Achene Fatty Acid Composition in the Tribe Anthemideae (Asteraceae)

Received for publication, July 17, 2014 Accepted, August 7, 2015

FAIK AHMET AYAZ*, HUSEYIN INCEER, SEMA HAYIRLIOGLU-AYAZ, NURSEN AKSU-KALMUK Karadeniz Technical University, Faculty of Science, Department of Biology, Trabzon, Turkey *Address correspondence to: Karadeniz Technical University, Faculty of Science, Department of Biology, 61080 Trabzon, Turkey. Tel.: +90462 337 3712; Fax.: +90462 337 3712; E-mail: [email protected]

Abstract Fatty acid composition of the achene of 44 taxa belonging to seven genera (Achillea, Anthemis, Artemisia, Glebionis, Matricaria, Tanacetum and Tripleurospermum) of the tribe Anthemideae (Asteraceae) from Turkey were analysed using gas chromatography. The presence of 16 fatty acids in the studied taxa was determined and quantified. The content of palmitic acid (C16:0) was 11.34%-30.20%, of oleic acid (C18:1) 4.17%-23.55, of linoleic acid (C18:2) 27.39%-65.52% and of linolenic acid (C18:3) 0.39%-21.53%. Total unsaturated fatty acids (UFAs) in the achene varied greatly among the studied taxa. As averaged, the highest total MUFA was 12.99%, while UFA 69.39% and PUFA 56.41%. The ratio of UFA/SFA was between 1.03 and 5.70. Results are discussed in terms of nutritional importance of these species as potential candidates for inclusion in botanically designated pastures to contribute to improving milk quality.

Keywords: achene, Anthemideae, fatty acid, Turkey

1. Introduction The health promoting effects of biologically active compounds in chronic diseases have received wide attention in the past two decades. Plants represent the unique source of biologically active compounds. Numerous plant species, mostly spices and aromatic herbs, have been examined for their nutrient and non-nutrient characteristics in many human age-related degenerative diseases. Many edible plants are capable of producing natural chemopreventive compounds which have no synthetic counterparts and which play a protective role in human health maintenance. Fatty acids (FAs) and their chemo-preventive effects are pharmacologically active in chronic or degenerative diseases. Research has proved that diets rich in saturated fatty acid (SFA) are a risk factor for cardiovascular diseases (CVDs) due to their raising serum low density lipoprotein (LDL) cholesterol concentrations. In contrast, it has been suggested that diets rich in monounsaturated fatty acids and in some polyunsaturated fatty acids (PUFAs), especially oleic acid (OA), linoleic acid (LA) and linolenic acid (LN), reduce or inhibit such CVDs (VASSILIOU & al. [1]; KRIS-ETHERTON & al. [2]; URSIN [3]). These acids are therefore essential to human health (ROS and MATAIX [4]). The human body is unable to synthesize these FAs. Known as essential FAs, neither linoleic acid (C18:2n-6) nor -

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Achene Fatty Acid Composition in the Tribe Anthemideae (Asteraceae)

linolenic acid (C18:3n-3) can be endogenously synthesized by humans. They therefore need to be supplied by diet, such as from seed oil from safflower, sunflower, pine nuts, sesame, flaxeed, walnuts, soybean, canola etc., or from fish, whale, seal, walrus, cod-liver, herring, salmon, etc., (KRIS-ETHERTON & al. [2]; URSIN [3]; ROS and MATAIX [4]; CONNOR [5]; INNIS [6]). Mammals are not able to introduce double bonds between the terminal methyl group and the ninth carbon atom in the FA chain (MCDONALD & al. [7]), while plants have the ability to synthesise C18:3n-3 acid de novo (DEWHURST & al. [8]). Efforts have therefore been made to introduce new oil seeds, with high nutritional and pharmaceutical values (GOLI & al. [9]). The tribe Anthemideae (Asteraceae) has a worldwide distribution, although the taxa are concentrated in Central Asia, the Mediterranean region and South Africa. Some members of the subtribes Ursinniinae, Artemisiinae, Chrysantheminae, Leucantheminae, Anthemidinae and Matricarinae are pernicious weeds, such as some of the Ursinia species introduced in Australia and New Zealand. Species of Achillea, Anthemis, Artemisia, Glebionis, Leucanthemum, Matricaria and Tripleurospermum are widespread weeds in both the northern and southern hemispheres (BREMER and HUMPRIES [10], OBERPRIELER & al. [11]). However, most taxa have discrete ranges and very obvious areas of endemism (BREMER and HUMPRIES [10]). Members of the tribe have for long been used as medicinal herbs in folk and alternative medicines. Much research has been conducted into essential oil compositions of the taxa in the tribe (BREMER and HUMPRIES [10]). However, little information about the FA composition in achenes of several species of the tribe has been reported (GOLI & al. [9]; PALIĆ & al. [12]; TSEVEGSUREN & al. [13]; WARNER & al. [14]). This scarcity led us to profile the FA composition in the achenes of 44 taxa belonging to seven genera (Achillea, Anthemis, Artemisia, Glebionis, Matricaria, Tanacetum and Tripleurospermum) of the tribe which are native to Turkey.

2. Materials and Methods 2.1. Experimental details and treatments 2.1.1. Experimental material Mature achenes of 44 taxa of the tribe Anthemidea were used for chemical analysis. Achenes, including mature achenes, belonging to 5-7 capitula of each taxon were collected from natural bulk populations in Turkey (Table 1). Plant vouchers are deposited in the herbarium at the Karadeniz Technical University, Department of Biology (KTUB). The mature achenes were removed from the fruit debris and used for extraction.

Table 1. Locality and voucher number of the investigated species

Taxon Locality Voucher

Achillea biebersteinii Hub.-Mor. A7 Gümüşhane: Köse Mountain, 1800 m a.s.l., 23.vii.2001. Inceer 142 A. bisserata M. Bieb. A7 Gümüşhane: Zigana Mountain, between Zigana Pass and Torul, Inceer 668 1200-1300 m a.s.l., 06.vii.2006. A. multifida (DC.) Griseb. A2 Bursa: Uludağ, 1820 m a.s.l., 27.vi.2007. Inceer 357 A. wilhelmsii K. Koch A3 Bolu: Near Abant Lake, 1331 m a.s.l., Inceer 575 12.vi. 2008. Anthemis cotula L. A2 Bursa: Uludağ-Bursa road, 1050 m a.s.l., 28.vi.2007. Inceer 368

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A. macrotis (Rech. f.) Oberpr. & Vogt C1 Muğla: Between Muğla and Köyceğiz, 11 m a.s.l., 10.iv.2008. Inceer 496 Artemisia annua L. A7 Trabzon: Near KTU, Kanuni Campus, 100 m a.s.l., 02.x.2008. Inceer 708 A. austriaca Jacq. A7 Gümüşhane: Köse Mountain, 1800 m a.s.l., 23.vii.2001. Inceer 140 A. santonicum L. A9 Erzurum: Near Olur, 983 m a.s.l., 19.vii.2007. Inceer 445 Glebionis coronaria (L.) Spach C1 Aydın: Kuşadası, 10 m a.s.l., 08.iv.2008. Inceer 483 G. segetum L. Fourr. A1 Çanakkale: From Bayramiç to Çanakkale, 250 m a.s.l., 12.v.2007. Inceer 337 Matricaria aurea (Loefl.) Sch. Bip. C1 Gaziantep/Şanlıurfa: Between Nizip-Birecik, near Dutlu, 440 m Inceer 322 a.s.l., 08.v.2007. M. chamomilla L. var. chamomilla C1 Muğla: between Milas and Bodrum, 10 m a.s.l., 08.iv.2008. Inceer 486 M. chamomilla L. var. recutita (L.) Fiori B1 İzmir: From Torbalı to Aydın, 64 m a.s.l., 07.iv.2008. Inceer 481 M. matricarioides (Less.) Porter A9 Kars: Kars-Ardahan, Göle Road, 1800 m a.s.l., 18.vii.2007. Inceer 420 Tanecetum vulgare L. A7 Giresun: Between Şehitler Pass and Şebinkarahisar, 1583 m a.s.l., Inceer 662 21.vii.2008. Tripleurospermum baytopianum E. Hossain A1 (E) Tekirdağ: From Malkara to Şarköy, 141 m a.s.l., 16.iv.2008. Inceer 506 T. callosum (Boiss & Hedlr) E. Hossain A7 Gümüşhane: Keçikaya Village, 1618 m a.s.l., 04.vii.2007. Inceer 382a T. caucasicum (Willd.) Hayek A8 Rize: Ayder, Yukarı Kavrun, 2278 m a.s.l., 23.vii.2008. Inceer 672 T. caucasicum (Willd.) Hayek A8 Rize: Ayder, Yukarı Kavrun, 2000 m a.s.l., 11.vii.2009. Inceer 765 T. conoclinium (Boiss. & Bal.) Hayek B2 İzmir: Bozdağ, 1178 m a.s.l., 14.iv.2007. Inceer 264 T. corymbosum E. Hossain B9 Ağrı: Suluçem (Musun), Balık Gölü, 2098 m a.s.l., 11.vii.2008. Inceer 612 T. decipiens (Fisch. & C. A. Mey.) Bornm. B3 Eskişehir: Midas road, 1290 m a.s.l., 28.vi.2007. Inceer 375 T. elongatum (DC.) Bornm. A7 Giresun: From Şehitler Pass to Şebinkarahisar, 1317 m a.s.l., Inceer 664 21.vii.2008. T. fissurale (Sosn.) E. Hossain A8 Artvin: Between Ispir and Yusufeli, 10 km to Yusufeli, 653 m a.s.l., Inceer 533 31.v.2008. T. heterolepis (Freyn & Sint.) Bornm. A7 Gümüşhane: Keçikaya Village, 1618 m a.s.l., 04.vii.2007. Inceer 382b T. hygrophilum (Bornm.) Bornm. B1 İzmir: Yamanlar Mountain, 887 m a.s.l., 15.iv.2007. Inceer 273 T. inodorum (L.) Sch. Bip. A9 Erzurum: Between Pasinler and Horasan, near Horasan, Köprü Inceer 600 Village, 1600 m a.s.l., 11.vii.2008. T. kotschyi (Boiss.) E. Hossain C5 Niğde: Ulukışla, Bolkar Mountains, near Karagöl, 2600 m a.s.l., Inceer 702 29.vii.2008. T. melanolepis (Boiss. & Buhse) Pobed. A9 Artvin: Şavşat, near Çamlıbel pass, 2550 – 2600 m a.s.l., Inceer 741 20.vi.2009. T. microcephalum (Boiss.) Bornm. B8 Muş: Near fallow fields, 1323 m a.s.l., 09. vii.2008. Inceer 594 T. monticolum Bornm. B8 Erzurum: Palandöken Mountain, 2907 m a.s.l., 13.vii.2008. Inceer 639 T. oreades var. oreades (Boiss.) Rech. f. A7 Giresun: Kümbet Plateau, 1719 m a.s.l., 21.vii.2008. Inceer 658 var. oreades T. oreades (Boiss.) Rech. f. var. A9 Artvin: Şavşat, 2185 m a.s.l., 17.vii.2007. Inceer 414 tchihatchewii (Boiss.) E. Hossain T. parviflorum (Willd.) Pobed C2 Denizli: Between Pamukkale and Denizli, cultivated fields, 931 m Inceer 499 a.s.l., 10.iv.2008. T. pichleri (Boiss.) Bornm. A2 Bursa: Uludağ, near hotels, 1828 m a. s. l., 11.vi.2008. Inceer 553 T. repens (Freyn&Sint.) Bornm. A7 Gümüşhane: From Gezge Village to Gezge, 1987 m a.s.l., Inceer 385 08.vii.2007. T. rosellum (Boiss. & Orph.) Hayek var. A1 Çanakkale: Gökçeada, between centrum and Dereköy, 220 m a.s.l., Inceer 719 album E. Hossain 17.iv.2009. T. rosellum (Boiss. & Orph.) Hayek var. A8 Rize: Between İkizdere and Cimil, 1800 – 1850 m a.s.l., Inceer 766 album E. Hossain 23.vii.2009. T. sevanense (Manden.) Pobed. B3 Eskişehir: Çatacık, 1304 m a.s.l., 27.vi.2007. Inceer 369b T. subnivale Pobed. A8 Rize: Ayder, Yukarı Kavrun, 2278 m a.s.l., 23.vii.2008. Inceer 671 T. tenuifolium (Kit.) Freyn ex Freyn A2 Bursa: Uludağ, near hotels, 1690 m a.s.l., 27.vi.2007. Inceer 353 T. transcaucasicum (Manden.) Pobed A9 Kars: Yalnızçam Mountains, from Göle to Olur, 980 m a.s.l., Inceer 438 19.vii.2007. T. ziganaense Inceer & Hayirlıoglu-Ayaz A7 Gümüşhane: Zigana Dağı, between Zigana Pass and Torul, 1300 m Inceer 666 a.s.l., 22.vii.2008.

2.1.2. Lipid extraction and fatty acid analysis Total lipids were extracted according to the method described by FOLCH & al. [15] with modifications. Briefly, approximately 0.5 g of achene sample in triplicate was extracted with 20mL of chloroform/methanol (2:1, v/v) at 4°C for 18h. The extracted lipids in the chloroform phase were separated from the aqueous phase by shaking and partitioning with

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4mL of 0.9% (w/v) NaCl, and were then collected and evaporated under a stream of nitrogen gas. The lipids were then redissolved with 5mL of chloroform. The fatty acid methyl esters were extracted with hexane and analysed and quantified using a Perkin Elmer Auto System XL gas chromatography equipped with a flame-ionization detector and a fused-silica capillary column (SP-2330, 30m  0.25mm, i.d., film thickness 0.20μm, Supelco, Bellefonte, PA, USA). Helium was used as the carrier gas. The injector was set at 240°C and the detector at 260°C. The temperature of the oven was initially 120°C. This was then raised to 220°C at 5°C/min increments and maintained for 10 min. Fatty acids were identified by comparison with the retention times of a standard mixture of fatty acid methyl esters (Supelco 37 components FAME Mix, SUPELCO Bellefonte, PA, USA). Quantification was carried out by internal standardization with triheptadecanoin (Sigma, St. Louis, MO, USA). The coefficient of variation ranged from 1.5% to 5.0% for the fatty acids (FOLCH & al. [15]).

3. Results and Conclusions This study investigated the FA composition of achenes of 44 taxa belonging to seven different genera (Achillea, Anthemis, Artemisia, Glebionis, Matricaria, Tanacetum and Tripleurospermum) in the tribe Anthemideae. Sixteen FAs were identified and quantified in the achenes, and their total FAs were evaluated in terms of total saturated (SFA), unsaturated (UFA), monounsaturated (MUFA) and polyunsaturated (PUFA) fatty acids (Tables 2 and 3). In the achenes of the studied taxa, palmitic acid in SFA (Table 2) and oleic acid (OA), linoleic acid (LA) and linolenic acid (LN) in UFA (Table 3) were determined in the greatest quantities. For instance, palmitic acid (C16:0) as the major component of the SFA, varied between 11.34% and 30.20%, averaging 21.10% among the studied species. It exhibited the highest content level in M. chamomilla var. chamomilla (30.20%). Stearic acid, averaged 3.67%, was highest in A. wilhelmsii (12.59%) and lowest in T. sevanense (1.15%). Other SFAs with their minor contents, such as C10:0, C12:0, C14:0, C15:0, C20:0, C22:0 and C24:0, averaged between 0.08% and 1.28% (Table 2). The oil achene of all the studied taxa was rich in linoleic, linolenic and oleic acid contents. Considerable variation was determined among the species in terms of the contents of the three major UFAs. Linoleic acid was found in the highest proportion in the achenes of the studied species. Its content (ave. 46.27%) was highest in T. heterolepis (65.52%). With its low average content (8.73%), linolenic acid was the predominant FA in T. caucasicum (Rize pop.-Inceer 765) (21.53%). Oleic acid was the second averaged FA (11.76%) and was highest in T. elongatum (23.55%). The contents of the remaining four unsaturated FAs, C15:1, C16:1, C20:4n-6 and C22:6n-3 were relatively low or sometimes not detected, at 0.55%, 0.68%, 0.98% and 0.43%, respectively (Table 3). Total UFAs varied greatly among the 44 taxa studied. The highest total MUFA, averaged 12.99%, was estimated for T. elongatum (23.55%) and the highest UFA (averaged 69.39%) for T. heterolepis (85.09%) and T. callosum (85.05%). PUFA, averaged 56.41%, was highest for T. callosum (68.41%) (Table 3). The ratio of UFA/SFA ranged between 1.03 in A. austriaca and 5.70 in T. heterolepis. In the studied taxa, A. cotula had the lowest (0.81) and T. callosum the highest (4.56) PUFA/SFA ratios (Table 3). The present findings are comparable with the data in the literature. Study of the tribe has largely concentrated on its essential oil components, due to the way members of the tribe are used in folk and alternative medicine (BREMER and HUMPRIES [10]). No satisfactory data have been reported in the literature with which to evaluate the FA composition of the tribe Anthemideae with the present findings. Several reports have been published, but these contain one to three species. WARNER & al. [14] Romanian Biotechnological Letters, Vol. 21, No. 3, 2016 11579

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recently reported low amounts of C16:0 (4%), C18:2 (5.6%) and C18:3 (10.68%) content in A. millefolium during the growing season. A significant variation in the content of C18:2 and C18:3 in three Achillea species was reported by PALIĆ & al. [12]. They reported 24.3%- 25.9%, 21.6%-16.8% and 41.3%-7.4% C18:2 and C18:3 in A. linguata, A. nobilis and A. crithmifolia, respectively (PALIĆ & al. [12]). In contrast, we found a higher mean concentration of C18:2 in four Achillea taxa used, ranging between 42.20% and 64.43%, than that of the above group. In addition, extremely high C18:2 (69.42%) and low C18:3 (1.73%) contents were reported for an Iranian A. tenuifolia by GOLI & al. [9]. In our Achillea specimens, the range of the C18:2 and C18:3 FAs varied between 32.40% and 64.43% and between 2.72% and 16.45%. Our Achillea taxa had relatively low levels of these three acids, such as A. bisserata (32.40%) and A. multifida (2.72%), although the general situation showed that the Achillea specimens collected from the alpines of Turkey contained these FAs in considerable amounts. Our results also revealed a wide range of C18:2 content, varying between 37.47% and 51.0%, among four Matricaria taxa. A varietal difference in the content of the acid was apparent between M. chamomilla var. chamomilla and M. chamomilla var. recutita, consisting of 37.47% and 46.03% of the total FA. The majority of this abundant fatty acid was also confirmed by BARROS & al. [16] in M. recutita (44.83%). Similarly, we determined approximately the same amount of fatty acid at level of 46.03% in M. chamomilla var. recutita.

Table 2. Saturated fatty acid composition (%, meanSD, n = 6 ) in the achenes of the tribe Anthemideae

C10:0 C 12:0 C14:0 C 15:0 C16:0 C18:0 C20:0 C22:0 C24:0 SFA* Taxon Achillea 0.680.02 N.D.** 2.940.01 N.D. 22.270.17 3.200.01 1.290.00 1.520.43 1.150.04 33.05 biebersteinii A. bisserata 1.740.04 N.D. 1.280.03 N.D. 21.680.33 3.740.04 2.510.03 0.830.08 1.940.17 33.72 A. multifida 0.200.03 N.D. 0.630.02 N.D. 12.340.10 4.190.53 1.810.07 0.410.02 0.640.07 20.22 A. wilhelmsii N.D. 0.370.01 3.720.46 0.800.16 17.690.18 12.590.06 3.800.02 1.770.14 1.190.10 41.93 Anthemis cotula 2.410.00 0.710.10 3.850.01 0.480.08 27.390.07 7.950.37 3.560.04 1.770.01 2.43007 50.55 A. macrotis 0.720.00 N.D. 0.380.01 N.D. 24.910.74 2.320.32 N.D. N.D. 1.760.22 30.09 Artemisia annua N.D. 0.400.03 0.650.05 0.800.16 20.430.61 2.730.13 1.790.06 1.220.30 1.190.02 29.21 A. austriaca 0.840.02 N.D. 3.150.07 N.D. 27.950.97 5.470.13 1.760.04 3.190.06 5.070.14 47.43 A. santonicum 0.570.04 N.D. 4.250.03 N.D. 23.560.18 5.180.06 4.260.02 2.950.41 2.930.02 43.70 Glebionis 1.380.04 N.D. 2.630.01 N.D. 21.810.04 7.530.13 3.220.10 2.540.10 3.640.35 42.75 coronaria

G. segetum 0.720.06 N.D. 0.380.10 1.920.09 20.760.30 4.240.07 3.450.25 5.420.11 1.370.39 38.26

Matricaria aurea 0.400.02 N.D. N.D. N.D. 30.190.52 2.240.15 0.320.32 0.120.01 1.490.07 34.76 M. chamomilla 1.370.15 N.D. 0.630.27 N.D. 30.200.33 7.410.47 N.D. 0.510.14 1.720.19 41.84 var. chamomilla M. chamomilla 1.560.23 N.D. N.D. N.D. 22.570.29 3.760.30 N.D. N.D. 2.840.06 30.73 var. recutita

M. matricarioides 0.640.12 N.D. N.D. N.D. 21.400.21 3.140.31 N.D. N.D. 1.640.16 26.82 Tanecetum N.D. N.D. 0.730.41 N.D. 12.690.28 3.650.20 0.630.01 2.860.03 1.270.06 21.83 vulgare Tripleurospermu 0.970.07 N.D. N.D. N.D. 18.820.47 2.620.66 1.580.30 N.D. 1.640.04 25.63 m baytopianum

T. callosum N.D. N.D. 0.720.05 N.D. 12.980.20 1.310.18 N.D. N.D. N.D. 15.01 T. caucasicum (Rize pop.-Inceer 0.230.15 N.D. 1.360.10 N.D. 30.160.67 4.410.37 0.870.03 N.D. 1.570.10 38.60 672) T. caucasicum (Rize pop.-Inceer N.D. 1.750.13 0.750.08 N.D. 17.740.27 1.920.05 0.790.04 2.740.05 2.500.15 28.19 765) T. conoclinium 0.740.01 N.D. 0.410.03 N.D. 17.710.40 2.680.21 0.420.02 N.D. N.D. 21.96

T. corymbosum N.D. N.D. 0.790.09 N.D. 16.230.14 1.760.45 N.D. N.D. 1.990.30 20.77

T. decipiens N.D. N.D. 0.330.02 N.D. 17.370.28 1.700.26 0.410.06 N.D. N.D. 19.81

T. elongatum N.D. N.D. N.D. N.D. 20.030.33 3.350.39 N.D. N.D. N.D. 23.38

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1.290.0 T. fissurale N.D. N.D. 1.310.26 N.D. 28.240.38 3.210.06 1.270.05 N.D. 35.32 4 T. heterolepis N.D. N.D. 0.670.13 N.D. 11.340.37 2.090.21 0.250.04 0.580.09 N.D. 14.93

T. hygrophilum 0.800.13 N.D. 0.380.05 N.D. 20.250.32 2.060.21 N.D. N.D. N.D. 23.49 1.360.3 T. inodorum N.D. N.D. 0.760.76 N.D. 24.600.33 3.860.08 N.D. N.D. 30.58 4 T. kotschyi 1.330.06 N.D. 0.160.07 N.D. 28.310.23 2.530.19 0.300.11 N.D. N.D. 32.63 1.800.2 T. melanolepis 0.830.04 N.D. 3.830.05 N.D. 26.140.39 6.180.11 1.850.03 0.870.02 41.50 4 T. microcephalum 2.040.07 N.D. 0.470.10 N.D. 25.720.67 1.840.03 0.580.07 N.D. N.D. 30.65

T. monticolum N.D. N.D. 1.340.40 N.D. 17.850.18 2.200.28 N.D. N.D. N.D. 21.39 0.910.0 T. oreades var. oreades N.D. N.D. 1.300.05 N.D. 18.110.29 2.720.22 N.D. N.D. 23.04 5 T. oreades var. 0.560.08 N.D. N.D. N.D. 16.240.21 1.740.25 N.D. N.D. N.D. 18.54 tchihatchewii 1.400.1 T. parviflorum 19.460.55 N.D. N.D. N.D. N.D. 2.330.50 0.460.08 N.D. 23.65 4 1.240.1 T. pichleri 1.050.02 N.D. 2.930.17 N.D. 25.160.67 3.090.08 1.340.009 N.D. 34.81 4 T. repens N.D. N.D. N.D. N.D. 19.640.40 2.910.51 0.500.10 N.D. N.D. 23.05

T. rosellum var. album 1.550.2 (Çanakkale pop.-Inceer 1.900.05 0.430.01 3.850.04 N.D. 23.790.17 4.630.27 0.980.11 0.670.24 37.80 7 719)

T. rosellum var. album 0.650.0 1.290.02 N.D. 0.470.02 N.D. 12.770.66 2.390.02 0.830.02 0.790.10 19.19 (Rize pop.-Inceer 766) 5

0.570.0 T. sevanense 0.500.12 N.D. 0.570.11 N.D. 21.300.18 1.150.16 0.160.0 0.550.11 24.80 3 0.230.0 T. subnivale 0.740.04 N.D. 1.230.05 N.D. 28.160.11 3.170.35 N.D. N.D. 33.53 8 2.940.1 T. tenuifolium 1.250.10 N.D. N.D. N.D. 25.490.18 4.560.26 N.D. 2.010.13 36.25 3 1.390.3 T. transcaucasicum N.D. N.D. N.D. N.D. 25.640.24 5.420.46 0.920.35 N.D. 33.37 3 1.170.3 T. ziganaense 0.940.09 N.D. 2.760.29 N.D. 20.950.62 4.240.12 1.110.03 6.740.32 37.91 3 Achillea biebersteinii 1.940.00 0.390.39 10.610.43 45.370.44 8.580.26 N.D. N.D. 12.94 53.95 66.89

A. bisserata 4.560.30 N.D.**** 8.950.03 32.400.24 16.450.23 N.D. 0.860.06 13.51 49.71 63.22

A. multifida 0.330.01 0.470.01 11.440.04 64.430.35 2.720.17 N.D. N.D. 11.04 67.15 78.19

A. wilhelmsii 0.880.08 N.D. 8.860.20 42.200.24 5.130.04 N.D. 0.870.08 9.74 48.20 57.94

Anthemis cotula N.D. N.D. 13.540.21 27.390.16 10.730.04 N.D. 2.700.13 13.54 40.82 54.36

A. macrotis N.D. N.D. 7.820.58 56.260.50 5.480.06 0.370.06 N.D. 7.82 62.11 69.93

Artemisia annua 0.500.11 0.400.11 13.090.29 51.800.30 4.160.14 N.D. 0.920.11 13.99 56.88 70.87

A. austriaca 3.060.08 N.D. 6.590.09 30.170.59 7.330.09 N.D. 1.870.12 9.65 39.37 49.02

A. santonicum 1.200.04 N.D. 7.060.03 35.380.33 10.600.14 N.D. 2.090.03 8.26 48.07 56.33

Glebionis coronaria 1.610.11 N.D. 14.870.09 32.970.16 5.600.07 N.D. 2.230.07 16.48 40.80 57.28

G. segetum N.D. N.D. 8.100.33 40.490.58 7.390.05 N.D. 5.800.20 8.10 53.68 61.78

Matricaria aurea N.D. 4.160.11 6.480.56 45.440.51 8.040.74 1.130.11 N.D. 10.64 54.61 65.25 M. chamomilla var. N.D. N.D. 5.060.78 37.470.54 14.060.13 1.580.08 N.D. 5.06 53.11 58.17 chamomilla M. chamomilla var. N.D. N.D. 11.260.16 46.030.49 9.530.23 2.460.30 N.D. 11.26 58.02 69.28 recutita M. matricarioides N.D. N.D. 13.260.14 51.000.53 7.320.36 1.620.14 N.D. 13.26 59.94 73.20

Tanecetum vulgare 2.850.10 0.910.02 10.310.16 58.030.29 5.390.05 N.D. 0.710.06 14.07 64.13 78.20 Tripleurospermum N.D. 0.830.06 14.490.55 43.080.67 15.990.93 N.D. N.D. 15.32 59.07 74.39 baytopianum T. callosum N.D. 1.440.10 15.200.38 62.300.61 5.320.33 0.790.09 N.D. 16.64 68.41 85.05

T. caucasicum (Rize N.D. 1.380.06 7.030.22 38.470.42 11.040.23 3.500.18 N.D. 8.41 53.01 61.42 pop.-Inceer 672)

T. caucasicum (Rize 0.580.05 N.D. 5.580.12 35.950.23 21.530.16 N.D. 0.760.12 6.16 58.24 64.40 pop.-Inceer 765)

T. conoclinium N.D. N.D. 15.710.21 48.200.37 13.680.21 0.470.09 N.D. 15.71 62.35 78.06

T. corymbosum N.D. N.D. 11.590.45 57.370.27 8.870.36 1.910.16 N.D. 11.59 68.15 79.74

T. decipiens N.D. 1.020.10 14.850.56 55.190.38 5.850.59 3.290.37 N.D. 15.87 64.33 80.20

T. elongatum N.D. N.D. 23.551.00 51.160.16 0.390.39 1.540.22 N.D. 23.55 53.09 76.64 64.69 T. fissurale N.D. N.D. 6.880.30 51.000.39 4.780.10 2.030.04 N.D. 6.88 57.81

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T. heterolepis N.D. 0.780.05 17.510.43 65.520.44 1.280.15 N.D. N.D. 18.29 66.80 85.09 T. hygrophilum N.D. 3.420.30 5.620.71 48.360.29 17.850.55 1.270.05 N.D. 9.04 67.48 76.52 T. inodorum N.D. 2.760.57 17.170.52 42.140.11 5.420.54 1.740.06 N.D. 19.93 49.30 69.23

T. kotschyi N.D. N.D. 7.310.19 50.430.50 9.510.43 0.150.06 N.D. 7.31 60.09 67.40

T. melanolepis 1.031.03 N.D. 8.410.14 36.371.18 12.700.87 N.D. N.D. 9.44 49.07 58.51

T. microcephalum N.D. 4.620.60 18.400.14 37.560.48 6.160.78 2.720.27 N.D. 23.02 46.44 69.46

T. monticolum N.D. N.D. 17.320.46 53.180.37 5.760.13 2.360.50 N.D. 17.32 61.30 78.62 T. oreades var. N.D. N.D. 16.090.50 55.600.51 4.370.41 0.910.12 N.D. 16.09 60.88 76.97 oreades T. oreades var. N.D. 0.250.17 14.530.24 56.710.47 9.650.14 0.340.35 N.D. 14.78 66.70 81.48 tchihatchewii T. parviflorum N.D. N.D. 13.400.43 48.290.19 13.860.52 0.810.19 N.D. 13.40 62.96 76.36

T. pichleri N.D. 1.480.07 9.100.33 47.440.48 5.59v0.96 1.600.04 N.D. 10.58 54.63 65.21

T. repens N.D. 1.180.06 15.790.72 56.370.53 1.140.07 2.470.06 N.D. 16.97 59.98 76.95 T. rosellum var. album (Çanakkale 1.810.03 0.350.07 5.740.03 34.980.57 17.970.59 N.D. N.D. 7.90 52.95 60.85 pop.-Inceer 719) T. rosellum var. album (Rize pop.- 0.760.02 0.270.05 6.914.03 61.990.46 10.900.39 N.D. N.D. 7.94 72.89 80.83 Inceer 766) T. sevanense N.D. 1.790.08 19.260.34 48.210.23 5.960.36 N.D. N.D. 21.05 54.17 75.22

T. subnivale N.D. 1.140.07 20.690.36 32.890.48 8.000.10 3.770.19 N.D. 21.83 44.66 66.49

T. tenuifolium N.D. N.D. 4.170.45 41.210.78 18.380.49 N.D. N.D. 4.17 59.59 63.76 T. N.D. N.D. 18.640.53 42.760.52 1.010.22 4.220.81 N.D. 18.64 47.99 66.63 transcaucasicum T. ziganaense 3.150.46 0.850.05 9.040.49 36.380.58 12.670.73 N.D. N.D. 13.04 49.05 62.09 *MUFA; total of monounsaturated fatty acids, as is sum of individual mono unsaturated fatty acids **PUFA; total of polyunsaturated fatty acids, as is sum of individual poly unsaturated fatty acids ***UFA; total of both mono- and polyunsaturated fatty acids, as is sum of individual mono- and poly unsatuarted fatty acids ****N.D.; not detected, 0.001

In general, there is considerable variation between the data reported in the literature and our findings. These differences may be due to geographical region, climate, soil type, light, day/night regimes, etc., which directly affect the lipid composition of plants. Recent reports have revealed that plants of the Compositae family such as chicory (Cichorium intybus L.), safflower (Carthamus tinctorius L.) have potential as forage for ruminants (LANDAU & al. [17]). It is widely agreed that the concentration of five major FAs in forage also depends on the botanical composition of grassland sward. Anthemideae species represent a large part of the plants representing forage in alpine regions, together with grasses (BREMER and HUMPRIES [10]; LANDAU & al. [17]). However, it is not easy to prove the direct effect of plant species on the chemical composition of milk, especially on the FA profile of milk fat. However, forming botanical pastures using taxa of the tribe studied can be recommended. It should also be underlined that milk FA composition (approximately 70% saturated, 25% monosaturated and 5% polyunsaturated FAs) is mainly related to the FA composition of animals’ feed, and to the effect of forage and feeding systems (ELGERSMA & al. [18], ARVIDSSON [19]). Results of many studies indicate that milk and meat produced from grassland, particularly from botanically diverse pastures rich in PUFA in seeds, leaves etc., have higher concentrations of FA and antioxidants, which are known to benefit human health (ARVIDSSON [19], MOLONEY & al. [20]). In terms of the concentration of UFAs in the studied taxa, mainly linoleic acid, feeding ruminants, especially cows, with fresh or conserved dried forms of these Anthemideae species may result in high concentrations of PUFA rich in ω-3. This recommendation was also confirmed by ELGERSMA & al. [18] and MOLONEY & al. [20]. Since linoleic acid (C18:2) and linolenic acid (C18:3) in feed are the precursors of conjugated linoleic acid (CLA) in milk and meat (ELGERSMA & al. [18], 11582 Romanian Biotechnological Letters, Vol. 21, No. 3, 2016

Achene Fatty Acid Composition in the Tribe Anthemideae (Asteraceae)

MOLONEY & al. [20]), the linolenic acid content in forage also increases the content of ω-3 fatty acids in milk and milk products (MOLONEY & al. [20]). The beneficial effect of unsaturated fats against CVDs is widely agreed. It may be predicted that in the future a special premium for using the taxa of tribe will be paid to those farmers who produce milk from grazed herbs rich in PUFA. Dietary C18:3 n-3 is the best precursor of C18:3n-3 in milk, and conjugated linolenic acids (CLAs) derive from both C18:2 n-6 and C18:3 n-3. There has therefore been growing interest in determining the concentration and composition of UFAs in forage. Since many metabolic factors affecting the conversion of PUFA in the rumen have not yet been identified, it is important to quantify the variations in FA concentration in herbs of meadows and grasslands. According to DEWHURST & al. [8], this will help to identify breeding goals and management strategies to increase PUFA in forages and animal-derived products. In conclusion, plant sources cited above that are rich in such unsaturated fatty acids beneficial for the health in the prevention of chronic diseases, can be obtained from the milk, meat etc., of grazing animals. In the near future, experts will provide special forage preparations for farmers who produce milk from grazed herbs rich in PUFA. The present taxa may bring an enrichment in PUFA content for such production policies. Although it was difficult to see any direct correlation between grazing with herbs that are rich in UFAs and milk fat quality, the studies cited above partly confirm the relationship. The average UFA for the present Anthemideae species was estimated at 69.39%, mostly consisting of linoleic acid. In particular, the PUFA content of the taxa of Achillea, Anthemis, Tanacetum and most of the Tripleurospermum was 60% or higher, up to 72.89% (T. rosellum var. album-Rize pop., Inceer 766). It may be concluded that if the botanical composition of forages in grasslands or meadows that are open to grazing animals is formed with these taxa, such botanically designated pastures will contribute to improving the milk quality of ruminants.

4. Acknowledgements The authors wish to thank Dr. Melahat OZCAN and Murat BAL for collecting some living species, and the Scientific and Technological Research Council of Turkey, TUBITAK- Project number: 106T162, for providing financial support.

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