Phytotaxa 394 (1): 071–078 ISSN 1179-3155 (print edition) https://www.mapress.com/j/pt/ PHYTOTAXA Copyright © 2019 Magnolia Press Article ISSN 1179-3163 (online edition) https://doi.org/10.11646/phytotaxa.394.1.4 Morphological, anatomical, palynological and ecological data on the local endemic Dianthus vanensis (Caryophyllaceae) from Turkey MEHMET CENGIZ KARAİSMAİLOĞLU1*, MEHMET EMRE EREZ1, SÜLEYMAN MESUT PINAR2, MEHMET FİDAN1 & HÜSEYIN EROĞLU3 1 Siirt University, Faculty of Arts and Sciences, Department of Biology, 56100, Kezer-Siirt, Turkey; e-mail: [email protected] 2 Yüzüncü Yıl University, Van School of Health, 65090, Tuþba-Van, Turkey. 3 Yüzüncü Yıl University, Faculty of Sciences, Department of Biology, 65090, Tuşba-Van, Turkey. *author of correspondence Abstract A first detailed taxonomical, palynological, and ecological study of the Turkish endemic Dianthus vanensis is shown based on field data collected at locus classicus. A revised morphological description, including data of seed and pollen by using Scanning Electron Microscope, is provided. Seed surface displays elongated cells, which are lobate with 14−28 teeth, I, S shaped or irregular, and papillate. Pollen grains are radially symmetric, apolar, pantoporate and spheroidal shape (size: 40.38 ± 0.64 × 37.98 ± 0.36 μm), with scabrate ornamentation, and 9–12 pores elliptical (different margin), and operculate (size: 6.17 ± 0.23 × 4.17 ± 0.39 μm). The anatomy of root, stem and leaf are also investigated. On the outer surface of the root is placed a multi-layered periderm. The cortex consists of multi-layered parenchymatic cells under periderm, and its thick- ness is 176.49 ± 7.85 μm. In the stem cross-sections, it is noticed one epidermis layer, comprising of flat cells in outermost. Its thickness ranged between 28.13 and 35.29 μm. The type of vascular bundle is hadrocentric. In the abaxial and adaxial surfaces of the leaf are located 1-layer epidermis cells, comprising of the flat or rectangular cells. The leaf is equifacial. Leaves have collateral vascular type. IUCN category of the taxon is assessed CR (Critical Endangered) based on criterion B2a,b(iii). Keywords: Anatomy, Dianthus, Irano-Turanian endemics, IUCN, palynology, SEM, seed morphology Introduction Caryophyllaceae Jussieu (1789: 299) (Caryophyllales Jussieu ex Bercht. & J. Presl) includes about 100 genera and 3000 species mainly distributed in the Holarctic region with a center of diversity in the Mediterranean and Irano-Turanian areas (see e.g., Hernández-Ledesma et al. 2015). Three subfamilies can be considered in Caryophyllaceae: subfam. Caryophylloideae, subfam. Alsinoideae (Candolle 1815: 445) Fenzl (1840: 963), and Paronychioideae (Jussieu 1789: 299) Meissner (1838: 96) (cfr. Rabeler & Bittrich 1993). Caryophyllaceae taxa usually occurs in dry and open habitats, and some of them are restricted in mountainous areas (see e.g., Pax & Hoffman 1934, Bittrich 1993, Johnson & Wilson 1993, Fior et al. 2006, Ilçim et al. 2013). Dianthus Linnaeus (1753: 409) comprises about 600 taxa mainly distributed in Europe, Mediterranean area and Asia, and it is critical from the taxonomic and nomenclatural points of views as highlighted by several authors (see e.g. Güner et al. 2000, Dequan & Turland 2001, Bacchetta et al. 2010, Iamonico et al. 2015). 74 Dianthus taxa occur in Turkey where the genus is of the most diversity (Hamzaoğlu et al. 2011, Yılmaz et al. 2011). The genus has also a significance for its use as medicinal or ornamental plant (Bakshi 1984). In the literature review, it has been run into some researches on Dianthus species such as morphology of the some Turkish (Kostak & Tan 1999, Yıldız 2002, Meriç & Güler 2008, Hamzaoğlu & Koç 2015) and cytology of some species (Jafari & Behoozian 2010, Şahin et al. 2016, Altay et al. 2017), palynology of some species (Yıldız 2002, Sahreen et al. 2008, Kızılpınar et al. 2010) and molecular data such as SSR and RAD marker (Yagi et al. 2017), nuclear DNA sequence (Farsi et al. 2013), mRNA (Shibuya et al. 2002), RAPD and RFLP (Maki & Horie 2008) to Accepted by Manuel B. Crespo: 17 Feb. 2019; published: 25 Feb. 2019 71 clarify the phylogenetic relationships within Caryophyllaceae. Aside from these, it is not possible to find out any work directly correlated to the genus. Owing to the lack of data on the taxa of this genus, multidisciplinary studies should be performed. Dianthus vanensis Behcet & Ilcim (2013: 219−224) is an endemic species from Eastern Anatolia which grows on steppe and rocky areas. Macro- and micro-morphological, palynological, anatomical, and ecological data, as well as IUCN risk assessment of this taxon have been studied in detail for the first time, and here presented. Materials and methods The research was carried out by field surveys in type locality in 2016 (Van: Çatak, Konalga village, Tanrıverdi hamlet, around Zevviçal, steppe, 2340 m, 37º51’09”N, 43º09’44”E; Fig. 1). Plants collected were identified in accordance with Ilçim et al. (2013) and exsiccata deposited at VANF (Van Yüzüncü Yıl University Herbarium). Macromorphological data on leaves, fruits and seeds were obtained using a Leica EZ4 binocular microscope with a HD camera. Pollen analysis was made using Olympus CX31 light microscope, Olympus SC30 camera and stream start software. Seeds and pollen grains were taken from specimens and arranged for scanning electron microscopy (SEM) by mounting to table with silver adhesive, coated with gold and platinum, and analyzed with JEOL Neoscope- 5000 Scanning Electron Microscope (Karaismailoğlu 2015a,b). Terminology follows Stearn (1985). Concerning anatomical investigations, cross sections were obtained with an automatic microtome (Thermo Shonda Met Finesse) from root, stem, and leaf. Later, they were passed through a variety of alcohol and xylene series and stained with haematoxylin and Eosin Y or safranin in a staining device (ASC 720 Medite) and were enclosed by Entellan to examine anatomical structures (Karaismailoğlu 2016, Karaismailoğlu & Erol 2018). Anatomical characters were detected by Olympus CX21FS1 microscope and Kameram Imaging Software. FIGURE 1. Dianthus vanensis. a) flower; b) general habit in nature; c) opened capsule with seeds; d) detail of seed. Results and Discussion Dianthus vanensis includes perennial plants, usually with an extensive creeping woody rootstock. Stem ascending- erect, up to 25 cm long, branched or not, 1–3-flowered, glabrous. Basal leaves few, 10–25 × 1–2 mm, linear to narrowly linear-lanceolate, acute, glabrous. Cauline leaves linear, acuminate, ± rigid, curved divaricate, canaliculate, 2–4 pairs, 15–40(–45) × 1–2 mm, usually shorter than internodes, rarely equal; sheet 1–3 mm, membranous. Bracts 4– 72 • Phytotaxa 394 (1) © 2019 Magnolia Press KARAİSMAİLOĞLU ET AL. 6, with scarious margin; outer 28–30(–34) mm long, almost equal, or slightly shorter to calyx length, curved divaricate narrowed from just above the middle into a long acuminate-aristate apex, inner shorter, (13–)15–17(–19) mm long, partly purplish at base, narrower than outer. Calyx 30–35 × 5–6 mm, glabrous, greenish to purplish, cylindrical, teeth 7–10(–12) mm, linear-lanceolate, mucro 1 mm long, with a narrow scarious, slightly ciliate at margin. Petal 35–37 mm long; limb 15 × 11 mm, barbulate, pinkish-purple, fimbriate for less than 1/3 its length. Seeds creamy brown or light brown, broadly ovate, 3.5–4 × (2–)2.5–3 mm. Our results are mainly congruent with the description reported in Ilçim et al. (2013), excepting for some features, i.e. the dimensions of stem, cauline leaves, bracts, calyx, calyx teeth and seed features, which seem to be more variable than previously reported (Table 1). TABLE 1. Comparison of morphological characters of Dianthus vanensis. Morphological characters Ilçim et al. (2013) This study Stem length (cm) 9–19 up to 25 Cauline leaves (mm) 15–40 × 1–2 up to 45 Bracts (mm) 15–30 up to 34 Calyx (mm) 30–32 × 5–5.5 30–35 × 5–6 Calyx teeth length (mm) 7–10 up to 12 Seed colour brown creamy brown or light brown shape broadly elliptic-oblong broadly ovate size (mm) 3–4 × 2–2.5 3.5–4 × (2–)2.5–3 As regards of the micromorphological structures on surfaces of seeds and fruits, they have high taxonomic value (see e.g., Bernard 2000). Heywood (1971) highlighted the importance of SEM for systematic issues. Many researchers reported that seed microstructures are beneficial characters to use in discriminating taxa within the family Caryophyllaceae (Yıldız 2002, Minuto et al. 2006). Dianthus vanensis displays seed surface cells elongated, lobate, with 14–28 lobes, I, S shaped or irregular, rough secondary sculpture and papillate. Furthermore, anticlinal cell walls in the studied species are sunken and undulate, periclinal cell walls are convex or flat as well. These outcomes show parallels with the other Caryophyllaceae representatives examined, such as some taxa of the genera Arenaria Linnaeus (1753: 423), Cerastium Linnaeus (1753: 437), Moehringia Linnaeus (1753: 359) and Stellaria Linnaeus (1753: 421) (cfr. Wyatt 1984, Minuto et al. 2006). Pollen grains of Dianthus vanensis are radially symmetric, apolar, pantoporate and spheroidal (Fig. 2). Their lengths are 40.38 ± 0.64 μm, widths are 37.98 ± 0.36 μm. The pores are elliptical, with different margin, operculate. The pore length is 6.17 ± 0.23 μm, pore width is 4.17 ± 0.39 μm. The pore number is between 9 and 12. The distance between the pores is 8.97 ± 0.45 μm. The exine thickness is 1.58 ± 0.14 μm. Furthermore, the intine thickness is 0.37 ± 0.04 μm. The pollen ornamentation is scabrate. According to Yıldız (2002), who worked on 45 species (15 genera) of Caryophyllaceae, 10 types of exine ornamentation have been recorded: Arenaria type, Stellaria type, Cerastium type, Dianthus type, Gypsophila repens type, Lychnis viscaria type, Silene vulgaris type, Silene caryophylloides subsp. subulata type, Silene conica type, and Agrostemma githago type. Dianthus type is scabrate as in Yıldız’s study.