© 2018 The Japan Mendel Society Cytologia 83(4): 407–414

Karyological Investigation of Sixteen Mill. (, Cardueae) Taxa from Turkey

Yasar Kiran*

Firat University, Science Faculty, Biology Department, Elazig, Turkey

Received June 27, 2018; accepted July 18, 2018

Summary The chromosome number and karyotype analysis of sixteen taxa of genus Cirsium Mill. distributed in Turkey were analyzed. This taxa are, Cirsium steriolepis Petr., C. bulgaricum DC., C. polycephalum DC., C. trachylepis Boiss., C. sommieri Petr., C. osseticum (Adams) Petrak, C. leucocephalum (Willd.) Spreng ssp. tenuilobum (K. Koch) Greuter, C. haussknechtii Boiss., C. ekimianum Yildiz & Dirmenci, C. handaniae Yildiz, Dirmenci & Arabacı, C. davisianum Kit Tan & Sorger, C. hypoleucum DC., C. pseudopersonata Boiss & Bal. ssp. pseudopersonata, C. pseudopersonata Boiss. & Bal. ssp. kuznezowianum (Somm. & Lev.) Petrak, C. canum (L.) All., C. obvallatum (Bieb.) Fischer, and nine of them are endemic species (C. steriolepis, C. polycephalum, C. trachylepis, C. sommieri, C. leucocephalum ssp. tenuilobum, C. davisianum, C. handaniae, C. ekimianum and C. pseudopersonata ssp. pseudopersonata). The somatic chromosome number of all studied taxa were deter- meined as 2n=2x=34. Total chromosome lengths varied 68.00 to 147.92 µm among species. Karyotype asymme- try indices other parameters were also calculated. Karyotype analysis indicated that Cirsium taxa generally have median (m and M) chromosomes. In addition, only C. davisianum species has one submedian (sm) chromosome. The chromosome number and karyotype of C. polycephalum, C. handaniae, C. davisianum and C. ekimianum are newly reported.

Key words Asteraceae, Cirsium, Karyotype, Chromosome number, Turkey.

The genus Cirsium Mill. (thistle) of the family Astera- 102 are either very rare or doubtful (Bures et al. 2004). ceae includes perennial, biennial, and annual members Chromosome studies of the Turkish Cirsium species re- and comprises more than 250 species distributed in Eu- ported that the taxa have 2n=32, 34, 60, and 68 diploid rope North Africa Asia and North and Central America. chromosome numbers (Ozcan et al. 2008, 2011, Yuksel Native species of Cirsium range from sea level to alpine et al. 2013). and occur in many habitats including swamps, mead- Cirsium is a taxonomically complex genus with ows, forests, prairies, sands, screes, steppes, and deserts many hybrids. Several multidisciplinary studies such (Charadze 1963, Davis and Parris 1975, Petrak 1979, as anatomy, palynology, molecular and karyological are Kadereit and Jeffrey 2007). needed to clarify the of the genus. Therefore, According to recent studies of the Turkish Cirsium, the karyology of Turkish Cirsium species is currently be- the genus is represented in Turkey by 66 species (78 ing studied in order to clarify their taxonomy and make taxa), of which 27 species (32 taxa) of them are endemic contributions to other multidisciplinary studies on the species, belonging to three sections; Epitrachys DC., genus. Cirsium DC., and Cephalonoplos (Neck.) DC. (Davis and Parris 1975, Davis et al. 1988, Guner et al. 2000, Materials and methods Daskin et al. 2006, Yildiz and Dirmenci 2008, Yildiz et al. 2009a, b, 2011, Arabacı and Dirmenci 2011, Yildiz materials 2012, Yuksel et al. 2013). In Turkey, this 66 species oc- All plant specimens were collected from natural cur, mainly distributed in east and north-east Anatolia, habitats during the fruiting season in 2006 and 2007 as with the main center of diversity between Trabzon and a part of a taxonomic revision of the genus Cirsium in Artvin provinces (Davis 1971). Turkey. The examined taxa, localities and voucher num- Approximately 69% of species of the genus Cirsium bers are given in Table 1. The specimens were identified are diploid 2n=34, 10% are tetraploid 2n=68, and 5% according to the pertinent reference works given in the are 2n=30 or 2n=32, while several chromosome num- literature (Davis and Parris 1975, Davis et al. 1988, bers such as 2n=16, 18, 20, 22, 24, 26, 28, 26, 51, and Guner et al. 2000, Yildiz et al. 2009a, Yildiz 2012). The voucher specimens were deposited at the Balıkesir and * Corresponding author, e-mail: ykiran@firat.edu.tr Inönü University Herbarium, Turkey. DOI: 10.1508/cytologia.83.407 408 Y. Kiran Cytologia 83(4)

Table 1. Localities and voucher numbers of the studied Cirsium taxa.

Taxa Locality Collecter (voucher number)

C. steriolepis A1 Tekirdag: Inecik, Ormanli-Guzel Village, Quercus area, 500–600 m, B. Yildiz & T. Dirmenci (BY 16507) 01.08.2007 C. bulgaricum B6 Yozgat: Akdag Mine, Olukozu Village, 1 km west, 1200 m, 11.08.2006 B.Yildiz & T. Dirmenci (BY 16232) C. polycephalum A1 Kırklareli: Saray, between Beyceler-Sinekli, 3–4 km, Quercus area, 170 m, B. Yildiz & T. Dirmenci (BY 16530) 02.08.2007 C. trachylepis A8 Rize: Ikizdere, Bas Village, Cicekli plateau, stony and meadow area, B. Yildiz & T. Arabacı (BY 16663) 2100–2300 m, 15.09.2007 C. sommieri B8 Bingol: Between Karlıova-Bingol, 15 km, 1767 m, 11.08.2007 T. Dirmenci & T. Arabacı (TD 3515) C. osseticum A7 Trabzon: Between Caykara, Karacakaya (Karacam) Village-Soganlı Gorge, B. Yildiz (BY 16755) 1900–2000 m, 21.09.2007 C. leucocephalum ssp. tenuilobum A8 Erzurum: Askale, Kop Mountain, steppe, 1750 m, 21.09.2007 B. Yildiz (BY 16746) C. haussknechtii B7 Malatya: Akcadag, Dedeyazı Village, Canakci location, 1450 m, 10.08.2007 T. Dirmenci & T. Arabacı (TD 3503) C. ekimianum A8 Erzurum: Askale, Kop Mountain, steppe, 1750 m, 21.09.2007 B. Yildiz (BY 16745) C. handaniae A9 Ardahan: Between Turkozu-Posof 3 km, 1500 m, 19.09.2007 B. Yildiz & T. Arabacı (BY 16731) C. davisianum B8 Erzurum: Askale, Kop Mountain, stony area, 2450 m, 04.08.2008 B. Yildiz (BY 16850) C. hypoleucum A2 Bursa: Between Keles-Inegöl, 15 km, Pinus forest, 1300 m, 21.08.2007 B. Yildiz, T. Arabacı (BY 16593) C. pseudopersonata ssp. pseudopersonata B7 Bayburt: Soganlı Mountain, steppe, 2200 m, 20.09.2007 B. Yildiz (BY 16754) C. pseudopersonata ssp. kuznezowianum A9 Artvin: Between Borcka-Macahel 34 km, forest openings, 1450 m, 16.09. B. Yildiz & T. Arabacı (BY 16682) 2007 C. canum A3 Bolu: Between Abant Lake-Akcaalan Village, water edge, 1250 m, 22.08.2007 B. Yildiz & T. Arabacı (BY 16610) C. obvallatum A8 Artvin: Ardanuc, Bilbilan Plateau, 2500 m, 19.09.2007 B. Yildiz & T. Arabacı (BY 16724)

Chromosome analysis asymmetry for karyotypes (A) (Huziwara 1962, Arano Approximately 100 seeds belong to 10 plant speci- 1963, Greilhuber and Speta 1976, Watanabe et al. 1999, mens were used for each taxon. Thirty roots of each spe- Venora et al. 2002). While the intrachromosomal asym- cies were used from seeds germinated on a moist filter metry index (A1) and the interchromosomal asymmetry paper in petri dishes at 25°C. The actively growing root index (A2) was calculated according to the formula pro- tips were pretreated with 0.05% colchicine for 3–3.5 h at posed by Romero Zarco (1986). Also, karyotype sym- room temperature. The material was fixed with Carnoy metry nomenclature was determined based on Stebbins' fixative (1 : 3 glacial acetic acid–absolute ethanol) for classification (Stebbins 1971). at least 24 h at 4°C, hydrolyzed in 1 M HCl at 60°C for 15 min, then rinsed in tap water for 3–5 min, stained in Results Feulgen staining or reaction for 1 h, and mounted in 45% acetic acid (Elci 1982). Digital microphotographs from C. steirolepis Petrak is an endemic species and in at least seven well-spread metaphase plates were taken montane woods (Davis and Parris 1975). The somatic using an Olympus BX51 microscope and were recorded chromosome number of this species was found to be with an Olympus Camedia C-4000 digital camera. 2n=2x=34. The chromosome length ranges 5.00 to The chromosome numbers of the species were iden- 8.93 µm and the total karyotype length was 113.9 µm. It tified and checked using the Flora of Turkey (Davis consists of 17 m chromosomes. Arm ratios are 1.07–1.43 and Parris 1975) and supplements (Davis et al. 1988, and relative lengths are 4.38–7.84% (Table 2). Somatic Guner et al. 2000). Also, the online chromosome num- metaphase chromosomes and idiograms are given in ber databases, Index to Plant Chromosome Numbers Figs. 1a and 2a. (http://www.tropicos.org/Project/IPCN) and Index to C. bulgaricum DC. İs growing in Pinus vegatation by Chromosome Numbers in Asteraceae (http://www.lib. a roadside, in Abies area, siliceous substrate, between kobe-u.ac.jp/infolib/meta_pub/G0000003asteraceaesre- altitudes of 1200 and 1800 m asl (Davis and Parris sult-en) were checked. 1975). The somatic chromosome number was found to The number of somatic chromosomes, chromosome be 2n=2x=34. The chromosome length ranges 3.60 to length range, haploid chromosome length, arm ratio, 6.40 µm and the total karyotype length was 83.80 µm. It relative length, and karyotype formula were deter- consists of 1M and 16 m chromosomes. Arm ratios are mined. Karyotype formula was determined based on 1.00–1.50 and relative lengths are 4.29–7.63% (Table centromere position (Levan et al. 1964). The ideograms 2). Somatic metaphase chromosomes and idiograms are of these taxa were arranged in decreasing lengths ac- given in Figs. 1b and 2b. cording to the metaphase chromosome size (Martin C. polycephalum DC. is an endemic species that et al. 2009). Karyotype asymmetry was determined by grows on dry places by roadsides, in fields, dry hills, using various techniques including total form percent- open places, near sea level, at altitudes of 50 m (Da- age (TF%), percentage karyotype asymmetry index vis and Parris 1975). The seeds were collected from (AsK%), index of karyotype symmetry (Syi), index of Kırklareli Province (Table 1). The somatic chromosome chromosome size resemblance (Rec) and the degree of number of this species was found to be 2n=2x=34. The 2018 Karyology of Cirsium Species from Turkey 409

Table 2. Karyological features of the studied Cirsium taxa.

Chromosome length (µm) Haploid Arm ratio Relative length (%) Karyotype Taxa 2n chromosome formulae Min Max length (µm) Min Max Min Max

C. steirolepis 34 5.00 8.93 113.90 1.07 1.43 4.38 7.84 17m C. bulgaricum 34 3.60 6.40 83.80 1.00 1.50 4.29 7.63 M+16m C. polycephalum 34 6.31 11.54 137.69 1.00 1.28 4.58 8.38 M+16m C. trachylepis 34 6.00 13.07 147.92 1.12 1.66 4.05 8.83 17m C. sommieri 34 3.00 5.60 73.90 1.10 1.33 4.05 7.57 17m C. osseticum 34 5.08 8.77 109.77 1.08 1.53 4.62 7.98 17m C. leucocephalum ssp. tenuilobum 34 5.25 10.31 127.13 1.12 1.50 4.12 8.10 17m C. haussknechtii 34 3.40 6.60 81.90 1.13 1.45 4.15 8.05 17m C. ekimianum 34 4.16 7.62 97.26 1.00 1.31 4.27 7.83 M+16m C. handaniae 34 5.54 8.92 120.59 1.00 1.52 4.59 7.39 M+16m C. davisianum 34 5.16 11.62 139.32 1.00 1.78 3.70 8.34 M+15m+sm C. hypoleucum 34 4.10 6.60 84.90 1.10 1.38 4.82 7.77 17m C. pseudopersonata ssp. pseudopersonata 34 2.70 5.30 68.00 1.00 1.43 3.97 7.79 2M+15m C. pseudopersonata ssp. kuznezowianum 34 5.69 10.00 118.44 1.05 1.35 4.80 8.44 17m C. canum 34 4.62 7.39 100.27 1.00 1.56 4.60 7.37 M+16m C. obvallatum 34 4.93 9.23 117.89 1.05 1.50 4.18 7.82 17m

Fig. 1. Metaphase chromosomes of 16 Cirsium taxa. a. C. steirolepis, b. C. bulgaricum, c. C. polycephalum, d. C. trachylepis, e. C. sommieri, f. C. osseticum, g. C. leucocephalum ssp. tenuilobum, h. C. haussknechtii, i. C. ekimianum, j. C. handaniae, k. C. davisianum, l. C. hypoleucum, m. C. pseudopersonata ssp. pseudopersonata, n. C. pseudopersonata ssp. kuzne- zowianum, o. C. canum, p. C. obvallatum. Scale bars=10 µm. chromosome length ranges from 6.31 to 11.54 µm and cies. Somatic metaphase chromosomes and idiograms the total karyotype length was 137.69 µm. It consists of are given in Figs. 1c and 2c. 1M and 16m chromosomes. Arm ratios are 1.00–1.28 C. trachylepis Boiss. is a perennial endemic species and relative lengths are 4.58–8.38% (Table 2). To our grows in woods at altitudes of 1700–2200 m (Davis and knowledge, this is the first count for this endemic spe- Parris 1975). The somatic chromosome number of this 410 Y. Kiran Cytologia 83(4)

Fig. 2. Haploid idiograms of 16 Cirsium taxa. a. C. steirolepis, b. C. bulgaricum, c. C. polycephalum, d. C. trachylepis, e. C. sommieri, f. C. osseticum, g. C. leucocephalum ssp. tenuilobum, h. C. haussknechtii, i. C. ekimianum, j. C. handaniae, k. C. davisianum, l. C. hypoleucum, m. C. pseudopersonata ssp. pseudopersonata, n. C. pseudopersonata ssp. kuznezowi- anum, o. C. canum, p. C. obvallatum. species was found to be 2n=2x=34. The chromosome from 5.08 to 8.77 µm and the total karyotype length was length ranges from 6.00 to 13.07 µm and the total karyo- 109.77 µm. It consists of 17 m chromosomes. Arm ratios type length was 147.92 µm. It consists of 17m chromo- are 1.08–1.53 and relative lengths are 4.62–7.98% (Table somes. Arm ratios are 1.12–1.66 and relative lengths are 2). Somatic metaphase chromosomes and idiograms are 4.05–8.83% (Table 2). Somatic metaphase chromosomes given in Figs. 1f and 2f. and idiograms are given in Figs. 1d and 2d. C. leucocephalum (Willd.) Spreng ssp. tenuilobum C. sommieri Petr. is a perennial endemic Irano-Tura- (K. Koch) Greuter is an endemic, Irano-Turanian Ele- nian Element. Its distributed on the forest (basalt rock), ment and synonym with C. lappaceum (Bieb.) Fisher roadside, rocky slopes, cultivated land and waste places, ssp. tenuilobum (C. Koch) Davis & Parris. Its grown in between 1170 and 2450 m asl (Davis and Parris 1975). edges of cultivated fields, fallow fields, steppe, between The somatic chromosome number of this species was 1520 and 2300 m asl (Davis and Parris 1975). The so- found to be 2n=2x=34. The chromosome length ranges matic chromosome number of this species was found from 3.00 to 5.60 µm and the total karyotype length was to be 2n=2x=34. The chromosome length ranges from 73.90 µm. It consists of 17m chromosomes. Arm ratios 5.25 to 10.31 µm and the total karyotype length was are 1.10–1.33 and relative lengths are 4.05–7.57% (Table 127.13 µm. It consists of 17m chromosomes. Arm ratios 2). Somatic metaphase chromosomes and idiograms are are 1.12–1.50 and relative lengths are 4.12–8.10% (Table given in Figs. 1e and 2e. 2). Somatic metaphase chromosomes and idiograms are C. osseticum (Adams) Petrak is growing on rocky given in Figs. 1g and 2g. slopes in Picea forest, coniferous forests, lush pasture, C. haussknechtii Boiss. is a perennial species and at altitudes of 950–1720 m asl (Davis and Parris 1975). Irano-Turanian Element, that grows on field drain, dry The somatic chromosome number of this species was waste ground, at altitudes of 1140–1950 m (Davis and found to be 2n=2x=34. The chromosome length ranges Parris 1975). The somatic chromosome number of this 2018 Karyology of Cirsium Species from Turkey 411 species was found to be 2n=2x=34. The chromosome of this species was found to be 2n=2x=34. The chromo- length ranges from 3.40 to 6.60 µm and the total karyo- some length ranges from 2.70 to 5.30 µm and the total type length was 81.90 µm. It consists of 17m chromo- karyotype length was 68.00 µm. It consists of% 2M and somes. Arm ratios are 1.13–1.45 and relative lengths are 15 m chromosomes. Arm ratios are 1.00–1.43 and rela- 4.15–8.05% (Table 2). Somatic metaphase chromosomes tive lengths are 3.97–7.79 (Table 2). Somatic metaphase and idiograms are given in Figs. 1h and 2h. chromosomes and idiograms are given in Figs. 1m and C. ekimianum Yildiz & Dirmenci is biennial and 2m. perennial herb. Its grown on the steppe, at altitudes of C. pseudopersonata Boiss. & Bal. ssp. kuznezowi- 1800–2000 m (Yildiz and Dirmenci 2008). The seeds anum (Somm. & Lev.) Petrak is a perennial species and were collected from Erzurum–Aşkale Province (Table Euxine-Element, that distributed on forests of Picea ori- 1). The somatic chromosome number of this species was entalis, shady banks and stream sides, alpine meadows, found to be 2n=2x=34. The chromosome length ranges at altitudes of 1480–2290 m (Davis and Parris 1975). from 4.16 to 7.62 µm and the total karyotype length was The somatic chromosome number of this species was 97.26 µm. It consists of 1M and 16m chromosomes. Arm found to be 2n=2x=34. The chromosome length ranges ratios are 1.00–1.31 and relative lengths are 4.27–7.83% from 5.69 to 10.00 µm and the total karyotype length (Table 2). To our knowledge, this is the first count for was 118.44 µm. It consists of 17m chromosomes. Arm this endemic species. Somatic metaphase chromosomes ratios are 1.05–1.35 and relative lengths are 4.80–8.44% and idiograms are given in Figs. 1i and 2i. (Table 2). Somatic metaphase chromosomes and idio- C. handaniae Yildiz, Dirmenci & Arabacı is peren- grams are given in Figs. 1n and 2n. nial, endemic Euro–Siberian Element. Its grown on the C. canum (L.) All. is perennial and Euro–Siberian deep-soiled steppe, where grasses are dominant. It is Element. Its grown on water meadows, marshy places, known only from the type locality (Yildiz et al. 2009a). springs and ditches, at altitudes of 150–1620 m (Davis The somatic chromosome number of this species was and Parris 1975). The somatic chromosome number of found to be 2n=2x=34. The chromosome length ranges this species was found to be 2n=2x=34. The chromo- from 5.54 to 8.92 µm and the total karyotype length was some length ranges from 4.62 to 7.39 µm and the total 120.59 µm. It consists of 1M and 16m chromosomes. karyotype length was 100.27 µm. It consists of 1M Arm ratios are 1.00–1.52 and relative lengths are 4.59– and 16m chromosomes. Arm ratios are 1.00–1.56 and 7.39% (Table 2). To our knowledge, this is the first count relative lengths are 4.60–7.37% (Table 2). Somatic meta- for this endemic species. Somatic metaphase chromo- phase chromosomes and idiograms are given in Figs. 1o somes and idiograms are given in Figs. 1j and 2j. and 2o. C. davisianum Kit Tan & Sorger is an endemic, C. obvallatum (Bieb.) Fischer is grown on ravines, Irano-Turanian Element. Its distributed on stony places, rocky igneous slopes near streams, at altitudes of 2050– steppe, between 2500 and 2800 m asl (Davis et al. 1988). 2800 m (Davis and Parris 1975). The somatic chromo- The somatic chromosome number of this species was some number of this species was found to be 2n=2x=34. found to be 2n=2x=34. The chromosome length ranges The chromosome length ranges from 4.93 to 9.23 µm from 5.16 to 11.62 µm and the total karyotype length and the total karyotype length was 117.89 µm. It consists was 139.32 µm. It consists of 1M, 15m and 1sm chromo- of 17m chromosomes. Arm ratios are 1.05–1.50 and somes. Arm ratios are 1.00–1.78 and relative lengths are relative lengths are 4.18–7.82% (Table 2). Somatic meta- 3.70–8.34% (Table 2). To our knowledge, this is the first phase chromosomes and idiograms are given in Figs. 1p count for this endemic species. Somatic metaphase chro- and 2p. mosomes and idiograms are given in Figs. 1k and 2k. C. hypoleucum DC. is a perennial species and Euxine- Discussion Element, that growing in forests (coniferous or decidu- ous), on shady banks and rock ledges, 2000 m (Davis and The genus Cirsium is a taxonomically complex ge- Parris 1975). The somatic chromosome number of this nus with many hybrids. Therefore, in this study, we species was found to be 2n=2x=34. The chromosome focused on this genus. In this study analyzed sixteen length ranges from 4.10 to 6.60 µm and the total karyo- taxa, nine of them are endemic species in Turkey. The type length was 84.90 µm. It consists of 17m chromo- somatic chromosome number of all studied taxa were somes. Arm ratios are 1.10–1.38 and relative lengths are determined as 2n=2x=34. Different chromosome counts 4.82–7.77% (Table 2). Somatic metaphase chromosomes have been reported in the genus Cirsium. In addition to and idiograms are given in Figs. 1l and 2l. these chromosome numbers, other chromosome counts C. pseudopersonata Boiss. & Bal. ssp. pseudoper- such as 2n=16, 18, 20, 22, 51, 56, 102 are also known sonata is an endemic, Euxine-Element. Its distributed (Aishima 1934, Arano 1957, 1963, Frankton and Moore on forests of Picea orientalis, shady banks and stream 1961, 1963, Fedorov 1969, Moore and Frankton 1962, sides, alpine meadows, at altitudes of 1480–2290 m (Da- 1963, 1966, 1967, 1969, 1974, Tonian 1981, 1982, vis and Parris 1975). The somatic chromosome number Goldblatt 1981, 1984, 1985, 1988). However, the most 412 Y. Kiran Cytologia 83(4)

Table 3. Values of asymmetry indices of the studied Cirsium taxa.

Taxa TF% As K% Syi Rec A A1 A2 Stebbins’s classification

C. steirolepis 44.67 55.32 80.73 75.02 0.10 0.18 0.15 1A C. bulgaricum 43.61 56.38 77.35 77.02 0.12 0.22 0.14 1A C. polycephalum 46.60 53.39 87.28 70.18 0.06 0.12 0.18 1A C. trachylepis 41.96 58.03 72.32 66.57 0.16 0.27 0.22 1B C. sommieri 44.99 55.00 81.79 77.62 0.09 0.17 0.15 1A C. osseticum 44.91 55.08 81.52 73.62 0.10 0.18 0.14 1A C. leucocephalum ssp. tenuilobum 44.19 55.80 79.18 72.53 0.11 0.20 0.21 1A C. haussknechtii 43.71 56.28 77.65 72.99 0.12 0.21 0.17 1A C. ekimianum 46.14 53.85 85.68 75.08 0.07 0.13 0.18 1A C. handaniae 45.65 54.34 83.99 79.52 0.09 0.16 0.14 1A C. davisianum 43.79 56.20 77.90 70.52 0.12 0.20 0.22 1B C. hypoleucum 44.40 55.59 79.87 75.66 0.10 0.19 0.12 1A C. pseudopersonata ssp. pseudopersonata 45.44 54.55 83.28 75.47 0.09 0.16 0.18 1A C. pseudopersonata ssp. kuznezowianum 46.42 53.57 86.66 69.67 0.06 0.12 0.16 1A C. canum 45.12 54.87 82.24 79.81 0.09 0.17 0.15 1A C. obvallatum 45.95 54.04 85.04 75.13 0.07 0.14 0.19 1A

frequent chromosome number within the genus Cirsium while the longest chromosome length was seen in C. is diploid 2n=2x=34 and tetraploid 2n=4x=68. Nou- trachylepis. The present study showed that the smallest roozi et al. (2010) were reported 2n=34 in C. osseticum, arm ratio was determined in C. bulgaricum, C. poly- C. haussknechtii and C. obvallatum. The chromosome cephalum, C. ekimianum, C. handaniae, C. davisianum, number of these species was also found to be 2n=34 in C. pseudopersonata ssp. pseudopersonata, C. canum our study. (1.00) and the longest arm ratio was deternined in C. da- Somatic chromosome number is 2n=34 of C. canum visianum (1.78), whereas the lowest relative length was were reported (Nilsson and Lassen 1971, Kuzmanov observed in C. davisianum (3.97) and the highest relative and Anchev 1973, Uhrikova 1978, Májovský 1978, length was found in C. trachylepis (8.83). Karyotype Javůrková-Jarolímová 1992, Albers and Pröbsting 1998). analysis indicated that chromosomes of Cirsium taxa Our investigations confirm these data, the determined generally have karyotypes composed of most m and a chromosome number is 2n=34. few M chromosomes. In addition, only C. davisianum Chromosome number of C. steirolepis was reported species has one sm chromosome (Table 2). as 2n=34 (Snogerup 1995). According to previous stud- Karyotype asymmetry was determined by using sev- ies by Ozcan et al. (2008, 2011) on Turkish Cirsium eral methods; TF%, AsK%, Syi, Rec and the degree of species, chromosome numbers of 36 taxa established as asymmetry A, A1 and A2. Also, the Stebbins classifica- 2n=32, 34, 36, and 68. This study reported chromosome tion of the Cirsium was determined in this study. The re- numbers (2n=34) of C. trachylepis, C. bulgaricum, C. sults of this study show that TF% between 41.96–46.60, sommieri, C. osseticum, C. hypoleucum, C. pseudoper- ASK% values between 53.39–58.03, Syi values between sonata ssp. pseudopersonata, C. pseudopersonata ssp. 72.32–87.28 and Rec values between 66.57-79.81. While kuznezowianum and C. obvallatum confirm these dates. A1 indexes between 0.12–0.27, A2 indexes between Yuksel et al. (2013) analyzed of 10 Cirsium species 0.12–0.22 (Table 3). belongs to sect. Epitrachys distributed in Turkey. It has This study will play a positive role to enlighten the been reported that the number of chromosomes gener- taxonomically complex genus. On the other hand, ally of these species is 2n=34. to evaluate ultimately the taxonomic position of this Chromosome length is important because it is useful genus in Turkey, it is necessary to carry out further to single out individuals, samples, populations or spe- chromosomal studies on Cirsium taxa. A combination cies, besides being an indirect indicator of the total DNA of detailed karyotype analysis with molecular studies amount. However, it has the disadvantage of being af- is needed in order to overcome taxonomic problems of fected by the degree of chromatin condensation and by Cirsium . the pretreatment method (Chiarini et al. 2018). Chromo- some lengths show wide variations. The smallest mea- Acknowledgements surement in terms of length of the haploid chromosome is C. pseudopersonata ssp. pseudopersonata (68.00 µm), The authors want to thank TUBITAK due to the and the largest haploid chromosome has a C. trachylepis financial support our researches (Project No: 106T167) (147.92 µm) taxon. The mean chromosome length varies and to Prof. Dr. Bayram YILDIZ, Prof. Dr. Tuncay between 2.70 and 13.07 µm. The smallest chromosome DIRMENCI and Prof. Dr. Turan ARABACI for helps length is in C. pseudopersonata ssp. pseudopersonata during the field studies. 2018 Karyology of Cirsium Species from Turkey 413

References Czech Vascular Plants. Academia, Praha. Kadereit, J. W. and Jeffrey, C. 2007. The Families and Genera of Vascular Plants, Vol. 8. Flowering Plants. : . Aishima, T. 1934. Chromosome numbers in the genus Cirsium I. Bot. Springer, Berlin. Mag. 48: 150–151. Kuzmanov, B. A. and Anchev, M. E. 1973. In: Löve, Á. (ed.). IOPB Albers, F. and Pröbsting, W. 1998. In: Wisskirchen, R. and Haeupler, chromosome number reports XLI. Taxon 22: 459–464. H. (eds.). Standardliste der Farn und Blütenpflanzen Deutschlands. Levan, A., Fredga, K. and Sanberg, A. A. 1964. Nomenclature for Bundesamt für Naturschutz und Verlag Eugen Ulmer, Stuttgart. centromeric position on chromosomes. Hereditas 52: 201–220. Arabacı, T. and Dirmenci, T. 2011. Cirsium yildizianum (Asteraceae: Martin, E., Dinç, M. and Duran, A. 2009. Karyomorphological study ), a new species from East Anatolia, Turkey. Ann. Bot. of eight Centaurea L. taxa (Asteraceae) from Turkey. Turk. J. Fenn. 48: 503–506. Bot. 33: 97–104. Arano, H. 1957. The karyotype analysis and its karyotaxonomic con- Májovský, J. (ed.) 1978. Index of chromosome numbers of Slovakian siderations in some genera of subtribe Carduinae. Jpn. J. Genet. flora (Part 6). Acta Fac. Rerum Nat. Univ. Comen. Bot. 26: 1–42. 32: 323–332. Moore, R. J. and Frankton, C. 1962. Cytotaxonomic studies in the Arano, H. 1963. Cytological studies in subfamily Carduoideae (Com- tribe Cynareae (Compositae). Can. J. Bot. 40: 281–293. positae) of Japan. IX. The karyotype analysis and phylogenetic Moore, R. J. and Frankton, C. 1963. Cytotaxonomic notes on some consideration on Pertya and Ainsliaea. Bot. Mag. 76: 32–39. Cirsium species of the Western . Can. J. Bot. 41: Bures, P., Wang, Y. F., Horova, L. and Suda, J. 2004. Genome size 1553–1567. variation in Central European species of Cirsium (Compositae) Moore, R. J. and Frankton, C. 1966. An evaluation of the status of C. and their natural hybrids. Ann. Bot. 94: 353–363. pumilum and C. hillii. Can. J. Bot. 44: 581–595. Charadze, A. L. 1963. Cirsium Mill. In: Bobrov. G., G. and Cherepa- Moore, R. J. and Frankton, C. 1967. Cytotaxonomy of foliose thistles nov, S. K. (eds.). Flora of the USSR XXVIII. Izdatel’stvo Akad. (Cirsium spp. aff. C. foliosum) of western . Can. J. Nauk SSSR, Moscow/Leningrad. pp. 63–270. Bot. 45: 1733–1749. Chiarini, F., Sazatornil, F. and Bernardello, G. 2018. Data reassess- Moore, R. J. and Frankton, C. 1969. Cytotaxonomy of some Cirsium ment in a phylogenetic context gives insight into chromosome species of the eastern United States with a key to eastern species. evolution in the giant genus Solanum (Solanaceae). Syst. Biodiv- Can. J. Bot. 47: 1257–1275. ers. 16: 397–416. Moore, R. J. and Frankton, C. 1974. The Cirsium arizonum complex Daskin, R., Yilmaz, O. and Kaynak, G. 2006. Presence of Cirsium er- of the southwestern United States. Can. J. Bot. 52: 543–551. iophorum (L.) Scop. (Asteraceae) in Turkey. Turk. J. Bot. 30: 1–5. Nilsson, Q. and Lassen, P. 1971. Chromosome numbers of vascular Davis, P. H. 1971. Distribution Patterns in Anatolia with Particular plants from Austria, Mallorca and Yugoslavia. Bot. Notiser. 124: References to Endemism. In: Davis, P. H., Harper, C. P. and 270–276. Hedge, I. C. (eds.). Plant Life of SW Asia. Botanical Society of Nouroozi, M., Sheidai, M., Attar, F. and Noormohammadi, Z. 2010. Edinburgh, Edinburgh. pp. 15–27. Contribution to cytotaxonomy of Iranian Cirsium (Asteraceae). Davis, P. H. and Parris, S. B. 1975. Cirsium Mill. In: Davis, P. H. Cytologia 75: 119–127. (ed.). Flora of Turkey and the East Aegean Islands Vol. 5. Edin- Ozcan, M., Hayırlıoglu-Ayaz, S. and Inceer, H. 2008. Chromosome burgh UniversityPress, Edinburgh. pp. 370–412. counts of some Cirsium (Asteraceae, Cardueae) taxa from Tur- Davis, P. H., Tan, K. and Mill, R. R. (eds.) 1988. Flora of Turkey and key. Caryologia 61: 375–382. the East Aegean Islands Vol. 10 (Suppl. I). Edinburgh University Ozcan, M., Hayırlıoglu-Ayaz, S. and Inceer, H. 2011. Chromosome Press, Edinburgh. pp. 164–165. reports in some Cirsium (Asteraceae, Cardueae) taxa from north- Elci, S. 1982. Observations in Cytogenetics and Research Methods. east Anatolia. Caryologia 64: 55–66. Fırat University, Science Literature Biology, Elazig. Petrak, F. 1979. Cirsium Mill. In: Rechinger K. H, (ed.). Flora Irani- Fedorov, A. A. 1969. Chromosome Numbers of Flowering Plants. ca. Compositae III̶Cynareae: Akademische Druck-u Verlag- Academy of Sciences of the USSR, VL Komarov Botanical Insti- sanstalt, Graz 139a: 231–285. tute, Moscow. Romero Zarco, C. 1986. A new method for estimating karyotype Frankton, C. and Moore, R. 1961. Cytotaxonomy, phylogeny and Ca- asymmetry. Taxon 35: 526–530. nadian distribution of Cirsium undulatum and Cirsium flodmanii. Snogerup, S. 1995. Reports 491–517. In: Kamari, G., Blanche, C. and Can. J. Bot. 39: 21–33. Garbari, F. (eds.). Mediterranean chromosome number reports-5. Frankton, C. and Moore, R. J. 1963. Cytotaxonomy of Cirsium muticum, Fl. Medit. 5: 331–334. Cirsium discolor and Cirsium altissimum. Can. J. Bot. 41: 73–84. Stebbins, G. L. 1971. Chromosomal Evolution in Higher Plants. Ed- Goldblatt, P. 1981. Index to plant chromosome numbers 1975–1978. ward Arnold Ltd. London. Monogr. Syst. Bot. Missouri Bot. Gard 6: 1–553. Tonian, T. R. 1981. New chromosome numbers of species of Cirsium Goldblatt, P. 1984. Index to plant chromosome numbers 1979–1981. Mill. grown in the Armenian SSR. Bio. Z. Armenii 34: 417–420, Monogr. Syst. Bot. Missouri Bot. Gard. 8: 1–427. 769–772. Goldblatt, P. 1985. Index to plant chromosome numbers 1982–1983. Tonian, T. R. 1982. New chromosome numbers of the species of Cir- Monogr. Syst. Bot. Missouri Bot. Gard. 13: 1–224. sium in Armenia. Ucenye Zapinski Erevan Univ. 3: 115–120. Goldblatt, P. 1988. Index to plant chromosome numbers 1984–1985. Uhrikova, A. 1978. In index of chromosome numbers of Slovakian Monogr. Syst. Bot. Missouri Bot. Gard. 23: 1–264. flora part 6. Acta Fac. Rerum Nat. Univ. Comenianae Bot. 26: Greilhuber, J. and Speta, F. 1976. C-banded karyotypes in the Scilla 1–42. hohenackeri group, S. persica and Puschkinia (Liliaceae). Plant Venora, G., Blangiforti, S., Ruffini, C. M., Pignone, D., Losavio, F. Syst. Evol. 126: 149–188. and Cremonini, R. 2002. Chromatin organisation and computer Guner, A., Ozhatay, N., Ekim, T. and Baser, K. H. C. (eds.) 2000. aided karyotyping of Triticum durum Desf. cv. Timilia. Caryolo- Flora of Turkey and the East Aegean Islands Vol. 11 (Suppl. II). gia 55: 91–98. Edinburgh University Press, Edinburgh. pp. 161–163. Watanabe, K., Yahara, T., Denda, T. and Kosuge, K. 1999. Chromo- Huziwara, Y. 1962. Karyotype analysis in some genera of Composi- somal evolution in the genus Brachyscome (Asteraceae, As- tae. VIII. Further studies on the chromosomes of Aster. Am. J. tereae): Statistical tests regarding correlation between changes in Bot. 49: 116–119. karyotype and habit using phylogenetic information. J. Plant Res. Javůrková-Jarolímová, V. 1992. List of Chromosome Numbers of the 414 Y. Kiran Cytologia 83(4)

112: 145–161. Soc. 158: 669–673. Yildiz, B. 2012. Cirsium Mill. In: Güner, A., Aslan, S., Ekim, T., Yildiz, B., Dirmenci, T. and Arabacı, T. 2009a. Cirsium handaniae Vural, M. and Babaç, M. T. (eds.). Turkey Plant List (Vascular (Asteraceae), a new species from Turkey. Ann. Bot. Fenn. 46: Plants). Nezahat Gökyiğit Botanic Garden and Flora Research 239–243. Associations Publication, İstanbul. pp. 141–146 (in Turkish). Yildiz, B., Dirmenci, T. and Arabacı, T. 2009b. A new record for the Yildiz, B., Arabacı, T., Dirmenci, T. and Celenk, S. 2011. Cirsium siv- flora of Turkey: Cirsium candelabrum Griseb. (Cirsium Sect. asicum sp. nov. and C. peshmenianum sp. nov. (Asteraceae) and Cirsium, Asteraceae, Cynareae). Turk. J. Bot. 33: 47–51. their allies from Turkey. Nor. J. Bot. 29: 26–37. Yuksel, E., Kiran, Y., Sahin, A., Yildiz, B. and Arabacı, T. 2013. Yildiz, B. and Dirmenci, T. 2008. A new species of Cirsium section Karyological studies of 10 Cirsium sect. Epitrachys (Asteraceae) Epitrachys (Asteraceae: Cardueae) from Turkey. Bot. J. Linn. species from Turkey. Turk. J. Bot. 37: 1085–1092.