Comparative morphological, karyological and ...

Botany Letters

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Comparative morphological, karyological and palynological investigations of three endemic Centaurea L. species (Asteraceae) in Turkey Sukru Hayta, Neslihan Tasar, Yasar Kiran & Birol Baser To cite this article: Sukru Hayta, Neslihan Tasar, Yasar Kiran & Birol Baser (2017) Comparative morphological, karyological and palynological investigations of three endemic Centaurea L. species (Asteraceae) in Turkey, Botany Letters, 164:3, 241-251, DOI: 10.1080/23818107.2017.1357497 To link to this article: http://dx.doi.org/10.1080/23818107.2017.1357497

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Date: 10 September 2017, At: 13:07

Botany Letters, 2017 VOL. 164, NO. 3, 241–251 https://doi.org/10.1080/23818107.2017.1357497

Comparative morphological, karyological and palynological investigations of three endemic Centaurea L. species (Asteraceae) in Turkey Sukru Haytaa, Neslihan Tasarb, Yasar Kiranc and Birol Baserd a

Faculty of Engineering and Architecture, Department of Environmental Engineering, Bitlis Eren University, Bitlis, Turkey; bTunceli Vocational School, Tunceli University, Tunceli, Turkey; cFaculty of Science, Department of Biology, Firat University, Elazig, Turkey; dFaculty of Arts and Science, Department of Biology, Bitlis Eren University, Bitlis, Turkey

Downloaded by [159.146.26.122] at 13:07 10 September 2017

ABSTRACT

Centaurea derderiifolia Wagenitz., Centaurea drabifolia Sm. subsp. floccosa (Boiss.) Wagenitz & Greuter and Centaurea kotschyi (Boiss. & Heldr.) Hayek var. floccosa (Boiss.) Wagenitz, which are endemic taxa from Turkey belonging to Centaurea sect. Cheirolepis, were subjected to morphological, karyological and palynological investigations. The somatic chromosome numbers, pollen morphology and detailed morphometric properties were examined. The somatic chromosome numbers were determined as 2n =2x = 36 in C. derderiifolia, C. kotschyi var. floccosa, and C. drabifolia subsp. floccosa. Karyotype analysis indicated that chromosomes of these Centaurea taxa generally have median region (m), submedian region (sm) and rarely median point (M) karyotypes. The pollen morphology of these endemic taxa of the genus Centaurea was investigated by light and scanning electron microscopy. The pollen grains of the examined species are mainly isopolar and radially symmetric, subprolate, tricolporate. Tectum ornamentation is scabrate and scabrate-perforate, amb triangular. The pollen grains of all taxa studied are 3-zonocolporate. The colpus is usually long, acute at ends and with smooth membrane. Pollen grains belonging to Cheirolepis section reveal homogeneity in their characters.

Introduction The genus Centaurea L. (Asteraceae) comprises more than 600 species worldwide, and is mostly distributed in the Mediterranean region and in West Asia (GarciaJacas et al. 2000). Centaurea has one of the highest rates of endemism in Turkey, with 112 endemics among 181 species (Güner et al. 2000). This high endemism ratio shows that Turkey is one of the centres of diversity of this genus (Celik, Uysal, and Menemen 2008). Centaurea is the third largest genus in terms of species number after Astragalus and Verbascum in Flora of Turkey (Güner et al. 2000). When the recently described species are added to previous Turkish floristic treatements (Wagenitz 1975; Davis, Mill, and Tan 1988; Güner et al. 2000), there are ca. 195 Centaurea species growing in Turkey. Those species have been described by Wagenitz and Hellwig 1997 [Centaurea yozgatensis]; Wagenitz, Ertugrul, and Dural 1998 [Centaurea hadimensis]; Duran and Duman 2002 [Centaurea antalyense]; Türkoğlu, Akan, and Civelek 2003 [Centaurea yıldızii]; Uzunhisarcıklı, Teksen, and Dogan 2005 [Centaurea marashica]; Aytaç and Duman 2005 [Centaurea goeksuense]; Wagenitz et al. 2006 [Centaurea werneri and Centaurea ulrichiorum]; Vural et al. 2006 [Centaurea tuzgoluensis]; Uzunhisarcıklı, Doğan, and Duman 2007 [Centaurea kizildaghensis]; Kaya and Vural 2007 [Centaurea elazigensis]; Uysal et al. 2007 [Centaurea CONTACT Sukru Hayta

[email protected]

© 2017 Société botanique de France

ARTICLE HISTORY

Received 7 April 2016 Accepted 17 July 2017 KEYWORDS

Chromosome; Centaurea; Cheirolepis; pollen; Turkey

glabro-auriculata]; Uysal 2008 [Centaurea ertugruliana]; Uysal and Köse 2009 [Centaurea dursunbeyensis]; Hamzaoğlu and Budak 2009 [Centaurea aksoyi]; Daşkın and Yılmaz 2009 [Centaurea kaynakiae]; Köse and Alan 2013 [Centaurea baseri]; Bancheva, Kaya, and Binzet 2014 [Centaurea aytugiana]; Negaresh, Kaya, and Rahiminejad 2015 [Centaurea sennikoviana]; Bona 2015 [Centaurea goksivriensis]; Bona 2016 [Centaurea amanosensis]; Yüzbaşıoğlu, Bona, and Genç 2015 [Centaurea ziganensis]; Uysal and Hamzaoğlu 2016 [Centaurea mersinensis]; Uysal et al. 2016 [Centaurea nallihanense]; and Uysal, Dural, and Tugay 2017 [Centaurea sakariyaensis]. This genus is known in Turkey as “peygamber çiçeği”, “zerdali dikeni”, “çoban kaldiran” and “Timur dikeni” (Baytop 1999). In Turkey, Centaurea generally grows on stony calcareous cliffs, vineyards, roadsides, seashores, gypsum fields, open woodlands and shrublands, waste places, steppe, fallow fields, maquis, sandy beaches, forests, dry meadows, rocky slopes and on maritime limestone cliffs. Centaurea species are potentialy important medicinal plants considering their biologically active lignan-type metabolites (Boldizsár et al. 2010; Szokol-Borsodi et al. 2012) and biological activities (Csupor et al. 2010). In Turkish folk medicine, aerial flowering parts or flower heads of some Centaurea species are used to alleviate a wide range of symptoms (Yesilada et al. 2004; Gürbüz and Yesilada 2007).


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The biggest and the most important problem of this genus is the taxonomy, which has been changed several times over the years. Taxonomically, Centaurea L. has been considered an unnatural grouping and recent approaches have split this taxon into several genera: Centaurea s.s., Rhaponticoides Vaill., Psephellus Cass. and Cyanus Mill. (Wagenitz and Hellwig 2000; Greuter 2003a, 2003b; Hellwig 2004). Recent molecular analyses of the genus and of subtribe Centaureinae, together with studies of morphology, palynology and karyology, have enabled the natural limits of Centaurea to be established with greater confidence (Susanna et al. 1995; Wagenitz and Hellwig 1996; Garcia-Jacas et al. 1998, 2000, 2001). Karyological characteristics such as chromosome number, location of centromeres, and number and position of satellites have been used in recent taxonomy studies to clarify the relationships between Centaurea species (Carr et al. 1999; Martin, Dinç, and Duran 2009; Yıldız, Minareci, and Cırpıcı 2009; Gunjan and Roy 2010; Tabur et al. 2012). However, only a few cytological studies have been published on the Centaurea genus in Turkey. The aims of this study is to carry out, on three endemic Centaurea species, a detailed study of the main morphological, karyological and palynological characters that are used for the sectional classification. Thus, this work aims to contribute to academic research on the Centaurea genus, particularly on its members which chromosome number, morphological and palynological characteristics are not very well known.

Materials and methods Plant material Plant materials were collected from natural habitats during fruiting season between 2011 and 2012. The voucher specimens were deposited at the Firat University Herbarium. The localities’ geographical position, altitude, and the voucher number of the specimen, are presented in Table 1. The distribution areas of the three endemics Centaurea derderiifolia, C. drabifolia subsp. floccosa and C. kotschyi var. floccosa in Turkey are indicated on the maps in Figure 1. Morphological analysis The morphological observations and morphometric analysis were carried out on both fresh and herbarium specimens. Some descriptive and diagnostic

characteristics such as leaves, flowers, involucre, pappus, achenes, and appendage shape were analysed and used for the description of species. Palynological analysis Pollen was removed from mature anthers of the three endemic taxa collected from natural habitats, as shown in Table 1. Pollen slides were prepared using the Wodehouse (1935) technique. Pollen grains were stained with glycerine-jelly plus basic-fuchsin with slight heating of the slide, then a coverslip was placed on it. These preparations were investigated and measured under the light microscope. The following parameters were measured: P, polar axis; E, equatorial diameter; P/E, ratio of polar axis and equatorial diameter; Clg, colpus length; Clt, colpus diameter; Plg, pore length; Plt, pore width – costae width. The photographs were taken with the Olympus BX51 digital photomicrograph system. Measurements were based on at least 30 pollen grains per specimen. For scanning electron microscopy (SEM) observation, dried pollen grains were transferred onto stubs and then coated with gold. They were observed and photographed with a JEOL JSM 7001F scanning electron microscope. Ten intact pollen grains from each taxon were scored on SEM. microphotography for the following parameters: number of scabrae in 5 μm2, scabrae width (dt) and scabrae length (dh). The terminologies of Faegri and Iversen (1992), Pinar and Inceoglu (1996), Punt et al. (2007), Punt and Hoen (2009), and Hesse et al. (2009) were followed. The pollen shape class, based on the ratio of polar axis to equatorial axis (P/E), was identified using Erdtman’s system (Erdtman 1969). Karyological studies The karyological studies were conducted on the meristematic cells of root tips. The seeds were germinated on moist filter paper in Petri dishes at 25 °C. The actively growing root tips were pretreated with aqueous colchicine (0.05%) for 3–3.5 h at room temperature. Subsequently, the root tips were fixed with Carnoy (1: 3 glacial acetic acid–absolute ethanol) for at least 24 h at 4 °C, hydrolysed in 1 N HCl at 60 °C for 15 min, then rinsed in tap water for 3–5 min. Finally, they were stained in Feulgen for 1 h and mounted in 45% acetic acid. Digital microphotographs from at least five well-spread metaphase plates were taken using an Olympus BX51 microscope,

Table 1. Localities and voucher numbers of Centaurea taxa investigated. Taxa C. drabifolia subsp. floccosa C. kotschyi var. floccosa C. derderiifolia

Locality Elazığ: Baskil, Haroğlu mountain 1950 m, 22.07.2011 Elazığ: Baskil in upward Kuluşağı village, 1350 m, 26.08.2012 Elazığ: Palu, Baltaşı village,back station. 1450 m, 22. 07. 2011.

Voucher number Tasar 1025 Tasar 1026 Tasar 1024


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Figure 1. Distribution map of C. derderiifolia, C. drabifolia subsp. floccosa and C. kotschyi var. floccosa in Turkey (Babac 2004).

and were recorded with an Olympus Camedia C-4000 digital camera. The short arm (s), long arm (l) and total lengths (tl) of each chromosome were measured and the relative lengths, arm ratios, and centromeric indices were determined from images of selected cells. Chromosomes were classified according to the nomenclature of Levan, Fredga, and Sandberg (1964). The chromosome numbers of the species were checked using the Flora of Turkey (Davis 1975) and supplements (Davis, Mill, and Tan 1988; Güner et al. 2000). Also, relevant literatures and the online chromosome number databases, Index to Plant Chromosome Numbers (IPCN) (https://www.tropicos.org/Project/ IPCN) and Index to Chromosome Numbers in Asteraceae (Info-Lib Index to Chromosome Numbers in Asteraceae, https://www.lib.kobe-u.ac.jp/infolib/meta_ pub/G0000003asteraceaesresult-en) were checked.

Results Morphological results Centaurea derderiifolia (Figure 2) 300–100 cm, perennial, with woody root stock and sterile shoots. Stem simple or branched; erectherbs. Leaves entire, linear-lanceolate, lateral veins not prominent, tomentose, margins scabrous; Basal leaves lanceolate, 4–7 × 0.8–1 cm, not withered at flowering time; stem leaves similar to basal leaves but reduced upwards, upper not enveloping capitula. Involucre ovoid, 25–30 × 16–23 mm, bracts large, lanceolate and hairy. Appendage orbiculate, 2–5 × 2–3 mm, including lateral fimbriae, concealing basal parts of the bracts, reddish-coloured with brownish centre, very weakly hooded

in the middle, laterally fimbriate with short fimbriae 2–5 mm, apical fimbria reduced in a mucro, 3–8 mm. Flowers yellow, marginal; not radiant, with nectar, lobes with stria brownish red longitudinally. Achenes lanceolate, 4–9 × 1–3 mm, blackish brown with lighter stripes, with creamish stria. Pappus 14–20 mm, simple, barbellate, brown, easily deciduous. Flowering: June, Mature fruit: July. Steppe with rocky areas, alt. 1000–1900 m. Centaurea drabifolia subsp. floccosa (Figure 3) 31–40 cm, perennial, with woody root stock and sterile shoots. Stem simple or branched; erectherbs. Leaves entire, linear-lanceolate, lateral veins not prominent, tomentose, margins scabrous; Basal leaves lanceolate, 8–9 × 0.5–1.2 cm, not withered at flowering time; stem leaves similar to basal leaves but reduced upwards, upper not enveloping capitula. Involucre ovoid, 15–22 × 8–20 mm, bracts large, lanceolate and hairy. Appendage orbiculate, 2–4 × 1–4 mm, including lateral fimbriae, concealing basal parts of the bracts, reddish-coloured with brownish centre, very weakly hooded in the middle, laterally fimbriate with short fimbriae 2–5 mm, apical fimbria reduced in a mucro, 3–5 mm. Achenes lanceolate, 2–4 × 1–2 mm, blackish-brown with lighter stripes, with creamish stria. Pappus 10–15 mm, simple, barbellate, brown, easily deciduous. Flowering: June, Mature fruit: July. Rocky areas, alt. 800–1900 m. Centaurea kotschyi var. floccosa (Figure 4) 26–45 cm, perennial, with woody root stock and sterile shoots. Stem simple or branched; erectherbs. Leaves entire, linear-lanceolate, lateral veins not prominent, tomentose, margins scabrous; Basal leaves lanceolate,

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4–9 × 0.5–1.5 cm, not withered at flowering time; stem leaves similar to basal leaves but reduced upwards, upper not envoloping capitula. Involucre ovoid, 8–15 × 8–15 mm, bracts large, lanceolate and hairy. Appendage orbiculate, 3–6 × 2–3 mm, including lateral fimbriae, concealing basal parts of the bracts, reddish-coloured with brownish centre, very weakly hooded in the middle, laterally fimbriate with short fimbriae 1–2, 5 mm, apical fimbria reduced in a mucro, 3–12 mm. Flowers yellow, marginal; not radiant, with nectar, lobes with stria brownish-red longitudinally. Achenes lanceolate, 8–15 × 3–5 mm, blackish-brown with lighter stripes, with creamish stria. Pappus 9–15 mm, simple, barbellate, brown, easily deciduous. Flowering: July, Mature fruit: August. Steppe with rocky areas, alt. 1500–2300 m.


Karyological results The number of somatic chromosomes, morphometric parameters, karyotype formula were determined for each taxon (Table 2)

Figure 2. Image of Centaurea derderiifolia.

Centaurea drabifolia subsp. floccosa The chromosome numbers were found to be 2n = 36 in specimens here studied. Metaphase chromosome length ranges from 1.79 to 4.42 μm, and the total length of the haploid set was 49.63 μm. There were 2 median points (M), 11 median regions (m) and 5 submedian regions (sm) chromosomes. (Tables 2–3; Figures 5–6) Centaurea kotschyi var. floccosa The chromosome numbers were found to be 2n = 36 in specimens here collected (Table 1). Metaphase chromosome length ranges from 1.44 to 4.93 μm, and the total length of the haploid set was 50.36 μm. There were 4 M, 11 m and 3 sm chromosomes (Tables 2–3; Figures 5–6).

Figure 3. Image of Centaurea drabifolia subsp. floccosa.

Centaurea derderiifolia The chromosome number was determined as 2n = 36 for this taxon. The chromosome length ranges from 1.13 to 2.48 μm, and the total length of the haploid set was 32.27 μm. There were 2 M, 13 m 3 sm chromosomes. (Tables 2–3; Figures 5–6). The taxa of genus Centaurea showed a wide range of chromosome numbers as 2n = 2x = 18, and 36 in this study. Analysis of the karyotype formulae indicated that chromosomes of three studied Centaurea taxa have m, sm and M, karyotypes (Tables 2–3). The taxa have x = 9 and 18 basic chromosome numbers and all of them are diploids. Palynological results

Figure 4. Image of Centaurea kotschyi var. floccosa.

The pollen of the three species examined was mainly isopolar and radially symmetric, subprolate, tricolporate with ornamentation scabrate and scabrateperforate, amb triangular. The pollen of all taxa studied were 3-zonocolporate. The colpus is usually long, acute

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Table 2. Somatic chromosome number (2n), ploidy level, karyotype formula, ranges of chromosome length and total karyotype length (TKL) for the studied Centaurea taxa. Taxa C. drabifolia subsp. floccosa C. kotschyi var. floccosa C. derderiifolia

Ploidy level 2x 2x 2x

2n 36 36 36

Karyotype formula 2 M + 11 m + 5sm 4 M + 11 m + 3sm 2 M + 14 m + 2sm

Chromosome length range (μm) 4,42–1,79 4,93–1,44 2,48–1,13

TKL (μm) 49.63 50.36 32.27


Table 3. Karyomorphological parameters of three Centaurea taxa belonging to Cheirolepis section. Pair No. RL C. kotschyi var. floccosa I 4.93 II 3.81 III 3.81 IV 3.26 V 3.1 VI 3 VII 2.9 VIII 2.85 IX 2.74 X 2.56 XI 2.41 XII 2.37 XIII 2.37 XIV 2.3 XV 2.22 XVI 2.22 XVII 2.07 XVIII 1.44 Centaurea drabifolia subsp. floccosa I 4.42 II 3.84 III 3.84 IV 3.37 V 3.28 VI 3.06 VII 3 VIII 2.89 IX 2.76 X 2.84 XI 2.54 XII 2.21 XIII 2.13 XIV 2.03 XV 1.95 XVI 1.89 XVII 1.79 XVIII 1.79

AR

CI

Type

Pair No.

2.41 2.03 1.45 1 1.38 1.5 1.64 1.57 1.64 1.65 1.24 1 1 1.21 1.14 2.33 1 1.6

29.32 33.01 40.78 50 41.94 40 37.93 38.96 37.84 37.68 44.62 50 50 45.16 46.67 30 50 38.46

sm sm m M m m m m m m m M M m m sm M m

1 1.92 1.92 2.37 2.05 1.53 1.48 1.62 1.39 2.38 1.41 1 1.25 1.26 1.47 1.25 1.13 1.27

50 34.25 34.25 29.69 32.8 39.59 40.35 38.18 41.9 29.63 41.45 50 44.44 44.16 40.54 44.44 47.06 44.12

M sm sm sm sm m m m m sm m M m m m m m m

I II III IV V VI VII VIII IX X XI XII XIII XIV XV XVI XVII XVIII

RL

AR Centaurea derderiifolia 2.48 1.44 2.4 1.4 2.3 1.55 2.1 1.66 2.06 1 2.07 1.29 1.98 1.56 1.9 1.31 1.85 1.45 3.42 1.95 1.67 1.62 1.54 1.2 1.52 2.13 1.47 1.02 1.35 1.33 1.35 1.33 1.29 1 1.13 0.84

CI

Type

41.04 41.67 39.28 37.61 50 43.72 39.02 43.22 40.87 33.85 38.22 45.54 31.91 49.45 42.86 42.86 50 54.29

m m m m M sm m m m sm m m sm m m m M m

Abbreviations: RL = relative length; AR = arm ratio (L/S); CI = centromeric index (100S/TL); m = median region; sm = submedian region; M = median point. Chromosome pairs are assigned roman numerals.

at ends and with smooth membrane. All of the morphological parameters investigated are shown in Tables 4 and 5 and Figure 7. The pollen grains of the three taxa have columellae on one layer and exine acaveate. Ectexine/endexine thickness values in thickness were calculated at 2/1 for these species. Centaurea drabifolia subsp. floccosa The pollen grains are of Wagenitz’s Jacea-type (1955), subprolate, with the ratio polar axis/equatorial diameter equal to 1.14. The exine sculpture is scabrate-perforate, with density of the sculpture elements around 6–7 per 5 μm² and the height of the microspines is 0.34 μm. Polar axis = 38.01 μm, equatorial axis = 33.08 μm; length of the colpus = 30.40 μm, breadth of the colpus = 2.75 μm (Tables 4, 5, Figure 7 a, b).

Centaurea kotschyi var. floccosa The pollen grains are of Wagenitz’s Jacea-type (1955), subprolate, with the ratio polar axis/equatorial diameter equal to 1.15. The exine sculpture is scabrate, with a density of the sculpture elements around 5–6 per 5 μm² and the height of the microspines is 0.41 μm. Polar axis = 46.99 μm, equatorial axis = 41.17 μm, length of the colpus = 37.43 μm. breadth of the colpus = 3.91 μm. (Tables 4, 5, Figure 7 c, d). Centaurea derderiifolia The pollen grains are of Wagenitz’s Jacea-type (1955), subprolate, with the ratio polar axis/equatorial diameter equal to 1.16. The exine sculpture is scabrate-perforate, with a density of the sculpture elements around 3–4 per 5 μm2 and the high of the microspines is 1.75 μm. Polar


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Figure 5. Somatic metaphases of three Centaurea taxa belonging to Sect. Cheirolepis. (1) C. derderiifolia (2n = 36), (2) C. drabifolia subsp. floccosa (2n = 36), (3) C. kotschyi var. floccosa (2n = 36).

axis = 46.99 μm, equatorial axis = 43.40 μm, length of the colpus = 40.53 μm, breadth of the colpus = 4.38 μm (Tables 4, 5, Figure 7 e, f).

Discussion The present study aimed to survey the morphological, karyological and palynological of three taxa belonging to three endemic Centaurea L. species growing naturally in Turkey, and to compare them with previous morphological, karyological and palynological investigations in the genus. There has been several studies employing light microscopy (LM) or scanning electron microscopy (SEM) to evaluate the pollen morphology of Centaurea species (Wagenitz 1955; Avetisjan 1964; Inceoglu and Karamustafaoglu 1977; Kaya 1985, 1986; Pehlivan 1994, 1995, 1996; Pinar and Inceoglu 1996; Wagenitz and Hellwig 1996; Kaya, Başaran, and Akkemik 2000; Vıllodre and Garcia-Jacas 2000; Celik, Uysal, and Menemen 2005; Celik et al. 2005; Uysal, Celik, and Menemen 2005a, 2005b; Jaffari and Ghanbarian 2007; Celik, Uysal, and Menemen 2008; Özler, Kaya, and

Pehlivan 2009; Gömürgen, Erkara, and Altınözlü 2010; Shabestari et al. 2013). In Turkey, the first detailed morphological analyses of pollen grains in Centaurea were made by Wagenitz (1955) and Avetisjan (1964) in an attempt to solve phylogenetic problems in this genus. Wagenitz (1955) classified Centaurea pollen grains by morphological features and recognized eight pollen types. He used the following features to group acetolyzed pollen: pollen shape, exine ornamentation, internal and external layers of columellae, length of colpus pore shape and costae. Serratula, Crupina and Centaurium pollen types were recognized as the most primitive, Dealbata, Cyanus and Montana as intermediate types, and Scabiosa and Jacea as the most advanced types. Some authors have asserted that the presence of single or double layers of columellae of the pollen wall can be used as principal criteria in taxonomic arguments (Wagenitz 1955; Skvarla et al. 1977). The results of this study on Centaurea sect. Cheirolepis species from Turkey indicate that the pollen of the three taxa here investigated are morphologically similar. Pollen grains of the three taxa analyzed in this study belongs


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Figure 6. Haploid idiograms of three Centaurea taxa belonging to Sect. Cheirolepis. (1) C. derderiifolia (2n=36), (2) C. drabifolia subsp. floccosa (2n=36), (3) C. kotschyi var. floccosa (2n=36). Table 4. Pollen morphological parameters of three Centaurea taxa belonging to Cheirolepis section (Mean μm ± standard deviation). E (μm) Mean ± standard deviation 33.08 ± 1.56 41.17 ± 2.82 43.40 ± 1.18 Plt (μm) Mean ± standard deviation 12.89 ± 1.09 13.53 ± 1.58 15.25 ± 0.89

Max

P/E Pollen shape

35.70 43.86 47.94 Max

Perprolate Perprolate Perprolate Min

15.36 17.28 18.24

1.44 1.68 1.92

Clg (μm) Min 28.56 29.58 33.66 Exine (μm) Mean ± standard deviation 2.26±0.23 2.08±0.26 2.51±0.40

Max

Mean ± standard deviation 30.40 ± 1.30 37.43 ± 2.87 40.53 ± 1.56 Max

Min

3.84 3.60 4.80

0,24 0.48 0.68

32.64 44.88 46.92

Clt (μm) Min 1.78 2.55 2.29 Intine (μm) Mean ± standard deviation 0.55 ± 0.11 0.61 ± 0.18 0.78 ± 0.27

Max

Mean ± standard deviation 2.75 ± 0.41 3.91 ± 0.50 4.38 ± 0.73 Max

Min

1.24 0.72 1.24

1.20 1.68 1.44

4.59 5.10 6.12

Costae (μm) Mean ± standard deviation 1.80 ± 0.20 1.77 ± 0.21 1.92 ± 0.11

Max

orn

2.40 2.64 3.36

S-Per S S-Per

Abbreviations: P = polar axis; E = equatorial axis; L = equatorial countour diameter; clg = length of the colpus; clt = width of the colpus plg = length of the porus; plt = width of the porus; costae = endexine thickening around the apertures; S = scabrate (spine lenght c 1pin); Om. (ornamentation) = sculpture of pollen wall; exine = at the thickest area, In = intine, Per = perforate.

Table 5. SEM characteristics of three Centaurea taxa belonging to Cheirolepis section. Taxon C. drabifolia subsp. floccosa C. kotschyi var. floccosa C. derderiifolia

Number of scabrae in 5 μm² 6–7

Skabrae width (dt) μm 1.16

Skabrae length (dh) μm 0.34

dh/dt 0.29

5–6

1.39

0.41

0.30

3–4

1.75

0.60

0.34

to Wagenitz’s Jacea pollen type based on the following characteristics: spheroidal-subprolate, costae, scabrate and microechinate ornamentation, long colpus, and the presence of cavea (Tables 4, 5; Figure 7).

The chromosome number of Centaurea drabifolia was reported previously from the west Asia population as 2n = 54 by (Garcia-Jacas et al. 1997). According to our data the diploid chromosome number is 2n = 36 in specimens collected from Turkish localities. In light of these informations, C. drabifolia seems to be tetraploid while it was expected to be hexaploid on the basis Garcia-Jacas and colleagues’ data (Garcia-Jacas et al. 1997). This result may be due to the fact that plant samples were collected from different locations. However, we agree on the basic chromose number for that taxa (x = 9). The basic Chromosome number of Centaurea derderiifolia was first reported by Uysal et al. (2009) with


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Figure 7. SEM microphotographs of pollen grains of three Centaurea taxa belonging to Sect. Cheirolepis (a–b) Centaurea drabifolia subsp. floccosa; general view (X1800), ornamentations (X7500). (c–d) C. kotschyi var. floccosa; general view (X2000), ornamentations (X10000). (e–f) C. derderiifolia; general view (X1800), ornamentations (X8000).

a basic chromosome number of x = 9. According to our study, the diploid chromosome number is established as 2n = 36. The chromosome number and morphology of Centaurea kotschyi var. floccosa are newly reported here. Karyological characteristics, such as chromosome number, the location of centromeres, and the number and position of satellites, are used in modern Centaurea taxonomy to clarify the relationships between species. With a basic chromosome number x = 9, Centaurea kotschyi var. floccosa matches others members of Cheirolepis section, and, broadly, members of the Cheirolepis, Pseudoseridia, Pteracantha, Plumosipappus complex of Garcia-Jacas et al. (2006). A close correlation among karyology, pollen morphology and systematic in the subtribe Centaureinae has been demonstrated, where basic chromosome numbers are considered a key character for sectional classification (Romaschenko 2004; Garcia-Jacas et al. 2006). For the three studied taxa, palynological data matches Wagenitz’s Jacea pollen type, while karyological data (particularly basic chromosome number) matches with a clade composed of members of Cheirolepis, Pseudoseridia, Pteracantha and Plumosipappus Sections in molecular phylogenies (Garcia-Jacas et al. 2006).

Conclusion The genus Centaurea presents taxonomic and nomenclatural problems. Palynological studies, together with recent molecular approaches and karyological studies, are very helpful in solving these problems. This paper gives detailed information on the morphological, karyological and pollen morphology of three Turkish endemic Centaurea; further morphological, karyological, palynological and molecular studies will help solve problems related to the taxonomy of these genus.

Disclosure statement No potential conflict of interest was reported by the authors.

Notes on contributors Sukru Hayta graduated from the Department of Biology at Firat University in 2002. He obtained his MS degree in 2006, and his PhD in 2012, from the same department and the same university in 2006. He is currently an associate professor in the Department of Environmental Engineering at Bitlis University. His research interests include plant taxonomy, plant morphology and anatomy, and essential oils.

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Neslihan Tasar is an assistant professor of Tunceli University. The main area of her research expertise is the plant caryology and anatomy. She collected plant material and established the karyotype analysis with Dr. Kiran. Yasar Kiran is an assistant professor of Firat University, and a leading specialist in the field of plant caryology. He has also contributed to the caryological studies in this paper. Birol Baser is an assistant professor of Bitlis Eren University, Department of Biology. He is a specialist in plant morphology and anatomy and pollen morphology and contributed to the palinological analysis in this study.


References Avetisjan, E. M. 1964. “K palynosystematike nekotorykh rodov triby Centaureinae semeistva Asteraceae. [Palynosystematics of the Tribe Centaureinae, Asteraceae].’’ Trudy Botanicheskogo Instituta Akademii Nauk Armyanskoi S.S.R. 14: 31–47. Aytaç, Z., and H. Duman. 2005. “A New Species of Centaurea L. (Compositae) from Turkey.” Pakistan Journal of Botany 37 (3): 563–566. Babac, M. T. 2004. “Possibility of an Information System on Plants of South-West Asia with Particular Reference to the Turkish Plants Data Service (TÜBİVES).” Turkish Journal of Botany 28: 119–127. Bancheva, S., Z. Kaya, and R. Binzet. 2014. “Centaurea aytugiana (Asteraceae), a New Species from North Anatolia, Turkey.” Novon 23 (2): 133–138. doi: 10.3417/2011026. Baytop, T. 1999. Türkiye’de Bitkiler ile Tedavi, Geçmiste ve Bugün [Therapy with Medicinal Plants in Turkey (Past and Present)]. 2nd ed. Istanbul: Nobel Tip Kitapevleri. Boldizsár, I., Zs. Füzfai, F. Tóth, É. Sedlák, L. Borsodi, and I. Molnár-Perl. 2010. “Mass Fragmentation Study of the Trimethylsilyl Derivatives of Arctiin, Matairesinoside, Arctigenin, Phylligenin, Matairesinol, Pinoresinol and Methylarctigenin: Their Gas and Liquid Chromatographic Analysis in Plant Extracts.” Journal of Chromatography A 1217 (10): 1674–1682. doi: 10.1016/j.chroma.2010.01.019. Bona, M. 2015. “Centaurea goksivriensis (Asteraceae), a New Species from Turkey.” Phytotaxa 203 (1): 63–68. doi: 10.11646/phytotaxa.203.1.6. Bona, M. 2016. “Centaurea amanosensis (Asteraceae), a New Species from Turkey.” Plant Biosystems 150 (5): 1083–1086. doi: 10.1080/11263504.2014.1001465. Carr, G. D., R. M. King, A. M. Powell, and H. Robinson. 1999. “Chromosome Number in Compositae XVIII.” American Journal of Botany 86 (7): 1003–1013. Celik, S., I. Uysal, and Y. Menemen. 2005. “Centaurea Species in Turkey (A): Centaurea odyssei Wagenitz (Asteraceae) in Kazdagi (Mt. Ida) National Park.” International Journal of Biodiversity Science and Management 1 (2): 113–120. doi: 10.1080/17451590509618085. Celik, S., I. Uysal, Y. Menemen, and E. Karabacak. 2005. “Morphology, Anatomy, Ecology, Pollen and Achene Structure of Centaurea consanguinea DC. (Sect. Acrolophus) in Turkey.” International Journal of Botany 1 (1): 85–89. doi: 10.3923/ijb.2005.85.89. Celik, S., I. Uysal, and Y. Menemen. 2008. “Morphology, Anatomy, Ecology and Palynology of two Centaurea Species from Turkey.” Bangladesh Journal of Botany 37 (1): 67–74. doi: 10.3329/bjb.v37i1.1566. Csupor, D., G. Blazsó, A. Balogh, and J. Hohmann. 2010. “The Traditional Hungarian Medicinal Plant Centaurea

249

sadleriana Janka Accelerates Wound Healing in Rats.” Journal of Ethnopharmacology 127 (1): 193–195. doi: 10.1016/j.jep.2009.09.049. Daşkın, R., and Ö. Yılmaz. 2009. “Centaurea kaynakiae (Asteraceae), a New Species from Turkey.” Annales Botanici Fennici 46 (5): 474–478. Davis, P. H. 1975. Flora of Turkey and the East Aegean Islands. vol. 5. Edinburgh: Edinburgh University Press. Davis, P. H., R. R. Mill, and K. Tan. 1988. “Centaurea L.” In Flora of Turkey and the East Aegean Islands, edited by P. H. Davis, R. R. Mill, and K. Tan, vol. 10, 166–169. Edinburgh: Edinburgh University Press. Duran, A., and H. Duman. 2002. “Two New Species of Centaurea (Asteracae) from Turkey.” Annales Botanici Fennici 39: 43–48. Erdtman, G. 1969. Handbook of Palynology. Copenhagen: Munksgaard, 486pp. Faegri, K., and J. Iversen. 1992. Textbook of Pollen Analysis. New York: Wiley. Garcia-Jacas, N., A. Susanna, R. Ilarslan, and H. Ilarslan. 1997. “New Chromosome Counts in the Subtribe Centaureinae (Asteraceae, Cardueae) from West Asia.” Botanical Journal of the Linnean Society 125 (3): 343–349. doi: 10.1111/j.1095-8339.1997.tb02263.x. Garcia-Jacas, N., A. Susanna, R. Vılatersana, and M. Guara. 1998. “New Chromosome Counts in the Subtribe Centaureinae (Asteraceae, Cardueae) from West Asia II.” Botanical Journal of the Linnean Society 128 (4): 403–412. doi: 10.1111/j.1095-8339.1998.tb02129.x. Garcia-Jacas, N., A. Susanna, R. Mozaffarian, and R. Ilarslan. 2000. “The Natural Delimitation of Centaurea (Asteraceae: Cardueae): ITS Sequence Analysis of the Centaurea jacea Group.” Plant Systematics and Evolution 223: 185–199. doi: 10.1007/BF00985278. Garcia-Jacas, N., A. Susanna, T. Garnatje, and R. Vilatersana. 2001. “Generic Delimitation and Phylogeny of the Subtribe Centaureinae (Asteraceae): A Combined Nuclear and Chloroplast DNA Analysis.” Annals of Botany 87: 503–515. doi: 10.1006/anbo.2000.1364. Garcia-Jacas, N., T. Uysal, K. Romashchenko, V. N. SuárezSantiago, K. Ertuğrul, and A. Susanna. 2006. “Centaurea Revisited: A Molecular Survey of the Jacea Group.” Annals of Botany 98 (4): 741–753. doi: 10.1093/aob/mcl157. Gömürgen, A. N., I. P. Erkara, and H. Altınözlü. 2010. “Chromosome and Pollen Morphology of the Rare Endemic Centaurea lycopifolia Boiss. & Kotcshy.” Bangladesh Journal of Botany 39 (2): 223–228. doi: 10.3329/bjb.v39i2.7484. Greuter, W. 2003a. “The Euro+Med Treatment of Cardueae (Compositae) – Generic Concepts and Required New Names.” Willdenowia 33 (1): 49–61. doi: 10.3372/ wi.33.33104. Greuter, W. 2003b. “The Euro+Med Treatment Senecioneae and the Minor Compositae Tribes – Generic Concepts and Required New Names, with an Addentum to Cardueae.” Willdenowia 33 (2): 245–250. doi: 10.3372/wi.33.33203. Güner, A., N. Ozhatay, T. Ekim, and K. H. C. Baser. 2000. Flora of Turkey and the East Aegean Islands (Supplement 2). vol. 11. Edinburgh: Edinburgh University Press. Gunjan, K., and B. K. Roy. 2010. “Karyotype Studies İn Dominant Species of Aloe from Eastern India.” Caryologia 63 (1): 41–49. doi: 10.1080/00087114.2010.10589707. Gürbüz, I., and E. Yesilada. 2007. “Evaluation of the Antiulcerogenic Effect of the Sesquiterpene Lactones from Centaurea solstitialis L. ssp. solstitialis by using Various in vivo and Biochemical Techniques.” Journal of Ethnopharmacology 112 (2): 284–295. doi: 10.1016/j. jep.2007.03.009.


250

S. HAYTA ET AL.

Hamzaoğlu, E., and U. Budak. 2009. “Centaurea aksoyi sp. nov. (Asteraceae: Cardueae) from Turkey and a Contribution to the Sectional Taxonomy.” Nordic Journal of Botany 27 (1): 16–20. doi: 10.1111/j.1756-1051.2009.00192.x. Hellwig, F. H. 2004. “Centaureinae (Asteraceae) in the Mediterranean-history of Ecogeographical Radiation.” Plant Systematics and Evolution 246 (3): 137–162. doi: 10.1007/s00606-004-0150-2. Hesse, M., H. Halbritter, M. Weber, R. Buchner, A. FroschRadivo, S. Ulrich, and R. Zetter. 2009. Pollen Terminology. An Illustrated Handbook. Vienna: Springer. Inceoglu, O., and F. Karamustafaoglu. 1977. “The Pollen Morphology of Plants in Ankara Region: 1. Compositae.” Communications de la Faculté de Sciences de l’Université d’Ankara, série C2, Botanique 21(5): 77–100. Jaffari, E., and G. Ghanbarian. 2007. “Pollen Morphological Studies on Selected Taxa of Asteraceae.” Journal of Plant Science 2 (2): 195–201. doi: 10.3923/jps.2007.195.201. Kaya, Z. 1985. Endemik iki Centaurea türü üzerinde taksonomik, ekolojik ve palinolojik araştırmalar [Palynological, Ecological and Taxonomical Research on Two Endemic Species of Centaurea]. PhD diss., Marmara University, Istanbul. Kaya, Z. 1986. “Endemik iki Centaurea türü üzerinde palinolojik araştırmalar [Palynological Studies on Two Endemic Centaurea L. Species].” İstanbul Üniversitesi Orman Fakültesi Dergisi 36 (2): 155–163. Kaya, Z., S. E. Başaran, and U. Akkemik. 2000. “Palynological Research on some Endemic Species of Centaurea L. in Turkey.” BIOS, Scientific Annals of the Scholl of Biology, Aristotle University 5: 27–34. Kaya, Z., and M. Vural. 2007. “A New Species of Centaurea Sect. Acrocentron (Asteraceae) from Turkey.” Novon: A Journal for Botanical Nomenclature 17 (2): 198–201. doi: 10.3417/1055-3177(2007)17[198:ANSOCS]2.0.CO;2. Köse, Y. B., and S. Alan. 2013. “Centaurea baseri (Compositae), a New Species from Turkey.” Phytotaxa 117 (1): 23–29. doi: 10.11646/phytotaxa.117.1.3. Levan, A., K. Fredga, and A. A. Sandberg. 1964. “Nomenclature for Centromeric Position on Chromosomes.” Hereditas 52 (2): 201–220. doi: 10.1111/j.1601-5223.1964.tb01953.x. Martin, E., M. Dinç, and A. Duran. 2009. “Karyomorphological Study of Eight Centaurea L. taxa (Asteraceae) from Turkey.” Turkish Journal of Botany 33 (2): 97–104. doi: 10.3906/bot-0703-16. Negaresh, K., Z. Kaya, and M. R. Rahiminejad. 2015. “Centaurea sennikoviana (Asteraceae, Cardueae), a New Species from Central Anatolia, Turkey.” Annales Botanici Fennici 52(5-6): 321–327. doi: 10.5735/085.052.0507. Özler, H., Z. Kaya, and S. Pehlivan. 2009. “Pollen Morphology of some Centaurea L., Psephellus Cass. and Cyanus Miller taxa.” Acta Biologica Cracoviensia Series Botanica 51 (2): 53–66. Pehlivan, S. 1994. “Scanning Electron Microscopic Studies of the Polen Grains of some Turkish Endemic Centaurea.” Journal of Faculty of Pharmacy of Gazi University 11 (2): 205–211. Pehlivan, S. 1995. “Pollen Morphology of some Turkish Endemic Centaurea L.” Grana 34 (1): 29–38. doi: 10.1080/00173139509429030. Pehlivan, S. 1996. “Light Microscopic Studies in the Pollen Morphology of some Endemic Turkish Centaurea Species.” Turkish Journal of Botany 20 (4): 311–321. Pinar, M. N., and O. Inceoglu. 1996. “A Comparative Study on the Pollen Morphology of Centaurea triumfettii All. groups. A, B and C with Light and Electron Microscopy.” Turkish Journal of Botany 20 (5): 395–399.

Punt, W., and P. P. Hoen. 2009. “The Northwest European Pollen Flora, 70: Asteraceae–Asteroideae.” Review of Palaeobotany and Palynology 157 (1–2): 22–183. doi: 10.1016/j.revpalbo.2008.12.003. Punt, W., P. P., Hoen, S. Blackmore, S. Nilsson, and Le Thomas A. 2007. “Glossary of Pollen and Spore Terminology.” Review of Palaeobotany and Palynology 143 (1–2): 1–81. doi: 10.1016/j.revpalbo.2006.06.008. Romaschenko, K. 2004. “New Chromosome Counts in the Centaurea jacea group (Asteraceae, Cardueae) and some related taxa.” Botanical Journal of Linnean Society 145 (3): 345–352. doi: 10.1111/j.1095-8339.2004.00292.x. Shabestari, E. S. B., F. Attar, H. Riahi, and M. Sheidai. 2013. “Pollen Morphology of Centaurea L. (Asteraceae) in Iran.” Acta Botanica Brasilica 27 (4): 669–679. doi: 10.1590/ S0102-33062013000400004. Skvarla, J. J., B. L. Turner, V. C. Patel, and A. S. Tomb. 1977. “Pollen Morphology in the Compositae and in Morphologically Related Families.” In The Biology and Chemistry of the Compositae, edited by V. H. Heywood, J. B. Harborne, and B. L. Turner, 141–248. London: Academic Press. Susanna, A., N. Garcia-Jacas, D. E. Soltis, and P. S. Soltis. 1995. “Phylogenetic Relationships in Tribe Cardueae (Asteraceae) Based on ITS Sequences.” American Journal of Botany 82 (8): 1056–1068. Szokol-Borsodi, L., A. Sólyomváry, I. MolnárPerl, and I. Boldizsár. 2012. “Optimum Yields of Dibenzylbutyrolactone-type Lignans from Cynareae Fruits, During their Ripening, Germination and Enzymatic Hydrolysis Processes, Determined by On-line Chromatographic Methods.” Phytochemical Analysis 23 (6): 598–603. doi: 10.1002/pca.2360. Tabur, S., S. Civelek, S. Öney, S. B. Y. Ergun, M. Kursat, and I. Turkoglu. 2012. “Chromosome Counts and Karyomorphology of some Species of Artemisia (Asteraceae) from Turkey.” Turkish Journal of Botany 36 (3): 235–246. doi: 10.3906/bot-1010-98. Türkoğlu, I., H. Akan, and S. Civelek. 2003. “A New Species of Centaurea (Asteraceae: sect. Psephelloideae) from Turkey.” Botanical Journal of Linnean Society 143 (2): 207–212. doi: 10.1046/j.1095-8339.2003.00205.x. Uysal, T. 2008. “Centaurea ertugruliana (Asteraceae), a New Species from Turkey.” Annales Botanici Fennici 45 (2): 137–142. doi: 10.5735/085.045.0208. Uysal, I., S. Celik, and Y. Menemen. 2005a. “Centaurea Species in Turkey (B): Comparative Studies of two Closely Related Species, C. kurdica Reichardt and C. sclerolepis Boiss.” International Journal of Biodiversity Science and Management 1 (2): 121–127. doi: 10.1080/17451590509618086. Uysal, I., S. Celik, and Y. Menemen. 2005b. “Morphology, Anatomy, Ecology, Pollen and Achene features of Centaurea polyclada DC. (Sect. Acrolophus) in Turkey.” Journal of Biological Science 5 (2): 176–180. doi: 10.3923/ jbs.2005.176.180. Uysal, T., H. Demirelma, K. Ertugrul, N. Garcia-Jacas, and A. Susana. 2007. “Centaurea glabro-auriculata (Asteraceae), a New Species from Turkey.” Annales Botanici Fennici 44 (3): 219–222. Uysal, T., H. Dural, and O. Tugay. 2017. “Centaurea sakariyaensis (Asteraceae), a New Species from Turkey.” Plant Biosystems 151 (1): 126–130. doi: 10.1080/11263504.2015.1108940. Uysal, T., K. Ertugrul, A. Susana, and N. Garcia-Jacas. 2009. “New Chromosome Counts in the Genus Centaurea (Asteraceae) from Turkey.” Botanical Journal of the


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Linnean Society 159 (2): 280–286. doi:10.1111/j.10958339.2008.00939.x. Uysal, T., and E. Hamzaoğlu. 2016. “A New Centaurea L. (Asteraceae) Species from Turkey.” Plant Biosystems: 1–9: doi: 10.1080/11263504.2016.1211196. Uysal, T., E. Hamzaoğlu, K. Ertuğrul, and M. Bozkurt. 2016. ‘‘A New Species of Centaurea (Asteraceae) from Turkey.’’ Phytotaxa 275(2): 149–158. doi: 10.11646/ phytotaxa.275.2.6. Uysal, T., and Y. B. Köse. 2009. “A New Centaurea L. (Asteraceae) Species from Turkey.” Turkish Journal of Botany 33 (1): 41–46. doi: 10.3906/bot-0803-1. Uzunhisarcıklı, M. E., E. Doğan, and H. Duman. 2007. “A New Species of Centaurea L. (Cardueae: Asteraceae) from Turkey.” Botanical Journal of the Linnean Society 153 (1): 61–66. doi: 10.1111/j.1095-8339.2007.00578.x. Uzunhisarcıklı, M. E., M. Teksen, and E. Dogan. 2005. “Centaurea marashica (Asteraceae), a New Species from Turkey.” Annales Botanici Fennici 42 (4): 309–312. Villodre, J. M., and N. Garcia-Jacas. 2000. “Pollen Studies in Subtribe Centaureinae (Asteraceae): The Jacea Group Analysed with Electron Microscopy.” Botanical Journal of the Linnean Society 133 (4): 473–484. doi: 10.1006/ bojl.1999.0330. Vural, M., H. Duman, Z. Aytac, and N. Adıgüzel. 2006. “Saponaria karapinarensis, Senecio salsuginea and Centaurea tuzgoluensis, Three New Species from Central Anatolia, Turkey.” Belgian Journal of Botany 139 (2): 252– 260. Wagenitz, G. 1955. “Pollen Morphologie und Systematik in der Gattung Centaurea L. s.l [Pollen Morphology and Systematic in Centaurea L. s.l. group].” Flora 142: 213–279. Wagenitz, G. 1975. “Genus Centaurea L.” In Flora of Turkey and the East Aegean Islands. vol. 5, edited by P. H. Davis, 465–585. Edinburgh: Edinburgh University Press. Wagenitz, G., K. Ertugrul, and H. Dural. 1998. “A New Species of Centaurea sect. Psephelloidea (Compositae)

251

from SW Turkey.” Willdenowia 28 (1-2): 157–161. doi: 10.3372/wi.28.2815. Wagenitz, G., and F. H. Hellwig. 1996. ‘‘Evolution of Characters and Phylogeny of Centaureinae.’’ In Compositae: Systematics. Proceedings of the International Compositae Conference, Kew, vol. 1, edited by D. J. N. Hind, and H. J. Beentje, 491–510. Kew: Royal Botanic Gardens. Wagenitz, G. and F. H. Hellwig. 1997. “Eine neue und eine verschollene Centaurea-Art aus der Türkei und eine neue Volutaria-Art (Compositae, Cardueae) [A New and a Lost Centaurea Species from Turkey and a New Volutaria Species].” Annalen des Naturhistorisches Museum Wien Serie B für Botanik und Zoologie 98 (Suppl.): 175–186. Wagenitz, G., and F. H. Hellwig. 2000. “The Genus Psephellus Cass. (Compositae, Cardueae) Revisited with a Broadened Concept.” Willdenowia 30 (1): 29–44. doi: 10.3372/ wi.30.30102. Wagenitz, G., F. H. Hellwig, P. Gerald, and M. Ludwıg. 2006. “Two New Species of Centaurea (Compositae, Cardueae) from Turkey.” Willdenowia 36 (1): 423–435. doi: 10.3372/ wi.36.36139. Wodehouse, R. P. 1935. Pollen Grains. Their Structure, İdentification and Significance in Science And Medicine. New York: McGraw-Hill. Yesilada, E., I. Gürbüz, E. Bedir, I. Tatli, and I. A. Khan. 2004. “Isolation of Antiulcerogenic Sesquiterpene Lactones from Centaurea solstitialis L. ssp. solstitialis through BioassayGuided Fractionation Procedures in Rats.” Journal of Ethnopharmacology 95 (2–3): 213–219. doi: 10.1016/j. jep.2004.07.021. Yıldız, K., E. Minareci, and A. Cırpıcı. 2009. “A Karyotypic Study on Silene sect. Sclerocalycinae in Turkey.” Nordic Journal of Botany 27 (6): 503–509. doi: 10.1111/j.17561051.2009.00506.x. Yüzbaşıoğlu, I. S., M. Bona, and I. Genç. 2015. “A New Species of Centaurea sect. Pseudoseridia (Asteraceae) from North-eastern Turkey.” Phytokeys 53: 27–38. doi: 10.3897/ phytokeys.53.5250.