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Anatomical Structure of Cladodes of Ruscus L. Taxa (Liliaceae) in Turkey

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Anatomical Structure of Cladodes of Ruscus L. Taxa (Liliaceae) in Turkey FABAD J. Pharm. Sci., 36, 119-128, 2011 RESEARCH ARTICLE Anatomical structure of cladodes of Ruscus L. taxa (Liliaceae) in Turkey Ayşegül GÜVENÇ*°, Maksut COŞKUN*, Okan ARIHAN** Anatomical structure of cladodes of Ruscus L. taxa Türkiye’deki Ruscus L. taksonlarının (Liliaceae) Kladot (Liliaceae) in Turkey Anatomisi Summary Özet The genus “Ruscus” of Liliaceae family is represented by four Liliaceae familasında yer alan “Ruscus” cinsi, Türkiye’de R. species (5 taxa) in Turkey; namely R. aculeatus L., R. hy- aculeatus L., R. hypoglossum L., R. colchicus P. F. Yeo ve poglossum L., R. colchicus P. F. Yeo, and R. hypophyllum L. R. hypophyllum L. olmak üzere 4 tür (5 takson) ile temsil Two varietes of R. aculeatus (R. aculeatus var. aculeatus and edilmektedir. R. aculeatus (R. aculeatus var. aculeatus ve R. R. aculeatus var. angustifolius Boiss.) are also listed in the aculeatus var. angustifolius Boiss.) iki varyete ile Türkiye Flora of Turkey. Flora’sında yer almaktadır. Microscopical properties of transverse sections from the Her bir taksonun kladotlarından alınan enine kesitlerin cladodes of each taxa were illustrated and photographed. In mikroskobik özellikleri, çizim ve fotoğraflarla verilmiştir. the cross section, it was not possible to differentiated upper Kladotların enine kesitlerinde alt ve üst epiderma farklılığını and lower epiderma of cladodes. In the mesophyll of cladodes belirlemek mümkün değildir. Kladot mezofilinde palizat palisade and spongy parenchyma are not seen. There are a ve sünger parenkiması görülmemiştir. Üst epidermanın few layers of parenchyma cells beneath the upper epidermis altında ve alt epidermanın üstünde çok dar parenkimatik and above the lower epidermis. Cross section of the midrib is hücre tabakası yer almaktadır. Orta damar, enine kesitte, clearly convex in the lower surface. Vascular bundles have alt yüzde hafifçe konvekstir. İletim demetleri sklerenkimatik been surrounded by sclerenchymatic bundles. The cladode is bir halka ile kuşatılmıştır. Kladotlar isobilateraldir. Her iki isobilateral and both epiderma are similar in appearance be- epiderma genellikle uzun hücrelerden oluşmuş ve birbirine ing composed of generally long cells with, straight or slightly benzer görünüşte, düz veya hafifçe girintili çıkıntılı sinuous walls. Numerous paracytic stomata are present on çeperlidir. Her iki yüzde de çok sayıda parasitik tipte stoma both surfaces. görülmektedir. Key Words: Ruscus aculeatus, Ruscus hypoglossum, Ruscus Anahtar Kelimeler: Ruscus aculeatus, Ruscus hypoglossum, colchicus, Ruscus hypophyllum, cladodes, anatomy, Liliaceae. Ruscus colchicus, Ruscus hypophyllum, kladot, anatomi, Liliaceae. Received: 01.10.2012 Revised: 09.01.2013 Accepted: 11.02.2013 * Ankara University, Faculty of Pharmacy, Department of Pharmaceutical Botany, Ankara, Turkey ** Yüzüncü Yıl University, Faculty of Arts and Sciences, Department of Biology, Zeve Campus-Van, Turkey ° Corresponding Author E-mail: [email protected] 119 Güvenç, Coşkun, Arıhan INTRODUCTION new steroidal saponins. The amounts of steroidal Ruscus species are members of the Liliaceae saponins have also been determined by HPTLC, family, which are perennial, rhizomatus and HPLC, HPLC-ESI-MS, and UPLC methods (6-18). evergreen shrubs. The genus Ruscus is native to the Mediterranean, Southern and Western Europe. Five Stem leaves in Ruscus species, are scale-like and taxa of Ruscus grow naturally in Turkey. These are R. membranous, additionally, stems are reduced to aculeatus var. aculeatus, R. aculeatus var. angustifolius, photosynthetic phylloclades. Being a metamorphic R. hypoglossum, R. colchicus and R. hypophyllum (1). form of stem, anatomical structure of the phylloclades should be very interesting. In a study, Sachs et al. The underground parts of the Ruscus aculeatus investigated variable development and cellular have been used medicinally as diuretic and anti- patterning in the epidermis of R. hypoglossum. This inflamatory agents, as well as for hemorroids study has concerned the presence and the meaning treatments and atherosclerosis and circulatory of immature stomata in the epidermis of it. The insufficiency prevention. As a treatment for diseases distribution of the stomata of Ruscus was remarkably of circulatory system, R. aculeatus has a long tradition variable (19). Although a lot of work has been done on of proven success in Europe. A written record of R. aculeatus, especially morphological characteristics, its use as a phlebotherapeutic agent dates back at phylloclade development and systematic (20-22), least 2000 years. These species are called as “sılcan, a few data are available in the literature related to tavşanmemesi, tavşan kirazı, diken kökü, yandak anatomical structure of the genus (19, 23). We studied dikeni, kandak, or sıçan dikeni”. A decoction of roots cross sections obtained from cladodes of R. aculeatus of R. aculeatus is widely used as a diuretic and for which is in European Pharmacopoeia 2008 (24) and the treatment of urinary system disorders, as well the other three species growing in Turkey. We used as kidney stones in folk medicine in the vicinities of the term of cladodes, because this term is used in the Eskişehir, Hatay, Mersin in Turkey (2-4). The roots of Flora of Turkey. these species, which is exported around 200-1000 tons per year from Turkey, either fresh or as dried roots. Materials and methods Because of this reason, R. aculeatus is vulnerable in Research materials were collected from various Turkey (5). locations in Turkey. Voucher specimens were deposited in AEF (the Herbarium of Ankara In the chemical studies conducted on various parts of University, Faculty of Pharmacy) Locations of the the Ruscus species, flavonoids and steroidal saponins investigated plant samples are given in Table 1. particularly ruscine, ruscoside, neoruscogenin and ruscogenin as well as many new steroidal saponins Plants materials were preserved in 70% ethanol. have been identified. Intensive chemical studies have Cross sections of the cladodes of the Ruscus taxa were been carried on the Ruscus species and isolated many performed by hand from the preserved materials in chloral hydrate solution and Sartur reagent. Sartur Table 1. Locations of the studied Ruscus taxa Species Locality Ruscus aculeatus var. aculeatus A3 Sakarya: Karasu, 22.9.2001, M. Coşkun, A. Güvenç & C. S. Erdurak (AEF 22940). R. aculeatus var. angustifolius A6 Samsun: Terme, Sivaslar village, 14.3.2002, M. Coşkun & O. Arıhan (AEF 22939). R. colchicus A6 Ordu: Ünye, 15.3.2002, M. Coşkun & O. Arıhan (AEF 22934). R. hypoglossum A7 Giresun: Tirebolu, İnköy roadside, 15.3.2002, M. Coşkun & O. Arıhan (AEF 22930); R. hypophyllum A7 Giresun: Tirebolu to Espiye, 16.3.2002, M. Coşkun & O. Arıhan (AEF 22933); 120 FABAD J. Pharm. Sci., 36, 119-128, 2011 reagent contains KI-I, aniline, sudan III, lactic acid, alcohol and water (25). Illustrations were made using a Leitz drawing prism attached to a Leitz- Wetzlar (45o) microscope. The cross sections were photographed with an automatic camera attached to Olympus BX 50 microscope. Results The anatomical features were observed in cross sections of the cladodes (Figures 1-10) and the comparisons of the anatomical characters of cladodes of taxa are given in Table 2. Cross section of cladode blade In the cross-section, it was not possible to differentiate upper and lower surfaces of the cladodes; after a thick cuticle, epidermis is preceded by parenchymatic cells. In the mesophyll of cladodes, palisade and spongy parenchymas were not seen. There were a few layers of parenchyma cells beneath the upper epidermis and above the lower epidermis. This parenchyma contains chloroplast and they exist instead of palisade parenchyma. Between the two parenchyma layers, one or two layers of large Figure 1. Transverse sections of the cladodes of Ruscus and colourless cells are present, differing from one species growing in Turkey. A- R. aculeatus var. aculeatus; B- R. aculeatus var. angustifolius; C- R. colchicus; D- R. species to other. This cell layer exists instead of hypoglossum; E- R. hypophyllum. spongy parenchyma (Fig. 2 A, B; Fig.3 A; Fig. 4 A; Fig. 5 A, B; Fig. 6 A, B; Fig. 9 A; Fig. 10 A). collateral and surrounded by sclerenchyma. The Cross section of the midrib number of vascular bundles differ from species Cross section of the midrib is clearly convex in to species; there are numerous vascular bundles the lower surface (Fig. 1). Vascular bundles are in R. colchicus, R. hypoglossum, and R. hypophyllum Table 2. Comparison of the cross sections of different Ruscus taxa native in Turkey. Epidermal Upper Lower Colorless Vascular bundles in the Section of Species cell parenchyma parenchyma parenchyma midrib cladode margin R. aculeatus var. Square or 3-4 (-5) layers 3-5 layers 2 layers 1, pith sclerenchymatic rectangular aculeatus rectangular R. aculeatus var. rectangular 3-4 layers 3- (-4) layers 2 layers 1, pith sclerenchymatic rectangular angustifolius 6-7,pith sclerenchymatic R. colchicus rectangular 3-5 layers 2-4 layers 1- (-2) layers acute and parenchymatic 10-11 (-13), pith R. hypoglossum rectangular 2-3 layers 2-3 layers (1-) -2 layers sclerenchymatic and rhombic parenchymatic 10-11 (-13), pith R. hypophyllum rectangular 2-4 layers 2-3 layers 1- (-2) layers sclerenchymatic and acute parenchymatic 121 Güvenç, Coşkun, Arıhan Figure 2. A-Cross sections of the cladodes margins of R. aculeatus var. aculeatus, A1- Surface of upper epidermis, A2-Surface of lower epidermis; B- R. aculeatus var. angustifolius; B1-Surface of upper epidermis B2-Surface of lower epidermis. a- cuticle, b- epidermis, c- upper parenchyma, d- colorless parenchyma, e- lover parenchyma, f- sclerenchyma, g- phloem, h- xylem. however few in the R. aculeatus (Fig. 3B; Fig. 4B; Fig. Surface epidermis 7; Fig. 8; Fig. 10B). Upper and lower epiderma are similar in appearance being composed of generally long cells, with straight Vascular bundles other than midrib the cross section or slightly sinuous walls. Numerous paracytic of the vascular bundles of the cladode blade, phloem stomata are present on both surface (Fig. 2 A1, A2, B1, is at the upper surface. B2; Fig. 5 A1, A2, B1, B2; Fig. 9 A1, A2). 122 FABAD J. Pharm. Sci., 36, 119-128, 2011 Figure 3.
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