Morphological and Leaf Anatomical Investigations on 2 Yellow Flowered Endemic Taxa of Crocus L. (Crocus ancyrensis, Crocus siehenaus) from Turkey

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1 International Journal of Agriculture, Forestry and Fisheries 2015; 3(3): Published online May 10, 2015 ( Morphological and Leaf Anatomical Investigations on 2 Yellow Flowered Endemic Taxa of Crocus L. (Crocus ancyrensis, Crocus siehenaus) from Turkey Feyza Candan Department of Biology, Faculty of Arts and Science, Celal Bayar University, Manisa, Turkey address feyzacandan2002@yahoo.com To cite this article Feyza Candan. Morphological and Leaf Anatomical Investigations on 2 Yellow Flowered Endemic Taxa of Crocus L. (Crocus ancyrensis, Crocus siehenaus) from Turkey. International Journal of Agriculture, Forestry and Fisheries. Vol. 3, No. 3, 2015, pp Abstract Crocus ancyrensis (Herbert) Maw and Crocus sieheanus Barr ex Burtt are close taxa which can be seen in Flora of Turkey consecutively. Their morphologies are smilar but have some differences when examined carefully. However, this genus shows useful leaf anatomical characteristics that are significantly different from each other. Leaves show useful taxonomic characters according to carina, vascular bundles and arms of the leaf. In this study, it is aimed to compare morphology and cross sections as regards mature leaf anatomy of Cocus ancyrensis and Crocus sieheanus in a detailed way. As a result, it has observed that morphological and anatomical features show taxonomical importance for their systematics. Keywords Morphology, Anatomy, Crocus ancyrensis, Crocus sieheanus 1. Introduction The Crocus which is known as Saffron genus, is one of the members of the family Iridaceae. As is known, the styles and stigmas are collected and dried to be used mainly as a seasoning and colouring agent in food. Saffron, long among the world's most costly spices by weight. On the other hand, Crocus species are being growth for delicating parks and gardens. İn the point of its agricultural importance, this genus has a commercially importance in the world (Mathew, 1982; Mathew, 2000). The plants in this family are herbs with rhizomes, corms or bulbs. Iridaceae is a large and diverse family of about 92 genera and 1800 species and mainly distributed in the Southern hemisphere continents (Mathew, 1982; Mathew, 2000). Crocus genus is represented by 40 species and 70 taxa of which 40 are endemics and 1 hybrid in Turkey (Mathew 1982, 1984,1998, 2002). After the book The Crocus written (Mathew 1982), different Crocus species have received attention on some ecological, anatomical, cytological, palynological and molecular studies (Kerndorff and Pasche 2004, 2006, 2011; Yüzbaşıoğlu and Varol 2004; Erol 2010, 2011; Işık and Oybak Dönmez 2006; Candan 2007; Candan and Kesercioğlu, 2007; Candan et al. 2009a, 2009b; Kandemir 2009; Şık et al, 2008; Şık and Candan 2009; Coşkun et al. 2010; Candan and Özhatay 2013). There are also some investigations made about some Crocus taxa distributed all over the World (Collins 1937, Karasawa 1942, Shorina 1975, Rudall 1990, 1992, Rudall and Mathew 1990, Satıl ve Selvi 2007). In these studies, especially according to Rudall and Mathew (1990), characteristic features and general importance of leaf blade cross sections were determined. The leaves of Crocus species hold keel at the center of the leaf and two arms next to the keel which is a unique shape regarding cross sections for the genus (Rudall and Mathew, 1990). As is seen, there aren t any detailed study on C. ancyrensis (Herbert) Maw and C. sieheanus Barr ex Burtt. Therefore, the initial objective of this study was to investigate the comparison of morphology and leaf anatomy as regards populations of these two endemic taxa naturally distributed in Anatolia. 2. Material and Method C. ancyrensis and C. sieheanus materials were collected from natural populations in flowering time. The populations of the plants examined are given below with the locality they deploy. The expressions A3 and C5 that are located before

2 International Journal of Agriculture, Forestry and Fisheries 2015; 3(3): the locality are given in accordance with the taxa order and grid square system used in Flora of Turkey (Mathew, 1984). C. ancyrensis (Herbert) Maw *A3 Bolu, Abant Lake between Mudurnu, near Çepni village, 1048 m. *A3 Bolu, Bolu-Gerede road, 13 km to Gerede, about Kayıkiraz village 1190 m. *A3 Bolu, Bolu-Kıbrısçık road, 10 km to Bolu, 1485 m. *A3 Bolu, Bolu-Seben road, 30 km to Seben, Karacasu plateau, 1485 m. *A3 Bolu, Bolu-Ankara road, 25 km to Kızılcahamam, Yünlü plateau, 1330 m. C. sieheanus Barr ex Burtt *C5 Adana, Adana-Pozantı road, Aladağlar, Fındıklı village, 1510 m. Plant material were collected from natural populations in flowering time. The localities of the samples are given below. Specimens were preserved in the herbarium of Celal Bayar University, Faculty of Arts and Science, Department of Biology. Taxonomical description of the plant taxon followed Flora of Turkey (Mathew, 1984; Mathew 2000). For anatomical studies plant specimens were fixed in 70 % alcohol. However, hand-cut sections were made for anatomical observation and stained with sartur reagent (Candan 2007). 3. Results 3.1. C. ancyrensis (Herbert) Maw a) Morphological Peculiarities Corm subglobose, 0,8-1,6x0,7-2,0 cm, tunic reticulatefibrous. Few rings with distinct tooth like projections. Cataphyll creamy-yellow, 3-4. Prophyll absent. Bract and bracteol nearly equal, membraneus. Leaves 3-7, synanthous, shorter or longer than flowers in flowering time, greyish green, 0,7-0,9 mm wide, glabrous, rarely papillose. Flowers 1-3, yellow or yellowish orange, diameter 1,0-3,4 cm. Perianth tube yellow rarely purple, 2-8 cm length, under ground part of the tube at flowering time (0,4)1,0-4,5(6,0) cm. Throat glabrous or papillose, pubescent. Tepals obovate or obtuse to subacute. Outer tepals 0,5-1,0x2,0-3,3 cm, inner tepals 0,4-1,2x1,1-3,0 cm. Filament yellow, yellowish orange, (2,8)3-7 mm, puberulent. Anhters yellow, (5)7,5-12 mm, basifixed, extrorse. Style longer than stamens or not and divided into 3 fimbriate or expanded branches, yellowish orange, orange, rarely red, 1,5-18,5 mm. Capsule ellipsoid, 0,7x1-1,5 cm diameters. Seeds reddish brown, (1,4)1,5-1,9(2)x2,5-3,7 mm diameters, usually with a distinct raphe and caruncle, ellipsoid (Fig.1a, b, c, d, e; Table 1). b) Leaf Anatomical Peculiarities Revolute view is seen on croos sections. Cross section is nearly symmetric. Cuticle is distinct. Epidermal cells are rectangular, rarely square shaped, more bigger on arms twisting parts. The leaves have a central rectangular keel at the base and two long lateral arms, with their margins recurved towards the keel. Palisade parenchyma cells are 2-3 layered and rectangular shaped. Spongy cells are 3-4 layered, isodiametrik, small at arms and big at tha carina. Vascular bundles are located in one row and below the abaxial epidermis. Major bundles occur at angles of keel and towards arm margins. Stomas are kseromorphic, rarely mesomorphic. Leaves rarely with papillas, without any hair. Vascular bundles are symmetric (Fig.2f, g; Table 2) C. sieheanus Barr ex Burtt a) Morphological Peculiarities Corm ovoid or subglobose, 0,8-1,5x0,8-1,7 cm, tunic membranous-coriaseus, fibrious at the base. Few rings with distinct long tooth like projections. Cataphyll 3 at young plants without flowers, 4 at plants with flowers, creamy yellow, membranous. Prophyll absent. Bract and bracteol nearly equal. Leaves 3-5, synanthous, shorten than flowers in flowering time, grey-green, (0,5)0,6-1,1(1,2) mm width, glabrous, rarely papillose. Flowers 1-3, pale yellow, 0,9-2,50 cm diameters, perianth tube yellow, 3-6,5 cm length, underground part of the tube at flowering time 2-3 cm, glabrous. Tepals oblanseolate or eliptic-obtuse. Outer tepals 0,4-0,5x2,1-2,5 cm, inner tepals 0,30-0,50x1,80-2,40 cm. Filaments pale yellow creamy yellow, 4-5 mm, glabrous, rarely papillose. Anther yellow, (6,2)7,2-11,2 mm, basifixed, extrors. Style longer than stamens and divided into 3 fimbriate or expanded branches, yellowish orange, orange, 12,8-16,5(20,0) mm. Capsula 13,5-16,0 mm. Seed pale reddish brown, (1,3)1,5-1,9(2,1)x2,7-3,5 mm, raphe and caruncla distinct, elipsoid (Fig.2a, b, c, d, e; Table 1). b) Leaf Anatomical peculiarities Revolute view is seen on croos sections. Cross section is nearly symmetric. Cuticle is distinct. Epidermal cells are rectangular, rarely square shaped, more bigger on arms twisting parts. The parenchymatous cells are present in the keel which lack chloroplast and break down to from an air space. Mesophyll located in the lateral arms have chloroplasts and differentiated into palisade and spongy parenchyma. Palisade parenchyma cells are 1-2 layered and rectangular shaped. Spongy cells are isodiametrik, small at arms and big at tha carina. Stomas are kseromorphic. Leaves without any papillas and any hair. Vascular bundles are nearly symmetric (Fig. 2f, g; Table 2). 4. Discussion It is aimed to compare the morphological and anatomical characters as ragards endemic two taxa Crocus ancyrensis and Crocus sieheanus with this investigation. Most of the Crocus species has characteristic leaves with central square or rectangular keel and two lateral arms with their margins generally received towards the keel (Karasawa 1942, Shorina 1975, Mathew 1982, Rudall and Mathew 1990). Crocus species conserve the main characteristics of leaf structure, however the exception leaf structures are reported by Rudall and Mathew (1990) in some species (C. carpetanus Boiss. and Reut., C. scharojanii Rupr., C. scardicus Kosanin). According to this study, two taxa has the main type of

3 95 Feyza Candan: Morphological and Leaf Anatomical Investigations on 2 Yellow Flowered Endemic Taxa of Crocus L. (Crocus ancyrensis, Crocus siehenaus) from Turkey characteristic leaves of Crocus genus. Figure 1. A.General view of C. ancyrensis, b.close view of flower, c.close view of corm, d.close view of fruit, e.close view of seed, f.cross section of leaf (scale bar 100 µm), g.close section of leaf (scale bar 10 µm), (E:Epidermis, P:Palisade, S:Spongy cells,i:investigated vascular bundles, St:Stoma)

4 International Journal of Agriculture, Forestry and Fisheries 2015; 3(3): Figure 2. A.General view of C. sieheanus, b.close view of flower, c.close view of corm, d.close view of fruit, e.close view of seed, f.cross section of leaf (scale bar 100 µm), g.close section of leaf (scale bar 10 µm), (E:Epidermis, P:Palisade, S:Spongy cells, i:investigated vascular bundles, St:Stoma)

5 97 Feyza Candan: Morphological and Leaf Anatomical Investigations on 2 Yellow Flowered Endemic Taxa of Crocus L. (Crocus ancyrensis, Crocus siehenaus) from Turkey Table 1. Comparison of the data obtained in this study and data in Flora of Turkey (1984) for C. ancyrensis and C. sieheanus Morphological peculiarities C. ancyrensis C. sieheanus Morphological Morphologicalmeas Morphological measurements Measurements of plant parts measurements in urements in Flora in this study Flora Of Turkey Of Turkey Morphological measurements in this study Corm measurements (cm) 1-1,5 0,8-1,6x0,7-2, x8-17 Cataphyll number Leaf number (2-) Leaf width (mm) 0,5-1(-1,5) 0,7-0,9 0,5-1,5 (0,5)0,6-1,1(1,2) Flower number 1-2(-3) Flower measurement (cm) - 1-3,4-0,9-2,5 Perianth tube(cm) ,5 Perianth tube s under ground part - (cm) (0,4)1,0-4,5(6,0) Tepal (outer tepal)0,5-1,0(1,2) (outer tepal) 0,4-0,5 (0,7-)0,9-1,3 0,7-1,1 width(cm) (inner tepal) 0,4-1,2 (inner tepal) 0,3-0,5 length (cm) (outer tepal) 2,0-3,3 (outer tepal) 2,1-2,5 (1,3-)1, (inner tepal) 1,1-3,0 (inner tepal) 1,8-2,4 Filament (mm) 2-4 (2,8) Anther (mm) 8-13 (5)7, (6,2)7,2-11,2 Stylus (mm) - 1,5-18,5-12,8-16,5(20,0) Fruit width (cm) length (cm) Seed width (mm) length (mm) 0,7 0,7 1-1,5 1,35-1,6 - (1,3)1,5-1,9(2,1) 1-1,5 1-1, ,7-3,5-3-4 (1,4)1,5-1,9(2) 2,5-3,7 Table 2. Leaf anatomical data in this study as regards C. ancyrensis and C. sieheanus (1,3)1,5-1,9(2,1) 2,7-3,5 Taxon Cuticle thickness Upper epidermis Lower epidermis Palisade Spongy widh length C. ancyrensis C. sieheanus width length width length The most detailed measurements were given in the point of these taxa with Flora of Turkey (1984). Nevertheless, these knowledge are not clariifying the taxa about their reproduction parts in a detailed way. Periods of flowering times and fruiting times, outer-inner tepals, anthers length in the comparison of stylus, fruits and seeds are given with more measurements and photographs with this investigation firstly. However, this is the first study about giving all the morphological measurements by comparing the measurments of Flora of Turkey (1984) and with detailed coloured photographs. On the other hand, all these measurements were given with a Table 1 in the point of comparison of the two endemic and similar taxa. According to the Table 1, it can be seen all compared characters measurements of Flora of Turkey accept number of leaves, perianth tube length, underground perianth tube length, detailed measurments of tepals-anthers-stylus-capsulas-seeds. Some of the morphological measurements are in accordance with the Flora of Turkey (1984), but more of the avarage measurements are different (Table 1). According to all measurements, both two taxa are very similar plants. However, C. sieheanus can be distinguished from C. ancyrensis by its slender paralel fibrous corms and with its very pale yellow flowers. It is observed that, leaf widthof plants is efected by weather conditions with this study. Because of that reason, it is thought that it is not a distictable taxonomic character. This information shows accordance with the area studies. According to the morphological part of this study, it can be said that, more of the differences are stem from seasonal changes because of global warming and the soil as ragards this study and the Flora of Turkey (1984), According to anatomical studies, it can be seen that leaves of two taxa have unique cells and distinctive revolute shapes (Table 2). On the other hand, it can be clearly seen that the symmetry of cross section and symmetry of vascular bundles at two taxa. But, the anatomic cross sections difference can be used as a distinctable taxonomic character for both taxa (Fig 1f, Fig 2f). As a result, it can be clearly said, anatomical structures of the leaves are very important for Crocus taxonomy. In all the studied taxa according to this investigation, some differences with accaptable taxonomical significance were found in the leaf and keel shape, arrangement, size and number of vascular bundles.

6 International Journal of Agriculture, Forestry and Fisheries 2015; 3(3): References [1] Candan, F Morphological, anatomical, cytological and palynological investigations on C. ancyrensis, C. sieheanus, C. chrysanthus and C. flavus taxa of the genus Crocus L. PhD thesis. Natural and Applied Sciences Institute, Celal Bayar University, Manisa, Turkey. [2] Candan, F.; Kesercioğlu, T Cytaxonomic Investigations on the Species Crocus chrysanthus (Herbert) Herbert. - International Conference on Environment:Survival and Sustainability, Proceedings, Vol. 2, February, Near East University, Nicosia-Northern, Cyprus. [3] Candan, F.; Şık, L.; Kesercioğlu, T. 2009a. Cytotaxonomical Studies on some Crocus L. Taxa in Turkey. African Journal of Biotechnology, Vol. 8(18), [4] Candan, F.; Kesercioğlu, T.; Şık, L. 2009b. Micromorphological Investigations on Pollen Samples of Four Yellow Flowered Taxa of Crocus L. (Iridaceae) from Turkey. Journal of Applied Biological Sciences, 3(2): [5] Candan, F.; Özhatay N, Crocus chrysanthus s. Lato (Iridaceae) in Turkey, Annales Botanici Fennici, 50 (6): [6] Collins E J Leaf Form and Structure, In:The Genus Crocus Proc. Linn. Soc., London (Abstract), 149: [7] Coşkun, F.; Selvi, S.; Satıl, F Phylogenetic relationships of some Turkish Crocus (Iridaceae) taxa based on morphological and anatomical characters. Turkish Journal of Botany, Vol. 34(3), [8] Erol, O.; Koçyiğit M.; Şık L.; Özhatay N; Küçüker O Crocus antalyensis subsp. nov. (Iridaceae) from southwest Anatolia. Nordic Journal of Botany,28: [9] Erol, O.; Şık, L.; Kaya, H. B.; Tanyolaç, B.; Küçüker, O Genetic diversity of Crocus antalyensis B. Mathew (Iridaceae) and a new subspecies from southern Anatolia. Plant Systematics and Evolution 294: [10] Işık, S., Oybak Dönmez, E Pollen Morphology of Some Turkish Crocus L. (Iridaceae) Species. Acta Biologica Cracoviensia, Series Botanica 48/1: [11] Karasawa K On the leaf structures of Crocus observed in transverse Sections and their chromosome numbers, Bot. Mag. Tokyo, 56: [12] Kandemir, N Morphology, Anatomy and Ecology of Critically Endangered Endemic Crocus pestalozzae Boiss. (Iridaceae) in North-West Turkey. Bangladesh Journal of Botany, Vol. 38, Issue:2, [13] Kerndorff, H.; Pasche, E Crocus biflorus in Anatolia, part two. Plantsman (new ser.) 3: [14] Kerndorff, H.; Pasche, E Crocus biflorus in Anatolia, part tree. Linzer Biol. Beitr. 38: [15] Kerndorff, H.; Pasche, E Two new taxa of Crocus (Liliiflorae, Iridaceae) from Turkey. Stapfia 95:2-5. [16] Mathew, B The Crocus: A revision of the genus Crocus (Iridaceae). -B.T. Batsford LTD. [17] Mathew, B., Crocus: -In Davis, P.H. (ed.), The Flora of Turkey and The East Aegean Islands, 8: , Edinburgh University. [18] Mathew, B Crocus kerndorffiorum. Curtis s Bot. Mag. 15: [19] Mathew, B Crocus. In Güner, A. et al. (eds), Flora of Turkey and the East Aegean Islands. Edinburgh Univ. Pres, 11: [20] Mathew, B Crocus Up-date. The New Plantsman Vol.1(1): [21] Rudall P Comparative leaf morphogenesis in Iridaceae, Bot. Jahrb. Syst., 112: [22] Rudall P., Mathew B Leaf Anatomy in Crocus (Iridaceae), Kew Bulletin, 45: [23] Rudall P Anatomy of Flowering Plants, Cambridge University Press, 1992 [24] Satıl F., Selvi S An anatomical and ecological study of some Crocus L. taxa (Iridaceae) from the west part of Turkey, Acta Bot. Croat. 66 (1), [25] Shorina N. I Leaf Structure in some saffrons in association with the evolution of the genus Crocus L. Byull. Mosk. Obsch. Ispyt. Prir., Otdel Biol., 80: [26] Şık, L.; Candan, F.; Soya, S.; Karamenderes, C., Kesercioğlu, T., Tanyolaç, B Genetic variation among Crocus L. species from Western Turkey as revealed by RAPD and ISSR markers. Journal of Applied Biological Sciences 2(2): [27] Şık, L.; Candan, F Ecological properties of some Crocus taxa in Turkey. African Journal of Biotechnology, Vol. 8(9), [28] Yüzbaşıoğlu, S., Varol, Ö., A new Autumn-flowering Crocus from SW Turkey. The Plantsman, New Series, Vol. 3, Part 2:

An anatomical and ecological study of some Crocus L. taxa (Iridaceae) from the west part of Turkey

Acta Bot. Croat. 66 (1), 25 33, 2007 CODEN: ABCRA25 ISSN 0365 0588 An anatomical and ecological study of some Crocus L. taxa (Iridaceae) from the west part of Turkey FATİH SATIL*, SELAMİ SELVİ Department