© 2018 The Japan Mendel Society Cytologia 83(4): 407–414 Karyological Investigation of Sixteen Cirsium Mill. (Asteraceae, 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 taxonomy 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 Plant 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.