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A Karyomorphological Study on the Subgenus Leopoldia of the Genus Muscari Growing in Turkey

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A Karyomorphological Study on the Subgenus Leopoldia of the Genus Muscari Growing in Turkey © 2020 The Japan Mendel Society Cytologia 85(1): 79–83 A Karyomorphological Study on the Subgenus Leopoldia of the Genus Muscari Growing in Turkey Yasar Kiran1, Süleyman Mesut Pınar2, Gulden Dogan1* and Hüseyin Eroğlu3 1 Department of Biology, Faculty of Science, Firat University, Elazig, Turkey 2 Van School of Health,Van Yüzüncü Yıl University, Tuşba-Van, Turkey 3 Department of Biology, Faculty of Sciences, Van Yüzüncü Yıl University, Tuşba-Van, Turkey Received November 2, 2019; accepted November 28, 2019 Summary The chromosome numbers and karyotypes of 11 species the subgenus Leopoldia of the genus Muscari distributed in Turkey were analyzed. These taxa are M. comosum (L.) Miller, M. weissii Freyn, M. cau- casicum (Griseb.) Baker, M. tenuiflorum Tausch, M. babachii Eker & Koyuncu, M. erdalii Özhatay & Demirci, M. longipes Boiss., M. massayanum Grunert, M. mirum Speta, M. elmasii Yıldırım, and M. haradjiani Briq. ex Rech., and six of them (M. babachii, M. erdalii, M. massayanum, M. mirum, M. elmasii, M. haradjiani) are en- demic species in Turkey. The somatic chromosome number of all studied taxa were determeined as 2n=2x=18. Haploid chromosome lengths varied 44.06 (M. massayanum) to 67.50 µm (M. babachii) among species. Karyo- type analysis indicated that Muscari taxa generally have median (m), submedian (sm) and subterminal (st) chromosomes. In addition, only M. comosum has one terminal (t) chromosome. Satellites were observed in five species, M. tenuiflorum, M. erdalii, M. longipes, M. mirum and M. haradjianii. This study is the first in terms of revealing the karyotypes of all taxa of subgenus Leopoldia on the genus Muscari. Keywords Muscari, Leopoldia, Karyotype, Chromosome number, Turkey. The genus Muscari Miller belonging to Asparagaceae, thus show a ploidy series ranging from 2x to 8x (Garbari has a natural distribution from the Mediterranean region 2003, Suárez-Santiago et al. 2007). and temperate Europe to Central Asia (Kubitzki 1998), Muscari is a taxonomically complex genus with four and is globally represented by 72 species (75 taxa). Due subgenera. Several multidisciplinary studies such as to its rich topographic and ecological heterogeneity, anatomy, palynology, molecular phylogeny, and karyol- Anatolia is an important diversification center for Mus- ogy are needed to clarify the taxonomy of the genus. cari. Indeed, more than half of taxa accepted worldwide Therefore, the karyology of subgenus Leopoldia species are native to Anatolia. According to recent studies con- of Muscari is currently being studied in order to clarify ducted in Turkey, the genus Muscari is represented by their taxonomy and make contributions to other multi- 44 species, of which 29 endemics to Turkey (Demirci disciplinary studies on the genus. et al. 2019, Dizkırıcı et al. 2019, Dogu and Uysal 2019, Eker 2019a, b, Eker et al. 2019). Materials and methods Infrageneric taxonomy, specifically boundaries of the subgenera have not been unambiguously delimited till All plant specimens were collected from natural habi- now. First of all, four subgenera, Muscari, Leopoldia, tats in 2016–2018 as a part of a taxonomic revision of Moscharia (Baker) Chouard (=Muscarimia), and Pseu- the genus Muscari in Turkey. Species names, localities domuscari (Losinsk.) D. C. Stuart, were accepted for the (geographical position, altitude) and voucher number are genus (Davis and Stuart 1980, Davis 1984, Speta 1989, given in Table 1. The voucher specimens were deposited 1998). Karlén (1991) grouped the genus into four sub- at the Yuzuncuyil University Herbarium, Turkey. genera as Botryanthus (Kunth) Zahar., Leopoldia (Parl.) The karyological studies are conducted on the meri- Rouy, Muscari, and Pseudomuscari Stuart. stematic cells of bulbs. The bulbs were germinated at Muscari is characterized by the base chromosome 25°C. The actively growing root tips were pretreated number x=9, with bimodal karyotype (Ruiz Rejón and with 0.05% colchicine for 3–3.5 h at room temperature. Oliver 1981). Species of subgenus Leopoldia are gener- Afterwards the root tips were fixed with Carnoy fixa- ally diploids (Nersesian 2001, Jafari and Maassoumi tive (1 : 3 glacial acetic acid–absolute ethanol) for at least 2011) while species of subgenera Muscari and Botryan- 24 h at 4°C, hydrolysed in 1 M HCl at 60°C for 3–5 min, then rinsed in tap water for 3–5 min. Finally they were stained in the Feulgen reagent for 1 h and mounted in * Corresponding author, e-mail: gdogan@firat.edu.tr DOI: 10.1508/cytologia.85.79 45% acetic acid (Elci 1982). Digital microphotographs With Supplement files of Tables S1–S11. from at least five well-spread metaphase plates were tak- 80 Y. Kiran et al. Cytologia 85(1) Table 1. Localities and voucher numbers of the studied Muscari taxa. Collecter Taxa Locality (voucher number) M. comosum C2 Muğla: Fethiye, between Fethiye and Korkuteli, Karabel Pass., scrub yards, 1284 m, 14.05.2016. H. Eroğlu (1269) M. weissii C3 Antalya, Manavgat, Sorgun, hotels districy, Pinus pinea forest, dunes, 28 m, 28.03.2018. H. Eroğlu (1350) M. caucasicum C4 Konya: Bozkır, between Karabayır and Kovanlık villages, Quercus yards, steppe, 1532 m, 10.05.2018. H. Eroğlu (1376) M. tenuiflorum B6 Adana: Tufanbeyli, Doğanbeyli, Burunoluk Hill, 1650 m, 16.06.2018. H. Eroğlu (1401) M. babachii C6 Hatay: Antakya, Kisecik Village, Radar road, stony rocky scrub yards, 1430 m, 12.06.2016. H. Eroğlu (1286) M. erdalii C4 İçel: Mut, south of İbrahimli Village, scrub yards, marly places, 900 m, 02.05.2016. H. Eroğlu (1255) M. longipes B6 Sivas: Hafik, west of Durulmuş village, marly hills, 1312 m, 30.05.2017. H. Eroğlu (1327) M. massayanum C5 Adana: Pozantı, above Hamidiye Village, serpentine Pinus yards, 1357 m, 01.05.2016. H. Eroğlu (1253) M. mirum C2 Denizli: Çameli, Denizli-Fethiye road, 4 kilometers before Aliveren village, Pinus yards, 1475 m, H. Eroğlu (1259) 04.05.2016. M. elmasii C2 Muğla: Dalaman, above Gürleyik Village, Çal Mountain, Pinus yards, serpentine places, 1271 m, H. Eroğlu (1270) 27.06.2016. M. haradjianii C6 Gaziantep: Nurdağı, between Gaziantep and Nurdağı, east of Atalar village, field edge, 1100 m, 13.05.2018. H. Eroğlu (1388) Fig. 1. Metaphase chromosomes of Muscari taxa. a. M. comosum. b. M. weissii. c. M. caucasicum. d. M. tenuiflorum. e. M. ba- bachii. f. M. erdalii. g. M. longipes. h. M. massayanum. i. M. mirum. j. M. elmasii. k. M. haradjianii. Scale bars=10 µm. en using an Olympus BX51 microscope and an Olympus mined based on centromer position (Levan et al. 1964). Camedia C-4000 digital camera. While the intrachromosomal asymmetry index (A1) and the interchromosomal asymmetry index (A2) was Results calculated according to the formula proposed by Romero Zarco (1986) was calculated (Table 2). The ideograms of The number of somatic chromosomes, chromosome these taxa are arranged in decreasing lengths according length range, haploid chromosome length, arm ratio, rel- to the chromosome size in the metaphase (Fig. 2). The ative length and karyotype formula were determined in examined taxa and characteristics of somatic chromo- this study were measured. Karyotype formula was deter- somes are given below. 2020 Karyology of Muscari 81 Table 2. Karyological features of the studied Muscari taxa. Chromosome Haploid chromosome Arm ratio Relative length (%) Taxa 2n length (µm) Karyotype formula A1 A2 length (µm) Min Max Min Max Min Max M. comosum 18 2.15 21.62 50.35 1.03 10.31 5.73 33.15 14m+2sm+2t 0.33 0.79 M. weisii 18 3.47 10.44 51.49 1.05 3.54 6.73 20.27 12m+2sm+4st 0.33 0.45 M. caucasicum 18 3.21 13.32 54.59 1.07 5.23 5.88 24.41 14m+4st 0.32 0.61 M. tenuiflorum 18 3.81 10.29 55.53 1.11 3.56 6.86 18.53 8m+2mSAT+6sm+2st 0.38 0.39 M. babachii 18 4.20 12.63 67.50 1.22 2.83 6.23 18.72 10m+8sm 0.42 0.37 M. erdalii 18 3.51 10.82 60.24 1.04 3.19 5.83 17.96 8m+2mSAT+6sm+2st 0.35 0.42 M. longipes 18 3.62 9.90 54.12 1.28 4.28 6.69 18.29 8m+2mSAT+6sm+2st 0.42 0.40 M.massayanum 18 3.28 7.93 44.06 1.02 2.87 7.46 18.00 10m+8sm 0.32 0.32 M. mirum 18 3.17 10.40 53.22 1.13 4.23 5.96 19.54 10m+2mSAT+4sm+2st 0.38 0.45 M. elmasii 18 3.46 12.12 57.46 1.17 4.01 6.03 21.10 14m+4st 0.38 0.50 M. haradjianii 18 3.36 10.27 56.68 1.07 4.75 5.93 18.12 6m+2mSAT+8sm+2st 0.41 0.41 See Tables S1–11 for mesurement data of each species. Fig. 2. Haploid idiograms of Muscari taxa. a. M. comosum. b. M. weissii. c. M. caucasicum. d. M. tenuiflorum. e. M. babachii. f. M. erdalii. g. M. longipes. h. M. massayanum. i. M. mirum. j. M. elmasii. k. M. haradjianii. M. comosum: The somatic chromosome number of this species was found to be 2n=18 (Fig. 1d). The karyo- this species was found to be 2n=18 (Fig. 1a). type fomula was 8m+2mSAT+6sm+2st (Fig. 2d, Tables The karyotype fomula was 14m+2sm+2t (Fig. 2a, 2, S4). Satellites were observed at the short arm of small Tables 2, S1). The chromosome length ranges 2.15 to m. The chromosome length ranges 3.81 to 10.29 µm 21.62 µm and the total length of the haploid complement and the total length of the haploid complement was was 50.35 µm.
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