0.8 _ _ 0.7 0.1

993 997 1002 1010 1015 1017 1020

_ 0.1 0.9 0.2 0.2 0.1 _

_ t 1.1 0.1 0.3 _ 0.2

_ t 1.4 0.1 _ 0.1 _

_ _ 0.2 _ _ _ _

_ _ 0.3 _ _ _ _

0.4 0.1 1.9 0.3 0.3 _ _

_ 0.1 0.9 _ 0.5 0.1 _

_ t 1.1 _ 0.4 _ _

_ 0.1 0.3 0.7 0.1 0.1 _

_ 0.2 1.7 1.5 0.1 0.2 _

1021

0.3

_

t

_

0.3

_

_

_

0.2

0.8

1032 1036 1038 1040 1041 1043 1048 1049 1054 1059 1060 1061 1062 1063 1072 1073 1078 1082

0.8 0.3 _ 0.2 _ _ _ _ t 0.4 _ _ 0.8 _ _ _ 0.1 _

0.2 0.2 _ _ 0.1 _ _ _ _ _ _ _ _ _ 0.1 _ 0.1 _

t 0.1 _ _ _ _ t _ _ 0.2 t _ _ _ _ t t _

t _ _ t _ _ _ _ _ _ _ _ _ _ _ _ t _

0.1 _ _ 0.2 _ _ _ _ _ _ _ _ _ 0.1 _ 0.2 0.1 _

0.8 _ _ 0.3 0.1 _ _ _ _ _ _ _ _ _ _ _ _ _

0.6 _ _ 0.1 _ _ _ _ _ _ _ _ _ _ _ _ _ _

0.6 _ _ 0.1 _ _ _ _ _ _ _ _ _ _ _ _ _ _

0.2 _ _ 0.3 0.1 0.3 _ 0.2 _ _ _ 0.6 _ _ 0.1 _ t 0.1

0.3 _ 0.8 _ 0.6 _ _ 0.4 _ _ _ 2.1 _ _ _ 0.1 0.5 _

(continued on next page) 31

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Table 2 (continued) Component

RI

L1

L2

H

Q

S

BJ

G

BL

F

K

α-Terpineol cis-4-Decenal Verbenone Myrtenol Berbenone Nopol Isoborneol trans-Carveol Hexyl butanoate Fenchyl acetate Palegone β-Citronellol Hexyl isovalerate Isopentyl hexanoate cis-4-Decen-1-ol trans-Anethone Bornyl acetate 2-Undecanone trans-Pinocarvyl acetate (E)-L-Formyl-6-methyl-3-(1-propylidene)-1-cyclohexene 2,4-Decadienal 2,4-Decadie-1-ol trans-Carvyl acetate δ-Elemene 2-Carene α-Cubebene trans-Sobrerol Hexyl hexanoate α-Copaene α-Bourbonene Hexyl caproate β-Bourbonene β-Cubebene β-Elemene Zingiberene β-Cedrene trans-Caryophyllene Widdrene Thujopsene γ-Elemene Soledene α-Humulene β-Farnesene Italicene epiBicyclosequiphellan drene ar-Curcumene Germacrene D α-Amorphene β-Selinene Cadinene Epizonaren α-Selinene Cuparene α-Muurolene Germacrene A α-Elemene γ-Cadinene γ-Selinene Ylangene depi-α-Cedrene δ-Cadinene α-Cadinene β-Himachalene β-Panasinene Ledene trans-γ-Bisabolene δ-Selinene Selina-3,7 (11)-diene Elemol Nerolidol Germacrene B Caryophyllene oxide Unknown

1083 1085 1087 1089 1093 1095 1096 1107 1113 1117 1118 1121 1138 1143 1148 1160 1169 1177 1178

0.2 _ _ 0.4 _ _ 0.1 0.1 _ _ _ _ 0.7 _ 0.1 _ 0.6 _ _

0.1 0.1 0.2 _ _ 0.1 _ 0.1 _ _ _ 0.1 0.2 _ 0.1 _ 0.9 _ 0.1

0.1 _ 0.1 _ _ – _ t _ _ _ t 0.2 _ t _ 0.2 _ _

_ _ 0.1 _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _

_ _ 0.4 _ _ _ _ _ _ _ _ _ _ _ _ _ 0.3 _ _

_ _ 0.4 _ 0.6 _ _ 0.1 0.2 _ _ _ 2.4 _ 0.7 _ 0.1 _ _

0.1 _ 0.1 _ _ _ _ _ 0.4 _ _ _ 0.8 0.3 t _ 0.1 _ _

_ _ _ _ _ _ _ _ _ _ _ _ 0.2 _ _ _ 0.1 _ _

_ 0.1 0.2 _ _ 0.1 _ 0.1 _ _ 0.1 0.1 0.4 0.1 0.6 0.1 0.4 0.1 _

0.3 _ _ _ _ _ 0.2 0.2 _ 0.1 _ _ 0.7 _ 0.4 _ 1.3 _ _

1186 1187 1193 1217 1234 1237 1245 1249 1262 1275 1292 1293 1294 1301 1302 1305 1316 1320 1327 1328 1330 1339 1342 1347 1350 1351 1358

0.2 _ _ _ 0.3 _ 0.1 _ _ 0.1 _ _ _ _ 0.4 _ 1.2 0.4 0.3 _ 0.5 0.2 _ 0.3 0.1 _ _

_ 0.1 0.2 t 0.9 _ _ _ _ 0.2 0.1 _ _ _ 0.8 _ _ 1.3 _ _ 0.9 _ 0.4 _ _ 0.2 _

0.1 _ _ _ 0.2 _ t _ 0.2 _ _ _ _ _ 0.2 0.1 3.1 _ _ 0.7 0.2 0.2 _ 0.8 0.3 0.1 _

_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ 0.1 _ _ _ _ _ _ _ _ _ _

_ _ _ _ _ _ _ 5.1 _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _

_ _ 0.1 _ 0.5 _ 0.1 _ 0.4 0.4 _ _ _ 0.1 0.6 _ _ 1.5 _ _ 0.5 _ 0.5 _ _ 0.3 _

_ _ _ _ 0.1 _ t _ 0.5 0.1 _ _ _ _ 0.1 _ _ 0.3 _ _ t 0.1 _ _ _ t 0.2

_ _ _ _ 0.2 _ t _ _ 0.1 _ _ _ _ 0.1 _ _ 0.3 _ _ 0.1 0.1 t _ _ t _

_ 0.1 0.2 _ _ _ _ _ _ 0.3 _ 0.3 _ _ 0.4 0.1 4.3 2.1 1.2 _ 0.5 _ 0.5 _ _ _ _

_ _ _ _ _ 0.4 0.2 _ _ _ _ _ 0.7 _ 0.7 _ 4.7 6.9 1.1 _ 1.2 _ 5.5 0.9 0.3 _ _

1359 1361 1363 1366 1367 1369 1371 1372 1373 1376 1379 1380 1381 1382 1388 1389 1391 1392 1393 1394 1395 1399 1400 1411 1412 1425 1439

0.5 0.2 0.2 0.1 _ 0.4 _ 0.2 _ _ 0.8 _ _ _ 1.2 _ _ _ 0.3 _ 0.1 _ 0.3 _ 1.1 0.1 _

2.3 1.4 0.1 _ _ _ _ 0.6 0.2 _ _ 0.2 _ _ 2.3 0.2 _ _ _ _ _ _ 0.7 0.1 3.9 _ _

0.6 0.2 0.2 0.1 _ _ _ 0.2 _ 0.1 _ _ _ 0.8 1.4 _ 0.2 _ _ 0.6 _ 0.3 _ _ 1.3 _ 1.3

_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _

_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _

2.4 2.5 0.3 _ 0.4 0.5 _ 1.1 _ _ 0.2 _ t _ 4.1 _ _ _ _ _ _ 0.3 _ _ 3.2 _ _

_ 0.1 0.1 _ _ 0.1 _ t _ _ 0.2 _ _ _ 0.2 _ _ t _ _ t t _ _ 0.2 _ _

0.2 t t _ _ _ _ 0.1 _ _ 0.1 _ _ _ 0.2 _ _ _ _ _ t t _ _ 0.2 _ _

0.9 0.6 0.3 _ _ _ 1.4 0.5 _ _ 3.4 _ _ 0.3 2.8 0.2 _ _ _ 0.5 _ 0.5 _ _ 2.1 0.9 _

2.1 _ 0.5 _ _ _ _ _ _ _ _ _ 0.1 _ _ _ _ _ _ 1.0 _ _ 0.8 _ 2.6 0.5 _

(continued on next page) 32

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Table 2 (continued) Component

RI

L1

L2

H

Q

S

BJ

G

BL

F

K

Spathulanol endo-1Bourbonanol α-Cedrol α-Eudesmol β-Oplopenone Fonenol Neryl acetate 8-β-H-Cedran-8-ol γ-Eudesmol β-Eudesmol tau-Cadinol t-Cadinol t-Muurolol α-Cadinol δ-Gurjunene Unknown Methyl myristate Cedranyl acetate α-Gurjunene Methyl palmitate Hexadecanoic acid Manonyl oxide Isopimaradiene Sclarene Methyl oleate Methyl stearate 9-Octadecanoic acid Octadecanoic acid Di (2-ethylhexyl) phthalate

1440 1443 1451

_ _ 3.3

_ 2.2 _

_ _ 13.2

_ _ t

_ _ 1.4

_ 0.2 _

_ _ t

_ _ t

1.9 _ 29.5

_ _ 9.6

1461 1469 1471 1478 1480 1485 1491 1496 1497 1502 1507 1513 1531 1564 1581 1593 1755 1781 1792 1799 1856 1877 1899 1902 1921 2141

_ _ 0.1 _ _ 0.1 0.1 0.2 _ _ 0.2 _ _ t _ t t t _ t _ t t t t _

_ 0.1 _ 0.1 _ _ _ _ _ 0.7 0.8 0.1 0.6 _ _ 0.1 _ _ _ _ _ _ _ _ _ _

_ _ _ _ 0.2 _ _ _ _ _ 0.3 _ _ _ 0.1 _ _ _ _ t 0.1 _ _ _ _ _

_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _

_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _

_ _ _ _ _ _ _ _ _ 0.3 0.2 _ _ _ _ 0.8 _ _ _ _ _ _ _ _ _ _

_ _ _ _ _ _ t _ t t _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _

_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ t _ _ _ _ _ t _ _

_ _ _ _ _ _ 0.4 _ 0.6 0.6 _ _ _ _ _ _ _ 0.2 _ _ _ _ _ 0.3 _ 1.2

0.6 _ _ _ _ _ 0.6 _ _ _ _ _ _ _ _ _ _ _ t t _ _ _ _ _ _

RI = Retention Index; t = trace quantities (< 0.1).

β-bourbonene and some compounds were only in Fasa population, including limonene-10-ol, β-fenchyl alcohol, trans-anethone, 2-undecanone, di (2-ethylhexyl) phthalate, hexyl caproate, cuparene, spathulanol and palegone. These data confirm occurring of two separated species in Iran: J. polycarpos in north and J. seravschanica in SE Iran.

Samples from SW Iran have been recognized as hybrids between J. polycarpos in north and J. seravschanica in SE (Hojjati et al., 2018). Data presented here also confirms SW populations probable backcrossing with J. seravschanica as a parent.

Fig. 1. UPGMA cluster analysis of essential oils of ten populations of Juniperus excelsa complex in Iran based on the quantitative data using Euclidean distance.

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Acknowledgement

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