Karyological Investigation on Five Taxa of Bellevalia Lapeyr
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© 2019 The Japan Mendel Society Cytologia 84(2): 173–177 Karyological Investigation on Five Taxa of Bellevalia Lapeyr. (Asparagaceae) from Turkey Gulden Dogan1*, Yasar Kiran1 and M. Yavuz Paksoy2 1 Department of Biology, Faculty of Science, Firat University, Elazig, Turkey 2 Department of Enviromental Engineering, Faculty of Engineering, Munzur University, Tunceli, Turkey Received December 31, 2018; accepted February 9, 2019 Summary The somatic chromosome numbers and karyotypes were examined in five Bellevalia taxa includ- ing two endemic species of B. chrisii and B. gracilis distributed in Turkey. The chromosome numbers were determined as 2n=2x=8 in all studied taxa [B. chrisii Yıldırım & B. Şahin sp. nov., B. glauca (Lindl.) Kunth., B. gracilis Feinbrun, B. longipes Post, B. sarmatica (Pallas ex Georgi) Woronow]. The karyotype formula was de- termined as 2m+sm+st in B. chrisii and B. longipes, 3m+st in B. glauca, B. gracilis and B. sarmatica. The find- ings for each of the analyzed taxa are compared with the results of previous studies. Key words Asparagaceae, Bellevalia, Chromosome number, Karyotype, Turkey. Bellevalia Lapeyr. is a genus of bulbous plants be- The aim of this paper was to provide karyotypes data longing to Asparagaceae, subfamily Scilloideae, tribe to clearly define chromosome number and karyotype for Hyacintheae (APG III 2009, Chase et al. 2009). The five Turkish Bellevalia species including B. chrisii for genus is distributed over the Mediterranean region which no information about chromosomes is available. from Morocco and Algeria eastwards to the Caucasus and Iran (Ozhatay et al. 1991, Bareka 2008, Jafari and Materials and methods Maassoumi 2008, Borzatti von Loewenstern et al. 2013, Karabacak et al. 2014, 2015) and 74 species are listed Plant materials of five Bellevalia species, B. chrisii (Govaerts 1996). The largest number of species in the Yıldırım & B. Şahin sp. nov., B. glauca (Lindl.) Kunth., genus is found in the Irano–Turanian phytogeographi- B. gracilis Feinbrun, B. longipes Post, B. sarmatica (Pal- cal region (Feinbrun 1938). In Turkey, Bellevalia is las ex Georgi) Woronow, were collected from natural represented by 28 taxa, 17 of which are endemic to the habitats during the fruiting season between 2017 and country (Wendelbo 1984, Davis et al. 1988, Ozhatay 2018. Their localities, geographical position, altitude, and Kultur 2006, Ozhatay et al. 2009, 2011, Guner et al. and voucher number are presented in Table 1. The 2012, Uzunhisarcıklı et al. 2013, Gurdal et al. 2014, voucher specimens were deposited at the Herbarium of Karabacak et al. 2014, 2015). Firat University. The chromosome numbers (2n=8) of six Bellevalia The karyological studies are conducted on the meri- species were published in the first supplement of the stematic cells of bulbs. The bulbs were germinated at “Flora of Turkey” (Davis et al. 1988), five by Von Both- 25°C. The actively growing root tips were pretreated mer and Wendelbo (1981) and one by Wendelbo (1984). with 0.05% colchicine for 3–3.5 h at room temperature. The second supplement (Ozhatay et al. 2000) contained Afterward, the root tips were fixed with Carnoy fixative chromosome numbers of 17 species. The chromosome (1 : 3 glacial acetic acid–absolute ethanol) for at least 24 h numbers (2n=8, 12, 16, 20, 24, 31, 32, 35, e.g.) have at 4°C, hydrolyzed in 1 M HCl at 60°C for 10–15 min, been reported in nine Bellevalia species (Ozhatay et al. then rinsed in tap water for 3–5 min. Finally, they were 1991, Johnson 1994, Ozhatay and Johnson 1996, John- stained in the Feulgen reagent for 1 h and mounted in son and Brandham 1997, Dane 1999). Intraspecific poly- 45% acetic acid (Kiran et al. 2012). Digital micropho- ploidy with the basic chromosome number of x=4 was tographs from at least five well-spread metaphase plates also reported in B. edirnensis (Ozhatay et al. 1991). The were taken using an Olympus BX51 microscope and an polyploids of tetraploid (2n=16), hexaploid (2n=24) and Olympus Camedia C-4000 digital camera. octoploid (2n=32) and, also triploid (2n=12) and penta- The number of somatic chromosomes, chromosome ploid (2n=20) are reported in some members of Belleva- length range, haploid chromosome length, arm ratio, lia (Ozhatay et al. 1991). relative length, and karyotype formula were determined in this study were measured. Karyotype formula was * Corresponding author, e-mail: gdogan@firat.edu.tr determined based on the centromere position using a DOI: 10.1508/cytologia.84.173 system of Levan et al. (1964). The ideograms of these 174 G. Dogan et al. Cytologia 84(2) taxa are arranged in decreasing lengths according to the Results and discussion chromosome size in the metaphase (Martin et al. 2009). Karyotype asymmetry was determined by using vari- The characteristics of somatic chromosomes of exam- ous methods including total form percentage (TF%), ined five Bellevalia taxa are given below. percentage karyotype asymmetry index (AsK%), index B. chrisii Yıldırım & B. Şahin is a local endemic spe- of karyotype symmetry (Syi), index of chromosome cies that is restricted to the Pötürge district in Malatya, size resemblance (Rec) and the degree of asymmetry for eastern Anatolia. Plants collected was on mountain karyotypes (A) (Huziwara 1962, Arano 1963, Greilhu- steppe at 1900–2000 m (Table 1) belonging to the Irano– ber and Speta 1976, Venora et al. 2002, Watanabe et al. Turanian floristic region. The somatic chromosome 1999). While the intrachromosomal asymmetry index number of this species was found to be 2n=8 in all five (A1) and the interchromosomal asymmetry index (A2) plants (Fig. 1a). The chromosome length ranges 10.64 to was calculated according to the formula proposed by 17.04 µm and the total length of the haploid complement Romero Zarco (1986). Also, karyotype evolution was was 53.10 µm. The haploid karyotype consists of two speculated based on Stebbins’ classification (1971). metacentric (m), one submetacentric (sm) and one sub- telocentric (st) chromosomes. Arm ratios are 1.27–3.57 and relative lengths are 20.04–32.09% (Table 2). Idio- Table 1. Localities and voucher numbers of the studied Bellevalia taxa. Taxa Locality Voucher number B. chrisii B7/Malatya: Pütürge, Kubbe Mountain, 1900 m. Dogan, 2052 B. glauca B7/Malatya: Doğanşehir, 1900 m. Dogan, 2065 B. gracilis B7/Malatya: Pütürge, Nemrut-Büyüközü Village, 1600 m. Dogan, 2053 B. longipes B7/Malatya: Darende, Irmaklı-Söğütlü, 1400 m. Dogan, 2116 B. sarmatica B7/Malatya: Bölüklü Village, 1500 m. Dogan, 2112 Fig. 1. Metaphase chromosomes of five Bellevalia taxa. a. B. chrisii, b. B. glauca, c. B. gracilis, d. B. longipes, e. B. sarmatica. Scale bars=10 µm. 2019 Karyology of Bellevalia 175 gram is given in Fig. 2a. This species of karyology was haploid karyotype consists of three m and one st chro- determined for the first time. mosomes. Chromosome arm ratios are 1.21–3.46 and B. glauca (Lindl.) Kunth. is a new record of Turkey relative lengths are 19.32–32.36% (Table 2). Idiogram is and is restricted to the Kubbe Pass in Malatya, eastern given in Fig. 2c. Anatolia. It grows on mountain steppe at 2000 m. It is an B. longipes Post distributes fields, c. 1050 m (Table 1). element belonging to the Irano–Turanian floristic region. It is an element belonging to the Irano–Turanian floristic The somatic chromosome number of this species was region. The somatic chromosome number of this species found to be 2n=8 in all five plants (Fig. 1b) and sup- was found to be 2n=8 in all five plants (Fig. 1d) support- ported the previous report (Nersesian 2001). The chro- ing the previous report (Von Bothmer and Wendelbo mosome length ranges 10.66 to 18.38 µm, and the total 1981). The chromosome length ranges 9.97 to 18.46 µm, length of the haploid complement was 56.84 µm. The and the total length of the haploid complement was haploid karyotype consists of three m and one st chro- 54.00 µm. The haploid karyotype consists of two m, one mosomes. Arm ratios are 1.24–3.43 and relative lengths sm, and one st chromosomes. Arm ratios are 1.29–3.86 are 18.75–32.33% (Table 2). Idiogram is given in Fig. 2b. and relative lengths are 18.46–34.18% (Table 2). Idio- B. gracilis Feinbrun is an endemic, and one of the Ira- gram is given in Fig. 2d. no–Turanian element. This taxon distributed stony hill- B. sarmatica (Pallas ex Georgi) Woronow distributed tops, eroded shaley slopes in Pinus brutia forest, fields, fallow fields, banks, steppe, between 870 and 1900 m between 600 and 1500 m (Table 1). The somatic chromo- (Table 1). The somatic chromosome number was found some number of this species was found to be 2n=8 in all to be 2n=8 (Fig. 1e), as in many works of literature five plants (Fig. 1c). Our observation confirmed diploid (Zakhariyeva and Makushenko 1969, Von Bothmer and number but not tetraploid (Kayiran and Ozhatay 2017) Wendelbo 1981, Pogosyan and Torosian 1983, Ozhatay and B chromosomes (Ozhatay et al. 1991). The chro- et al. 1991, Johnson 2003). The chromosome length mosome length ranges 12.21 to 20.45 µm, and the total ranges 8.70 to 14.81 µm and the total length of the hap- length of the haploid complement was 63.19 µm. The loid complement was 45.95 µm. The haploid karyotype Table 2. Karyological features of the studied Bellevalia taxa. Chromosome length (µm) Haploid chromosome Arm ratio Relative length (%) Taxa 2n Karyotype formula Min Max length (µm) Min Max Min Max B. chrisii 8 10.64 17.04 53.10 1.27 3.57 20.04 32.09 2m+sm+st B.